This experiment tests the consequence of a mutation at the FatB gene in the wound-response of Arabidopsis. The FatB mutant allele (fatb KD J. (Plant Cell 2003, Vol 15, 1020-1033)) was obtained from Dr. Katayonn Dehesh, of California, Davis, Davis, CA. This allele is in the Ws background.The growth conditions are as follows: 1. Seeds (between 14 and 16) are sown on in 100 x 100 x 15mm square Falcon Petri Dishes (Fisher Scientific, catalogue Seeds were arranged on the plates in a single horizontal line at the 1-cm mark the top of the plate.2. Each plate contains between 20 and 25-ml of sterile MS containing 0.1% (w/v) sucrose.3. Prior to sowing, seeds were sterilized by for 1 minute at room temperature with a 300-l solution of 50% (v/v) ethanol, solution was removed and replaced with a 300-l solution consisting of 1% (v/v) 20 (Fischer BioReagents, catalogue #BP33750), and 50% (v/v) bleach solution and incubated at room temperature for 10-minutes. The seeds were then washed three changes of 0.3-ml of sterile water.
Lipidomics studies on NIDDK / NIST human plasma samples
STUDY_TYPE
MS analysis on human plasma
STUDY_SUMMARY
The National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) in with the National Institute of Standards (NIST) recently produced a human standard reference material (SRM 1950) for metabolite analysis. The SRM was by obtaining plasma samples from 100 individuals between 40 and 50 years of whose ethnicity was representative of the US population and that included an number of men and women. The intent of the NIDDK/NIST project was to provide a material that would be publically available to researchers and that could be by the clinical chemistry community to identify plasma metabolites for purposes. Signature metabolites could then be further probed for their as disease biomarkers. The LIPID MAPS Consortium has undertaken the task to this SRM by systematically identifying and quantifying the lipid molecular in the six main categories of mammalian lipids. The quantitative levels of over different lipids present in this reference human plasma sample are presented
Timecourse on RAW 264.7 cells treated with Kdo2-Lipid A and compactin
STUDY_TYPE
Timecourse experiment
STUDY_SUMMARY
Lipidomics studies on macrophages - RAW 264.7 cells treated with Kdo2-Lipid A compactin. Experiments were conducted with RAW264.7 cells fed 10% fetal calf 8-timepoint study: Measurements were taken at 0, 0.5,1,2,4,8, 12, and 24hrs (i) compactin, (ii) Kdo2-Lipid A, (iii) compactin + Kdo2-Lipid A. and (iv)
In this study, seventeen white wines including Chardonnays, Viogniers, Pinot gris, Rieslings and Sauvignon blancs (which were part of a M.S. study in the Viticulture & Enology Department on white wine mouthfeel properties), were analyzed by GC-TOF. Additionally, chemical data obtained will be mined with the sensory data collected to further investigate the chemical basis for mouthfeel properties in wine.
INSTITUTE
University of California, Davis
DEPARTMENT
Davis Genome Center
LABORATORY
Fiehn
LAST_NAME
Kind
FIRST_NAME
Tobias
ADDRESS
451 E. Health Sci. Drive, Davis, California 95616, USA
Bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo), rise to devastating crop losses in rice. Disease resistant rice cultivars are most economical way to combat the disease. The TP309 cultivar is susceptible to by Xoo strain PXO99. A transgenic variety, TP309_Xa21, expresses the pattern receptor Xa21, and is resistant. PXO99?raxST, a strain lacking the raxST gene, able to overcome Xa21-mediated immunity. We used a single extraction solvent to comprehensive metabolomics and transcriptomics profiling under sample limited and analyze the molecular responses of two rice lines challenged with either or PXO99?raxST. LCTOF raw data file filtering resulted in better within group of replicate samples for statistical analyses. Accurate mass match compound with molecular formula generation (MFG) ranking of 355 masses was achieved with METLIN database. GCTOF analysis yielded an additional 441 compounds after database processing, of which 154 were structurally identified by retention library matching. Multivariate statistics revealed that the susceptible and genotypes possess distinct profiles. Although few mRNA and metabolite were detected in PXO99 challenged TP309 compared to mock, many differential occurred in the Xa21-mediated response to PXO99 and PXO99?raxST. Acetophenone, fatty acids, alkaloids, glutathione, carbohydrate and lipid biosynthetic were affected. Significant transcriptional induction of several pathogenesis genes in Xa21 challenged strains, as well as differential changes to GAD, PAL, and Glutathione-S-transferase transcripts indicated limited correlation with changes under single time point global profiling conditions.
INSTITUTE
University of California, Davis
DEPARTMENT
Davis Genome Center
LABORATORY
Fiehn
LAST_NAME
Kind
FIRST_NAME
Tobias
ADDRESS
451 E. Health Sci. Drive, Davis, California 95616, USA
This experiment tests the effect of individual mutations on the metabolome of The standardized growth conditions are as follows: 1. Seeds (between 14 and 16) sown on media in 100 x 100 x 15mm square Falcon Petri Dishes (Fisher catalogue #08-757-11A). Seeds were arranged on the plates in a single line at the 1-cm mark from the top of the plate. 2. Each plate contains between and 25-ml of sterile MS media containing 0.1% (w/v) sucrose. 3. Prior to seeds were sterilized by treating for 1-minute at room temperature with a 300-l of 50% (v/v) ethanol, this solution was removed and replaced with a 300-l consisting of 1% (v/v) Tween 20 (Fischer BioReagents, catalogue #BP33750), and (v/v) bleach solution (Clorox), and incubated at room temperature for The seeds were then washed with three changes of 0.3-ml of sterile water. 4. sowing with seeds, the plates were wrapped with Micropore tape (3M Health Care, #1530-0), and then stored horizontally for 4-days at 4 °C, with of 1 mol/m2. 5. On the 5th day, plates were moved to the growth room, and held a vertical position in Plexi-glass holders for 17-days this growth room is labeled in Table I. 6. On 18th day Petri plates were opened and the aerial of these plants were harvested immediately upon plate opening. 7. Upon plant material was quenched by immersion in liquid nitrogen and stored at 70
INSTITUTE
University of California, Davis
DEPARTMENT
Davis Genome Center
LABORATORY
Fiehn
LAST_NAME
Kind
FIRST_NAME
Tobias
ADDRESS
451 E. Health Sci. Drive, Davis, California 95616, USA
To determine the basis of running capacity and health differences in outbread N/NIH rats selected for high capacity (HCR) and low capacity (LCR) running (a for VO2max) (see:Science. 2005 Jan 21;307(5708):418-20). Plasma collected at 12 of age in generation 28 rats after ad lib feeding or 40% caloric restriction at week 8 of age. All animals fasted 4 hours prior to collection between 5-8
Determine purity and quality of IROA labelled glucose
STUDY_TYPE
NMR and MS analysis
STUDY_SUMMARY
We demonstrate the global metabolic analysis ofCaenorhabditis elegansstress responses using a mass-spectrometry-based technique called isotopic ratio outlier analysis (IROA). In an IROA protocol, control and experimental samples are isotopically labeled with 95 and 5%13C, and the two sample populations are mixed together for uniform extraction, sample preparation, and LC-MS analysis.To illustrate the utility of IROA for global metabolomics, we exposed wild-type (N2) worms to a heat shock (30 min heat shock at 33 C), which causes significant, widespread changes in metabolism. We collected and analyzed material from the exometabolome (all material that worms release in the supernatant) and the endometabolome (homogenized total extracts from the worm bodies).
INSTITUTE
University of Florida
DEPARTMENT
Biochemistry & Molecular Biology
LAST_NAME
Stupp
FIRST_NAME
Gregory
EMAIL
stuppie@ufl.edu
STUDY_COMMENTS
Determine purity and quality of IROA labelled glucose
The goal of the study was to determine whether there is a set of metabolites that differentiate people who have knee OA and show radiographic disease progression over 18 months from those who have knee OA and do not show disease progression over the same time period.
This metabolomics pilot and feasibility (P & F) study was conducted to provide data to be used to gain a better understanding of metabolic alterations in people with knee osteoarthritis (OA) and to discover novel biomarkers of the disease. The goal of the metabolomics study was to determine if metabolic differences, detected by a comprehensive metabolomics analysis, can be used to distinguish people who will develop symptomatic knee OA from those who will not. For this metabolomics study, individuals participating in T1 or T1* with 5-year follow-up at T2 were selected. At T2 subjects were on average 68.1(9.12) years old with an average BMI of 31.4(7.01) with 32% men and one-third African American. All had weight-bearing posterior-anterior knee films obtained with the Synaflexer positioning device at both time points and read paired for Kellgren-Lawrence grade and minimum joint space. Urine samples (second morning void) collected from 36 overweight or obese participants in the JoCo at T1 or T1* were selected from two subgroups (a group that developed radiographic osteoarthritis (n=16) and an age, race, sex, and BMI matched group that did not develop osteoarthritis (n=20). Radiographic knee OA was defined as Kellgren-Lawrence grade 2-4 at T2 in a person with Kellgren-Lawrence grade 0 or 1 at T1 or T1*.
Metabolomics Analysis of Thermally Challenged Mayfly Larvae (GCMS analysis)
STUDY_TYPE
Metabolomic analysis of mayflies
STUDY_SUMMARY
The purpose of this study was to examine the metabolic profiles of mayfly triangulifer) larvae subjected to thermal challenge. This species is unusual in of its ease of culture, and its suitability as a laboratory test organism. Our here was to examine how an environmentally realistic thermal challenge affects physiology of this organism. In this study, we obtained several types of insect and we were able to show that GC-MS Metabolomics could be used to distinguish the different types of larvae.
Metabolomics Involved in Early Life Antibiotic Exposures(DuraSTAT-Cecal)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the DuraSTAT sub-study, a total of 18 samples from 8 week old, female C57BL/6 mice; comprised of 6 urine samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were given STAT penicillin and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a high fat diet.
Metabolomics Involved in Early Life Antibiotic Exposures(DuraSTAT-Liver)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the DuraSTAT sub-study, a total of 18 samples from 8 week old, female C57BL/6 mice; comprised of 6 urine samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were given STAT penicillin and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a high fat diet.
Metabolomics Involved in Early Life Antibiotic Exposures(DuraSTAT-Urine)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the DuraSTAT sub-study, a total of 18 samples from 8 week old, female C57BL/6 mice; comprised of 6 urine samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were given STAT penicillin and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a high fat diet.
Metabolomics Involved in Early Life Antibiotic Exposures(TranSTAT-Cecal)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the TranSTAT sub-study, a total of 18 samples from 8 week old, female Swiss webster mice; comprised of 6 serum samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were inoculated with cecal contents from STAT mice and 3 mice/matrix were inoculated with cecal contents from Control mice. The mice were housed with conventional bedding and fed a high fat diet.
Metabolomics Involved in Early Life Antibiotic Exposures(TranSTAT-Liver)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the TranSTAT sub-study, a total of 18 samples from 8 week old, female Swiss webster mice; comprised of 6 serum samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were inoculated with cecal contents from STAT mice and 3 mice/matrix were inoculated with cecal contents from Control mice. The mice were housed with conventional bedding and fed a high fat diet.
Metabolomics Involved in Early Life Antibiotic Exposures(TranSTAT-Serum)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the TranSTAT sub-study, a total of 18 samples from 8 week old, female Swiss webster mice; comprised of 6 serum samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were inoculated with cecal contents from STAT mice and 3 mice/matrix were inoculated with cecal contents from Control mice. The mice were housed with conventional bedding and fed a high fat diet.
Metabolomics Involved in Early Life Antibiotic Exposures(NOD-Cecal)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the NOD sub-study, a total of 18 samples from 6 week old, male NOD/ShiLtj mice; comprised of 6 serum samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were exposed to PAT and 3 mice/matrix were non-exposed Controls. The mice were housed with SPF (Helicobacter neg/MNV neg) bedding and fed a normal diet.
Metabolomics Involved in Early Life Antibiotic Exposures(NOD-Liver)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the NOD sub-study, a total of 18 samples from 6 week old, male NOD/ShiLtj mice; comprised of 6 serum samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were exposed to PAT and 3 mice/matrix were non-exposed Controls. The mice were housed with SPF (Helicobacter neg/MNV neg) bedding and fed a normal diet.
Metabolomics Involved in Early Life Antibiotic Exposures(NOD-Serum)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the NOD sub-study, a total of 18 samples from 6 week old, male NOD/ShiLtj mice; comprised of 6 serum samples, 6 cecal content samples and 6 liver tissue samples were analyzed. Three mice/matrix were exposed to PAT and 3 mice/matrix were non-exposed Controls. The mice were housed with SPF (Helicobacter neg/MNV neg) bedding and fed a normal diet.
Metabolomics Involved in Early Life Antibiotic Exposures(EstroSTAT-Liver )
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the EstroSTAT sub-study, a total of 18 samples from 23 week old, female C57BL/6 mice; comprised of 6 urine samples, 6 serum samples and 6 liver tissue samples were analyzed. Three mice/matrix were given STAT penicillin and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a Low phyto-estrogen diet.
Metabolomics Involved in Early Life Antibiotic Exposures(EstroSTAT-Serum)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the EstroSTAT sub-study, a total of 18 samples from 23 week old, female C57BL/6 mice; comprised of 6 urine samples, 6 serum samples and 6 liver tissue samples were analyzed. Three mice/matrix were given STAT penicillin and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a Low phyto-estrogen diet.
Metabolomics Involved in Early Life Antibiotic Exposures(EstroSTAT-Urine)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the EstroSTAT sub-study, a total of 18 samples from 23 week old, female C57BL/6 mice; comprised of 6 urine samples, 6 serum samples and 6 liver tissue samples were analyzed. Three mice/matrix were given STAT penicillin and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a Low phyto-estrogen diet.
Metabolomics Involved in Early Life Antibiotic Exposures(VGSTAT-Cecal)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the VG STAT sub-study, a total of 18 samples from 7 week old, male C57BL/6 mice; comprised of 9 cecal content samples and 9 liver tissue samples were analyzed. Three mice/matrix were given penicillin VK, 3 were given penicillin G, and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a normal diet.
Metabolomics Involved in Early Life Antibiotic Exposures(VGSTAT-Liver)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
In the VG STAT sub-study, a total of 18 samples from 7 week old, male C57BL/6 mice; comprised of 9 cecal content samples and 9 liver tissue samples were analyzed. Three mice/matrix were given penicillin VK, 3 were given penicillin G, and 3 mice/matrix were non-treated Controls. The mice were housed with conventional bedding and fed a normal diet.
Human subjects were given high PUFA diet for 21 days then converted to high diet for another 21 days. Plasma samples were drawn during the feeding process 7 visits.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Plasma metabolomics: Comparison of non-diabetic controls with T1D patients
STUDY_TYPE
Drug effect study
STUDY_SUMMARY
Non-diabetic controls whose metabolites were compared to T1D patients with and insulin. Seven C-peptide?negative T1D subjects were studied on two occasions: during insulin treatment and the other following withdrawal of insulin for 8 h compared with matched healthy ND participants
Identification of altered metabolic pathways in Alzheimer's disease, mild impairment and cognitively normals using Metabolomics (plasma)
STUDY_TYPE
1
STUDY_SUMMARY
Criteria for the diagnosis of amnestic MCI included: (i) memory complaint by the patient and collateral source; (ii) impairment in 1 or more of the 4 domains (memory, executive functioning/attention, visuospatial, or language); essentially normal functional activities of daily living; and (iv) absence of In general, the amnestic MCI determination is made when the memory measures 1.0Ć¢??1.5 SD below the means for age and education appropriate individuals our community; however, rigid cutoffs on psychometric scores were not used to the diagnosis of amnestic MCI which was made on clinical grounds. The diagnosis dementia was made using DSM-IV criteria, and the diagnosis of AD was made using criteria. Subjects were considered to be CN if they performed within the range and did not meet criteria for MCI or dementia.
Identification of altered metabolic pathways in Alzheimer's disease, mild impairment and cognitively normals using Metabolomics (CSF)
STUDY_TYPE
MS
STUDY_SUMMARY
Criteria for the diagnosis of amnestic MCI included: (i) memory complaint by the patient and collateral source; (ii) impairment in 1 or more of the 4 domains (memory, executive functioning/attention, visuospatial, or language); essentially normal functional activities of daily living; and (iv) absence of In general, the amnestic MCI determination is made when the memory measures 1.0?1.5 SD below the means for age and education appropriate individuals in community; however, rigid cutoffs on psychometric scores were not used to the diagnosis of amnestic MCI which was made on clinical grounds. The diagnosis dementia was made using DSM-IV criteria, and the diagnosis of AD was made using criteria. Subjects were considered to be CN if they performed within the range and did not meet criteria for MCI or dementia.
Metabolomic & lipidomic profiles in response to exogenous insulin & GLP-1 during prolonged fasting (GCMS)
STUDY_TYPE
Timecourse
STUDY_SUMMARY
This application requests funding to access state-of-the-art metabolomics and platforms at the NIH West Coast Metabolomics Center to analyze plasma samples recent insulin and glucagon-like peptide-1 (GLP-1) infusion experiments in prolong-fasted elephant seals. This suite of studies was designed to better the mechanisms contributing to the onset of an insulin resistantlike condition by prolonged food deprivation/starvation in mammals. Because elephant seals evolved robust physiological mechanisms that have allowed them to naturally such protracted bouts of fasting, they provide an ideal model to address our hypothesis that increased lipid utilization late in the fast contributes to resistance in elephant seals. Insulin resistance is a common consequence of in mammals and, while the mechanisms by which it manifests are still unclear, a shift favoring increased mobilization and utilization of lipids during food deprivation may be a principal causative factor. Insulin resistance has a connotation due to its association with obesity and diabetes among humans, but has been suggested to be an adaptive response to food deprivation.
INSTITUTE
University of California, Davis
DEPARTMENT
GBSF
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
451 Health Sci Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
1-530-752-8258
NUM_GROUPS
5
TOTAL_SUBJECTS
117
STUDY_COMMENTS
5 general classes are designed in the experiment:#1 - A=No GLP1/ Late GTT#2 - (Low Dose)#3 - C=GLP1 (High Dose)#4 - IL- Early Fasting Insulin Infusion#5 - Late Fasting Insulin InfusionEach of the 5 classes has 6 timepoints (5x6):T1 - minT2 - 10 minT3 - 30 minT4 - 60 minT5 - 120 minEach timepoint had 5 animals (5 x 5 classes x 6 timepoints = 150 totalBecause certain animals and timepoints to be excluded 108 measurments remain.The experiment also contains 9 technical of pooled samples (pool)The total sample number is 108 (biological) + 9 pooled = 117
Metabolomics Analysis of Thermally Challenged Mayfly Larvae (NMR analysis)
STUDY_TYPE
Metabolomic analysis of mayflies
STUDY_SUMMARY
The purpose of this study was to examine the metabolic profiles of mayfly (Centroptilum triangulifer) larvae subjected to thermal challenge. This species is unusual in terms of its ease of culture, and its suitability as a laboratory test organism. Our purpose here was to examine how an environmentally realistic thermal challenge affects the physiology of this organism. In this study, we obtained several types of insect species and we were able to show that NMR Metabolomics could be used to distinguish among the different types of larvae.
Brophy and Askenazi hypothesize that postnatal renal maturation results in specific identifiable patterns of urinary metabolites and that these profiles will be altered in the presence of AKI. Their long-term goal is to identify novel metabolomics urinary profiles that can provide real-time evidence of evolving neonatal renal injury thereby allowing earlier diagnosis and treatment.This study includes 40 pre-term infants age 2 days. Twenty infants were identified as not having AKI (11 females, mean gestational age=182.8 days, mean birth weight=834.0 grams) and 20 infants were identified as having AKI (13 females, mean gestational age=183.9 days, mean birth weight=815.8 grams). There are no statistical differences between the two groups based on gender, gestational age, and birth weight.
INSTITUTE
RTI International
DEPARTMENT
RTI RCMRC
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 East Cornwallis Road, Research Triangle Park, NC 27709, USA.
Fetal metabolomic signature of exposure to iAs during pregnancy
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
This metabolomics pilot study was aimed to identify a fetal metabolomic signature of exposure to iAs during pregnancy. Fetal cord blood serum was collected immediately after delivery from a saline cleaned umbilical cord using an anticoagulant-free vacutainer tube, after clot formation the tube was centrifuged at 1200 rpm and the serum was collected and stored at -70Ā°C. A subset of 50 cord serum samples were selected from the larger BEAR cohort to represent a wide range of iAs exposure as determined by iAs in drinking water (DW-iAs). The exposure during pregnancy was confirmed using U-tAs. A total of 50 cord blood serum samples were used in the metabolomics analysis The samples included 25 newborns with lower maternal iAs exposure levels (DW < 25?g As/L, mean U-tAs=16 ?g/L) and 25 newborns with higher maternal iAs exposure levels (DW> 25?g As/L, mean U-tAs =107 ?g/L).
INSTITUTE
RTI International
DEPARTMENT
Systems and Translational Sciences (STS)
LABORATORY
RTI RCMRC
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 East Cornwallis Roadm Research Triangle Park, NC 27709, USA
Metabolic Microenvironments in Normal Breast and Breast Cancer (MS analysis)
STUDY_TYPE
Metabolomic analysis of normal breast tissue
STUDY_SUMMARY
The purpose of this study was to understand the metabolomic changes in a breast microenvironment at various stages of cancer development and progression (i.e. breast, DCIS, and invasive cancer). The goal of this metabolomics pilot and study was to apply a broad spectrum GC-MS metabolomics method to determine changes in normal tissue of women with cancer correlate with progression and or subtype of the disease.
Effect of kinase inhibitors on FLT3-ITD AML cell metabolomes
STUDY_TYPE
Metabolomics analysis on the effect of kinase inhibitors on FLT3-ITD AML cell to identify changes in cell signaling networks
STUDY_SUMMARY
FLT3-ITD AML cells (MR2) obtained from mice were treated with two MEK kinase (GSK and AZD) at 10 uM versus media only control. Conditioned media aliquots cellular fractions comprised of two aliquots (tecnical replicates) for LC-MS and one for Western blot analyses were collected at 0, 4, 24, and 48 hours. The was repeated three times. HPLC-MS data were acquired for 24 samples from one experiment.
Environmental impact on metabolomics and food allergy
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
This metabolomics pilot and feasibility (P&F) was conducted to determine if varying the levels of environmental LPS exposure and immunization with the vaccine adjuvant alum alters host metabolism. Additionally, the metabolomics will identify biomarkers that can predict allergic disease development and or disease severity after a peanut challenge in hypersensitive mice. Information gained from the proposed studies may allow for the identification of individuals who are Āat riskĀ or Ānot at riskĀ for the development of allergic disease. Furthermore, completion of these studies may lead to identification of biological targets that may be used to develop novel therapies to treat or prevent allergic disease via future funding.
Combined Metabolomics and Lipidomics of Type 1 Diabetes (GCMS)
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project granted to Bell Ā (University of Chicago) and collaborator Manami Hara uses a multi-omics to investigate the metabolome (primary metabolites), lipidome (complex lipids) signaling lipids (oxylipins) in the plasma of Non-obese Diabetic (NOD) mice progressed to Type 1 Diabetes Mellitus (T1D) and those that did not. NOD Mice were assessed as diabetic or non-diabetic based on their fasting (4hr) blood levels at sacrifice, which defined 30 hyperglycemic (glucose = 250 mg/dL) and normoglycemic animals. The primary objective of this study was to identify biomarkers associated with beta-cell destruction/survival and T1D progression.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Metabolite changes associated with methionine stress sensitivity of cancer (GC MS analysis)
STUDY_TYPE
timecourse study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project granted to Kaiser (UC Irvine),Ā aims to achieve understanding of a unique metabolic of cancer cells to explore development of novel unconventional therapeutic that exploit dependence of cancer cells on methyl-donor abundance. The past few have highlighted the role of altered metabolism in cancer. While mechanistic into changed metabolism in cancer is very limited, the importance of the pathway surrounding homocysteine and methionine for cancer cell proliferation been known for over 30 years.These findings, generally summarized as or methionine stressĀ sensitivity, describe the phenomenon that most cancer cannot proliferate in growth medium where the amino acid methionine is replaced its direct metabolic precursor homocysteine. Importantly, non-tumorigenic cells unaffected by replacing methionine with homocysteine in the growth medium. For past years we have been studying methionine dependence of breast and prostate and demonstrated that methionine-dependence is caused by insufficient flux this pathway to sustain synthesis of the downstream metabolite and the methyl-donor S-adenosylmethionine (SAM).We have isolated rare cell clones from breast cancer cells (referred to as MB468RES) that are no longer methionine and proliferate in homocysteine medium. Interestingly, MB468RES have lost their for anchorage independent growth, a hallmark of cancer. The MB468 and MB468RES line pair confirms other observations showing that methionine dependence is linked to tumorigenicity. Importantly, this cell line pair is an ideal model to metabolite signatures linked to cancer cell methionine dependence. We propose characterize the metabolic changes triggered by the shift from normal growth to homocysteine medium in MB468 breast cancer cells and the methionine stress MB468RES derivatives. In addition we have developed cancer cell lines with shRNAs targeting methionine adenosyltransferase (MAT), the enzyme catalyzing of SAM from methionine and ATP. Inducible knockdown of MAT allows us to reduce SAM synthesis. Our previous results suggest that SAM limitation is the trigger for cancer cell methionine dependence. Thus metabolite profiling using MAT knockdown system will provide an independent dataset that together with profiles from the MB468 and MB468RES cell line pair will define critical profiles related to cancer cell methionine dependence.Ā In the current untargeted analysis of primary metabolites and complex lipids, coupled with analysis of methionine pathway intermediates (folate and respective s-adenosylmethoinine, s-adenosylhomocysteine, choline, betaine) and metabolic will be conducted on MB468, MB468RES and MB468shRNA following the switch from containing media to homocysteine containing media over the course of 0, 2, 4, 12, 24 and 48 hours.Ā The primary objectives were to 1) characterize the response to methionine stress and SAM limitation and 2) correlate the metabolic with cancer cell proliferation arrest and death.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Metabolic Profiling of Visceral and Subcutaneous Adipose Tissue from Colorectal Patients: GC-TOF MS analyis of subcutaneous and visceral adipose tissue samples
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Johanna Lampe (Fred Hutchinson Cancer Research Center at Univ. of Washington, In the current investigation, unbiased profiling of the metabolome and lipidome adipose tissue samples (visceral(VAT) and subcutaenous (SAT)) and serum of 50 patients, including stages I-IV, from the Fred Hutchinson Cancer Center and the German Cancer Research Center (Heidelberg, Germany) was conducted. The and metabolome of adipose tissue (VAT/SAT) and serum were analyzed using UPLC-QTOFMS analysis and GC-TOFMS analyses, respectively.Ā The primary of this project were to 1) compare the metabolome and lipidome SAT adipose tissue of n=50 Colorectal Cancer Cell (CRC) patients, 2) the associations between the lipidome and metabolome in adipose tissue and serum of n=50 CRC patients and 3) test the associations between the of VAT and serum with the tumor stage of CRC patients.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Biomarkers for Depression in Human Plasma in a Population Sample
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Roel Ophoff (UC Los Angeles ) and Steve HorwathĀ (UC Los Angeles ) . Major disorder (MDD) is one of the most debilitating disorders in the United States a 12-month prevalence of 6.7% in the adult population. The disorder affects of Americans daily and is a major health concern with enormous economic cost society at large. Criteria for MDD diagnosis and treatment are based on various and symptoms not always fitting into strict diagnostic categories such as Despite various known risk factors (such as family history, age, and gender), markers supporting diagnosis or prediction of MDD are unavailable. We have cerebrospinal fluid (CSF) and peripheral blood of more than 600 subjects from general population. For each of the participants we also obtained biometric as well as behavioral trait measures. One of the measures is the Beck Inventory (BDI), a well-established questionnaire for measuring severity of Based on the BDI, roughly 5% of participants suffer from severe depressive while most of these subjects are not under treatment or receiving any for depressionĀ In the current investigation, untargeted analysis of primary was conducted onĀ age and genderĀ matchedĀ human cerebrospinal fluid (CSF) and from subjects suffering with MDDĀ (Ā n=50) and control subjects (n=50). were diagnosed as having MDD based on the Beck Depression Inventory.The primary of this study were to 1) identify metabolites which discriminate between with and without depression symptoms in the CSF and plasma and 2) how changes between CSF and plasma.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Biomarkers for Depression in Human Cerebrospinal Fluid in a Population Sample
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Roel Ophoff (UC Los Angeles ) and Steve HorwathĀ (UC Los Angeles ) . Major disorder (MDD) is one of the most debilitating disorders in the United States a 12-month prevalence of 6.7% in the adult population. The disorder affects of Americans daily and is a major health concern with enormous economic cost society at large. Criteria for MDD diagnosis and treatment are based on various and symptoms not always fitting into strict diagnostic categories such as Despite various known risk factors (such as family history, age, and gender), markers supporting diagnosis or prediction of MDD are unavailable. We have cerebrospinal fluid (CSF) and peripheral blood of more than 600 subjects from general population. For each of the participants we also obtained biometric as well as behavioral trait measures. One of the measures is the Beck Inventory (BDI), a well-established questionnaire for measuring severity of Based on the BDI, roughly 5% of participants suffer from severe depressive while most of these subjects are not under treatment or receiving any for depressionĀ In the current investigation, untargeted analysis of primary was conducted onĀ age and genderĀ matchedĀ human cerebrospinal fluid (CSF) and from subjects suffering with MDDĀ (Ā n=50) and control subjects (n=50). were diagnosed as having MDD based on the Beck Depression Inventory.The primary of this study were to 1) identify metabolites which discriminate between with and without depression symptoms in the CSF and plasma and 2) how changes between CSF and plasma.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Metabolic Profiling of Visceral and Subcutaneous Adipose Tissue from Colorectal Patients: GC-TOF MS analyis of serum samples
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Johanna Lampe (Fred Hutchinson Cancer Research Center at Univ. of Washington, In the current investigation, unbiased profiling of the metabolome and lipidome adipose tissue samples (visceral(VAT) and subcutaenous (SAT)) and serum of 50 patients, including stages I-IV, from the Fred Hutchinson Cancer Center and the German Cancer Research Center (Heidelberg, Germany) was conducted. The and metabolome of adipose tissue (VAT/SAT) and serum were analyzed using UPLC-QTOFMS analysis and GC-TOFMS analyses, respectively.Ā The primary of this project were to 1) compare the metabolome and lipidome SAT adipose tissue of n=50 Colorectal Cancer Cell (CRC) patients, 2) the associations between the lipidome and metabolome in adipose tissue and serum of n=50 CRC patients and 3) test the associations between the of VAT and serum with the tumor stage of CRC patients.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Metabolomic profiling of influenza: a 2009 pandemic H1N1 influenza in lean and mice (via blood and tissue)
STUDY_TYPE
Metabolomics analysis on the effec of diet-related obesity on the immune to pH1N1 infection
STUDY_SUMMARY
Lung samples were obtained at 15 weeks from 56 mice that had received either a low fat diet, or a high fat diet and that were uninfected, 4 days or 8 days post-infection.
Effect of diet and age on ovarian metabolome (via tissue)
STUDY_TYPE
Metabolomics analysis on the ovarian metabolome
STUDY_SUMMARY
Ovarian samples from twenty-one adult, female Cynomolgus monkeys were studied, of which were fed the Western #907 diet, and 13 of which were fed the Prudent diet. HPLC-MS data were acquired for the 21 samples.
Effect of diet and age on ovarian metabolome (serum metabolome compared to
STUDY_TYPE
Metabolomics analysis on the serum metabolome for comparison against the metabolome
STUDY_SUMMARY
Serum samples from twenty-one adult, female Cynomolgus monkeys were studied, of which were fed the Western #907 diet, and 13 of which were fed the Prudent diet. HPLC-MS data were acquired for the 21 samples.
Genetic effects of high fat diet on mouse fecal metabolomics
STUDY_TYPE
Metabolomics analysis on the effect of genetics and diet on the metabolism of GI tract and obesity
STUDY_SUMMARY
This study includes 72 female mice with 4 mice from each of the 18 mice Two mice from each strain were fed a high fat diet and two mice were fed a fat diet. The 36 mice fed a normal fat diet will serve as the controls. All are age 27.6 weeks or older at the time of sacrifice. UPLC-MS data was for all 72 samples.
Combined Metabolomics and Lipidomics of Type 1 Diabetes (QTOF)
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project granted to Bell Ā (University of Chicago) and collaborator Manami Hara uses a multi-omics to investigate the metabolome (primary metabolites), lipidome (complex lipids) signaling lipids (oxylipins) in the plasma of Non-obese Diabetic (NOD) mice progressed to Type 1 Diabetes Mellitus (T1D) and those that did not. NOD Mice were assessed as diabetic or non-diabetic based on their fasting (4hr) blood levels at sacrifice, which defined 30 hyperglycemic (glucose = 250 mg/dL) and normoglycemic animals. The primary objective of this study was to identify biomarkers associated with beta-cell destruction/survival and T1D progression.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Metabolite changes associated with methionine stress sensitivity of cancer (CSH MS analysis)
STUDY_TYPE
timecourse study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project granted to Peter Kaiser (UC Irvine), aims to achieve understanding of a unique metabolic dependence of cancer cells to explore development of novel unconventional therapeutic strategies that exploit dependence of cancer cells on methyl-donor abundance. The past few years have highlighted the role of altered metabolism in cancer. While mechanistic insight into changed metabolism in cancer is very limited, the importance of the metabolic pathway surrounding homocysteine and methionine for cancer cell proliferation has been known for over 30 years. These findings, generally summarized as methionine-dependence or methionine stress sensitivity, describe the phenomenon that most cancer cells cannot proliferate in growth medium where the amino acid methionine is replaced with its direct metabolic precursor homocysteine. Importantly, non-tumorigenic cells are unaffected by replacing methionine with homocysteine in the growth medium. For the past years we have been studying methionine dependence of breast and prostate cancer and demonstrated that methionine-dependence is caused by insufficient flux through this pathway to sustain synthesis of the downstream metabolite and the principal methyl-donor S-adenosylmethionine (SAM). We have isolated rare cell clones from MDA-MB468 breast cancer cells (referred to as MB468RES) that are no longer methionine dependent and proliferate in homocysteine medium. Interestingly, MB468RES have lost their ability for anchorage independent growth, a hallmark of cancer. The MB468 and MB468RES cell line pair confirms other observations showing that methionine dependence is tightly linked to tumorigenicity. Importantly, this cell line pair is an ideal model to identify metabolite signatures linked to cancer cell methionine dependence. We propose to characterize the metabolic changes triggered by the shift from normal growth medium to homocysteine medium in MB468 breast cancer cells and the methionine stress insensitive MB468RES derivatives. In addition we have developed cancer cell lines with inducible shRNAs targeting methionine adenosyltransferase (MAT), the enzyme catalyzing synthesis of SAM from methionine and ATP. Inducible knockdown of MAT allows us to specifically reduce SAM synthesis. Our previous results suggest that SAM limitation is the critical trigger for cancer cell methionine dependence. Thus metabolite profiling using the MAT knockdown system will provide an independent dataset that together with metabolite profiles from the MB468 and MB468RES cell line pair will define critical metabolic profiles related to cancer cell methionine dependence. In the current investigation, untargeted analysis of primary metabolites and complex lipids, coupled with quantitative analysis of methionine pathway intermediates (folate and respective derivatives, s-adenosylmethoinine, s-adenosylhomocysteine, choline, betaine) and metabolic flux will be conducted on MB468, MB468RES and MB468shRNA following the switch from methionine containing media to homocysteine containing media over the course of 0, 2, 4, 8, 12, 24 and 48 hours. The primary objectives were to 1) characterize the metabolic response to methionine stress and SAM limitation and 2) correlate the metabolic signatures with cancer cell proliferation arrest and death.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
Metabolic Profiling of Visceral and Subcutaneous Adipose Tissue from Colorectal Patients: UHPLC-QTOF MS analyis of subcutaneous and visceral adipose tissue
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Johanna Lampe (Fred Hutchinson Cancer Research Center at Univ. of Washington, In the current investigation, unbiased profiling of the metabolome and lipidome adipose tissue samples (visceral(VAT) and subcutaenous (SAT)) and serum of 50 patients, including stages I-IV, from the Fred Hutchinson Cancer Center and the German Cancer Research Center (Heidelberg, Germany) was conducted. The and metabolome of adipose tissue (VAT/SAT) and serum were analyzed using UPLC-QTOFMS analysis and GC-TOFMS analyses, respectively.ĆĀ The primary of this project were to 1) compare the metabolome and lipidome SAT adipose tissue of n=50 Colorectal Cancer Cell (CRC) patients, 2) the associations between the lipidome and metabolome in adipose tissue and serum of n=50 CRC patients and 3) test the associations between the of VAT and serum with the tumor stage of CRC patients.ĆĀ
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
NUM_GROUPS
2
TOTAL_SUBJECTS
59
STUDY_COMMENTS
Lipidomics profiles for studyThis is the SAT and VAT part of the studyLabel-ID conserved for all parts of the study---The samples had to be diluted for mode measurements (therefore 2 dilutions are measured for the same sample). are then combined and scaling factor is used for final results.
Metabolic Profiling of Visceral and Subcutaneous Adipose Tissue from Colorectal Patients: UHPLC-QTOF MS analyis of serum samples
STUDY_TYPE
biomarker study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Johanna Lampe (Fred Hutchinson Cancer Research Center at Univ. of Washington, In the current investigation, unbiased profiling of the metabolome and lipidome adipose tissue samples (visceral(VAT) and subcutaenous (SAT)) and serum of 50 patients, including stages I-IV, from the Fred Hutchinson Cancer Center and the German Cancer Research Center (Heidelberg, Germany) was conducted. The and metabolome of adipose tissue (VAT/SAT) and serum were analyzed using UPLC-QTOFMS analysis and GC-TOFMS analyses, respectively.Ā The primary of this project were to 1) compare the metabolome and lipidome SAT adipose tissue of n=50 Colorectal Cancer Cell (CRC) patients, 2) the associations between the lipidome and metabolome in adipose tissue and serum of n=50 CRC patients and 3) test the associations between the of VAT and serum with the tumor stage of CRC patients.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
NUM_GROUPS
1
TOTAL_SUBJECTS
59
STUDY_COMMENTS
Lipidomics profiles for studyThis is the serum part of the studyLabel-ID os for all parts of the study---
A Multi-Omic View of Host-Pathogen-Commensal Interplay in Salmonella-Mediated Infection
STUDY_TYPE
Timecourse of Infection
STUDY_SUMMARY
The potential for commensal microorganisms indigenous to a host (the or ĀmicrobiotaĀ) to alter infection outcome by influencing host-pathogen is largely unknown. We used a multi-omics ĀĀsystemsĀĀ approach, proteomics, metabolomics, glycomics, and metagenomics, to explore the molecular between the murine host, the pathogen Salmonella enterica serovar Typhimurium Typhimurium), and commensal gut microorganisms during intestinal infection with Typhimurium. We find proteomic evidence that S. Typhimurium thrives within the 129/SvJ mouse gut without antibiotic pre-treatment, inducing inflammation and the intestinal microbiome (e.g., suppressing Bacteroidetes and Firmicutes while growth of Salmonella and Enterococcus). Alteration of the host microbiome structure was highly correlated with gut environmental changes, including the of metabolites normally consumed by commensal microbiota. Finally, the less phase of S. TyphimuriumĀs lifecycle was investigated, and both proteomic and evidence suggests S. Typhimurium may take advantage of increased fucose to metabolize fucose while growing in the gut. The application of multiple measurements to Salmonella-induced intestinal inflammation provides insights complex molecular strategies employed during pathogenesis between host, and the microbiome.
Model-driven multi-omic data analysis elucidates metabolic immunomodulators of activation
STUDY_TYPE
growth condition, timecourse
STUDY_SUMMARY
Macrophages are central players in immune response, manifesting divergent to control inflammation and innate immunity through release of cytokines and signaling factors. Recently, the focus on metabolism has been reemphasized as signaling and regulatory pathways of human pathophysiology, ranging from cancer aging, often converge on metabolic responses. Here, we used genome-scale and multi-omics (transcriptomics, proteomics, and metabolomics) analysis to metabolic features that are critical for macrophage activation. A genome-scale network for the RAW 264.7 cell line was constructed to determine metabolic of activation. Metabolites well-known to be associated with immunoactivation and arginine) and immunosuppression (tryptophan and vitamin D3) were among the critical effectors. Intracellular metabolic mechanisms were assessed, a suppressive role for de-novo nucleotide synthesis. Finally, underlying mechanisms of macrophage activation were identified by analyzing multi-omic obtained from LPS-stimulated RAW cells in the context of our flux-based This study demonstrates that the role of metabolism in regulating activation be greater than previously anticipated and elucidates underlying connections activation and metabolic effectors. This submission corresponds to the data from this study.
Salmonella Modulates Metabolism during Growth under Conditions that Induce of Virulence Genes
STUDY_TYPE
growth conditions, timecourse
STUDY_SUMMARY
Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative that uses complex mechanisms to invade and proliferate within mammalian host To investigate possible contributions of metabolic processes to virulence in S. grown under conditions known to induce expression of virulence genes, we used a systems biology approach coupled with genome scale modeling. First, we distinct metabolite profiles associated with bacteria grown in either rich or media and report the most comprehensive coverage of the S. Typhimurium to date. Second, we applied an omics-informed genome scale modeling analysis of functional consequences of adaptive alterations in S. Typhimurium metabolism growth under our conditions. Modeling efforts highlighted a decreased cellular to both produce and utilize intracellular amino acids during stationary phase in virulence conditions, despite significant abundance increases for these as observed by our metabolomics measurements. Furthermore, analyses of omics in the context of the metabolic model indicated rewiring of the metabolic to support pathways associated with virulence. For example, cellular of polyamines were perturbed, as well as the predicted capacity for secretion uptake.
A study of changes in lipid metabolism of ovarian cancer cells co-cultured with GC-TOF MS analysis
STUDY_TYPE
timecourse study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Ernst Lengyel Ā (University of Chicago).The biology of ovarian cancer (OvCa) is distinct from that of most epithelial tumors, in that hematogenous metastases rare, and ovarian tumors remain confined to the peritoneal cavity. The omentum, large pad of fat tissue (20x13x3cm) covering the bowel, is the most common site OvCa metastasis. It consists primarily of adipocytes, which become the microenvironment for the OvCa cells. The underlying hypothesis for this is that, in the presence of adipocytes, the metabolism of OvCa cells is and shifts towards lipid utilization, which provides energy that facilitates growth and metastasis. Preliminary results suggest that primary human omental secrete cytokines which promote the metastasis of OvCa cells to the omentum and subsequent invasion. Once metastasis has occurred, OvCa cells induce lipolysis omental adipocytes, and use the energy derived from these lipids to study the metabolic changes in the tumor microenvironment we have established a organotypic culture of the human omentum using primary human cells established patient tissue. Metabolic studies will be performed on adipocytes and OvCa individually, on conditioned media and on adipocytes and OvCa cells co-cultured our 3D model, with the goal of arriving at a comprehensive analysis of primary and lipids in the tumor microenvironment.In the current investigation, analysis of primary metabolites and complex lipids were conducted on adipocytes OvCa cells individually, on conditioned media and on adipocytes and OvCa cells in our 3D model. Analysis of oxylipins was conducted on conditioned media. To better understanding of the dynamic regulation of metabolic pathways we will perform metabolic flux analysis using labeled cells (13C-glucose, in the 3D culture model.The primary objective of this study is to gain insight the dynamic interactions between OvCa cells and human adipocytes with the of elucidating targets of therapeutic intervention.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
NUM_GROUPS
2
TOTAL_SUBJECTS
14
STUDY_COMMENTS
Lipidomics profiles for studyFor the co-culture Human Adipocytes were grown in of SKOV3ip1 ovarian cancer cellsFor control samples the adipocytes were grown the absence of SKOV3ip1 ovarian cancer cells---Exp design 2 x 14Final result is by merging results from both files and applying dilution factor.Reason was high concentration in positive mode onlyRaw Data File (Positive Mode_TGs) Data File (Positive Mode_Non-TGs) (dilution2)
A study of changes in lipid metabolism of ovarian cancer cells co-cultured with UHPLC-QTOF MS analysis
STUDY_TYPE
timecourse study
STUDY_SUMMARY
This West Coast Metabolomics Center pilot and feasibility project was granted Ernst Lengyel Ā (University of Chicago).The biology of ovarian cancer (OvCa) is distinct from that of most epithelial tumors, in that hematogenous metastases rare, and ovarian tumors remain confined to the peritoneal cavity. The omentum, large pad of fat tissue (20x13x3cm) covering the bowel, is the most common site OvCa metastasis. It consists primarily of adipocytes, which become the microenvironment for the OvCa cells. The underlying hypothesis for this is that, in the presence of adipocytes, the metabolism of OvCa cells is and shifts towards lipid utilization, which provides energy that facilitates growth and metastasis. Preliminary results suggest that primary human omental secrete cytokines which promote the metastasis of OvCa cells to the omentum and subsequent invasion. Once metastasis has occurred, OvCa cells induce lipolysis omental adipocytes, and use the energy derived from these lipids to study the metabolic changes in the tumor microenvironment we have established a organotypic culture of the human omentum using primary human cells established patient tissue. Metabolic studies will be performed on adipocytes and OvCa individually, on conditioned media and on adipocytes and OvCa cells co-cultured our 3D model, with the goal of arriving at a comprehensive analysis of primary and lipids in the tumor microenvironment.In the current investigation, analysis of primary metabolites and complex lipids were conducted on adipocytes OvCa cells individually, on conditioned media and on adipocytes and OvCa cells in our 3D model. Analysis of oxylipins was conducted on conditioned media. To better understanding of the dynamic regulation of metabolic pathways we will perform metabolic flux analysis using labeled cells (13C-glucose, in the 3D culture model.The primary objective of this study is to gain insight the dynamic interactions between OvCa cells and human adipocytes with the of elucidating targets of therapeutic intervention.Ā
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility451 Health Sciences DriveDavis, CA
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
NUM_GROUPS
2
TOTAL_SUBJECTS
14
STUDY_COMMENTS
Lipidomics profiles for studyFor the co-culture Human Adipocytes were grown in of SKOV3ip1 ovarian cancer cellsFor control samples the adipocytes were grown the absence of SKOV3ip1 ovarian cancer cells---Exp design 2 x 14Final result is by merging results from both files and applying dilution factor.Reason was high concentration in positive mode onlyRaw Data File (Positive Mode_TGs) Data File (Positive Mode_Non-TGs) (dilution2)
Quantitative Metabolomics by 1H-NMR and LC-MS/MS Confirms Altered Metabolic Pathways in Diabetes
STUDY_TYPE
Pre- and Post- insulin study with matched controls
STUDY_SUMMARY
We obtained plasma samples from 7 c-peptide negative type 1 diabetic individuals (T1D) and 7 non-diabetic controls (Con) that were matched for age (T1D = 31.1Ā±2.9 yrs, Con = 30.2Ā±3.4 yrs), body mass (T1D = 80.2Ā±4.7kg, Con = 81.9Ā±7.4 kg) and BMI (T1D = 26.5Ā±1.2 kg/m2, Con = 25.2Ā±1.3 kg/m2). Type 1 diabetic people were studied while treated with insulin and also after 8 hours of insulin deprivation
A statistical analysis of the effects of urease pre-treatment on the of the urinary metabolome by gas chromatographyĀmass spectrometry
STUDY_TYPE
Analytical Comparison
STUDY_SUMMARY
Urease pre-treatment of urine has been utilized since the early 1960s to remove levels of urea from samples prior to further processing and analysis by gas spectrometry (GCĀMS). Aside from the obvious depletion or elimination of urea, effect, if any, of urease pre-treatment on the urinary metabolome has not been in detail. Here, we report the results of three separate but related that were designed to assess possible indirect effects of urease pre-treatment the urinary metabolome as measured by GCĀMS. In total, 235 GCĀMS analyses performed and over 106 identified and 200 unidentified metabolites were across the three experiments. The results showed that data from urease samples (1) had the same or lower coefficients of variance among reproducibly metabolites, (2) more accurately reflected quantitative differences and the ratios among different urine volumes, and (3) increased the number of identifications. Overall, we observed no negative consequences of urease In contrast, urease pre-treatment enhanced the ability to distinguish between and biological sample types compared to no treatment. Taken together, these show that urease pre-treatment of urine offers multiple beneficial effects that any artifacts that may be introduced to the data in urinary metabolomics
Metabolomics Analysis of Frontal Fibrosing Alopecia
STUDY_TYPE
Unaffected and Affected patient scalp biopsies
STUDY_SUMMARY
Scarring alopecia consists of a collection of disorders characterized by of hair follicles, replacement with fibrous scar tissue, and irreversible hair Alopecia affects men and women worldwide and can be a significant source of stress and depression for affected individuals. The purpose of this study was explore metabolic profiles in scalp tissue samples from normal control subjects and in matched samples obtained from affected (n=12) and unaffected (n=12) of the scalp in patients with lymphocytic Frontal Fibrosing Alopecia (FFA). fibrosing alopecia results from destruction of hair follicles by an lymphocytic infiltrate that is localized around the upper portion of the hair
INSTITUTE
Case Western Reserve University
DEPARTMENT
Dermatology
LABORATORY
Karnik Lab
LAST_NAME
Karnik
FIRST_NAME
Pratima
ADDRESS
-
EMAIL
psk11@case.edu
PHONE
216-368-0209
NUM_GROUPS
3 groups-Paired unaffected and affected (n=12),Normals(n=6)
TOTAL_SUBJECTS
Patients (N=12), Normals (N=6)
STUDY_COMMENTS
Affected scalp biopsies were obtained from frontal scalp, Unaffected from scalp. Normal scalp biopsies were obtained from the occipital scalp.
Dysfunctional lipid metabolism underlies the effect of perinatal DDT exposure the development of metabolic syndrome
STUDY_TYPE
Chemical dosage and feeding study
STUDY_SUMMARY
Targeted metabolomic analysis of bile acids was performed on 15 mouse liver collected from mice euthanized at 9 months following consumption of a high fat w/o perinatal DDT exposure. Funded by the National Institute of Health (R00 R03 DK082724, U24 DK092993, U24 DK097154, T32 ES007059, and P60 DK020541), the Diabetes Association, and USDA-ARS intramural Project 5306-51530-019-00D. were analyzed by UPLC-MS/MS using a Waters Acquity UPLC and detected on an API QTrap (AB Sciex, Framingham, MA, USA) by multiple reaction monitoring (MRM) negative mode electrospray ionization.
A study of changes in lipid metabolism of ovarian cancer cells co-cultured with adipocytes: UPLC-QTRAP MS analysis
STUDY_TYPE
Timecourse
STUDY_SUMMARY
The study investigated the interaction between omental adipocytes and OvCa cells, as a follow up to preliminary data indicating this leads to reprograming of metabolic (especially lipid) profiles in both adipocytes and OvCa cells as ovarian cancer cells (OvCa) readily metastasize to the omental fat pad in the abdomen and stimulate the release of fatty acids. In order to mimic the interaction between OvCa and omental adipocytes during metastasis, a coculture system was used that employed OvCa cells and primary human adipocytes isolated from omentum. Human primary adipocytes were isolated from omental explants from patients undergoing surgery for benign conditions. After surgical removal, omental tissue was digested with collagenase I, and primary cultures of adipocytes were established, characterized, and incorporated into the co-culture. The primary adipocytes were isolated and co-cultured with the OvCa cell line Skov3ip1. In this current submission, the the samples will be collected at 4, 18 and 24 hour time points post co-culture to determine the time dependent effect on lipid mediators, including oxylipins and ceramides. The study results included in this DRCC submission were the 18 hour time point data for oxylipins and ceramides from targeted metabolomic analysis of lipid mediators performed by the Newman lab.
Measurements were performed on serum using 6 month old C57/B10 control (n=6) and mdx (n=8) mice. 1D proton and carbon spectra were collected on these samples
INSTITUTE
University of Florida
DEPARTMENT
Department of Biochemistry and Molecular Biology / SECIM
LABORATORY
Arthur Edison
LAST_NAME
Edison
FIRST_NAME
Arthur
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry & Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245
Two groups of mixtures with 5 replicates each were each made using 20 synthetic metabolites. Some metabolites were at equal concentrations in both groups, and 10 metabolites differed between groups with half higher in group A, and half higher in group B.
INSTITUTE
University of Florida
DEPARTMENT
Department of Biochemistry and Molecular Biology /SECIM
LABORATORY
Arthur Edison
LAST_NAME
Arthur
FIRST_NAME
Edison
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry & Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245
Cold adaptation shapes the robustness of metabolic networks in Drosophila melanogaster
STUDY_TYPE
Time course during cold exposure for four genetic lines of flies
STUDY_SUMMARY
Metabolites of cold hardy versus cold susceptible flies were compared using N less than R-based metabolomics. We used 8 replicates per line (2 hardy lines, two susceptible lines), and sampled each line at three time points (before, during and after cold), giving rise to 96 samples total.
Factors for Epigenetic Silencing of Lung Cancer Genes
STUDY_TYPE
Metabolomic analysis of Plasma
STUDY_SUMMARY
To test the hypothesis that the propensity for silencing of tumor suppressor genes in the respiratory epithelium of chronic smokers by promoter hypermethylation is influenced by sequence variations that modify the activity of genes and microRNAĆs that directly or indirectly influence de novo methylation and chromatin remodeling.
This experiment is investigating the metabolome of men, healthy women, and with Polycystic Ovary Syndrome (PCOS) with and without Obstructive Sleep Apnea
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomic and lipidomic profiles in response to exogenous insulin and GLP-1 during prolonged fasting
STUDY_TYPE
Timecourse
STUDY_SUMMARY
Seventeen northern elephant seal pups constituting four different cohorts at Nuevo State Reserve were studied at two post-weaning periods: early (1-2 wk weaning; n=5) and late (6-7 weeks post weaning; n=12). Study #1: Prior to each protocol, plasma U/kg). Following the infusion, blood samples were collected at determine the effects of prolonged fasting on peripheral insulin activity and samples were collected. Ten fasting seal pups (n=5 early, n=5 late) were (i.v.) with a mass-specific dose of insulin (0.065 U/kg). Following the blood samples were collected at 10, 30, 60, 90, and 120 minutes. Study #2: late-fasted seal pups were administered either a low (LDG; 10 pmol/kg; n=3) or (HDG; 100 pmol/kg; n=4) dose of GLP-1 immediately following a glucose bolus g/kg) (i.v.) infused within 2 mins. the infusions, blood samples were collected 10, 30, 60, 90, 120, and 150 minutes.
SIRM Analysis of human P493 cells under hypoxia in [U-13C/15N] labeled medium (Positive ion mode FTMS)
STUDY_TYPE
tracer
STUDY_SUMMARY
SIRM Analysis of MYC inducible P493 cells under normoxia and hypoxia in labeled Glutamine medium
INSTITUTE
University of Kentucky
DEPARTMENT
Toxicology
LAST_NAME
Fan
FIRST_NAME
Teresa
ADDRESS
-
EMAIL
twmfan@gmail.com
PHONE
-
NUM_GROUPS
4
TOTAL_SUBJECTS
8
STUDY_COMMENTS
For protocols and additional information, see Le, A., Lane, A.N., Hamaker, M., S., Barbi, J., Tsukamoto, T., Rojas, C.J., Slusher, B.S., Zhang, H., Zimmerman. Liebler, D.C., Slebos, R.J.C., Lorkiewicz, P.K., Higashi, R.M., Fan, T.W-M., Dang, C.V. Cell Metabolism 15, 110 (2012). 13C/15N Gln was used as the tracer this study, even though the analysis did not explicity look for 15N
Impact of insulin deprivation and treatment on sphingolipid distribution in muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice
STUDY_TYPE
Insulin depravation
STUDY_SUMMARY
Experiments were conducted using 13-wk-old male C57BL/6J mice (Jackson Bar Harbor, ME). Mice were housed individually with free access to water and (TD.10112; Harlan Laboratories, Indianapolis, IN), with a 12:12-h light-dark and temperature and humidity control. Mice were acclimated for 1 wk prior to beginning of the experiment. The protocol was approved by the Mayo Clinic Animal Care and Use Committee. Following a 6-h fast, mice were given injections of STZ (125 mg/kg; in sodium acetate buffer, pH = 4.5) (67). were repeated on the following day. Control animals received intraperitoneal of vehicle. Only mice that displayed blood glucose ?300 mg/dl and an increase blood ketones (both values by Precision Xtra glucometer; Abbott Laboratories, Park, IL), hyperphagia, and polyuria and were positive for urine glucose via dipstick (Uristix, Bayer, Pittsburgh, PA) on day 7 after the first STZ dose included in the experiment. Animals that were positive for STZ diabetes LinBit subcutaneous insulin implant (LinShin Canada, Toronto, ON, Canada) (79) pentobarbital sodium anesthesia (Nebutal, 40 mg/kg of body wt) according to the protocol. Each animal received two subcutaneous implants (total dose: 0.2 U/24 for >30 days, 10 U/kg for 20-g mice). Insulin treatment was continued for 3 wk. animals (C; n = 13) received blank implants. Diabetic control was confirmed by measurements of blood and urinary glucose. In some cases, when urine glucose present and blood glucose was >288 mg/dl, the animal received a third implant. insulin treatment was continued until initially lower plasma glucose content in animals reached control values. Three weeks following implantation, diabetic were divided randomly into diabetic-treated (D + I; n = 13) and (D ? I; n = 13) groups. Insulin implants were removed from the D ? I group pentobarbital anesthesia, which led to the return of the diabetic phenotype 24 h. Animals from the D + I group continued on insulin treatment (Fig. 1). At age of 18 wk, animals from all groups were analyzed for body composition by an Body Composition Analyzer (EchoMRI, Houston, TX) and euthanized by decapitation wk after the initial STZ or vehicle dose. Figure 1 depicts the timeline of the and blood glucose profiles for each experimental group. Additional animals were for estimation of skeletal muscle insulin sensitivity by acute insulin The mice were divided into the C (n = 6), D ? I (n = 7), and D + I (n = 7) and followed appropriate experimental treatment, except for acute insulin 10 min prior to euthanization by pentobarbital overdose. Figure 1 of the PDF of the article summarizes the study design
Disruption of Zinc homeostasis can impair maternal glucocorticoid metabolism: on the developing fetus
STUDY_TYPE
steroid panel in pregnant rats
STUDY_SUMMARY
Steroids play a broad and vital role in regulation of gene expression, sexual characteristics, maturation, reproduction, and neurological functions; an imbalance in steroid metabolism is also linked to development and of many diseases including autism. Prenatal stress of different nature has been to affect both the mother and the offspring. Adverse nutritional conditions gestation can impair the maternal hypothalamic-pituitary-adrenal axis (HPA) and the fetus to high levels of glucocorticoids (GC). Evenwhen GC are required for brain development; an increased exposure of the fetus to GC as a consequence of stress can affect fetal hypothalamic-pituitary-gonad axis (HPG) development, neurogenesis, and have a long term impact on the offspringĀs mental health. zinc availability can occur during pregnancy as a consequence of different (nutritional deficiency, infections, diabetes, alcohol consumption, and to certain toxicants). Importantly, several of these gestational conditions been linked to autism. In fact, alterations in maternal zinc homeostasis upon to select environmental stressors (e.g. the phthalate plasticizer phthalate (DEHP)) that have become increasingly common since the industrial may underlie the recent rise in the incidence of autism.Alterations in maternal homeostasis could expose the fetus to high GC concentrations secondary to a maternal GC production and/or to a decreased capacity of the placenta to GC to inactive metabolites. The overall goal of this proposal is to investigate alterations in zinc homeostasis during gestation triggered by either a marginal nutrition or exposure to an environmental pollutant (the phthalate plasticizer phthalate (DEHP)) can impair maternal and fetal endocrine signaling leading to fetal brain development.
Impact of anesthesia and euthanasia on metabolomics of mammalian tissues: in a C57BL/6J mouse model
STUDY_TYPE
Anesthesia effect study
STUDY_SUMMARY
We examined the effect of several commonly-used methods of anesthesia and for collection of skeletal muscle, liver, heart, adipose and serum of C57BL/6J The data revealed tissue-specific impacts of different anesthesia and strategies. Based on these findings, we present a more optimal collection mammalian tissues and recommend that rodent tissues intended for metabolomics be collected under anesthesia rather than post-euthanasia.
INSTITUTE
University of Michigan
DEPARTMENT
Internal Medicine
LABORATORY
Evans Lab / Burant Lab
LAST_NAME
Charles
FIRST_NAME
Evans
ADDRESS
6321 Brehm Tower, 1000 Wall Street, Ann Arbor MI 48105
Monitoring In Vitro Response of Selenium-Treated Prostate Cells by 1H NMR Spectroscopy
STUDY_TYPE
Metabolite level response after treatment with organoselenium
STUDY_SUMMARY
Metabolomics analysis was performed on DU145 prostate cancer cells and PNT1A non-tumorigenic prostate cells after treatment with selenomethionine and Se-methylselenocysteine using 800 MHz Bruker NMR spectrometer on 18 cell samples.
Targeted lipids were quantified and compared to the quantities of other labs the nation. Comparisons were between pooled serum from humans who have consumed seed for the past month, pooled serums from humans who have consumed fish oil the last month, and pooled serum from humans who have not consumed either flax or fish oil for the past month. Pooled samples were prepared in triplicate.
INSTITUTE
University of Florida
DEPARTMENT
College of Medicine, Department of Pathology, Immunology, and Laboratory
Aliquots of Jurkat T-lymphocyte cells were washed with 5 different rinsing (0.3% amm. formate, 0.3% amm. acetate, 0.9% NaCl, 1 M PBS, 100 mM PBS) and ran triplicate to monitor the effect of ion suppression on the electrospray signal.
Metabolic analysis of Normal Mouse Lung Fiboblasts with/without TGFbeta treatment (Part 1)
STUDY_TYPE
Glycolysis/TCA/Nucleotide analysis (tissue/cells)
STUDY_SUMMARY
This study is a part of series performed for the same researcher through pilot/feasibility grant program, so the publication is relevant reference for other studies, see the project for this study. This specific experiment is a small pilot study to establish method performance, it includes four biological replicas of identical cell cultures after the identical treatment and a single tissue sample.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Labeling of cells was carried out in triplicate with each unstimulated sample containing 20e6 cells and stimulated cells containing 12.8e6 cells. Cells were deprived of glucose or glutamine for 1 h then resuspended in 10mM [13C1,2] D-glucose labeled medium. At this time cells were also activated with 1 ug/ml anti-CD3 and anti-CD28. Cells were labeled (and activated) for 12 h when 1 ml of spent medium was collected and cells were resuspended in 200 ul ice cold methanol. All samples were immediately stored in -80Ā°C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Cells were deprived of glucose or glutamine for 1 h then resuspended in 10mM [13C1,2] D-glucose labeled or 4 mM [13C5]-U-glutamine labeled medium, respectively. At this time cells were also activated with 5 ug/ml anti-CD3 and anti-CD28. Cells were labeled (and activated) for 12 h when samples were spun down, and cells were resuspended in 200 ul ice cold methanol
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Jurkat parental cells (JP6) or cells stably expressing the pCDNA Noxa vector (N5) cells were cultured to confluence and counted. 20E6 cells in triplicate were resuspended in glucose free or glutamine free medium for 2 hours. Cells were then pelleted and resuspended in 13C-1,2 Glucose (10mM) or 13C-U-glutamine (4mM) for 24 hours. Cells were pelleted and spent medium was collected. Cell pellets were washed 1X with ice cold PBS and cell pellets were resuspended in 400ul -20 methanol. Cells and media were snap frozen in liquid nitrogen and stored at -80.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
High Insulin Combined With Essential Amino Acids Stimulates Skeletal Muscle Protein Synthesis While Decreasing Insulin Sensitivity in Healthy Humans
STUDY_TYPE
High and low insulin with and without essential amino acids
STUDY_SUMMARY
Thirty participants were randomized to 3 groups of 10 each with each studied twice. Study groups comprised (1) low and high insulin, (2) low insulin and without EAAs, and (3) high insulin with and without EAAs.
Association of Metabolic Profile and Microbiome in Chronic Pressure Ulcer Wounds
STUDY_SUMMARY
Chronic, non-healing wounds contribute significantly to the suffering of patients with co-morbidities in the clinical population with mild to severely compromised immune systems. Normal wound healing proceeds through a well-described process. However, in chronic wounds this process seems to become dysregulated at the transition between resolution of inflammation and re-epithelialization. Bioburden in the form of colonizing bacteria is a major contributor to the delayed headlining in chronic wounds such as pressure ulcers. However how the microbiome influences the wound metabolic landscape is unknown. Here, we have used a Systems Biology approach to determine the association between the taxonomic and metabolomic profile of wounds colonized by bacteria. Pressure ulcer biopsies were harvested from primary chronic wounds and bisected into top and bottom sections prior to analysis of microbiome by pyrosequencing and analysis of metabolome using 1H nuclear magnetic resonance (NMR) spectroscopy. Bacterial taxonomy revealed that wounds were colonized predominantly by three main phyla, but differed significantly at the genus level. While taxonomic profiles demonstrated significant variability between wounds, metabolic profiles shared significant similarity based on the depth of the wound biopsy. Association between taxonomy and metabolic landscape indicated significant wound-to-wound similarity in metabolite enrichment sets and metabolic pathway impacts, especially with regard to amino acid metabolism. To our knowledge, this is the first demonstration of a statistically robust correlation between bacterial colonization and metabolic landscape within the chronic wound environment.
INSTITUTE
Montana State University
DEPARTMENT
Chemistry and Biochemistry
LABORATORY
Ammons
LAST_NAME
Ammons
FIRST_NAME
Mary Cloud
ADDRESS
103 CBB, Montana State University, Bozeman, MT 59717
Labeling of cells was carried out in triplicate with each sample containing Cells were deprived of glucose or glutamine for 1 h then resuspended in 10mM D-glucose labeled or 4 mM [13C5]-U-glutamine labeled medium, respectively. At time cells were also activated with 5 ug/ml anti-CD3 and anti-CD28. Cells were (and activated) for 16 h when samples were spun down, and cells were in 200 ul ice cold methanol. All samples were immediately stored in -80Ā°C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
The human acute lymphoblastic leukemia cell line (Jurkat) was used to isolate a clones based on Noxa expression. JP6 is low Noxa, JP3 is high Noxa. Furthermore, a Noxa expressing plasmid was tranfected and stable clones were selected for a Noxa high model (N5). 10E6 cells were starved of glucose (A) or glutamine (B&C) for 3 hours and then fed 13C 1,2 glucose (A), 15N alpha nitrogen glutamine (B) or 15N amide nitrogen glutamine (C) for 24 hours. Cells were washed 1X in ice cold PBS and resuspended in -20C methanol. Quenched cells were snap frozen and stored at -80. For experiment A we would like to observe the contribution of labeled glucose into the synthesis of amino acids, specifically glycine and serine. For experiment B we are most interested in the nitrogen incorporation into aspartate. SCB edits: This fluxomics study requires measuring amino acids for M, M+1, M+2, and M+3 prioritizing glycine, serine, aspartate, asparagine, ornithine, citrulline, then as many as possible using a diamond hydride column and LCMS method (see CEvans).
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Flies were fed a normal diet or high-sugar (30% sucrose) diet for 7 days. of flies fasted for 24h or sated were microdissected and immediately frozen in ice. The microdissection process from taking the fly out of the vial to the brain took less than a minute. Each brain microdissected piece contains 50K cells.We dissected 40 brains per condition (4 conditions total) and 4-5 replicates
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
13C mass isotopomer analysis (LCMS flux studies) MLL-AF9 (part II)
STUDY_TYPE
timecourse
STUDY_SUMMARY
How cancer cells adapt to metabolically adverse conditions in patients and to proliferate is a fundamental question in cancer biology. Here we show that protein kinase (AMPK), a metabolic checkpoint kinase, confers metabolic stress to leukemia-initiating cells (LICs) and promotes leukemogenesis. Upon dietary MLL-AF9-induced murine acute myeloid leukemia (AML) activated AMPK and leukemogenic potential. AMPK deletion significantly delayed leukemogenesis and LICs by reducing the expression of glucose transporter 1 (Glut1), compromising flux, and increasing oxidative stress and DNA damage. LICs were particularly on AMPK to suppress oxidative stress in the hypoglycemic bone marrow Strikingly, AMPK inhibition synergized with physiological metabolic stress by dietary restriction and profoundly suppressed leukemogenesis. Our results that AMPK protects LICs from metabolic stress and that combining AMPK with physiological metabolic stress potently suppresses AML by inducing stress and DNA damage.
INSTITUTE
University of Michigan
DEPARTMENT
Molecular and Human genetics (Baylor College of Medicine)
Human fecal bile acid profiles before and after fecal transplant
STUDY_TYPE
Understand the bile acid profiles from the feces of fecal microbiota transplant patients that successfully recover from recurrent C. difficile infection
STUDY_SUMMARY
Understand the bile acid profiles from the feces of fecal microbiota transplant patients that successfully recover from recurrent C. difficile infection
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
U13C-Glutamine and U13C-Glucose Flux Analysis (MFA SiHa B16F10)
STUDY_TYPE
SiHa_NT and SiHa_L are 2 experimental group, control and treatment incubated with U13Cglutamine before lysis for 2 hours. SiHa_WT and SiHa_F3 are different cell isogenic clones to incubated before lysis with U13C glucose for Similarly, B1610 and B16M4 are 2 cell lines incubated before lysis with U13C for 2hours
STUDY_SUMMARY
SiHa_NT and SiHa_L are 2 experimental group, control and treatment incubated with U13Cglutamine before lysis for 2 hours. SiHa_WT and SiHa_F3 are different cell isogenic clones to incubated before lysis with U13C glucose for Similarly, B1610 and B16M4 are 2 cell lines incubated before lysis with U13C for 2hours
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
This experiment was performed using the following cohorts: 1) healthy controls, patients with scleroderma at low risk for pulmonary hypertension, 3) pateints scleroderma at high risk for pulmonary hypertension. Whole blood was drawn into stop soilution (1:1 ratio) at rest and again at peak exercise for each subject.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomic analysis of normal and diabetic mouse bone marrow under PBS or treatment
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
Wild type and diabetic (MKR) mice were treated daily with intraperitoneal of 50 microliters of PBS (control) or metformin (200 mg/kg body weight) for 14 Their bone marrow cells were collected and compared by untargeted metabolomics.
INSTITUTE
New York University
DEPARTMENT
College of Dentistry, Basic Science and Craniofacial Biology
LABORATORY
Xin Li Laboratory
LAST_NAME
Xin
FIRST_NAME
Li
ADDRESS
345 East 24th Street, Room 901D, New York, NY 10010
EMAIL
xl15@nyu.edu
PHONE
1-(212)992-7009
NUM_GROUPS
4
TOTAL_SUBJECTS
15 samples
STUDY_COMMENTS
each sample with 3 technical replicates in each mode
Sparing of muscle mass and function by passive loading in an experimental intensive care unit model
STUDY_TYPE
time course + intervention
STUDY_SUMMARY
A unique experimental rat ICU model has been used allowing long-term (weeks) time-resolved analyses of the effects of standardized unilateral passive mechanical loading on skeletal muscle size and function and underlying mechanisms. Results show that passive mechanical loading alleviated the muscle wasting and the loss of force-generation associated with the ICU intervention, resulting in a doubling of the functional capacity of the loaded versus the unloaded muscles after a 2-week ICU intervention.
Cardiac Resynchronization Therapy Induces Adaptive Metabolic Transitions in the Metabolomic Profile of Heart Failure
STUDY_TYPE
intervention
STUDY_SUMMARY
This prospective study consisted of 24 patients undergoing CRT for advanced HF and 10 control patients who underwent catheter ablation for supraventricular arrhythmia but not CRT. Blood samples were collected before and 3 months after CRT. Metabolomic profiling of plasma samples was performed using gas chromatographyĀmass spectrometry and nuclear magnetic resonance.
The purpose of this study was to optimize conditions for cold storage of rat spheroids without freezing. Rat hepatocytes were isolated by a two-step method; hepatocyte spheroids were formed during 48 h of rocked culture in medium (SFM). Spheroids were then maintained in rocked culture at 37Ā°C condition) or cold stored at 4Ā°C for 24 or 48 h in six different cold storage SFM alone; SFM + 1 mM deferoxamine (Def); SFM + 1 ?M cyclosporin A (CsA); SFM + mM Def + 1 ?M CsA, University of Wisconsin (UW) solution alone, UW + 1 mM Def.
Effect of Insulin Sensitizer Therapy on Amino Acids and Their Metabolites
STUDY_TYPE
drug + time course
STUDY_SUMMARY
We previously reported the overall study design for the parent study [29]. The report primarily examines the effect of three months of insulin sensitizer on plasma concentrations of BCAA, AAA, and AA metabolites in overweight/obese with fasting hyperglycemia, defined as either impaired fasting glucose or diabetes [29]. Briefly, 25 drug naĆÆve, Northern European American participants fasting blood glucose concentrations of 108?180 mg/dL were randomized to either 45 mg of pioglitazone per day plus 1 g of metformin twice per day (n = or placebo (n = 13) for 12 weeks. We chose metformin based on its proven effect hepatic insulin sensitivity and pioglitazone based on its effect on peripheral sensitivity. Current use of hypoglycemic medications excluded participants from present study.
In this experiment 2 day old baby mice were exposed to hyperoxia (75% O2) for 14 days. Control baby mice were housed in room air (normoxia). Plasma and lavage fluid (BALF) were harvested after 14 days of exposure (on Day of life
STUDY_SUMMARY
In this experiment 2 day old baby mice were exposed to hyperoxia (75% O2) for 14 days. Control baby mice were housed in room air (normoxia). Plasma and lavage fluid (BALF) were harvested after 14 days of exposure (on Day of life
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
13C mass isotopomer analysis (LCMS flux studies) MLL-AF9 (part III)
STUDY_TYPE
Glycolysis/TCA/Nucleotide analysis (tissue/cells)
STUDY_SUMMARY
How cancer cells adapt to metabolically adverse conditions in patients and to proliferate is a fundamental question in cancer biology. Here we show that protein kinase (AMPK), a metabolic checkpoint kinase, confers metabolic stress to leukemia-initiating cells (LICs) and promotes leukemogenesis. Upon dietary MLL-AF9-induced murine acute myeloid leukemia (AML) activated AMPK and leukemogenic potential. AMPK deletion significantly delayed leukemogenesis and LICs by reducing the expression of glucose transporter 1 (Glut1), compromising flux, and increasing oxidative stress and DNA damage. LICs were particularly on AMPK to suppress oxidative stress in the hypoglycemic bone marrow Strikingly, AMPK inhibition synergized with physiological metabolic stress by dietary restriction and profoundly suppressed leukemogenesis. Our results that AMPK protects LICs from metabolic stress and that combining AMPK with physiological metabolic stress potently suppresses AML by inducing stress and DNA damage.
INSTITUTE
University of Michigan
DEPARTMENT
Molecular and Human genetics (Baylor College of Medicine)
THP-1 Human Monocyte cells (10^6) were seeded in RPMI 1640 with 0.5% FBS. The were incubated for 30 minutes in either no citrate, 1 mM citrate, or 6 mM Separately, cells were either incubated in 1 mM 13C6 citrate or 6mM 13C6 (Sigma-Aldrich #606081). After the 30 minute pre-incubation time, the cells harvested or stimulated with lipopolysaccharide (LPS) for 1 h or 30 min. Cells subsequently spun down and washed in 150mM ammonium acetate. Cell were respun the supernatant was discarded. Cells were flash frozen in liquid nitrogen and on dry ice.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Comparative metabolomics study of WT and iButOH tolerant E. coli
STUDY_SUMMARY
This study aims to elucidate metabolic mechanisms of tolerance to isobutanol in coli. Two strains are utilized: WT parental strain EcHW24, and isobutanol strain 38-20-4 created via multiplex genome engineering. The metabolic response growth with isobutanol (0.7% w/v) and without (0% w/v) on NG50 minimal media be compared for each strain. 3 replicates for each strain/iButOH concentration been submitted, excpet for WT/0.7%; we have observed high growth phenotype for this combination, and have corresponding submitted 5x replicates. Strain contains mutations in the TCA cycle (gltA SNP) and amino acid metabolism indels in glnE, gltD, and tnaA), thus our hypothesis is that tolerance arises rewiring of TCA cycle and amino acid metabolism, possibly resulting in intracellular ratio of glutamine:glutamate
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
The experiment was done on 4/2/2014 and consists of 7 experimental conditions x replicates per condition = 14 samples. The experimental duplicates are numbered (i.e. 1 and 2, 3 and 4, etc.). All samples are primary mouse macrophages from bone marrow. 12 million cells were plated in each 10 cm tissue culture plate (Corning) and the following day the cells were stimulated with LPS, CL097 and/or Gardiquimod in 10 mls of 2% FBS 25 mM HEPES IMDM. After 60 minutes medium was aspirated and the cells were rapidly wahsed with 15 mls of 150 mM acetate and after aspiration liquid nitrogen was poured on top of the cells and to -80 C freezer. Sample pairs 7 - 8 and 9-10 were treated identically except the rinse was done with ammonium acetate (7 and 8) or distilled water (9 and to evaluate the effect of the rinse solution in the quality of the obtained
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
T cell metabolism during graft-versus-host disease (CAB 307)-PART I
STUDY_TYPE
Glycolysis/TCA/Nucleotide analysis (tissue/cells)
STUDY_SUMMARY
T cells were injected into mice to model graft-versus-host disease. On day 7 after bone marrow transplantation (BMT), cells were recovered and CD8 T cells were purified to > 90% purity over magnetic columns. NaĆÆve T cells were used as control. Cells were then plated at 5x10^6 cells/ml onto plates coated with anti-CD3/CD28 antibodies in the presence of 300ĀµM 13C-palmitate conjugated to BSA (in PBS). Cells were incubated at 37oC for 60min, after which time they were removed from the plates, counted, and an equal number (2.5x10^6) were placed into a 1.7 ml micro-centrifuge tubes and spun down. Cells were washed once with ammonium chloride, residual volume was removed by a second brief spin, cell pellets were flash frozen in liquid nitrogen and stored at -80oC until analysis. Several mice lines were used in this study.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
We recently reported superior right ventricle (RV) performance in response to acute afterload challenge in lambs with a model of congenital heart disease with chronic left-to-right cardiac shunts. Compared with control animals, shunt lambs demonstrated increased contractility because of an enhanced Anrep effect (the slow increase in contractility following myocyte stretch). This advantageous physiological response may reflect preservation of a fetal phenotype, since the RV of shunt lambs remains exposed to increased pressure postnatally. Nitric oxide (NO) production by NO synthase (NOS) is activated by myocyte stretch and is a necessary intermediary of the Anrep response. The purpose of this study was to test the hypothesis that NO signaling is increased in the RV of fetal lambs compared with controls and shunt lambs have persistence of this fetal pattern. An 8-mm graft was placed between the pulmonary artery and aorta in fetal lambs (shunt). NOS isoform expression, activity, and association with activating cofactors were determined in fetal tissue obtained during late-gestation and in 4-wk-old juvenile shunt and control lambs. We demonstrated increased RNA and protein expression of NOS isoforms and increased total NOS activity in the RV of both shunt and fetal lambs compared with control. We also found increased NOS activation and association with cofactors in shunt and fetal RV compared with control. These data demonstrate preserved fetal NOS phenotype and NO signaling in shunt RV, which may partially explain the mechanism underlying the adaptive response to increased afterload seen in the RV of shunt lambs. Research is published, core data not used but project description is relevant: http://ajpheart.physiology.org/content/309/1/H157.long
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
This experiment was performed in wild type vs. CD39 knockout (KO) mice that had infusions of either saline or apyrase over the course of 2 weeks. Whole blood obtained via intracardiac puncture, and was drawn into a syringe prefilled with anti-enzyme stop solution.
STUDY_SUMMARY
This experiment was performed in wild type vs. CD39 knockout (KO) mice that had infusions of either saline or apyrase over the course of 2 weeks. Whole blood obtained via intracardiac puncture, and was drawn into a syringe prefilled with anti-enzyme stop solution.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Patients between age of 6 weeks and 16 years of age who were undergoing small resection were selected, including those who were either receiving enteral preoperatively (control group), or those who were without enteral nutrition and TPN (TPN group). At the time of operation, 100-500 microliters of effluent from small bowel lumen was aspirated during the small bowel resection. The samples immediately frozen in liquid nitrogen and stored at -80deg. After five control five TPN samples were collected, all samples were submitted for amino acid to quantify the relative amounts of amino acids in the small bowel effluent each patient. We hypothesize that the TPN group will contain higher levels of amino acids present in TPN, such as leucine, phenylalanine, and valine.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Samples 4-34 are sera and feces from an experiment to determine the effects of on SCFA in Ang II-induced hypertension. Samples 35-59 are plasma samples fron experiment to determine the effects of ACE2 activator, DIZE on SCFA in pulmonary hypertension. Plasma samples were collected using heparin
STUDY_SUMMARY
Samples 4-34 are sera and feces from an experiment to determine the effects of on SCFA in Ang II-induced hypertension. Samples 35-59 are plasma samples fron experiment to determine the effects of ACE2 activator, DIZE on SCFA in pulmonary hypertension. Plasma samples were collected using heparin
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
This is a time restricted feeding study (see M Hatori, et al. Cell Metabolism 15(6), 848-860)
STUDY_SUMMARY
This is a time restricted feeding study (see M Hatori, et al. Cell Metabolism 15(6), 848-860). Briefly, there are four groups of mice: (1) NA mice are on a chow ad libitum diet; (2) FA mice are on a high fat ad libitum diet (i.e. Diet obesity); (3) FT mice are on a high fat time restricted diet (i.e. only eat for hours/day and fast for 16); and (4) CDKO mice which are cry knock out mice on a chow ad libitum diet. We have collected stool from these four groups while they in the light and in the dark. Our expectation is that their stool metabolites be different at different times of the day.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Twelve weeks old WT and BAF60a Liver specfific knockout mice were fed with high diet for 8 weeks. Mice were sacrified after 4 h of starvation. Blood samples harvested by heart puncture for plasma. Liver were dissected and immediately in liquid nitrogen. All the samples were kept in -80oC freezer for long term
STUDY_SUMMARY
Twelve weeks old WT and BAF60a Liver specfific knockout mice were fed with high diet for 8 weeks. Mice were sacrified after 4 h of starvation. Blood samples harvested by heart puncture for plasma. Liver were dissected and immediately in liquid nitrogen. All the samples were kept in -80oC freezer for long term
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Lanthanide-mineral induced alteration of bile acid metabolism in a murine model steatohepatitis
STUDY_SUMMARY
120 mice (equal number of males and females) were randomized into 3 equal One group is on a HFWD alone, a second group is on HFWD supplemented with and calcium, and a third control group is supplemented with calcium alone. At termination (18 months) we will harvest hepatocytes and colonic enterocytes for of epithelial gene expression patterns. We will also harvest cecal contents and for microbial profiling by 16S rRNA pyrosequencing. For this small pilot we wish to add an untargeted metabolomic analysis component. We will harvest (right medial lobe with gall bladder), serum, and feces. We will assay liver/gall bladder samples (8 from the HFWD group and 8 from the supplemented group). Remaining liver samples and the serum and feces will be for future investigation. Liver is being targeted first since both and hepatocellular carcinoma were seen in our previous study in mice on HFWD Additionally, alterations of bile acid profiles and bile acid metabolism have associated with both steatohepatitis and hepatic cancers.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Understand the differences in Bile Acid composition between knockout and floxed at each intestinal segment. Also difference in metabolites between these two at each level of the intestine.
STUDY_SUMMARY
Understand the differences in Bile Acid composition between knockout and floxed at each intestinal segment. Also difference in metabolites between these two at each level of the intestine.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Murine gastrointestinal bile acid profiles before and after antibiotics
STUDY_SUMMARY
Mice were treated with cefoperazone for 10 days and then allowed 6 weeks to off of the antibiotic. Other antibiotic treatments included IP clindamycin the 6 week recovery, IP clindamycin alone, vancomycin alone, kanamycin alone metronidazole. Gut luminal contents of these mice were collected at the time of and included both ileal and cecal content. Paired samples will be used for both analysis and targeted bile acid analysis to define how gut bacteria alter bile profiles in the gut and in turn how this affects C. difficle spore germination outgrowth.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Liver and Plasma metaboites for 13C-glucose load in wild type, LIRKO and LIRFKO
STUDY_SUMMARY
Mice were fasted for 18 hr overnight then sacrificed or treated with (2 g/kg ip) and sacrificed 1 hr later by decapitation and liver was immediately and stored in liquid N2 and then at -80 C. Wild type (IR and IR/FoxO1 floxed) were sacrificed after fasting and 1 hr post-glucose treatment. Liver-specific receptor knockout (LIRKO) and insulin receptor/FoxO1 double knockout (LIRFKO) were sacrificed 1 hr post glucose treatment
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Tumor neurospheres were grown in culture until 90% confluent. To collect cells, were filtered through 0.45um nylon filters (Fischer Sci 099029) that were with methanol and ultrapure water using a microanalytical glass filter holder Sci 09753E). The cells were filtered through filtration apparatus, then washed 1mL of distilled water very quickly. Filter paper is taken immediatedly with and plunged cell side down into 6-well plate filled with liquid nitrogen placed dry ice. Plate is wrapped in aluminum foil and stored at -80C before the liquid evaporates. Samples are shipped on dry ice
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
THP-1 Human Monocyte cells (10^6) were seeded in RPMI 1640 with 0.5% FBS. The were incubated for 30 minutes in either no citrate, 1 mM citrate, or 6 mM Separately, cells were either incubated in 1 mM 13C6 citrate or 6mM 13C6 (Sigma-Aldrich #606081). After the 30 minute pre-incubation time, the cells harvested or stimulated with lipopolysaccharide (LPS) for 1 h or 30 min. Cells subsequently spun down and washed in 150mM ammonium acetate. Cell were respun the supernatant was discarded. Cells were flash frozen in liquid nitrogen and on dry ice.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Wild type and AMPK KO bone marrow derived macrophage were stimulated with LPS for various time period and plates were frozen in liquid nitrogen after washing sodium acetate (200uM) solution and kept in deep freezer for further analysis.
STUDY_SUMMARY
Wild type and AMPK KO bone marrow derived macrophage were stimulated with LPS for various time period and plates were frozen in liquid nitrogen after washing sodium acetate (200uM) solution and kept in deep freezer for further analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
T cell metabolism during graft-versus-host disease (CAB 307)-PART II
STUDY_SUMMARY
T cells were injected into mis-matched animals undergoing bone marrow On day 7 after BMT, cells were receovered and CD8 T cells were purified to > purity over magnetic column. Cells were washed once in PBS, then once in chloride, residual volume was removed and cell pellets were flash frozein in and stored at -80C
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Changes to bile acids in the murine gut after antibiotics
STUDY_SUMMARY
Changes to bile acids in the murine gut after antibiotics
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@umich.edu
PHONE
-
NUM_GROUPS
12
TOTAL_SUBJECTS
60
STUDY_COMMENTS
Samples were collected at the time of necropsy. Contents from the lumen of the tract of C57BL/6 mice were collected, flash frozen and stored in the -80C until for metabolomics assay. C57BL/6 mice treated with cefoperazone for 5 days with days of water (n=5) were harvested along with non-antibiotic treated mice Contents from the small intesine (duodenum, jejunum and ileum), the large (cecum, colon) and stool were collected for targeted bile acid analysis.
Liver and Plasma metaboites for 13C-glucose load in wild type, LIRKO GTT 1 mice
STUDY_TYPE
13C mass isotopomer analysis (flux studies)
STUDY_SUMMARY
Mice were fasted for 18 hr overnight then sacrificed or treated with 13C-U-glucose (2 g/kg ip) and sacrificed 1 hr later by decapitation and liver was immediately freeze-clamped and stored in liquid N2 and then at -80 C. Wild type (IR and IR/FoxO1 floxed) mice were sacrificed after fasting and 1 hr post-glucose treatment. Liver-specific insulin receptor knockout (LIRKO) and insulin receptor/FoxO1 double knockout (LIRFKO) mice were sacrificed 1 hr post glucose treatment. http://www.nature.com/ncomms/2015/150512/ncomms8079/full/ncomms8079.html
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Role of Microbiome in Psoriatic Arthritis (SCFA in PsA)
STUDY_SUMMARY
To investigate associations of gut and skin microbiota diversity with psoriatic pathogenesis.Characterize the abundance and diversity of gut microbiota in with never-treated, new-onset psoriatic arthritis (PsA). Methods: 16s rRNA pyrosequencing was utilized to compare the community composition of microbiota in PsA patients, psoriasis patients and healthy, matched controls. from patients with PsA, psoriasis of the skin (Ps), new-onset rheumatoid (NORA) and healthy controls were assessed for the presence and levels of fecal immunoglobulin A (sIgA) and various proteins and pro-inflammatory cytokines
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Comparison of normal, lung adenocarcinoma and SCLC tissue metabolomes
STUDY_SUMMARY
In addition to the generation and analysis of metabolomics data on cell lines, of normal lung tissue, adenocarcinoma lung tissue and small cell lung carcinoma (seven samples/group) were processed and evaluated metabolite profile under the scope of the pilot and feasibility study. These data can be to the metabolite profiles defined in the SCLC and NSCLC cell lines and with the ABPP-determined metabolic kinases to identify distinct metabolic or biomarkers (?oncometabolites?) that distinguish small cell lung cancer from cell lung cancer.
Normal plasma cells,Low proliferation multiple myeloma and High proliferation myeloma cells
STUDY_TYPE
differential metabolomics
STUDY_SUMMARY
CD138 sorted bone marrow plasma cell were obtained from a normal patient, a myelow with slow proliferation and a multiple myelow with rapid proliferation.
Bile acid targeted metabolomics of the small intestine in malnourished and mice
STUDY_TYPE
Targeted metabolomics
STUDY_SUMMARY
A total of 8 samples from 6 week old, female C57BL/6 mice, treated for 3 weeks a malnourished diet or a control-fed isocaloric diet. Samples were taken from small intestinal fecal content at the terminus of the ileum for targeted bile analysis.
INSTITUTE
University of Victoria
DEPARTMENT
The Uvic Proteomics and Metabolomics Innovation Centre
LAST_NAME
Borchers
FIRST_NAME
Christoph
ADDRESS
#3101-4464 Markham St Victoria, BC Canada, V8Z 7X8
EMAIL
christoph@proteincentre.com
PHONE
-
NUM_GROUPS
2
TOTAL_SUBJECTS
8*
STUDY_COMMENTS
*One sample failed quality control in the malnourished group, thus was not in the study
1) Characterize plasma metabolome in Barth Syndrome 2) To implement targeted, quantitative studies on prospective biomarkers and metabolites of interest derived from the non-targeted phase.
Vitamin targeted metabolomics of the small intestine in malnourished and mice
STUDY_TYPE
Targeted metabolomics
STUDY_SUMMARY
A total of 8 samples from 6 week old, female C57BL/6 mice, treated for 3 weeks a malnourished diet or a control-fed isocaloric diet. Samples were taken from small intestinal fecal content at the terminus of the ileum for targeted of vitamin concentrations.
INSTITUTE
University of Victoria
DEPARTMENT
The Uvic Proteomics and Metabolomics Innovation Centre
LAST_NAME
Borchers
FIRST_NAME
Christoph
ADDRESS
#3101-4464 Markham St Victoria, BC Canada, V8Z 7X8
Metabolomics Reveals that Aryl Hydrocarbon Receptor Activation by Environmental Chemicals Induces Systemic Metabolic Dysfunction in Mice (Part I)
STUDY_TYPE
drug dosage
STUDY_SUMMARY
12 mice were acclimatized for a week, taught to eat dough pills then half were treated with TCDF per pill per day for 5 days Also 10 AHR -/- mice were used and treated the same way
Concentrations of cocaine (5,3,2,1.5,1,0.75,0.5,0.25,0.125, 0.062, and 0.031 were spiked into whole human blood for MALDI-MS analysis. A 10 uL volume of mixture was allowed to dry on Whatman Grade 2 filter paper. A 1ppm Cocaine-d3 was used as an internal standard. Parameters of the MALDI system optimized, including laser energy, number of laser shots, and matrix. The standard was added on top of the dried blood spot sample. Wide isolation tandem was used to generate a calibration curve.
INSTITUTE
University of Florida
DEPARTMENT
Chemistry
LABORATORY
Chemistry Lab Building
LAST_NAME
Dhummakupt
FIRST_NAME
Elizabeth
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry and Molecular PO Box 100245, Gainesville, FL 32610-0245
EMAIL
emuffly@ufl.edu
PHONE
352-392-0515
NUM_GROUPS
11
TOTAL_SUBJECTS
154
STUDY_COMMENTS
11 concentrations were run as 7 samples/concentration for 2 days
Analysis of Dried Blood Spots by Paper Spray Ionization - MS
STUDY_TYPE
Analysis of Dried Blood Spots by Paper Spray Ionization - MS
STUDY_SUMMARY
Concentrations of cocaine (6, 5,3,2,1.5,1,0.75,0.5,0.25,0.125, 0.062, and 0.031 were spiked into whole human blood for PS-MS analysis. A 10 uL volume of mixture was allowed to dry on Whatman Grade 2 filter paper. A 1ppm Cocaine-d3 was used as an internal standard. Parameters of the paper spray were optimized, including spray voltage, capillary temperature and solvent. The standard was added on top of the dried blood spot sample. Wide isolation tandem was used to generate a calibration curve.
INSTITUTE
University of Florida
DEPARTMENT
Chemistry
LABORATORY
Chemistry Lab Building
LAST_NAME
Dhummakupt
FIRST_NAME
Elizabeth
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry and Molecular PO Box 100245, Gainesville, FL 32610-0245
EMAIL
emuffly@ufl.edu
PHONE
352-392-0515
NUM_GROUPS
11
TOTAL_SUBJECTS
154
STUDY_COMMENTS
11 concentrations were run as 7 samples/concentration for 2 days
Untargeted metabolomic analysis of the small intestinal content of malnourished
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
A total of 8 samples from 6 week old, female C57BL/6 mice, treated for 3 weeks a malnourished diet or a control-fed isocaloric diet. Samples were taken from small intestinal fecal content at the terminus of the ileum.
INSTITUTE
University of Victoria
DEPARTMENT
The Uvic Proteomics and Metabolomics Innovation Centre
LAST_NAME
Borchers
FIRST_NAME
Christoph
ADDRESS
#3101-4464 Markham St Victoria, BC Canada, V8Z 7X8
Metabolomics Reveals that Aryl Hydrocarbon Receptor Activation by Environmental Induces Systemic Metabolic Dysfunction in Mice (Part II)
STUDY_TYPE
Time course
STUDY_SUMMARY
12 C57BL/6J male mice were acclimatized for 1 week and then divided into two and trained to eat dough pills. One group was control and the other was exposed TCDF through the dough pills. The mice were treated for 5 days at a dosage of TCDF for a final concentration of 5Ī¼g/kg body weight. Also looked at were mice and they were treated the same way as the C57BL/6J mice.
INSTITUTE
Pennsylvania State University
DEPARTMENT
Department of Veterinary and Biomedical Sciences
LAST_NAME
Patterson
FIRST_NAME
Andrew
ADDRESS
101 Life Sciences, University Park, State college, 16802
Adult (14 weeks old) Sprague-Dawley rats showing at least three consecutive periods (4 day) of estrous cycles were randomly assigned to four groups: 1: 2: nicotine (6 mg/kg), 3: OC and 4: nicotine (6 mg/kg) + OC. Rats were exposed these treatments for a month. At the end of treatments, hippocampus was immediately frozen in liquid nitrogen. We are sending one side of hippocampi to Center for Integrated Metabolomics for analysis at the University of Florida.
Quick Comparison of Urine Metabolites in Human and SD Rats of Different Sex by UPLC-TOFMS and In-house Software Platform
STUDY_TYPE
Biomarker Discovery
STUDY_SUMMARY
Human urine samples were collected before breakfast from 14 male and 13 female post-graduate students, age from 23 to 29, on the morning of sample collection Male (n=8) and female (n=8) SD rats weighing between 220 and 250g were used. On morning of sample collection day, each rat was deprived of food and put in cage for 24h urine collection. All urine samples were frozen at -80Ā°C prior to
Quick Comparison of Serum Metabolites in SD Rats of Different Sex by Untargeted and In-house Software Platform
STUDY_TYPE
Biomarker Discovery
STUDY_SUMMARY
Male (n=8) and female (n=8) SD rats weighing between 220 and 250g were used. On morning of sample collection day, each rat was deprived of food and put in cage for 24h urine collection. Then a blood sample (3-5ml) was collected from aorta of the rat under anesthesia and centrifuged to obtain serum. All urine serum samples were frozen at -80Ā°C prior to analysis.
Sexual antagonism in exuded non-volatile metabolites in C. purpureus
STUDY_TYPE
male - female
STUDY_SUMMARY
The experimental approach seeks to test for sexual dimorphism in exuded metabolites in C. purpureus. The proposed research is creative and original in its inter-disciplinary approach and its use of a biochemically tractable to develop a much-needed link between natural selection for sexual dimorphism the molecular targets of that selection pressure.
Cyclobutene- and cyclobutane-functionalized fatty acids as novel biochemical of structure and function in HepG2 cells
STUDY_TYPE
Lipid analysis novel C18 fatty acid anologues in complex lipids
STUDY_SUMMARY
Human hepatoma HepG2 cells (American Type Culture Collection; HB-8065) were in 75 ml tissue cell culture flasks in Eagle's minimal essential medium (EMEM) with 10% fetal bovine serum at 37Ā°C in a humidified atmosphere with 5% CO2. treatment with fatty acids or analogues, the cells were seeded at a density of Ć 106 cells in a T25-cm2 flask for 24 h. Each FA or analogue was added to the medium as a fatty acid-bovine serum albumin (BSA) complex (2.5:1, FA:BSA) to the desired final concentration. The controls in these experiments were HepG2 with BSA alone. After 24 h treatment, the media was collected, cells were twice with PBS and cells were harvested for analysis. To each cell suspension to lipid extraction a standard mixture of 25 Āµg C15:0 PE, C17:0 PC and C71:1 was added as standards. Extraction of lipids was performed according to the method. For metabolomics analysis, the lipid extracts were resuspended in 2:1 (v/v).All analyses were carried out using an Agilent 1200 Series HPLC, ACE C8-300 column (2.1 x 100 mm) and linear gradient elution at a flow rate of 0.1 Mobile phase A and B consisted of 0.1% formic acid; 10 mM ammonium acetate in and 0.1% formic acid; 10 mM ammonium acetate in ACN/isopropanol (50/50; v/v), The injection volume was 4 ĀµL. Separation of metabolites was achieved at the gradient: T=0 min: 30% B; T=1 min: 30% B; T=25 min: 100% B; T=45 min: 100% B; min: 30% B; and T=60 min: 30% B (re-equilibration). The HPLC system was coupled to a Bruker Soalrix 70 Hybrid FTMS instrument equipped with ionization source (ESI) (Bruker Daltonics). The system was controlled by HyStar software. MS data was collected with resolving power of 78,000 (at m/z 400) in or negative mode under following conditions: a capillary voltage of (+/-) 4,500 and an end plate offset of -500 V. The dry temperature was set at 180Ā°C. Dry flow was maintained 4 L/min. Acquisition range was 244-1,800 m/z with 0.2 s ion time. LC-MS data was converted into mzXML format using CompassXport v. 3.0.6 processed by mzMine v.2.10 [25] or XCMS data analysis software. Data processing mass detection, chromatographic peak detection and deconvolution, isotopic grouping, normalization and peak alignment. Metabolite data were mean-centered unit-variance scaled to remove the offsets and adjust the importance of high low abundance metabolites to an equal level. Significantly altered metabolites defined by a fold change (FC) >2 and p<0.05. Principal component analysis (PCA) hierarchical clustering analysis (HCA) of signature metabolites altered in treated cells compared to control were performed in the Metaboanalyst web (www.metaboanalyst.ca).
INSTITUTE
University of Nebraska - Lincoln
DEPARTMENT
Biochemistry
LABORATORY
DiRusso Black FATTT Lab
LAST_NAME
DiRusso
FIRST_NAME
Concetta
ADDRESS
Department of Biochemistry, University of Nebraska-Lincoln, N241 Beadle Center Vine St.
EMAIL
cdirusso2@unl.edu
PHONE
402-472-6504 or 402-613-9293
NUM_GROUPS
8
TOTAL_SUBJECTS
24+24=48
STUDY_COMMENTS
8 groups in triplicate ran in both negative and positive mode
Whole unconditioned medium (Defined culture media, M199),Whole M1 medium,Whole medium
STUDY_TYPE
differential metabolomics
STUDY_SUMMARY
Media was flash frozen with N2 and stored at -80 C. Samples can be stored at C until use. ~1 ml aliquots. The following media has been provided to the core in biological triplicates.Complete (Whole) Media:1) Whole unconditioned (Defined culture media, M199)2) Whole M1 medium3) Whole M2 medium
Metabolomic-based investigation on the effects of antifungal agents in Candida
STUDY_TYPE
in vitro study/drug dosage
STUDY_SUMMARY
Our aim is to determine the metabolic effects of increasing doses of an agent on C. albicans metabolism (untargeted, steady state metabolomics). We culture in vitro Candida cells to the mid-logarithmic growth in liquid media at 37Ā°C and then inoculate biological replicates (1ml) onto 22mm filters under vocuum filtration in sterile conditions. Subsequently, isolates be cultivated to midlogarithmic phase of growth on the same agar (RPMI-1640) to the antifungal agent has been added at a range of concentrations to achieve equivalent to 0 MIC (no drug), 0.0625 MIC, 0.125 MIC, 0.25 MIC, 0.5 MIC and 1.0 at 37Ā°C. At mid-logarithmic phase of growth (12h) replicates will be quenched by immersion into a solvent mixture of 40% acetonitrile: 40% methanol: water precooled at -40Ā°C. The resulting quenched isolate/solvent mixtrue will mechanically lysed by bead-beating with 0.1mm Zirconia beads in a tissue and then centrifuged to seperate out cell wall components. Supernatants will be and stored at -80Ā°C until they will be sent to SECIM facility.
In this project we will investigate the feasibility of metabolomics and to the diversity of ?metabotypes? in the horse, towards discovery of markers and associated with obesity and insulin resistance in the equine model. The team of researchers assembled for this work have identified horses severely with Equine Metabolic syndrome, often characterized by obesity and These animals are all from the Arabian breed, to control for some genetic Horses are age, sex and farm of residence matched with a control animal possible. Carefully controlled collection protocols were utilized to ensure variability in sample age and quality. Blood plasma is submitted for both LC-MS analysis through the SECIM core facilities. This discovery-based approach begin to generate new targets for the development of novel therapeutic for the treatment and prevention of obesity, type-2 diabetes as well as related conditions in both humans and horses. Finally, as the first dataset of its kind the horse, we may also be able to highlight promising new biomarkers for diagnostic use.
Aliquots of Jurkat T-lymphocyte cells were extracted using the Folch or Matyash at different total volumes and run in triplicate to calculate the extraction for each method.
INSTITUTE
University of Florida
DEPARTMENT
Dept. of Chemistry/SECIM
LABORATORY
Biomedical Mass Spectrometry Lab
LAST_NAME
Ulmer
FIRST_NAME
Candice
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry and Molecular PO Box 100245, Gainesville, FL 32610-0245
Determining the metabolic profile of wildtype and nos mutant Staphylococcus grown in media lacking glucose using targeted LC/MS
STUDY_TYPE
Single time point
STUDY_SUMMARY
Whole cells from wildtype, nos mutant, SrrAB mutant, SrrAB nos double mutant, complement strains will be isolated for targeted metabolite analysis. In supernatants (extracellular metabolites) will also be analyzed for their profile.
Measurement of free amino acid (AA) in response to MYC
STUDY_TYPE
treatment
STUDY_SUMMARY
The proposed study will provide us information how the different AAs are by MYC. MYC is known to cause increased levels of glutamin due to import and but we hypothesize that the overall pool of free AA in the cells is reduced, when cells are starved.
Impact of recurrent hypoglycemia on brain metabolite profile
STUDY_TYPE
Insulin treatment in diabetic rats
STUDY_SUMMARY
The goal of this study is to determine the effect of mild / moderate on brain metabolism. To achieve this goal insulin-treated diabetic rats will be to recurrent mild / moderate hypoglycemia.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Dave
FIRST_NAME
Kunjan
ADDRESS
-
EMAIL
Kdave@med.miami.edu
PHONE
305-243-3590
NUM_GROUPS
2
TOTAL_SUBJECTS
40
STUDY_COMMENTS
Two groups (i) Insulin treated diabetic + Recurrent hypoglycemia, (ii) Insulin diabetic + Recurrent hypoglycemia (only insulin injection) + Glucose infusion maintain glucose to baseline levels.Total 40 samples. Description: Ten samples and 10 hypothalamus sample from ITD+RH group of rats.Similarly, Ten samples and 10 hypothalamus sample from ITD+RH+Glucose group of rats.
NIH WCMC Pilot & Feasibility Project: ĀMetabolomics of Neonatal Pulmonary HypertensionĀ
STUDY_TYPE
Treatment and feeding study
STUDY_SUMMARY
Targeted metabolomic analyses of oxylipins were performed on 16 rat plasma and lung samples collected from rats euthanized at 14 days following exposure to growth restriction and/or hyperoxia. Samples were analyzed by UPLC-MS/MS using a Waters Acquity UPLC and detected on an API 4000 QTrap (AB Sciex, Framingham, MA, USA) by multiple reaction monitoring (MRM) after negative mode electrospray ionization.
The role of microbial metabolites in experimental liver disease
STUDY_TYPE
Targeted Metabolomic Analysis of plasma samples
STUDY_SUMMARY
Aim 1: Our experimental approach is to understand the effect of drinking water supplemented bacterial metabolite, Indole-3-propionic Acid (IPA), in liver disease in an acute alcohol model. Aim 2: Determine the levels of IPA in plasma of conventional mice in a chronic alcohol model. Aim 3: Determine the levels of IPA in plasma of conventional WT (C57BL/6) and mutant SL (sublytic, which is a mouse with a point mutation in ATP4a) mice.
INSTITUTE
University of North Carolina
DEPARTMENT
Discovery Science Technology
LABORATORY
Sumner Lab
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
NIH WCMC Pilot & Feasibility Project: ĀMetabolomics of Neonatal Pulmonary in human
STUDY_TYPE
Case-control study
STUDY_SUMMARY
Targeted metabolomic analyses of oxylipins were performed on 40 human umbilical plasma samples collected from infants born prematurely (gestational age <37 Samples were analyzed by UPLC-MS/MS using a Waters Acquity UPLC and detected on API 4000 QTrap (AB Sciex, Framingham, MA, USA) by multiple reaction monitoring after negative mode electrospray ionization.
NIH WCMC Pilot & Feasibility Project: ĀMetabolite changes associated with lossĀ
STUDY_TYPE
Feeding study
STUDY_SUMMARY
Targeted metabolomic analyses of oxylipins, endocannabinoids, and ceramides performed on 18 mouse liver, adipose, hypothalamus, plasma, and muscle samples from mice euthanized at 18 weeks following consumption of three diet regimens induced lean, obese, and weight loss phenotypes. Samples were analyzed by using a Waters Acquity UPLC and detected on an API 4000 QTrap (AB Sciex, MA, USA) by multiple reaction monitoring (MRM) after negative mode electrospray
Metabolic contribution of pSymA and pSymB megaplasmid/chromid for multipartite meliloti cultured in minimal M9 medium
STUDY_TYPE
megaplasmid deletion
STUDY_SUMMARY
To understand the contribution of pSymA and pSymB to the metabolism of S. the intracellular metabolome was analyzed at five time points (exponential and growth phases) across the growth curve of strains with or without pSymA and/or grown in a defined, minimal medium (M9).
INSTITUTE
McMaster University
DEPARTMENT
Department of Biology
LAST_NAME
Finan
FIRST_NAME
Turlough
ADDRESS
Department of Biology, McMaster University, Hamilton, Canada L8S4K1
metabolic contriubtion of pSymA and pSymB megaplasmid/chromid for multipartite meliloti cultured in rich LBmc medium
STUDY_TYPE
megaplasmid deletion
STUDY_SUMMARY
We wished to evaluate the contribution of pSymA and pSymB towards the of various metabolites in a nutritionally complex environment and to examine S. meliloti influences its surrounding environment.
INSTITUTE
McMaster University
DEPARTMENT
Department of Biology
LAST_NAME
Finan
FIRST_NAME
Turlough
ADDRESS
Department of Biology, McMaster University, Hamilton, Canada L8S4K1
We analyzed metabolites in the brains of wild type mice and DJ-1 knockout mice. DJ-1 knockout mouse is a model of an inherited form of Parkinson's disease.
INSTITUTE
National Institute on Aging
DEPARTMENT
Laboratory of Neurogenetics
LABORATORY
Cell Biology and Gene Expression Section
LAST_NAME
Hauser
FIRST_NAME
David
ADDRESS
35 Lincoln Drive, BLDG 35, Room 1A-1012, Bethesda, MD 20892
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv1713
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
NUM_GROUPS
3 replicates X 3 timepoints X with/without GTP
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv1016
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
NUM_GROUPS
3 replicates X 3 timepoints
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3280
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
NUM_GROUPS
3 replicates X 3 timepoints
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3244
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
NUM_GROUPS
3 replicates X 3 timepoints
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3801
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
NUM_GROUPS
3 replicates X 3 timepoints
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3722c
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3311
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in M. tuberculosis genome with whole-cell lysate and assaying changes in levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might
INSTITUTE
Weill Cornell Medical College
DEPARTMENT
Microbiology and Immunology
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 Avenue, New York, NY
EMAIL
kyr9001@med.cornell.edu
PHONE
-
NUM_GROUPS
3 replicates X 3 timepoints
STUDY_COMMENTS
The timepoints in the datafiles refer the amount of time (in minutes) that each was incubated with lysate.The units for 'mzmed' are mass/charge.The units for are seconds (retention time).The units for all the other data columns in the are arbitrary; they are integrated peak counts.In this study, Rv1713 was with lysate with and without 1mM GTP added as a co-factor.
Mice were fed with a TFD for 8 or 24 weeks to induce NAFLD or NASH, Targeted analyses examined diacylglycerols and ceramides in 6-8 mice per group 4 groups including 8 week control, 24 week control, 8 week TFD, and 24 week Samples were analyzed via UHPLC-HRMS
INSTITUTE
University of Florida
DEPARTMENT
Chemistry
LABORATORY
Yost Laboratory
LAST_NAME
Patterson
FIRST_NAME
Rainey
ADDRESS
214 Leigh Hall, University of Florida, Gainesville, FL 32611
Serum samples from Mla patients responsive or not responsive to chemotherapy
STUDY_SUMMARY
In the current study we will perform global metabolic profiling on serum samples obtained at diagnosis from pediatric AML patients (n=20) treated under St. Jude AML02 clinical trial to identify potential biomarkers of clinical significance. These patients include 10 responders and 10 non responders. In a subset of patients (n=7), we have matched samples that were obtained at remission allowing us to determine the change in serum metabolome at diagnosis and after remission followed by investigation of the metabolome change analysis with clinical response.
Pyruvate isotopically labeled by 13C either at position 1 or 2 was used to carbon flux in 3T3-L1 mouse cells line in the presence of various combinations drugs
INSTITUTE
University of Michigan
DEPARTMENT
Molecular and Integrative Physiology (MCTP)
LABORATORY
Macdougald Lab (MCTP)
LAST_NAME
MacDougald
FIRST_NAME
Ormond
ADDRESS
University Michigan, 2900 Huron Parkway, Ann Arbor, MI 48105
To understand the contribution of adipose tissue fatty acid oxidation to metabolism, we generated mice with an adipose-specific knockout of carnitine 2 (CPT2A?/?), an obligate step in mitochondrial long-chain fatty acid CPT2A?/? mice became hypothermic after an acute cold challenge, and CPT2A?/? adipose tissue (BAT) failed to upregulate thermogenic genes in response to stimulation. The adipose-specific loss of CPT2 resulted in diet-dependent in adiposity but did not result in changes in body weight on low- or high-fat Additionally, CPT2A?/? mice had suppressed high-fat diet-induced oxidative and inflammation in visceral white adipose tissue (WAT); however, high-fat glucose intolerance was not improved. These data show that fatty acid oxidation required for cold-induced thermogenesis in BAT and high-fat diet-induced stress and inflammation in WAT.
INSTITUTE
University of Michigan
DEPARTMENT
Biological Chemistry(Johns Hopkins University School of Medicine)
LABORATORY
Wolfgang Lab(Johns Hopkins University School of Medicine)
LAST_NAME
Wolfgang
FIRST_NAME
Michael
ADDRESS
733 North Broadway, Suite G49 Baltimore, MD 21205-2196
EMAIL
mwolfga1@jhmi.edu
PHONE
443-287-7680
NUM_GROUPS
2
TOTAL_SUBJECTS
16
STUDY_COMMENTS
(CPT2A?/?) stands for Carnitine palmitoyltransferase 2 knock out
Short-chain fatty acid analysis in bronchoalveolar lavage fluid (BAL SCFA)
STUDY_TYPE
We will correlate the bacterial gene abundance with the metabolite concentration
STUDY_SUMMARY
The study is intended to find if correlation exists between the abundance of bacterial gene for pyruvate ferredoxin oxidoreductase (PFOR) and short-chain fatty acids concentration in bronchoalveolar lavage fluid from patients with HIV-Associated Bacterial Pneumonia.
INSTITUTE
University of Michigan; New York University
DEPARTMENT
Pulmonary Medicine
LABORATORY
Weiden Lab(New York University School of Medicine)
Lung contusion is a major risk factor for the development of acute respiratory syndrome. Hypoxia-inducible factor-1? is the primary transcription factor that responsible for regulating the cellular response to changes in oxygen tension. set to determine if hypoxia-inducible factor-1? plays a role in the of acute inflammatory response and injury in lung contusion.Nonlethal unilateral lung contusion was induced in a hypoxia reporter mouse model and 2 cell-specific hypoxia-inducible factor-1? conditional knockout mice. The mice killed at 5-, 24-, 48-, and 72-hour time points, and the extent of systemic and hypoxia was assessed. In addition, injury and inflammation were assessed by bronchoalveolar lavage cells (flow cytometry and cytospin), albumin injury), and cytokines (inflammation). Isolated type 2 cells from the factor-1? conditional knockout mice were isolated and evaluated for cytokines following lung contusion. Finally, the role of nuclear factor-?B and as intermediates in this interaction was studied.
INSTITUTE
University of Michigan
DEPARTMENT
Surgery
LABORATORY
Raghavendran Lab
LAST_NAME
Thomas
FIRST_NAME
Bivin
ADDRESS
Ann Arbor, MI
EMAIL
bthomas@umich.edu
PHONE
734-615-7142
NUM_GROUPS
4
TOTAL_SUBJECTS
8
STUDY_COMMENTS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4245055/ : HIF1 (+/+) stands for reporter mouse modelHIF1 (-/-) stands for type 2 cell-specific factor-1? conditional knockout
Metabolic analysis of Parp1 ko/wt Saline & Bleo Mouse Lung Fiboblasts and Human & Normal Lung Fiboblasts (Part 2)
STUDY_TYPE
Glycolysis/TCA/Nucleotide analysis. Ceramide analysis for parp1 wild type lung after saline or bleomycin treatment.
STUDY_SUMMARY
This study is a part of series performed for the same researcher through grant program, so the publication is relevant reference for other studies ST000183)This specific experiment is a small pilot study to establish method it includes four biological replicas of identical cell cultures after the treatment and a single tissue sample.
This study is a part of series performed for the same researcher through pilot/feasibility grant program, so the publication is relevant reference for other studies (ST000199)This specific experiment is a pilot study to compare the metabolism of cells transformed using empty vector against cells transformed with vector carrying short hairpin RNA (shRNA) targeted to silence isocitrate dehydrogenase-1 (IDH1) gene.
The goal of the study was to compare the influence of Ileal pouch-anal (IPAA) surgical treatment on patients with ulcerative colitis (UC) vs those familial adenomatous polyposis (FAP). Stool samples were obtained at the time clinic visit, immediately frozen and stored at -80oC until the assay. There no exclusion criteria.
Small airway epithelial cell (SAEC) (Lonza Walkersville, Inc., MD) were grown according to the manufacturer's instructions in growth medium (SAGM) containing 0.03 mg/ml bovine pituitary extract (BPE), 0.5 Āµg/ml hydrocortisone, 0.5 ng/ml hEGF, 0.5 Āµg/ml epinephrine, 10 Āµg/ml transferrin, 5 Āµg/ml insulin, 0.1 ng/ml retinoic acid, 6.5ng/ml triiodothyronine, 50 Āµg/ml gentamicin and 50ng/ml Amphotericin-B, and 0.5 mg/ml bovine serum albumin (BSA, fatty acid free). When SAE grew to around 80 to 90% confluence, the cells were put into basal medium not supplemented with growth factors 3hours. Then the cell monolayers were infected with naĆÆve hMPV at multiplicity of infection (MOI) of 3 to certain time. The supernatant was saved and cells were harvested and stored exactly according to the protocol provided by University of Michigan Metabolomics Core Facility for metabolomics analysis. 50mM ammonium acetate in water was used for rinse buffer.
Small airway epithelial cell (SAEC) (Lonza Walkersville, Inc., MD) were grown according to the manufacturer's instructions in growth medium (SAGM) containing 0.03 mg/ml bovine pituitary extract (BPE), 0.5 Āµg/ml hydrocortisone, 0.5 ng/ml hEGF, 0.5 Āµg/ml epinephrine, 10 Āµg/ml transferrin, 5 Āµg/ml insulin, 0.1 ng/ml retinoic acid, 6.5ng/ml triiodothyronine, 50 Āµg/ml gentamicin and 50ng/ml Amphotericin-B, and 0.5 mg/ml bovine serum albumin (BSA, fatty acid free). When SAE grew to around 80 to 90% confluence, the cells were put into basal medium not supplemented with growth factors 3hours. Then the cell monolayers were infected with naĆÆve hMPV at multiplicity of infection (MOI) of 3 to certain time. The supernatant was saved and cells were harvested and stored exactly according to the protocol provided by University of Michigan Metabolomics Core Facility for metabolomics analysis. 50mM ammonium acetate in water was used for rinse buffer.
Pilot Study 13C flux effects when RhoC or RhoA perturbed (13C BCs)
STUDY_TYPE
13C mass isotopomer analysis (flux studies)
STUDY_SUMMARY
Rho-GTPases are small GTP-binding proteins that contribute to the epithelial-to-mesenchymal transition by regulating several cellular processes including organization of the actin cytoskeleton, cell motility, transcription, and cell proliferation. Overexpression of RhoC-GTPases (RhoC) in breast cancer has been implicated in poor disease prognosis due to increased cancer cells invasion, migration, and motility, which warranted its consideration as a therapeutic target for inhibiting breast cancer metastasis. Using silencing RNA (siRNA) molecules to knockdown RhoC expression is a promising approach to inhibit breast cancer metastases.
INSTITUTE
University of Michigan
DEPARTMENT
Internal Medicine
LABORATORY
Merajver Lab
LAST_NAME
Wynn
FIRST_NAME
Michelle
ADDRESS
University Michigan, 2900 Huron Parkway, Ann Arbor, MI 48105
This experiment was performed using the following cohorts: 1) healthy controls, patients with scleroderma at low risk for pulmonary hypertension, 3) pateints scleroderma at high risk for pulmonary hypertension. Whole blood was drawn into stop soilution (1:1 ratio) at rest and again at peak exercise for each subject.
Colorectal Cancer Detection Using Targeted Serum Metabolic Profiling
STUDY_SUMMARY
Colorectal cancer (CRC) is one of the most prevalent and deadly cancers in the Despite an expanding knowledge of its molecular pathogenesis during the past decades, robust biomarkers to enable screening, surveillance, and therapy of CRC are still lacking. In this study, we present a targeted liquid mass spectrometry-based metabolic profiling approach for identifying biomarker that could enable highly sensitive and specific CRC detection using human serum In this targeted approach, 158 metabolites from 25 metabolic pathways of significance were monitored in 234 serum samples from three groups of patients CRC patients, 76 polyp patients, and 92 healthy controls). Partial least analysis (PLS-DA) models were established, which proved to be powerful for CRC patients from both healthy controls and polyp patients. Receiver operating curves generated based on these PLS-DA models showed high sensitivities (0.96 0.89, respectively, for differentiating CRC patients from healthy controls or patients); good specificities (0.80 and 0.88), and excellent areas under the (0.93 and 0.95) were also obtained. Monte Carlo cross validation (MCCV) was applied, demonstrating the robust diagnostic power of this metabolic profiling
Despite the fact that colorectal cancer (CRC) is one of the most prevalent and deadly cancers in the world, the development of improved and robust biomarkers to enable screening, surveillance, and therapy monitoring of CRC continues to be evasive. In particular, patients with colon polyps are at higher risk of developing colon cancer; however, noninvasive methods to identify these patients suffer from poor performance. In consideration of the challenges involved in identifying metabolite biomarkers in individuals with high risk for colon cancer, we have investigated NMR-based metabolite profiling in combination with numerous demographic parameters to investigate the ability of serum metabolites to differentiate polyp/CRC patients from healthy subjects. We also investigated the effect of disease risk on different groups of biologically related metabolites. Our study may explain some of the challenges and promise a novel avenue for future metabolite profiling methodologies.
In vitro study with AML cell lines that are treated with different concentrations of cytarabine (nucleoside analog). 8 AML cell lines were incubated for 24hr with 0uM, 1uM and 10uM ara-C. After 24hr the cells were washed and pellets were stored in -80Ā°C for genomic and metabolomic analysis.
Maize plants were transformed with endosperm-specific PPDK RNAi knockout constructs to alter starch/protein ratios. Metabolite pool comparisons will be examined from sibling kernels harvested from segregating ears.
Maize plants were grown under three different temperature regimes: 1) normal day / normal night; 2) hot day / normal night; 3) hot day / hot night. Kernels from developing ears were taken 14, 16, 18, 22, 26 and 40 days after pollination.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LABORATORY
Stewart
LAST_NAME
Stewart
FIRST_NAME
Jon D.
ADDRESS
102 Leigh Hall
EMAIL
jds2@chem.ufl.edu
PHONE
352-846-0743
NUM_GROUPS
3
TOTAL_SUBJECTS
47
STUDY_COMMENTS
Normal day / normal night (17 samples); Hot day / normal night (14 samples); Hot day / hot night (17 samples)
LC-MS Based Approaches to Investigate Metabolomic Differences in the Urine of Young Women after Drinking Cranberry Juice or Apple Juice
STUDY_TYPE
drug dosage
STUDY_SUMMARY
Eighteen healthy female college students between 21-29 years old with a normal BMI of 18.5-25 were recruited. Each subject was provided with a list of foods that contained significant amount of procyanidins, such as cranberries, apples, grapes, blueberries, chocolate and plums. They were advised to avoid these foods during the 1-6th day and the rest of the study. On the morning of the 7th day, a first-morning baseline urine sample and blood sample were collected from all human subjects after overnight fasting. Participants were then randomly allocated into two groups (n=9) to consume cranberry juice or apple juice. Six bottles (250 ml/bottle) of juice were given to participants to drink in the morning and evening of the 7th, 8th, and 9th day. On the morning of 10th day, all subjects returned to the clinical unit to provide a first-morning urine sample after overnight fasting. The blood sample was also collected from participants 30 min later after they drank another bottle of juice in the morning. After two-weeks of wash out period, participants switched to the alternative regimen and repeated the protocol. One human subject was dropped off this study because she missed part of her appointments. Another two human subjects were removed from urine metabolomics analyses because they failed to provide required urine samples after juice drinking.The present study aimed to investigate overall metabolic changes caused by procyanidins concentrates from cranberries and apples using a global LCMS based metabolomics approach. All plasma and urine samples were stored at -80oC until analysis.
LC-MS Based Approaches to Investigate Metabolomic Differences in the Plasma of Young Women after Drinking Cranberry Juice or Apple Juice
STUDY_TYPE
drug dosage
STUDY_SUMMARY
Eighteen healthy female college students between 21-29 years old with a normal BMI of 18.5-25 were recruited. Each subject was provided with a list of foods that contained significant amount of procyanidins, such as cranberries, apples, grapes, blueberries, chocolate and plums. They were advised to avoid these foods during the 1-6th day and the rest of the study. On the morning of the 7th day, a first-morning baseline urine sample and blood sample were collected from all human subjects after overnight fasting. Participants were then randomly allocated into two groups (n=9) to consume cranberry juice or apple juice. Six bottles (250 ml/bottle) of juice were given to participants to drink in the morning and evening of the 7th, 8th, and 9th day. On the morning of 10th day, all subjects returned to the clinical unit to provide a first-morning urine sample after overnight fasting. The blood sample was also collected from participants 30 min later after they drank another bottle of juice in the morning. After two-weeks of wash out period, participants switched to the alternative regimen and repeated the protocol. One human subject was dropped off this study because she missed part of her appointments. Another two human subjects were removed from urine metabolomics analyses because they failed to provide required urine samples after juice drinking.The present study aimed to investigate overall metabolic changes caused by procyanidins concentrates from cranberries and apples using a global LCMS based metabolomics approach. All plasma and urine samples were stored at -80oC until analysis.
Vitamin B6 supplementation with 10 mg/d pyridoxine-HCl for 28-d was given to oral contraceptive (OC) users who initially had vitamin B6 deficiency (PLP < 30 nmol/L). Samples are analyzed before and after supplementation. In addition samples from OC users with low (PLP < 30 nmol/L) , middle (PLP 31-99 nmol/L) and high (PLP > 100 nmol/L) vitamin B6 concentration are compared.
Mechanisms of Metabolic Cycles in Diapausing Flesh Fly by Metabolomics Approach
STUDY_TYPE
time course
STUDY_SUMMARY
Insects use diapause, a programmed period of dormancy, to avoid stressful times of the year and to exploit seasonal times of resource availability. Because most diapausing insects do not feed, they must live off their body reserves for several months and the proper use of metabolic reserves is critical for surviving diapause and performing after diapause termination. Across multiple insects, metabolic depression during diapause has been associated with a switch from aerobic metabolism to facultative anaerobic metabolism, despite insects not suffering environmental oxygen limitation. While metabolic rates are depressed during diapause overall to save energy, some insects show regular cyclical bouts of higher metabolic activity during diapause. The functional importance of these metabolic cycles and the mechanisms underlying these cycles are still unknown, but they may be critical for properly maintaining the balance between energy states and purge the accumulation of anaerobic metabolic byproducts. In the present study, we will test the hypothesis that periodic cycles of increased metabolism during insect diapause are associated with both regenerating organismal energetic states, particularly ATP that may decline during metabolic depression, and for purging metabolites associated with anaerobic metabolism. We will use a combination of non-targeted uHPLC-MS/MS metabolomics and targeted NMR-spectroscopy to identify and quantify metabolites that are altered during the cycles in diapausing pupae of the flesh fly, Sarcophaga crassipalpis. This work will allow us to propose specific biochemical and cellular hypotheses for the regulation of cyclic releases from metabolic depression in diapausing insects. Our work may not only reveal the physiological mechanisms regulating metabolic cycles during diapause in flesh fly, but also provide insight to understand the regulation of similar metabolic cycles in mammalian hibernators (i.e., periodic arousal), and also provide insights into how these cycles could be exploited to disrupt the diapause of insect pests.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Chen
FIRST_NAME
Chao
ADDRESS
Department of Entomology and Nematology, Bldg. 970, 1881 Natural Area Dr., Gainesville, FL 32611
Chronic mild stress and Lactobacillus experiments on mice
STUDY_SUMMARY
Mice were devided into three groups (Naive untreated, Stressed untreated, Stressed + Lacto). The stressed groups were subjected to unpredictable chronic mild stress for seven weeks. Three weeks into the protocol, the +Lacto groups were administered probiotic Lactobacillus daily.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Postprandial lipids are lower after vertical sleeve gastrectomy (VSG) surgery and this efect is independent of lipid absorption and chylomicron production. This poses a question of where the lipids are going. In order to test the hypothesis that intestinal oxidation of lipids is increased, Sham or VSG animals were gavaged with glycerol trioleate or water and sacrificed 2h later.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Conjugated linoleic acid (CLA) study in LDLR-/- mice
STUDY_TYPE
Acylcarnitine analysis
STUDY_SUMMARY
LDLR-/- mice were fed a high fat high sucrose diet with either 9,11 CLA or 10,12 CLA. Control groups included no supplementation or caloric restriction to mirror weight loss seen in CLA group. Mice were sacrificed and blood was collected into EDTA tubes. Isolated plasma was immediately frozen at -80C. Adipose tissue from gonadal fat (epididymal white adipose tissue, EWAT) and subcutaneous fat (inguinal white adipose tissue, IWAT) and liver were harvested, snap frozen in liquid nitrogen, and immediately frozen at -80C. An aliquot of thawed plasma was prepared for this project and frozen in an eppendorf tube. Small pieces of frozen tissue were cut and weighed on dry ice and packaged in individual foil packets for this project. **Note: tissue weight is approximate**
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Cecal samples were isolated directly from the ceca of dissected chickens that were either experimentally infected with C. jejuni DRH212 or mock-infected with PBS. Cecal samples were re-suspended in Life Technologies 1X PBS based on weight of sample (1 ml/100 mg = 10-1 dilution) and stored at -80C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
GBM cell lines were plated onto 10 cm dishes at 100,000-200,000 cells per dish and allowed to grow for 3-4 days until confluency reached 50-70%. Media for all cell lines was DMEM with 10% FBS including 25 mM glucose, 6.2 mM glutamine and 200 uM Oleic Acid (conjugated to BSA). 1-2 hours prior to tracer incubation, media was aspirated and fresh media was used. Immediately prior to tracer incubation, media was again aspirated and then cells were washed with warm PBS to remove unlabeled metabolites. Cells were then incubated with labeled tracer (DMEM with 10% FBS including 25 mM Uniformly labeled 13C-6 glucose, 6.2 mM unlabeled glutamine and 200 uM unlabeled oleic acid). After 2 hours of incubation with tracer, media was aspirated, cells were quickly washed with 15 mL DI water, quenched with liquid nitrogen and then stored at -80oC until analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Tumor neurospheres were grown in culture until ~90% confluent. Media was changed to contain 1mM acetate. After 24h, 0h time points were collected and media was changed on all other cells to 1mM 13C-acetate containing media. Cells were then collected at their various time points, 1, 3, 24, 48, or 72 hours. Cells were collected into 15mL tubes, spun down at 100xg for 1min and media aspirated. Pellet was washed (not resuspended) in 150mM ammonium acetate. This was then aspirated off and the cells snap frozen and stored at -80C until all time points complete to ship on dry ice. 0h, 1h, and 3h A, B, and C samples will have gTn by HILIC + TCA by GCMS done on them, while 0h, 1h, and 3h D, E, and F will have FAMES and DG & PC analysis done on them. 24h, 48h, and 72h A, B, and C samples will have FAMES analysis done on them.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Human fecal bile acid profiles before and after fecal transplant
STUDY_SUMMARY
Understand the bile acid profiles from the feces of fecal microbiota transplant FMT patients that successfully recover from recurrent C. difficile infection. Submitting fecal samples from patients prior to their FMT and post FMT. Interested in the bile acid profiles of the donor stool that is used in successful transplants. Bile acids are important for C. difficile spore germination and outgrowth.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Early in life exposure studies on human and mouse samples
STUDY_SUMMARY
Experiment1: 2 day old baby mice were exposed to hyperoxia (75% O2) continuously for 7 days. Control baby mice were housed in room air (normoxia). Plasma and bronchoalveolar lavage fluid (BALF) were harvested after 7 days of exposure (on Day of life 9). Experiment2: 2 day old baby mice were exposed to room air or hyperoxia for 14 days and subsequently treated with RV1 or sham.Plasma was collected 5 days after treatment. Human tracheal aspirates were collected from prematurely born infants undergoing mechanical ventilation for respiratory distress syndrome in the first week of life.Tracheal aspirate supernatants are submitted for the assay. We would like to measure adenosine, AMP, ADP and ATP levels.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Insulin-resistant subjects develop more severe and diffuse coronary artery atherosclerosis than insulin sensitive control but the mechanisms that mediate the atherosclerosis phenotype are unknown. The objective of this study is to investigate whether the severity of atherosclerosis is associated not only with lipoprotein concentrations, weight, blood pressure, biomarkers of inflammation and IR in an animal model but also changes in parameters that measure protein glycation. The experimental approach was to study normocholestrolemic pigs fed a high fat diet that also contained increased NaCl. The choice of pigs was driven by the fact that, like humans, they develop coronary artery and aortic atherosclerosis and insulin resistance. In addition, pigs have been used in many studies to define the mechanisms that mediate increased atherosclerosis in diabetes.
Metabolomics Approach to Identify Molecules and Pathways Involved in the Development of Atherosclerotic Coronary Artery Disease
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Genetics play major roles in the development of atherosclerotic coronary artery disease (CAD). Despite tremendous efforts worldwide invested to decipher the genetic components controlling the development of CAD, the genetic architecture of CAD remains largely unclear. As part of an on-going effort to identify molecules and pathways involved in the development of atherosclerotic CAD, we propose to use rigorous angiographic criteria to define CAD phenotype for genomics and metabolomics study. We identified two extreme groups, namely āyoung CADā group, who are very young individuals (age <= 40 years) proven to have severe CAD required revascularization, and āCAD-free elderlyā, who are at very advanced age (Age >= 80 years) but have no angiographically apparent CAD. Phenotypically, these two groups are in sharp contrary. Conventional risk factors account for small portion of different phenotypes. We hypothesize that there are genetically programmed pathways and molecules accelerating atherosclerotic pathogenesis, in the āyoung CADā patients and preventing the development of CAD in the āCAD-free elderlyā patients. We sought to combine genomics and metabolomics approaches to profile and identify these pathways and molecules. Both plasma and urine samples from patients in these two groups, and their age matched control groups, will undergo unbiased metabolomics profiling with high throughput quantitative nuclear magnetic resonance (NMR) and mass spectrometry (MS) technology in RTI metabolomics core facility. Comprehensive statistic and multi-variant analytic approaches will be used to identify pathways and molecules significance to the pathogenesis of atherosclerosis. These data will be integrated with genomics data from next generation sequencing of genetic materials from the same groups of patients to further explore the molecular mechanisms underlying atherosclerosis and CAD.
13C mass isotopomer analysis (LCMS flux studies) MLL-AF9 (part I)
STUDY_TYPE
1,2-13C2 Flux analysis
STUDY_SUMMARY
"How cancer cells adapt to metabolically adverse conditions in patients and strive to proliferate is a fundamental question in cancer biology. Here we show that AMP-activated protein kinase (AMPK), a metabolic checkpoint kinase, confers metabolic stress resistance to leukemia-initiating cells (LICs) and promotes leukemogenesis. Upon dietary restriction, MLL-AF9-induced murine acute myeloid leukemia (AML) activated AMPK and maintained leukemogenic potential. AMPK deletion significantly delayed leukemogenesis and depleted LICs by reducing the expression of glucose transporter 1 (Glut1), compromising glucose flux, and increasing oxidative stress and DNA damage. LICs were particularly dependent on AMPK to suppress oxidative stress in the hypoglycemic bone marrow environment. Strikingly, AMPK inhibition synergized with physiological metabolic stress caused by dietary restriction and profoundly suppressed leukemogenesis. Our results indicate that AMPK protects LICs from metabolic stress and that combining AMPK inhibition with physiological metabolic stress potently suppresses AML by inducing oxidative stress and DNA damage. Research is published: http://www.sciencedirect.com/science/article/pii/S1934590915003744 "
INSTITUTE
University of Michigan
DEPARTMENT
Molecular and Human genetics (Baylor College of Medicine)
LABORATORY
Nakada Lab (Baylor College of Medicine)
LAST_NAME
Saitoh
FIRST_NAME
Yusuke
ADDRESS
Houston, TX
EMAIL
Yusuke.Saitoh@bcm.edu
PHONE
713-798-1175
NUM_GROUPS
7
TOTAL_SUBJECTS
18
STUDY_COMMENTS
1. Collect leukemia cells from MLL-AF9 leukemia mouse bone marrow. 2. Leukemia cell were cultured in RPMI1640 medium for 16hr. 3. Cell pellets were resuspend in pre-warmed labeling medium containing 1,2-13C2-glucose(2g/L) and incubated for 5min (sample 5) or 60min(sample 60).
Metabolome of three Vancomycin-intermediate Staphylococcus aureus (VISA) mutants compared with the parent strain MM66
STUDY_SUMMARY
Vancomycin-intermediate Staphylococcus aureus (VISA) evolve in a strain-specific manner and acquire mutations that lead to alterations in cell wall metabolism that reduce susceptibility to vancomycin. We had earlier isolated several VISA mutant strains of the clinical hVISA strain MM66. This study is aimed at analyzing the metabolome of these mutants in comparison to the parent strain.
INSTITUTE
Oklahoma State University, Stillwater, OK
LAST_NAME
Gustafson
FIRST_NAME
John
ADDRESS
246C Noble Research Center Oklahoma State University Stillwater, OK 74078-3035
IDH1R132H activity in glioma cell lines and tumnor tissue (2HG)
STUDY_TYPE
Regular
STUDY_SUMMARY
We developed genetically engineered mice to generate brain tumors with especific genetic lessions. The animals were split in three groups: NRAS, P53 knockdown, IDH1-R132H and ATRX knock down (NPAID); NRAS, P53 knockdown, IDH1-R132H (NPI); NRAS, P53 knockdown, (NshP53). From these tumor we obtain and culture tumor cells growth like neurospheres and attached cells.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Muscle Clock knock out mice metabolic changes (iMSBmal1-Exp1)
STUDY_TYPE
Regular
STUDY_SUMMARY
My lab studies the function of the molecular clocks in skeletal muscle. We have an inducible genetic mouse model (C57Bl6 background) in which we knock out the core clock gene, Bmal1, only in adult skeletal muscle after treatment with tamoxifen. We have found that the mice maintain body mass but lose fat mass at 10 weeks after loss of Bmal1. We have done expression profiling on the skeletal muscles and gene expression changes (insulin signaling, CHO metabolism, fat metabolism) suggest significant changes in substrate metabolism. To analyze TCA, CHO metabolites we have collected gastrocnemius muscles from these mice following instructions from Dr. Burant. Mice were anaesthetized with isoflurane, the gastrocnemius muscle dissected and flash frozen with tongs cooled with liquid N2. They have been stored in cryovials in our -80 freezer for 2 months.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
NSAID treatment alters the metabolomics profile of liver, kidney, lung, and heart in an experimental mouse model of heat stroke
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
The objective of this study is to exploit broad spectrum metabolomic analysis to identify new biomarkers of multi-organ damage that will improve heat stroke (HS) diagnosis and treatment. The central hypothesis is that HS will lead to significant alterations in multi-organ metabolomics profiles that will serve as markers of HS severity, which will be shifted and intensified further by the acute use of NSAIDs. To test this hypothesis, we will be performing broad spectrum metabolomics to identify alternations in the metabolic signatures of key organs (heart, liver, kidney, and lung) in a highly validated rodent HS model leveraging implantable radiotelemetry. We will then compare these results with already completed histological gene/protein expression analysis to determine the best metabolic markers of HS induced organ damage. The results from this study will aid in the identification of preventative measures to reduce HS risk, as well as in developing therapeutics to treat multi-organ damage and facilitate recovery. The proposed study will provide the first metabolic assessment of HS severity and NSAID use, which will support future studies in HS patients to validate novel biomarkers that will improve clinical assessment of organ damage and recovery.
Metabolomics and Childhood Obesity: A Pilot and Feasibility Study With Multiple Phenotypic Anchors
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
āMetabolomicsā is a powerful new analytical approach for measuring and evaluating all small and intermediate sized metabolites in a variety of tissues or samples in conditions of health and disease. The purpose of this research is to determine if āmetabolomicsā can be used to address several important unanswered questions about obesity in children. First we will use metabolomics to identify patterns of metabolites in blood that are unique to obese children. We will then determine if these patterns are predictive of excessive weight gain and/or poor weight loss response in non-obese and obese children enrolled in an exercise program.
Comparison of Metabolites Variation and Antiobesity Effects of a Mixture of Cudrania tricuspidata, Lonicera caerulea, and the Soybean According to Fermentation in vitro and in vivo
STUDY_SUMMARY
We used ultra-performance-liquid-chromatography with quadrupole-time-of-flight mass spectrometry to study the changes in metabolites in the mixture of Cudrania tricuspidata, Lonicera caerulea, and soybean (CLM) during fermentation. Additionally, the antiobesity effects of CLM and fermented-CLM (FCLM) were studied based on the analysis of plasma from high-fat diet (HFD)-fed mice. The levels of cyanidin and the glycosides of luteolin, quercetin, and cyanidin derived from L. caerulea were decreased, whereas the levels of luteolin and quercetin were increased during fermentation. Isoflavone glycosides and soyasaponins originating from the soybean were decreased, whereas their aglycones such as daidzein, glycitein, and genistein were increased. As for prenylated flavonoids from C. tricuspidata, these metabolites were decreased at the early stage of fermentation, and were increased at end of the fermentation. In terms of the functional food product, various metabolites derived from diverse natural products in CLM had complementary effects and demonstrated higher antioxidant and pancreatic lipase inhibition activities by fermentation; these activities were closely related to flavonoid aglycones including genistein, daidzein, glycitein, luteolin, and quercetin. In vivo experiment, several clinical parameters affected by HFD were remarkably improved by the administration of either CLM or FCLM, but there was a difference in the antiobesity effects. The levels of lysoPCs with C20:4, C16:0, and C22:6 were significantly attenuated by CLM administration, while the attenuated levels of lysoPCs with C20:4 and C18:2 were significantly restored by FCLM administration. These metabolites may explain the above-mentioned differences in antiobesity effects. Although only the changes in plasma lysophospholipids could not fully explain antiobesity effects between non-fermented and fermented plant mixtures from our results, we suggest that metabolomics approach could provide a way to reveal the metabolite alterations in the complex fermentation process and understand the differences or changes in bioactivity according to fermentation.
INSTITUTE
Konkuk university
LAST_NAME
Suh
FIRST_NAME
Dong Ho
ADDRESS
Neong-Dong-ro 120, Seoul, Kwang-Gin-gu, 05029, Korea, South
Experiment to test how different growth mediums affect bacterial growth. The supernatants of 17 strains of bacteria (10 grown in one medium and 7 grown in another medium) were submitted for metabolite analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Allantoin differences in Synechococcus cells grown in high versus low lightgrowth
STUDY_SUMMARY
This experimented consisted of analysis of 500-1000ml of Synechococcus dense cell cultures grown in high versus low light. The goal was to see any differences in the metabolites between the two treatments, especially with respect to Allantoin.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Single treatment gene impact on Arabidopsis metabolites
STUDY_SUMMARY
This experiment aims to measure the impact of the genes that have been introduced into WT lines and compare the metabolic profiling of these plants with WT control plants. Compounds of particular intereset for this study include pyruvate, fumarate, malate, glyoxylate, anthocyanin, carotenes, and lipid compounds.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
This experiment tests the effects of alcoholism by examining the primary metabolites obtained from mouse stool. Stool was collected from 4 groups of mice with varying treatments. One group was not humanized, another was humanized from a healthy human, a third was humanized from a current drinker, and the final group was humanized from a current drinker but also given a treatment of LGG. Humanizations were done to create a model of human gut activity in the mice.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Progesterone level effects on primary metabolites in uterus, blood, and ovaries (Part 1:Plasma)
STUDY_SUMMARY
In this experiment a hormonal protocol was applied to control follicle growth to yield larger or smaller preovulatory follicle and CLs and consequently different circulating Progesterone (P4) concentrations during early diestrus. The two different animal's group are: high or low progesterone levels. The effects of these progesterone levels was tested in the blood of the cow.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Progesterone level effects on primary metabolites in uterus, blood, and ovaries (Part 2:Uterine flush)
STUDY_SUMMARY
In this experiment a hormonal protocol was applied to control follicle growth to yield larger or smaller preovulatory follicle and CLs and consequently different circulating Progesterone (P4) concentrations during early diestrus. The two different animal's group are: high or low progesterone levels. The effects of these progesterone levels was tested in the uterus of the cow.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Progesterone level effects on primary metabolites in uterus, blood, and ovaries (Part 3:Ovaries)
STUDY_SUMMARY
In this experiment a hormonal protocol was applied to control follicle growth to yield larger or smaller preovulatory follicle and CLs and consequently different circulating Progesterone (P4) concentrations during early diestrus. The two different animal's group are: high or low progesterone levels. The effects of these progesterone levels was tested in the ovaries (follicle fluid) of the cow.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic effects of metformin on mouse liver, intestine, and serum
STUDY_SUMMARY
Experiment to test the different metabolomic effects of two different doses of metformin (50mg vs 150 mg). A saline treatment group was used as a control. The effects were measured at the liver, intestine, and serum of the mouse.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Role of medium in bacterial growth (HILIC chromatography)
STUDY_SUMMARY
Experiment to test how different growth mediums affect bacterial growth. The supernatants of 17 strains of bacteria (10 grown in one medium and 7 grown in another medium) were submitted for metabolite analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Primary metabolites at different points along dog gastrointestinal tract
STUDY_SUMMARY
This experiment tests the primary metabolites at four different points along the gastrointestinal tract of a dog. The four points being tested were the duodenum, ileum, colon, and rectum.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Primary metabolites at different points along dog gastrointestinal tract (RP chromatography)
STUDY_SUMMARY
This experiment tests the primary metabolites at four different points along the gastrointestinal tract of a dog. The four points being tested were the duodenum, ileum, colon, and rectum.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
This experiment tested the affects of different diets on mice esophagus metabolites. The diets ranged from zinc sufficient to zinc deficient and a third group that included zinc deficient mice that were put back on zinc sufficient diets.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Effects of Zinc on GI tract metabolites (Part 2: Prostate)
STUDY_SUMMARY
This experiment tested the affects of different diets on mice prostate metabolites. The diets ranged from zinc sufficient to zinc deficient and a third group that included zinc deficient mice that were put back on zinc sufficient diets.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
This experiment aimed to see the effects of Giardia Intestinalis on the small intestine of mice. The metabolites of the proximal and distal ends of the small intestine of healthy mice were compared to those of mice who had been infected with Giardia intestinalis for 7 days.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomics of bovine uterine fluid at the onset of conceptus elongation
STUDY_TYPE
Prospective cohort study
STUDY_SUMMARY
The objective is to investigate changes in metabolomics of uterine lumen content of lactating dairy cows associated with the onset of conceptus (embryo and associated membranes) elongation. Lactating dairy cows had estrous cycles synchronized and were subjected to induced ovulation and timed artificial insemination (AI). The day of AI was considered study d 0. On d 15, uteri were flushed by transcervical catheterization and infusion of 20 mL of phosphate buffered solution with 0.1% of polyvinyl acetate. Recovered conceptuses were classified based on morphology/length as ovoid (OV; 1-4 mm), tubular (TUB; 5-19 mm) and filamentous (FIL; 20-85 mm). The first 20 mL infused in the uterus were recovered, placed in conical tubes and centrifuged at 2,000 Ć g at 4įµC. The supernatant was collect, aliquoted and stored at -80įµC for later analyses of fluid composition, including measurement of IFN-Ļ concentration. Cows with no conceptus recovered and no detection of IFN-Ļ in uterine flushing were considered as nonpregnant (NPREG). The experimental design was then considered a prospective cohort study with 4 independent groups (NPREG, OV, TUB, and FIL). The additional 5th group represents a specific physiological condition of cows within the study and it will be compared to TUB and FIL groups combined, working as a pilot study for future research.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LABORATORY
Laboratory of Reproduction and Nutrition of Dairy Cows
Targeted LC/MS of urine from boys with DMD and controls
STUDY_TYPE
Natural History
STUDY_SUMMARY
Quantify the urine levels of amino acids and organic acids in patients with DMD both with and without steroid treatment. Track the progression of DMD in patients who have provided multiple urine samples.
Metabolomics Approach to Allograft Assessment in Liver Transplantation
STUDY_TYPE
Retrospective analysis of biobanked liver tissue from organ donors
STUDY_SUMMARY
This pilot study is designed to apply several types of metabolomic analysis for liver allograft assessment with the aim of identifying candidate biomarkers for allograft function and to develop a methodology that could be applied to larger scale studies. The three main categories of metabolomic analysis are reflected in each of the specific aims, including a targeted profiling of central carbon metabolism, open lipidomic and metabolomic profiling for hypothesis generation and MALDI-IMS for tissue-based spatial analysis. Each of these approaches offers potential benefits that could be optimized in a protocol for larger scale studies depending the results of the pilot investigations.
INSTITUTE
Ochsner Multi-Organ Transplant Institute
DEPARTMENT
SECIM
LABORATORY
Abdominal Organ Transplantation
LAST_NAME
Seal
FIRST_NAME
John
ADDRESS
1514 Jefferson Highway, New Orleans, LA 70121
EMAIL
John.Seal@ochsner.org
PHONE
504-232-4253
NUM_GROUPS
3
TOTAL_SUBJECTS
Targeted analysis and open profiling: Control group - standard criteria donor (N=15); Experimental group - donation-after-cardiac-death donors (N=10). MALDI-IMS - standard criteria donor (N=10)
Unbiased profiling uncovers a crucial role for gut microbiome derived metabolites in modulating GI epithelial cell damage and mitigating GVHD.
STUDY_SUMMARY
Taxonomic alterations in the intestinal microbiota are being progressively associated with many diseases, including graft-versus host disease (GVHD). However, the impact of these alterations on microbial metabolites and by-products and their subsequent impact on disease processes, such as GVHD, are not known. Here we utilized a targetedn unbiased and blinded approach in a blinded fashion to identify novel alterations in the levels of microbial metabolites, specifically levels including the short chain fatty acid (SCFA) and endogenous histone deacetylase inhibitor (HDACi), butyrate, after allo-BMT. Surprisingly, alterations were observed only in intestinal epithelial cells (IECs) but not in the luminal contents. The reduced butyrate in IECs (CD326+) after allo-BMT resulted in decreased histone acetylation, which was restored upon local administration of exogenous butyrate. This resulted in improved IEC junctional integrity, increased anti-apoptotic proteins, decreased GVHD, and improved survival. Furthermore, alteration of endogenous microflora with 17 rationally selected strains of high butyrate producing Clostridia, also decreased GVHD and increased survival following allo-BMT in experiments performed at two different institutions. These data demonstrate an heretofore unrecognized role of microbial metabolites and suggests that local and specific alteration of microbial metabolites has direct salutary effects on GVHD target tissues and mitigates its severity.
This experiment aimed to discover the effects of metformin on mouse liver and kidney tissue. The effects were seen by comparing the liver of the metformin group to the liver of a control group of mice treated given saline solution.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Renal metabolic pathways indicating ischemic or inflammatory changes
STUDY_SUMMARY
Tissues were acquired from kidkeys that were deemed unsuitable for transplant and were then analyzed by the lab through normothermic machine perfusion. They were perfused with either whole blood perfusate or with packed red blood cell perfusate. These tissues' metabolic pathways were then analyzed for markers of ischemic or inflammatory responses in the renal tissue
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Modification of metabolites by gut microbiota in response to diet
STUDY_SUMMARY
This experiment is looking at effects of diets on rats. Specifically how those diets might alter metabolites that could be modified by gut microbiota and in particular indoles and bile salts.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Effects of dietary supplement on hamster metabolism
STUDY_SUMMARY
This experiment aims to analyze spent media from a protein over-expression system. The treatment was a lipid supplement given to hamsters. The spent media was then analyzed to see how the lipid supplement affected lipid metabolism.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolites detected from human bronchoalveolar lavage
STUDY_SUMMARY
This is a preliminary trial to determine how viable this system will be to use on a much larger number of samples (up to 150). We would like to determine the range, number of metabolite species, and relative concentrations than can be detected in human bronchoalveolar lavage. We would also like to determine how clear the distinction is between the 3 patient groups as this will inform us as to how many of the 150 samples we need to run in the study proper.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic analysis on samples from rats expressing human amylin (cardiac tissue).
STUDY_SUMMARY
Human amylin proteotoxicity impairs protein biosynthesis, and alters major cellular signaling pathways in the heart, brain and liver of humanized diabetic rat model in vivo. The 37 amino acid hormone Amylin is coĀsecreted with insulin from Ć cells in the pancreas. In preĀdiabetic and obese humans, chronic amylin hypersecretion parallels the course of disease and is involved in the pathophysiology of beta cell destruction in the pancreas. Recent studies in rats with transgenic expression of beta cell amylin (HIP), we have discovered that human amylin is prone to misfolding and has proteotoxic effects in vivo, resulting in the induction of cell death paralleling the pathophysiology of neurodegenerative disease. These misfolded proteotoxic amylin proteins are found to migrate to both the brain and heart to induce both neurologic deficits and cardiac dysfunction. In the present study, we use nonĀtargeted GCĀMS metabolomics analysis to investigate the metabolic consequences of amyloidogenic and cytotoxic amylin oligomers and diabetes in HIP heart, brain, liver, and plasma compared to wild type controls at 1 year of age. We identified that HIP hearts had 45 significantly altered metabolites by tĀtest (p<0.05) compared to wildtype control hearts (0.1Ā34.3 fold different,N=8/group). Similarly, we identified 30 metabolites significantly different in the HIP brain by tĀtest (p<0.05) compared to wildtype control brains (0.2Ā25.2 fold different (N=~10/group). HIP livers had 58 metabolites significantly altered by tĀtest (p<0.05) compared to wildtype livers (0.01Ā99.4 fold different,N=~10/group. Pathway enrichment analysis identified a systemic alteration in protein biosynthesis in the heart and brain of HIP rats compared to wild type controls. Alterations in phenylalanine metabolism and aminoacylĀtRNA biosynthesis were specifically affected in heart and plasma. Tyrosine metabolism is affected across organs, including decreased tyrosine (heart), phenylalanine (heart, liver, brain), and increased fumarate (heart, liver, brain). Increased urea and urea cycle were identified in heart and liver. As protein degradation is a major upĀregulator of the urea cycle in human rat diabetic models, these findings suggest a broader connection between amylin, diabetes, protein catabolism, and effects on the urea cycle, which may contribute to the increased morbidity and mortality in diabetics at a multiĀsystem level beyond the effects on glucose metabolism.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Ilaiwy
FIRST_NAME
Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Metabolomic analysis on samples from rats expressing human amylin (brain tissue).
STUDY_TYPE
Non targeted metabolomics-Brain tissue
STUDY_SUMMARY
Human amylin proteotoxicity impairs protein biosynthesis, and alters major cellular signaling pathways in the heart, brain and liver of humanized diabetic rat model in vivo. The 37 amino acid hormone Amylin is coĀsecreted with insulin from Ć cells in the pancreas. In preĀdiabetic and obese humans, chronic amylin hypersecretion parallels the course of disease and is involved in the pathophysiology of beta cell destruction in the pancreas. Recent studies in rats with transgenic expression of beta cell amylin (HIP), we have discovered that human amylin is prone to misfolding and has proteotoxic effects in vivo, resulting in the induction of cell death paralleling the pathophysiology of neurodegenerative disease. These misfolded proteotoxic amylin proteins are found to migrate to both the brain and heart to induce both neurologic deficits and cardiac dysfunction. In the present study, we use nonĀtargeted GCĀMS metabolomics analysis to investigate the metabolic consequences of amyloidogenic and cytotoxic amylin oligomers and diabetes in HIP heart, brain, liver, and plasma compared to wild type controls at 1 year of age. We identified that HIP hearts had 45 significantly altered metabolites by tĀtest (p<0.05) compared to wildtype control hearts (0.1Ā34.3 fold different,N=8/group). Similarly, we identified 30 metabolites significantly different in the HIP brain by tĀtest (p<0.05) compared to wildtype control brains (0.2Ā25.2 fold different (N=~10/group). HIP livers had 58 metabolites significantly altered by tĀtest (p<0.05) compared to wildtype livers (0.01Ā99.4 fold different,N=~10/group. Pathway enrichment analysis identified a systemic alteration in protein biosynthesis in the heart and brain of HIP rats compared to wild type controls. Alterations in phenylalanine metabolism and aminoacylĀtRNA biosynthesis were specifically affected in heart and plasma. Tyrosine metabolism is affected across organs, including decreased tyrosine (heart), phenylalanine (heart, liver, brain), and increased fumarate (heart, liver, brain). Increased urea and urea cycle were identified in heart and liver. As protein degradation is a major upĀregulator of the urea cycle in human rat diabetic models, these findings suggest a broader connection between amylin, diabetes, protein catabolism, and effects on the urea cycle, which may contribute to the increased morbidity and mortality in diabetics at a multiĀsystem level beyond the effects on glucose metabolism.
INSTITUTE
Duke University
DEPARTMENT
Sarah W. Stedman Nutrition and Metabolism Center
LABORATORY
Metabolomics lab
LAST_NAME
Ilaiwy
FIRST_NAME
Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
EMAIL
amroilaiwy@gmail.com, monte_willis@med.unc.edu
PHONE
210-596-0171
STUDY_COMMENTS
Brain tissue-Metabolomic analysis was performed at Duke Metabolomics lab
Metabolomic analysis on samples from rats expressing human amylin (hepatic tissue).
STUDY_SUMMARY
Human amylin proteotoxicity impairs protein biosynthesis, and alters major cellular signaling pathways in the heart, brain and liver of humanized diabetic rat model in vivo. The 37 amino acid hormone Amylin is coĀsecreted with insulin from Ć cells in the pancreas. In preĀdiabetic and obese humans, chronic amylin hypersecretion parallels the course of disease and is involved in the pathophysiology of beta cell destruction in the pancreas. Recent studies in rats with transgenic expression of beta cell amylin (HIP), we have discovered that human amylin is prone to misfolding and has proteotoxic effects in vivo, resulting in the induction of cell death paralleling the pathophysiology of neurodegenerative disease. These misfolded proteotoxic amylin proteins are found to migrate to both the brain and heart to induce both neurologic deficits and cardiac dysfunction. In the present study, we use nonĀtargeted GCĀMS metabolomics analysis to investigate the metabolic consequences of amyloidogenic and cytotoxic amylin oligomers and diabetes in HIP heart, brain, liver, and plasma compared to wild type controls at 1 year of age. We identified that HIP hearts had 45 significantly altered metabolites by tĀtest (p<0.05) compared to wildtype control hearts (0.1Ā34.3 fold different,N=8/group). Similarly, we identified 30 metabolites significantly different in the HIP brain by tĀtest (p<0.05) compared to wildtype control brains (0.2Ā25.2 fold different (N=~10/group). HIP livers had 58 metabolites significantly altered by tĀtest (p<0.05) compared to wildtype livers (0.01Ā99.4 fold different,N=~10/group. Pathway enrichment analysis identified a systemic alteration in protein biosynthesis in the heart and brain of HIP rats compared to wild type controls. Alterations in phenylalanine metabolism and aminoacylĀtRNA biosynthesis were specifically affected in heart and plasma. Tyrosine metabolism is affected across organs, including decreased tyrosine (heart), phenylalanine (heart, liver, brain), and increased fumarate (heart, liver, brain). Increased urea and urea cycle were identified in heart and liver. As protein degradation is a major upĀregulator of the urea cycle in human rat diabetic models, these findings suggest a broader connection between amylin, diabetes, protein catabolism, and effects on the urea cycle, which may contribute to the increased morbidity and mortality in diabetics at a multiĀsystem level beyond the effects on glucose metabolism.
INSTITUTE
University of North Carolina
LABORATORY
Multiple Centers
LAST_NAME
Ilaiwy
FIRST_NAME
Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Metabolomic analysis on samples from rats expressing human amylin (plasma).
STUDY_SUMMARY
Human amylin proteotoxicity impairs protein biosynthesis, and alters major cellular signaling pathways in the heart, brain and liver of humanized diabetic rat model in vivo. The 37 amino acid hormone Amylin is coĀsecreted with insulin from Ć cells in the pancreas. In preĀdiabetic and obese humans, chronic amylin hypersecretion parallels the course of disease and is involved in the pathophysiology of beta cell destruction in the pancreas. Recent studies in rats with transgenic expression of beta cell amylin (HIP), we have discovered that human amylin is prone to misfolding and has proteotoxic effects in vivo, resulting in the induction of cell death paralleling the pathophysiology of neurodegenerative disease. These misfolded proteotoxic amylin proteins are found to migrate to both the brain and heart to induce both neurologic deficits and cardiac dysfunction. In the present study, we use nonĀtargeted GCĀMS metabolomics analysis to investigate the metabolic consequences of amyloidogenic and cytotoxic amylin oligomers and diabetes in HIP heart, brain, liver, and plasma compared to wild type controls at 1 year of age. We identified that HIP hearts had 45 significantly altered metabolites by tĀtest (p<0.05) compared to wildtype control hearts (0.1Ā34.3 fold different,N=8/group). Similarly, we identified 30 metabolites significantly different in the HIP brain by tĀtest (p<0.05) compared to wildtype control brains (0.2Ā25.2 fold different (N=~10/group). HIP livers had 58 metabolites significantly altered by tĀtest (p<0.05) compared to wildtype livers (0.01Ā99.4 fold different,N=~10/group. Pathway enrichment analysis identified a systemic alteration in protein biosynthesis in the heart and brain of HIP rats compared to wild type controls. Alterations in phenylalanine metabolism and aminoacylĀtRNA biosynthesis were specifically affected in heart and plasma. Tyrosine metabolism is affected across organs, including decreased tyrosine (heart), phenylalanine (heart, liver, brain), and increased fumarate (heart, liver, brain). Increased urea and urea cycle were identified in heart and liver. As protein degradation is a major upĀregulator of the urea cycle in human rat diabetic models, these findings suggest a broader connection between amylin, diabetes, protein catabolism, and effects on the urea cycle, which may contribute to the increased morbidity and mortality in diabetics at a multiĀsystem level beyond the effects on glucose metabolism.
INSTITUTE
University of North Carolina
LABORATORY
Multiple Centers
LAST_NAME
Ilaiwy
FIRST_NAME
Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Determining the metabolic profile of wildtype, lrgAB, and atlA mutant Steptococcus mutans grown aerobically and anaerobically
STUDY_TYPE
Single time point, aerobic vs. anaerobic cultures
STUDY_SUMMARY
Whole cells from both aerobic and anaerobic cultures of wild-type,lrgAB and atlA strain will be isolated for analysis of the whole cytosolic metabolome. Supernatants will be also analyzed for their metabolite profile in wild type and lrgAB cultures.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Ahn
FIRST_NAME
Sang-Joon
ADDRESS
1395 Center Drive, Room D5-29 Gainesville FL 32610
The Development of Metabolomic Markers in African Bermudagrass (C. transvaalensis) for Sting Nematode (Belonolaimus longicaudatus) Response
STUDY_TYPE
Disease response in terms of nematode reproduction and root weight
STUDY_SUMMARY
The objective of the proposed pilot study is to identify metabolites up- and down-regulated in African bermudagrass that are tolerant and sensitive to the sting nematode and develop metabolomic markers for the highest expressed metabolites associated with tolerance. Future work will include additional accessions and species of bermudagrass, and testing under field conditions. Bermudagrass accessions identified as tolerant or sensitive by Pang et al. (2011) will be assessed under controlled greenhouse conditions to identify metabolites linked to sting nematode tolerance. Nematode response will be quantified through determination of root length and weight and the number of nematodes present 136 days after inoculation. Higher root length and weight indicate tolerance or resistance. Higher nematode counts indicate greater reproduction (i.e. a susceptible plant), while lower counts indicate that the accession may have some resistance. Metabolites from root tissue of these accessions will be compared to identify those associated with tolerance/resistance, and those that are associated with nematode infestation by comparing inoculated plants to uninoculated controls. Metabolomic markers will then be developed for the metabolites associated with tolerance/resistance. These markers will be used to guide future screening of bermudagrass accessions for breeding nematode-tolerant or -resistant varieties.
Metabolite comparison of mouse gastric tissue and glands
STUDY_SUMMARY
The goal of this project was to compare the metabolite profiles of the: mouse gastric antrum and the mouse gastric corpus, the mouse gastric antrum and the mouse gastric antrum isolated glands, and the mouse gastric corpus and the mouse gastric corpus isolated glands.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Broad Spectrum MS analysis of mouse hypothalmus from Anxiety Prone HSV-Latently Infected Obese Mice
STUDY_TYPE
Broad Spectrum LCMS
STUDY_SUMMARY
Brain tissue samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics.
Broad Spectrum MS analysis of mouse hippocampus from Anxiety Prone HSV-Latently Infected Obese Mice
STUDY_TYPE
Broad Spectrum LCMS
STUDY_SUMMARY
Brain tissue samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics.
Broad Spectrum MS analysis of mouse microglia cells from Anxiety Prone HSV-Latently Infected Obese Mice
STUDY_TYPE
Broad Spectrum LCMS
STUDY_SUMMARY
A mouse model of obese HSV-1 latent infection was developed. Broad spectrum metabolomics analysis was performed to better understand the metabolomic profile of microglia cells and to compare this metabolomics profile with that of the hippocampus, hypothalamus, and peripheral blood mononuclear cells. Microglia cell samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and microglia cell samples were collected and processed for metabolomics.
Characterization and Plasticity of the Metabolome in Peripheral Cells in Bipolar I Disorder
STUDY_TYPE
Broad Spectrum LCMS
STUDY_SUMMARY
Patients with Bipolar I Disorder (BPI) and matched non-affected controls were recruited. Males and females from all races and ethnicities between the ages of 19-65 participated in the research. Skin biopsies were obtained and fibroblasts were isolated from each of these biopsies. A total of 1 x ć10ć^7 fibroblasts cells were used for metabolomics. Cell pellets were flash frozen in liquid nitrogen and shipped on dry ice to the NIH Eastern Regional Comprehensive Metabolomic Resource Core at RTI International in North Carolina. Metabolomics will be determined using a broad spectrum metabolomics protocol by RTI
Associations between 69 Sphingolipids and Emphysema, Chronic Bronchitis, Exacerbations, and FEV1/FVC
STUDY_TYPE
Association Study
STUDY_SUMMARY
One hundred twenty-nine current and former smokers from the COPDGene cohort had 69 distinct sphingolipid species detected in plasma by targeted mass spectrometry. Of these, 23 were also measured in 131 plasma samples (117 independent subjects) using an untargeted platform in an independent laboratory. Regression analysis with adjustment for clinical covariates, correction for false discovery rate, and metaanalysis were used to test associations between COPD subphenotypes and sphingolipids. Peripheral blood mononuclear cells were used to test associations between sphingolipid gene expression and plasma sphingolipids.
Broad Spectrum MS analysis of mouse peripheral blood mononuclear cells (PBMC) from Anxiety Prone HSV-Latently Infected Obese Mice
STUDY_TYPE
Broad Spectrum LCMS
STUDY_SUMMARY
Mononuclear cell samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and mononuclear cell samples were collected and processed for metabolomics.
Zebrafish Metabolomics: Model for Environmental Metal Toxicity
STUDY_SUMMARY
This metabolomics seed project will test the hypothesis that zebrafish can provide mechanistic insights into the human health effects of developmental exposure to Cd and Pb. We will use broad spectrum metabolomics of zebrafish larvae after exposure to Cd, Pb, and Cd and Pb compared to controls. Activity observed at 5 days post fertilization is will be used to determine if there is a correlation between biological pathways implicated by these metabolic profiles and cardiovascular, metabolic (obesity), and neurological phenotypes. The behavioral phenotypes have been quantified in zebrafish previously and have been measured in the Newborn Epigenetic STudy (NEST), a NIH-funded project that is investigating how environmental exposures and nutrition, in the womb and during childhood, affect how genes work and how these exposures developed into obesity and other diseases, disorders, and conditions. The results from this study will demonstrate the power of using zebrafish as a model for mechanism discovery in exposure using metabolomics to advance understanding of early life exposure to Cd and Pb and serve as preliminary data for opportunities.
INSTITUTE
University of North Carolina
DEPARTMENT
Discovery Sciences
LABORATORY
Sumner Lab
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Metabolomics analysis of colon adenoma in African Americans
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
This NMR Metabolomics analysis was performed on feces samples derived from healthy (n = 10) and adenoma (n = 10) African American subjects with the goal of identifying perturbations in metabolomics profiles in colon cancer.
Distinctly perturbed metabolic networks underlie differential tumor tissue damages induced by immune modulator b-glucan in a two-case ex vivo non-small cell lung cancer study
STUDY_TYPE
tracer
STUDY_SUMMARY
[U-13C]-Glu SIRM study of ex vivo tissue slices in matched-pair tumor/non-tumor ex vivo tissue slices from two human patients with and withou beta-glucan.
INSTITUTE
University of Kentucky
DEPARTMENT
CESB
LAST_NAME
Fan
FIRST_NAME
Teresa
ADDRESS
Rm 516 Biopharm Complex, 789 S. Limestone St.,Univ. of Kentucky, Lexington, KY 40536
Investigation of metabolomic blood biomarkers for detection of adenocarcinoma lung cancer
STUDY_TYPE
Lung cancer case control biomarker discovery
STUDY_SUMMARY
Recently, the National Lung Cancer Screen Trial (NLST) demonstrated that low-dose CT (LDCT) screening could reduce mortality due to lung cancer by 20%. However, LDCT screening is largely hindered by high false-positive rates (96%), particularly in high-risk populations (heavy smokers), due to the low prevalence rates (less than 2%) of malignant tumors and high incidence of benign lung nodules. Consequently, complementary biomarkers that can be used in conjunction with LDCT screening to improve diagnostic capacities and reduce false-positive rates are highly desirable. Preferably, such complementary tools should be noninvasive and exhibit high sensitivity and specificity. The application of ā-omicā sciences (genomics, transcriptomics, proteomics, and metabolomics) represents valuable tools for the discovery and validation of potential biomarkers that can be used for detection of NSCLC. Of these omic sciences, metabolomics has received considerable attention for its application in cancer. Metabolomics is the assessment of small molecules and biochemical intermediates (metabolites) using analytic instrumentation. Metabolites in blood are the product of all cellular processes, which are highly responsive to conditions of disease and environment, and represent the final output products of all organs forming a detailed systemic representation of an individual's current physiologic state. In this study, we used an untargeted metabolomics approach using gas chromatography time-of-flight mass spectrometry (GCTOFMS) to analyze the metabolome of serum and plasma samples both collected from the same patients that were organized into two independent caseācontrol studies (ADC1 and ADC2). In both studies, only NSCLC adenocarcinoma was investigated. The overall objectives were to (i) determine whether individual or combinations of metabolites could be used as a diagnostic test to distinguish NSCLC adenocarcinoma from controls and (ii) to determine which, plasma or serum, provides more accurate classifiers for the detection of lung cancer. We developed individual and multimetabolite classifiers using a training test from the ADC1 study and evaluated the performance of the constructed classifiers, individually or in combination, in an independent test/validation study (ADC2). This study shows the potential of metabolite-based diagnostic tests for detection of lung adenocarcinoma. Further validation in a larger pool of samples is warranted.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
SS = Sigma sample and is used as a quality control The first set (ADC1) used for biomarker development consisted of serum and plasma samples obtained from 52 stages IāIV NSCLC adenocarcinoma patients (52 plasma and 49 serum), and 31 healthy controls (31 pairs of serum and plasma) for a total of 163 samples. This set was regarded as the training set for biomarker discovery and classifier development. A second, independent caseācontrol study (ADC2) consisting of serum and plasma samples collected from 43 stage IāIV NSCLC adenocarcinoma patients and 43 healthy controls (total 172 samples) was used as an independent test set for biomarker evaluation. A limitation of this study is the relatively small sample size for each cohort (52 cases, 31 controls for ADC1, and 43 cases and 43 controls for ADC2) because patient variability can be a big factor in smaller studies.
Investigation of metabolomic blood biomarkers for detection of adenocarcinoma lung cancer (part II)
STUDY_TYPE
Lung cancer case control biomarker discovery
STUDY_SUMMARY
Recently, the National Lung Cancer Screen Trial (NLST) demonstrated that low-dose CT (LDCT) screening could reduce mortality due to lung cancer by 20%. However, LDCT screening is largely hindered by high false-positive rates (96%), particularly in high-risk populations (heavy smokers), due to the low prevalence rates (less than 2%) of malignant tumors and high incidence of benign lung nodules. Consequently, complementary biomarkers that can be used in conjunction with LDCT screening to improve diagnostic capacities and reduce false-positive rates are highly desirable. Preferably, such complementary tools should be noninvasive and exhibit high sensitivity and specificity. The application of ā-omicā sciences (genomics, transcriptomics, proteomics, and metabolomics) represents valuable tools for the discovery and validation of potential biomarkers that can be used for detection of NSCLC. Of these omic sciences, metabolomics has received considerable attention for its application in cancer. Metabolomics is the assessment of small molecules and biochemical intermediates (metabolites) using analytic instrumentation. Metabolites in blood are the product of all cellular processes, which are highly responsive to conditions of disease and environment, and represent the final output products of all organs forming a detailed systemic representation of an individual's current physiologic state. In this study, we used an untargeted metabolomics approach using gas chromatography time-of-flight mass spectrometry (GCTOFMS) to analyze the metabolome of serum and plasma samples both collected from the same patients that were organized into two independent caseācontrol studies (ADC1 and ADC2). In both studies, only NSCLC adenocarcinoma was investigated. The overall objectives were to (i) determine whether individual or combinations of metabolites could be used as a diagnostic test to distinguish NSCLC adenocarcinoma from controls and (ii) to determine which, plasma or serum, provides more accurate classifiers for the detection of lung cancer. We developed individual and multimetabolite classifiers using a training test from the ADC1 study and evaluated the performance of the constructed classifiers, individually or in combination, in an independent test/validation study (ADC2). This study shows the potential of metabolite-based diagnostic tests for detection of lung adenocarcinoma. Further validation in a larger pool of samples is warranted.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
SS = Sigma sample and is used as a quality control The first set (ADC1) used for biomarker development consisted of serum and plasma samples obtained from 52 stages IāIV NSCLC adenocarcinoma patients (52 plasma and 49 serum), and 31 healthy controls (31 pairs of serum and plasma) for a total of 163 samples. This set was regarded as the training set for biomarker discovery and classifier development. A second, independent caseācontrol study (ADC2) consisting of serum and plasma samples collected from 43 stage IāIV NSCLC adenocarcinoma patients and 43 healthy controls (total 172 samples) was used as an independent test set for biomarker evaluation. A limitation of this study is the relatively small sample size for each cohort (52 cases, 31 controls for ADC1, and 43 cases and 43 controls for ADC2) because patient variability can be a big factor in smaller studies.
Labeling of cells was carried out in triplicate with each sample containing 35e6. Cells were deprived of glucose or glutamine for 1 h then resuspended in 10mM [13C1,2] D-glucose labeled or 4 mM [13C5]-U-glutamine labeled medium, respectively. At this time cells were also activated with 5 ug/ml anti-CD3 and anti-CD28. Cells were labeled (and activated) for 16 h when samples were spun down, and cells were resuspended in 200 ul ice cold methanol. For this part of the experiment, the spent medium was collected for analysis. All samples were immediately stored in -80oC.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research
LABORATORY
Core Facilities Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
There are 4 germfree controls and 4 germfree with bacteria. After treatment, the mouse stool samples were collected at different timepoints: day 0, day 10, day 26, day 47, and day 61 or 70. Place the stool samples into liquid nitrogen after they were collected, then stored in -80oC.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Measure change in metabolites based on diet and feeding status
STUDY_TYPE
Regular
STUDY_SUMMARY
Flies were fed a normal diet or high-sugar (30% sucrose) diet for 10 days. Flies of each diet were flash frozen in liquid nitrogen as sated (refed for 3hrs on 400mM D-glucose) or after fasting for 24hrs. Heads and bodies were separated using a sieve-system and placed immediately on dry ice (total time app. 1min). We collected 50 heads per condition (4 conditions total) and 5 biological replicates
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Measure change in metabolites based on diet and feeding status (part II)
STUDY_TYPE
Regular
STUDY_SUMMARY
Flies were fed a normal diet or high-sugar (30% sucrose) diet for 10 days. Flies of each diet were flash frozen in liquid nitrogen as sated (refed for 3hrs on 400mM D-glucose) or after fasting for 24hrs. Heads and bodies were separated using a sieve-system and placed immediately on dry ice (total time app. 1min). We collected 50 heads per condition (4 conditions total) and 5 biological replicates
Mice were run for 15 minutes on a treadmill and then mice were induced into anesthesia at a dose of 5% isoflurane, then maintained by continuous inhalation of 2% isoflurane. Quadriceps muscle was collected first and flash frozen in liquid nitrogen. Then blood was collected from the left ventricle of the heart and placed into serum separating tubes. Blood was allowed to clot for 40 minutes and centrifuged, the supernatant was frozen in liquid nitrogen
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Facilities Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Mice were run for 15 minutes on a treadmill and then mice were induced into anesthesia at a dose of 5% isoflurane, then maintained by continuous inhalation of 2% isoflurane. Quadriceps muscle was collected first and flash frozen in liquid nitrogen. Then blood was collected from the left ventricle of the heart and placed into serum separating tubes. Blood was allowed to clot for 40 minutes and centrifuged, the supernatant was frozen in liquid nitrogen
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
We use ChAT(BAC)-EGFP mice, which express enhanced green fluorescent protein (EGFP) under the control of transcriptional regulatory elements for choline acetyltransferase (ChAT), the sole enzyme that catalyzes the biosynthesis of acetylcholine (ACh). These mice were treated with inducers of ChAT. Kidney were rapidly removed, frozen on liquid nitrogen and stored at -80C
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Rats were treated with donepezil (DPZ) after the induction of a model of glomerulonephritis (GN) to increase kidney ACh levels. Rats were euthanized at day 25 after the induction of the disease. Kidneys were rapidly removed, frozen in liquid nitrogen and stored at -80C
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
2 hydroxiglutarate with with IDH1-R132H inhibitor in neurospheres
STUDY_TYPE
Regular
STUDY_SUMMARY
Tumor neurospheres carrying genetic lession NPI (NRAS, shP53, IDH1-R132H) colony 1 and 2 (C1 and C2); NPAI (NRAS, shP53, shATRX, IDH1-R132H) colony 1 and 2 (C1 and C2); and NPA (NRAS, shP53, shATRX) (C1 and C2). Was treated with IDH1-R132H inhibitor. By 2HG assay we want to check the effect of the inhibitor in our cells.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Temporal metabolomic responses of cultured HepG2 liver cells to high fructose and high glucose exposures
STUDY_SUMMARY
High fructose consumption has been implicated with deleterious effects on human health, including hyperlipidemia elicited through de novo lipogenesis. However, more global effects of fructose on cellular metabolism have not been elucidated. In order to explore the metabolic impact of fructose-containing nutrients, we applied both GC-TOF and HILIC-QTOF mass spectrometry metabolomic strategies using extracts from cultured HepG2 cells exposed to fructose, glucose, or fructose + glucose. Cellular responses were analyzed in a time-dependent manner, incubated in media containing 5.5 mM glucose + 5.0 mM fructose in comparison to controls incubated in media containing either 5.5 mM glucose or 10.5 mM glucose. Mass spectrometry identified 156 unique known metabolites and a large number of unknown compounds, which revealed metabolite changes due to both utilization of fructose and high-carbohydrate loads independent of hexose structure. Fructose was shown to be partially converted to sorbitol, and generated higher levels of fructose-1-phosphate as a precursor for glycolytic intermediates. Differentially regulated ratios of 3-phosphoglycerate to serine pathway intermediates in high fructose media indicated a diversion of carbon backbones away from energy metabolism. Additionally, high fructoseconditions changed levels of complex lipids toward phosphatidylethanolamines. Patterns of acylcarnitines in response to high hexose exposure (10.5 mM glucose or glucose/fructose combination) suggested a reduction in mitochondrial beta-oxidation.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Temporal metabolomic responses of cultured HepG2 liver cells to high fructose and high glucose exposures
STUDY_SUMMARY
High fructose consumption has been implicated with deleterious effects on human health, including hyperlipidemia elicited through de novo lipogenesis. However, more global effects of fructose on cellular metabolism have not been elucidated. In order to explore the metabolic impact of fructose-containing nutrients, we applied both GC-TOF and HILIC-QTOF mass spectrometry metabolomic strategies using extracts from cultured HepG2 cells exposed to fructose, glucose, or fructose + glucose. Cellular responses were analyzed in a time-dependent manner, incubated in media containing 5.5 mM glucose + 5.0 mM fructose in comparison to controls incubated in media containing either 5.5 mM glucose or 10.5 mM glucose. Mass spectrometry identified 156 unique known metabolites and a large number of unknown compounds, which revealed metabolite changes due to both utilization of fructose and high-carbohydrate loads independent of hexose structure. Fructose was shown to be partially converted to sorbitol, and generated higher levels of fructose-1-phosphate as a precursor for glycolytic intermediates. Differentially regulated ratios of 3-phosphoglycerate to serine pathway intermediates in high fructose media indicated a diversion of carbon backbones away from energy metabolism. Additionally, high fructoseconditions changed levels of complex lipids toward phosphatidylethanolamines. Patterns of acylcarnitines in response to high hexose exposure (10.5 mM glucose or glucose/fructose combination) suggested a reduction in mitochondrial beta-oxidation.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
This West Coast Metabolomics Center sponsored pilot grant goal is to identify and validate interstitial cystitis/painful bladder syndrome (IC/PBS)-associated urinary metabolites. Our central hypothesis is that IC/PBS-associated metabolites in the urine of IC/PBS patients can segregate patients from control subjects, and that their levels are correlated with clinical symptoms. To test this hypothesis, we will identify IC/PBS-associated metabolites in urine using two independent platforms, GC-MS and quadrupole time-of-flight (Q-TOF) mass spectrometry under the collaboration with UC Davis WCMC scientists. We believe that this study will provide a significant potential clinical impact because results may lead to clinical methods to increase diagnostic accuracy and an improved understanding of the molecular basis of IC/PBS and its relationship to urologic conditions with overlapping symptoms.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
This West Coast Metabolomics Center sponsored pilot grant goal is to identify and validate interstitial cystitis/painful bladder syndrome (IC/PBS)-associated urinary metabolites. Our central hypothesis is that IC/PBS-associated metabolites in the urine of IC/PBS patients can segregate patients from control subjects, and that their levels are correlated with clinical symptoms. To test this hypothesis, we will identify IC/PBS-associated metabolites in urine using two independent platforms, GC-MS and quadrupole time-of-flight (Q-TOF) mass spectrometry under the collaboration with UC Davis WCMC scientists. We believe that this study will provide a significant potential clinical impact because results may lead to clinical methods to increase diagnostic accuracy and an improved understanding of the molecular basis of IC/PBS and its relationship to urologic conditions with overlapping symptoms.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
Health Sciences Drive, Davis, California, 95616, USA
Plasma Metabolomic Profiles Reflective of Glucose Homeostasis in Non-Diabetic and Type 2 Diabetic Obese African-American Women
STUDY_SUMMARY
Insulin resistance progressing to type 2 diabetes mellitus (T2DM) is marked by a broad perturbation of macronutrient intermediary metabolism. Understanding the biochemical networks that underlie metabolic homeostasis and how they associate with insulin action will help unravel diabetes etiology and should foster discovery of new biomarkers of disease risk and severity. We examined differences in plasma concentrations of >350 metabolites in fasted obese T2DM vs. obese non-diabetic African-American women, and utilized principal components analysis to identify 158 metabolite components that strongly correlated with fasting HbA1c over a broad range of the latter (r?=??0.631; p<0.0001). In addition to many unidentified small molecules, specific metabolites that were increased significantly in T2DM subjects included certain amino acids and their derivatives (i.e., leucine, 2-ketoisocaproate, valine, cystine, histidine), 2-hydroxybutanoate, long-chain fatty acids, and carbohydrate derivatives. Leucine and valine concentrations rose with increasing HbA1c, and significantly correlated with plasma acetylcarnitine concentrations. It is hypothesized that this reflects a close link between abnormalities in glucose homeostasis, amino acid catabolism, and efficiency of fuel combustion in the tricarboxylic acid (TCA) cycle. It is speculated that a mechanism for potential TCA cycle inefficiency concurrent with insulin resistance is āanaplerotic stressā emanating from reduced amino acid-derived carbon flux to TCA cycle intermediates, which if coupled to perturbation in cataplerosis would lead to net reduction in TCA cycle capacity relative to fuel delivery.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
G/g and g/a are polymorphisms in the promoter region of the UCP3 gene that leads to the gene being enhanced and an increased chance of obesity in those with this polymorphism.
PUBLICATIONS
Plasma Metabolomic Profiles Reflective of Glucose Homeostasis in Non-Diabetic and Type 2 Diabetic Obese African-American Women
Metabolomic profiles in P. gingivalis cells treated with pABA
STUDY_SUMMARY
Many human infections are polymicrobial in origin, and synergistic interactions among community inhabitants control colonization and pathogenic potential (Murray et al., 2014). However, few interspecies interactions have been functionally dissected at the molecular level or characterized on a systems level. Periodontitis, which is the sixth most prevalent infectious disease worldwide (Kassebaum et al., 2014), is associated with a dysbiotic microbial community, and the keystone pathogen Porphyromonas gingivalis forms synergistic communities with the accessory pathogen Streptococcus gordonii (Lamont and Hajishengallis, 2015). P. gingivalis and S. gordonii communicate through co-adhesion and metabolite perception, and close association between P. gingivalis and S. gordonii results in significant changes in the expressed proteomes of both organisms (Kuboniwa et al., 2012, Hendrickson et al., 2012). Here we show that streptococcal 4 aminobenzoate/para-amino benzoic acid (pABA) is required for maximal accumulation of P. gingivalis in communities with S. gordonii. Exogenous pABA upregulates production of fimbrial interspecies adhesins and of a tyrosine phosphorylation-dependent signaling system in P. gingivalis. Consequently, fimbrial-dependent attachment and invasion of epithelial cells by P. gingivalis is also increased by pABA. Moreover, trans-omics studies performed by proteomics and metabolomics showed that pABA induces metabolic shifts within P. gingivalis, predominantly folate derivative biosynthesis. In a murine oral infection model, pABA increased colonization and survival of P. gingivalis, but did not increase virulence. The results establish streptococcal pABA as a major component of the interspecies S. gordonii-P. gingivalis interaction which regulates distinct aspects of polymicrobial synergy.
Study1 Investigation of metabolomic blood biomarkers for detection of adenocarcinoma lung cancer (training set)
STUDY_SUMMARY
Recently, the National Lung Cancer Screen Trial (NLST) demonstrated that low-dose CT (LDCT) screening could reduce mortality due to lung cancer by 20%. However, LDCT screening is largely hindered by high false-positive rates (96%), particularly in high-risk populations (heavy smokers), due to the low prevalence rates (less than 2%) of malignant tumors and high incidence of benign lung nodules. Consequently, complementary biomarkers that can be used in conjunction with LDCT screening to improve diagnostic capacities and reduce false-positive rates are highly desirable. Preferably, such complementary tools should be noninvasive and exhibit high sensitivity and specificity. The application of ā-omicā sciences (genomics, transcriptomics, proteomics, and metabolomics) represents valuable tools for the discovery and validation of potential biomarkers that can be used for detection of NSCLC. Of these omic sciences, metabolomics has received considerable attention for its application in cancer. Metabolomics is the assessment of small molecules and biochemical intermediates (metabolites) using analytic instrumentation. Metabolites in blood are the product of all cellular processes, which are highly responsive to conditions of disease and environment, and represent the final output products of all organs forming a detailed systemic representation of an individual's current physiologic state. In this study, we used an untargeted metabolomics approach using gas chromatography time-of-flight mass spectrometry (GCTOFMS) to analyze the metabolome of serum and plasma samples both collected from the same patients that were organized into two independent caseācontrol studies (ADC1 and ADC2). In both studies, only NSCLC adenocarcinoma was investigated. The overall objectives were to (i) determine whether individual or combinations of metabolites could be used as a diagnostic test to distinguish NSCLC adenocarcinoma from controls and (ii) to determine which, plasma or serum, provides more accurate classifiers for the detection of lung cancer. We developed individual and multimetabolite classifiers using a training test from the ADC1 study and evaluated the performance of the constructed classifiers, individually or in combination, in an independent test/validation study (ADC2). This study shows the potential of metabolite-based diagnostic tests for detection of lung adenocarcinoma. Further validation in a larger pool of samples is warranted.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
SS = Sigma sample and is used as a quality control The first set (ADC1) used for biomarker development consisted of serum and plasma samples obtained from 52 stages IāIV NSCLC adenocarcinoma patients (52 plasma and 49 serum), and 31 healthy controls (31 pairs of serum and plasma) for a total of 163 samples. This set was regarded as the training set for biomarker discovery and classifier development. A second, independent caseācontrol study (ADC2) consisting of serum and plasma samples collected from 43 stage IāIV NSCLC adenocarcinoma patients and 43 healthy controls (total 172 samples) was used as an independent test set for biomarker evaluation. A limitation of this study is the relatively small sample size for each cohort (52 cases, 31 controls for ADC1, and 43 cases and 43 controls for ADC2) because patient variability can be a big factor in smaller studies.
Study 2 Investigation of metabolomic blood biomarkers for detection of adenocarcinoma lung cancer (test/validation)
STUDY_SUMMARY
Recently, the National Lung Cancer Screen Trial (NLST) demonstrated that low-dose CT (LDCT) screening could reduce mortality due to lung cancer by 20%. However, LDCT screening is largely hindered by high false-positive rates (96%), particularly in high-risk populations (heavy smokers), due to the low prevalence rates (less than 2%) of malignant tumors and high incidence of benign lung nodules. Consequently, complementary biomarkers that can be used in conjunction with LDCT screening to improve diagnostic capacities and reduce false-positive rates are highly desirable. Preferably, such complementary tools should be noninvasive and exhibit high sensitivity and specificity. The application of ā-omicā sciences (genomics, transcriptomics, proteomics, and metabolomics) represents valuable tools for the discovery and validation of potential biomarkers that can be used for detection of NSCLC. Of these omic sciences, metabolomics has received considerable attention for its application in cancer. Metabolomics is the assessment of small molecules and biochemical intermediates (metabolites) using analytic instrumentation. Metabolites in blood are the product of all cellular processes, which are highly responsive to conditions of disease and environment, and represent the final output products of all organs forming a detailed systemic representation of an individual's current physiologic state. In this study, we used an untargeted metabolomics approach using gas chromatography time-of-flight mass spectrometry (GCTOFMS) to analyze the metabolome of serum and plasma samples both collected from the same patients that were organized into two independent caseācontrol studies (ADC1 and ADC2). In both studies, only NSCLC adenocarcinoma was investigated. The overall objectives were to (i) determine whether individual or combinations of metabolites could be used as a diagnostic test to distinguish NSCLC adenocarcinoma from controls and (ii) to determine which, plasma or serum, provides more accurate classifiers for the detection of lung cancer. We developed individual and multimetabolite classifiers using a training test from the ADC1 study and evaluated the performance of the constructed classifiers, individually or in combination, in an independent test/validation study (ADC2). This study shows the potential of metabolite-based diagnostic tests for detection of lung adenocarcinoma. Further validation in a larger pool of samples is warranted.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
SS = Sigma sample and is used as a quality control The first set (ADC1) used for biomarker development consisted of serum and plasma samples obtained from 52 stages IāIV NSCLC adenocarcinoma patients (52 plasma and 49 serum), and 31 healthy controls (31 pairs of serum and plasma) for a total of 163 samples. This set was regarded as the training set for biomarker discovery and classifier development. A second, independent caseācontrol study (ADC2) consisting of serum and plasma samples collected from 43 stage IāIV NSCLC adenocarcinoma patients and 43 healthy controls (total 172 samples) was used as an independent test set for biomarker evaluation. A limitation of this study is the relatively small sample size for each cohort (52 cases, 31 controls for ADC1, and 43 cases and 43 controls for ADC2) because patient variability can be a big factor in smaller studies.
Changes in the metabalome and lipidome in response to exercise training
STUDY_TYPE
HERITAGE (HEalth, RIsk factors, exercise Training And GEnetics) family study
STUDY_SUMMARY
The overall objective of the Heritage Family Study is to study the role of the genotype in cardiovascular, metabolic, and hormonal responses to aerobic exercise training and the contribution of regular exercise to changes in several cardiovascular disease and diabetes risk factors. The study cohort used in this analysis was derived from the pool of 473 Caucasian subjects (230 male and 243 female) from 99 nuclear families who completed ā„58 of the prescribed 60 exercise-training sessions. Utilizing a subsample of this Caucasian cohort, we selected family members from the Quebec center (N=125) to assess the metabolome and lipidome of circulating plasma under two well-defined environmental conditions, the pre- and post-training conditions.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
Health Sciences Drive, Davis, California, 95616, USA
Serum phosphatidylethanolamine levels distinguish benign from malignant solitary pulmonary nodules and represent a potential diagnostic biomarker for lung cancer (part I)
STUDY_SUMMARY
Recent computed tomography (CT) screening trials showed that it is effective for early detection of lung cancer, but were plagued by high false positive rates. Additional blood biomarker tests designed to complement CT screening and reduce false positive rates are highly desirable. In the current study, we expand upon our initial experimental findings as part of the discovery phase by evaluating metabolites in serum from subjects with benign or malignant SPNs using a combined approach of gas chromatography time-of-flight mass spectrometry (GC-TOFMS) and hydrophilic liquid chromatography accurate mass quadrupole time-of-flight mass spectrometry (HILIC-qTOFMS). Furthermore, we evaluated serum collected pre-diagnosis and at-diagnosis of lung cancer in addition to samples obtained post-surgical intervention from subjects with malignant SPNs (post-diagnosis). We hypothesize that our systems biology approach to identify candidate metabolomics biomarkers will ultimately lead to improved early detection of lung cancer and can be used in as a companion blood test to LDCT screening.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Serum phosphatidylethanolamine levels distinguish benign from malignant solitary pulmonary nodules and represent a potential diagnostic biomarker for lung cancer (part II)
STUDY_SUMMARY
Recent computed tomography (CT) screening trials showed that it is effective for early detection of lung cancer, but were plagued by high false positive rates. Additional blood biomarker tests designed to complement CT screening and reduce false positive rates are highly desirable. In the current study, we expand upon our initial experimental findings as part of the discovery phase by evaluating metabolites in serum from subjects with benign or malignant SPNs using a combined approach of gas chromatography time-of-flight mass spectrometry (GC-TOFMS) and hydrophilic liquid chromatography accurate mass quadrupole time-of-flight mass spectrometry (HILIC-qTOFMS). Furthermore, we evaluated serum collected pre-diagnosis and at-diagnosis of lung cancer in addition to samples obtained post-surgical intervention from subjects with malignant SPNs (post-diagnosis). We hypothesize that our systems biology approach to identify candidate metabolomics biomarkers will ultimately lead to improved early detection of lung cancer and can be used in as a companion blood test to LDCT screening.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic markers of altered nucleotide metabolism in early stage adenocarcinoma (part I)
STUDY_SUMMARY
Lung cancer has been the leading cause of cancer death in the United States and worldwide for many decades. Low dose spiral computerized tomography (LDCT) is likely to become the first approved screening and early detection test in the upcoming year, but it is plagued by a high false-positive rate. There is a need to develop complementary screening and early detection tools. A blood-based "lung cancer" signature is an attractive solution. Given that our knowledge of the molecular biology of smoking-induced lung cancer has dramatically increased over the past few years, this approach is plausible. To date, this effort has been focused on the identification of genomic and proteomic signatures with limited success. A broader strategy that incorporates additional cancer traits is needed. It is well recognized that wide coverage of cellular metabolism in cancer could help provide valuable diagnostic biomarkers and potentially identify molecular drivers of tumorigenesis. Recent advances in mass spectrometry have enabled comprehensive metabolomic analyses of lipids, carbohydrates, amino acids, and nucleotides within a variety of biologic matrices. Early evidence from metabolomic investigation of cancer has identified many altered biochemical profiles. However, to date, there have been few investigations of lung cancer, and most studies have looked at blood plasma or were limited by small sample sizes with mixed histologies. In the current investigation, gas chromatography time-offlight mass spectrometry (GC-TOF) was used to measure 462 lipid, carbohydrate, amino acid, organic acid, and nucleotide metabolites in 39 malignant and nonmalignant lung tissue pairs from current or former smokers with early stage adenocarcinoma. This study cohort represents patient characteristics and tumor histology most likely to be detected with LDCT screening. We hypothesize that identification of cancer-induced cellular and tissue level biochemical changes can offer a robust method for identification of candidate circulating biomarkers and improve our understanding of biochemical changes involved in adenocarcinoma tumorigenesis.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic markers of altered nucleotide metabolism in early stage adenocarcinoma (part II)
STUDY_SUMMARY
Lung cancer has been the leading cause of cancer death in the United States and worldwide for many decades. Low dose spiral computerized tomography (LDCT) is likely to become the first approved screening and early detection test in the upcoming year, but it is plagued by a high false-positive rate. There is a need to develop complementary screening and early detection tools. A blood-based "lung cancer" signature is an attractive solution. Given that our knowledge of the molecular biology of smoking-induced lung cancer has dramatically increased over the past few years, this approach is plausible. To date, this effort has been focused on the identification of genomic and proteomic signatures with limited success. A broader strategy that incorporates additional cancer traits is needed. It is well recognized that wide coverage of cellular metabolism in cancer could help provide valuable diagnostic biomarkers and potentially identify molecular drivers of tumorigenesis. Recent advances in mass spectrometry have enabled comprehensive metabolomic analyses of lipids, carbohydrates, amino acids, and nucleotides within a variety of biologic matrices. Early evidence from metabolomic investigation of cancer has identified many altered biochemical profiles. However, to date, there have been few investigations of lung cancer, and most studies have looked at blood plasma or were limited by small sample sizes with mixed histologies. In the current investigation, gas chromatography time-offlight mass spectrometry (GC-TOF) was used to measure 462 lipid, carbohydrate, amino acid, organic acid, and nucleotide metabolites in 39 malignant and nonmalignant lung tissue pairs from current or former smokers with early stage adenocarcinoma. This study cohort represents patient characteristics and tumor histology most likely to be detected with LDCT screening. We hypothesize that identification of cancer-induced cellular and tissue level biochemical changes can offer a robust method for identification of candidate circulating biomarkers and improve our understanding of biochemical changes involved in adenocarcinoma tumorigenesis.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Systemic Metabolomic Changes in Blood Samples of Lung Cancer Patients Identified by Gas Chromatography Time-of-Flight Mass Spectrometry
STUDY_SUMMARY
Lung cancer is a leading cause of cancer deaths worldwide. Metabolic alterations in tumor cells coupled with systemic indicators of the host response to tumor development have the potential to yield blood profiles with clinical utility for diagnosis and monitoring of treatment. We report results from two separate studies using gas chromatography time-of-flight mass spectrometry (GC-TOF MS) to profile metabolites in human blood samples that significantly differ from non-small cell lung cancer (NSCLC) adenocarcinoma and other lung cancer cases. Metabolomic analysis of blood samples from the two studies yielded a total of 437 metabolites, of which 148 were identified as known compounds and 289 identified as unknown compounds. Differential analysis identified 15 known metabolites in one study and 18 in a second study that were statistically different (p-values <0.05). Levels of maltose, palmitic acid, glycerol, ethanolamine, glutamic acid, and lactic acid were increased in cancer samples while amino acids tryptophan, lysine and histidine decreased. Many of the metabolites were found to be significantly different in both studies, suggesting that metabolomics appears to be robust enough to find systemic changes from lung cancer, thus showing the potential of this type of analysis for lung cancer detection.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
TOTAL_SUBJECTS
82
NUM_MALES
20
NUM_FEMALES
62
STUDY_COMMENTS
SS = Sigma sample and is used as a quality control
Long-Chain Fatty Acid Combustion Rate Is Associated with Unique Metabolite Profiles in Skeletal Muscle Mitochondria
STUDY_SUMMARY
Incomplete or limited long-chain fatty acid (LCFA) combustion in skeletal muscle has been associated with insulin resistance. Signals that are responsive to shifts in LCFA Ī²-oxidation rate or degree of intramitochondrial catabolism are hypothesized to regulate second messenger systems downstream of the insulin receptor. Recent evidence supports a causal link between mitochondrial LCFA combustion in skeletal muscle and insulin resistance. We have used unbiased metabolite profiling of mouse muscle mitochondria with the aim of identifying candidate metabolites within or effluxed from mitochondria and that are shifted with LCFA combustion rate. This proof-of-principle study establishes that large-scale metabolomics methods can be applied to organelle-level models to discover metabolite patterns reflective of LCFA combustion, which may lead to identification of molecules linking muscle fat metabolism and insulin signaling. Our results suggest that future studies should focus on the fate of effluxed TCA cycle intermediates and on mechanisms ensuring their replenishment during LCFA metabolism in skeletal muscle.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
The circadian oscillator in Synechococcus elongatus controls metabolite partitioning during diurnal growth
STUDY_SUMMARY
Cyanobacteria are increasingly being considered for use in large-scale outdoor production of fuels and industrial chemicals. Cyanobacteria can anticipate daily changes in light availability using an internal circadian clock and rapidly alter their metabolic processes in response to changes light availability. Understanding how signals from the internal circadian clock and external light availability are integrated to control metabolic shifts will be important for engineering cyanobacteria for production in natural outdoor environments. This study has assessed how āknowingā the correct time of day, via the circadian clock, affects metabolic changes when a cyanobacterium goes through a dark-to-light transition. Our data show that the circadian clock plays an important role in inhibiting activation of the oxidative pentose phosphate pathway in the morning. Synechococcus elongatus PCC 7942 is a genetically tractable model cyanobacterium that has been engineered to produce industrially relevant biomolecules and is the best-studied model for a prokaryotic circadian clock. However, the organism is commonly grown in continuous light in the laboratory, and data on metabolic processes under diurnal conditions are lacking. Moreover, the influence of the circadian clock on diurnal metabolism has been investigated only briefly. Here, we demonstrate that the circadian oscillator influences rhythms of metabolism during diurnal growth, even though lightādark cycles can drive metabolic rhythms independently. Moreover, the phenotype associated with loss of the core oscillator protein, KaiC, is distinct from that caused by absence of the circadian output transcriptional regulator, RpaA (regulator of phycobilisome-associated A). Although RpaA activity is important for carbon degradation at night, KaiC is dispensable for those processes. Untargeted metabolomics analysis and glycogen kinetics suggest that functional KaiC is important for metabolite partitioning in the morning. Additionally, output from the oscillator functions to inhibit RpaA activity in the morning, and kaiC-null strains expressing a mutant KaiC phosphomimetic, KaiC-pST, in which the oscillator is locked in the most active output state, phenocopies a ĪrpaA strain. Inhibition of RpaA by the oscillator in the morning suppresses metabolic processes that normally are active at night, and kaiC-null strains show indications of oxidative pentose phosphate pathway activation as well as increased abundance of primary metabolites. Inhibitory clock output may serve to allow secondary metabolite biosynthesis in the morning, and some metabolites resulting from these processes may feed back to reinforce clock timing.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
The first 4 samples were a test run to see how efficient the analysis was and were run on a lipidomics platform. The next 12 samples were the used in the paper and were the same as the original 4 samples, but they were split into 3 biological replicates and run on the GC platform.
The circadian oscillator in Synechococcus elongatus controls metabolite partitioning during diurnal growth (part II)
STUDY_SUMMARY
Cyanobacteria are increasingly being considered for use in large-scale outdoor production of fuels and industrial chemicals. Cyanobacteria can anticipate daily changes in light availability using an internal circadian clock and rapidly alter their metabolic processes in response to changes light availability. Understanding how signals from the internal circadian clock and external light availability are integrated to control metabolic shifts will be important for engineering cyanobacteria for production in natural outdoor environments. This study has assessed how āknowingā the correct time of day, via the circadian clock, affects metabolic changes when a cyanobacterium goes through a dark-to-light transition. Our data show that the circadian clock plays an important role in inhibiting activation of the oxidative pentose phosphate pathway in the morning. Synechococcus elongatus PCC 7942 is a genetically tractable model cyanobacterium that has been engineered to produce industrially relevant biomolecules and is the best-studied model for a prokaryotic circadian clock. However, the organism is commonly grown in continuous light in the laboratory, and data on metabolic processes under diurnal conditions are lacking. Moreover, the influence of the circadian clock on diurnal metabolism has been investigated only briefly. Here, we demonstrate that the circadian oscillator influences rhythms of metabolism during diurnal growth, even though lightādark cycles can drive metabolic rhythms independently. Moreover, the phenotype associated with loss of the core oscillator protein, KaiC, is distinct from that caused by absence of the circadian output transcriptional regulator, RpaA (regulator of phycobilisome-associated A). Although RpaA activity is important for carbon degradation at night, KaiC is dispensable for those processes. Untargeted metabolomics analysis and glycogen kinetics suggest that functional KaiC is important for metabolite partitioning in the morning. Additionally, output from the oscillator functions to inhibit RpaA activity in the morning, and kaiC-null strains expressing a mutant KaiC phosphomimetic, KaiC-pST, in which the oscillator is locked in the most active output state, phenocopies a ĪrpaA strain. Inhibition of RpaA by the oscillator in the morning suppresses metabolic processes that normally are active at night, and kaiC-null strains show indications of oxidative pentose phosphate pathway activation as well as increased abundance of primary metabolites. Inhibitory clock output may serve to allow secondary metabolite biosynthesis in the morning, and some metabolites resulting from these processes may feed back to reinforce clock timing.
INSTITUTE
UC Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
Health Sciences Drive, Davis, California, 95616, USA
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
The first 4 samples were a test run to see how efficient the analysis was and were run on a lipidomics platform. The next 12 samples were the used in the paper and were the same as the original 4 samples, but they were split into 3 biological replicates and run on the GC platform.
Recently, major efforts have been directed toward early detection of lung cancer through low-dose computed tomography (LDCT) scanning. Data from the National Lung Screening Trial (NLST) suggest that yearly screening with thoracic LDCT scanning for high-risk current and former smokers reduces lung cancer mortality by 20% and total mortality by 7%. However, issues including indeterminate nodules detected by LDCT and radiation exposure impact the practicality of LDCT-based screening on a national and global basis. A blood-based biomarker or multiplexed marker panel that could complement LDCT would represent a major advance in implementing lung cancer screening. Efforts to develop blood-based biomarkers for lung cancer early detection using a variety of methodologies are currently ongoing. Proteomic studies have led to the identification of several candidate markers including pro-surfactantproteinB(pro-SFTPB), a target of a lineage-survival oncogene in lung cancer, NKX2-1.Validation studies using blood samples collected at the time of LDCT screening for lung cancer substantiated the performance of pro-SFTPB. Multivariable logistic regression models were used to evaluate the predictive ability of pro-SFTPB. The area under the curve (AUC) values of the full model with and without pro-SFTPB were 0.741 (95% CI, 0.696 to 0.783) and 0.669 (95%CI, 0.620 to 0.717), respectively (difference in AUC, P_.001). Single markers are unlikely to have sufficient performance for implementation in a screening setting, hence the need to explore several discovery platforms to identify markers that provide complementary performance. Metabolomics represents a global unbiased approach to the profiling of small molecules and has been established as a platform for biomarker discovery for a variety of human biofluids and tissues. Here we used an untargeted liquid chromatography/mass spectrometry (MS) metabolomics approach to identify metabolites that distinguish human sera collected before the diagnosis of lung cancer from matched control sera in a prospective cohort of highrisk patients from the Beta-Carotene and Retinol Efficacy Trial (CARET).
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Long-term neural and physiological phenotyping of a single human
STUDY_TYPE
Longitudinal
STUDY_SUMMARY
The dynamics of human brain function are increasingly well understood at the short timescale of seconds/minutes (for example, through studies of learning) and the long timescale of years/decades (for example, through studies of development andageing), but almost nothing is known about how the human brainfunction varies across the range of days to months. This is a critical gap, because major psychiatric disorders show large fluctuations in brain function over this timescale. However, the kind of dense longitudinal phenotyping that is necessary to understand this question is extremely challenging with healthy human volunteers,who are unlikely to be sufficiently motivated to sustain frequent participation in a study over a long period. For this reason, the participation of motivated experimenters can be uniquely useful for demanding longitudinal studies. We investigated the long-range dynamics of brain function andtheir relation to a broad set of psychological and biological variables in a single healthy human (author R.A.P.) over the course of 532 days (along with several follow-up visits), representing one of the most intensive biological characterizations of a single individual ever performed (referred to hereafter as the MyConnectomestudy). The study was designed to measure the broadest possible range of human phenotypes (the phenomeā3,4) to allow the widespread assessment of relations between psychological, neural and metabolic function. The results of the present study demonstrate that healthy brain function shows rich dynamics over the course of 18 months, and that these dynamics are paralleled by ongoing fluctuations in psychological and physiological function as observed in behaviour,gene expression and metabolomic measurements. These findings provide a proof of concept for the dynamic longitudinal phenotyping of individuals, which we propose will be crucial togain a better understanding of the substantial fluctuations in psychological and neural function in individuals with major psychiatric disorders.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolic profiling of maternal urine can aid clinical management of Gestational Diabetes Mellitus (GDM)
STUDY_TYPE
Search for non-treated and treated GDM biomarkers in urine
STUDY_SUMMARY
NMR metabolomics study of maternal urine of 1) GDM women at the time of diagnosis and before treatment, to define the urine metabolic profile of untreated GDM, and of 2) GDM women treated using diet control alone or with the addition of insulin, to identify treatment-resistant and treatment-responsive metabolic pathways and, hence, evaluate treatment efficacy, and 3) GDM treatment prediction at the time of diagnosis, with the aim of finding potential predictive markers of future treatment requirements based on each individual metabotype.
INSTITUTE
University of Aveiro
DEPARTMENT
CICECO-Department of Chemistry, University of Aveiro
LAST_NAME
Gil
FIRST_NAME
Ana
ADDRESS
CICECO-Department of Chemistry, University of Aveiro
E.coli effects on growth and substrate uptake of green algae (part I - HILIC)
STUDY_SUMMARY
The purpose of this project was to quantify the exchange of thiamine between bacteria and algae. We previously observed that the model bacteria, Escherichia coli, enhanced the growth and substrate uptake of the green algae, Auxenochlorella protothecoides. We hypothesized that this growth enhancement was due to the secretion of thiamine derivatives or degradation products by E. coli followed by uptake of these compounds by A. protothecoides. Targeted and untargeted LCMS revealed the presence of thiamine dervatives in E. coli cell extracts. These LCMS methods were also used to quantify thiamine derivatives and two degradation products, HMP and THZ, present in E. coli medium after cell removal. The LCMS results along with culture studies were employed to show that thiamine derivatives and degradation products were the primary mechanism of symbiosis between E. coli and A. protothecoides.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
E.coli effects on growth and substrate uptake of green algae (part II - Reverse Phase)
STUDY_SUMMARY
The purpose of this project was to quantify the exchange of thiamine between bacteria and algae. We previously observed that the model bacteria, Escherichia coli, enhanced the growth and substrate uptake of the green algae, Auxenochlorella protothecoides. We hypothesized that this growth enhancement was due to the secretion of thiamine derivatives or degradation products by E. coli followed by uptake of these compounds by A. protothecoides. Targeted and untargeted LCMS revealed the presence of thiamine dervatives in E. coli cell extracts. These LCMS methods were also used to quantify thiamine derivatives and two degradation products, HMP and THZ, present in E. coli medium after cell removal. The LCMS results along with culture studies were employed to show that thiamine derivatives and degradation products were the primary mechanism of symbiosis between E. coli and A. protothecoides.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
Health Sciences Drive, Davis, California, 95616, USA
Inhibition of diamine oxidase promotes uptake of putrescine from rat small intestine
STUDY_SUMMARY
Metformin, a biguanide molecule, which is used as first line therapy for type 2 diabetes. In this study, we would like to investigate the inhibition of an enzyme called diamine oxidase (DAO) (also known as ABP1), by metformin. Based on our preliminary in vitro study using diamine oxidase enzyme, we saw increased level of putrescine with increasing metformin concentrations (see reference PMID: 26335661). This proposed in vivo study was to determine whether metformin could increase putrescine levels and other metabolites in mice. Aminoguanidine, a known inhibitor of DAO, in this study as positive control, following similar study design described in this paper (PMID: 8912017).
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Impact of glucose on the central metabolome of C. minutissima
STUDY_SUMMARY
Axenic Chlorella minutissima (UTEX 2341) was grown under mixotrophic and autotrophic conditions to compare metabolome differences. The purpose of this study was to understand how glucose impacted the central metabolome of C. minutissima.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
The proton channel HVCN1 is expressed in B cell malignancies at high levels but its role remains unclear. From initial experiments during which HVCN1 was downregulated in human multiple myeloma cell lines, we observed an increase in some glycolytic and TCA metabolites. We want to get a better idea if HVCN1 is playing a role in regulating energy metabolism in multiple myeloma.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic Profiling in Early Pregnancy using Pre-Diagnostic Sera from Women Who Developed Placental Abruption
STUDY_TYPE
Metabolomics Biocrates Panel
STUDY_SUMMARY
Placental abruption (PA) is an ischemic placental disorder that results from the premature separation of the placenta from the wall of the uterus before delivery of the fetus. This disorder is associated with pre-term delivery, fetal death, maternal hemorrhagic shock, and renal failure. Several physiologic disturbances, such as oxidative stress, carbohydrate/fatty acid metabolism, and mitochondrial dysfunction, have been associated with ischemic placental disorder as well as with placental abruption. This preliminary study proposes to identify metabolites associated with incident placental abruption using existing serum and clinical data from a previously studied cohort. Metabolomics analysis will be carried out on maternal serum (51 cases and 51 controls) collected in early pregnancy (early second trimester).
Noninvasive Recognition and Biomarkers of Early Allergic Asthma in Cats using Multivariate Statistics of NMR Spectra of Exhaled Breath Condensate
STUDY_TYPE
Statistical Analysis of NMR spectra of EBC samples
STUDY_SUMMARY
Asthma is prevalent in children and cats, and needs means of noninvasive diagnosis. We sought to distinguish noninvasively the differences in 53 cats before and soon after induction of allergic asthma, using NMR spectra of exhaled breath condensate (EBC). Statistical pattern recognition was improved by preprocessing the spectra with glog transformation. Classification of the 106 preprocessed spectra by principal component analysis, partial least squares with discriminant analysis (PLS-DA), and multi-level PLS-DA appears to be impaired by variances unrelated to eosinophilic asthma. By subtracting out confounding variances, orthogonal signal correction (OSC) PLS-DA greatly improved the separation of the healthy and early asthmatic states, attaining 94% specificity and 71% sensitivity in predictions. OSC-PLS-DA results highlight the elevation of acetone in two-thirds of the cats with early asthma. Asthma also decreased at least a dozen compounds, especially carboxylic acids such as short chain fatty acids and amino acids. These trends suggest that a majority of the cats with allergic asthma underwent alteration of metabolic fluxes through pyruvate and acetyl-CoA to promote ketosis. The noninvasive detection of early experimental asthma, its biomarkers in EBC, and metabolic rerouting invite further investigation of the diagnostic potential in humans.
INSTITUTE
University of Missouri-Columbia
DEPARTMENT
Department of Biochemistry, Department of Veterinary Medicine and Surgery
LABORATORY
Van Doren Lab & Reinero Lab
LAST_NAME
Van Doren
FIRST_NAME
Steven
ADDRESS
117 Schweitzer Hall, Biochemistry Department, University of Missouri-Columbia, Columbia, MO 65211
The goal of this study was to identify metabolic differences between 6 week old and 1 year old infants that have been potentially exposed to arsenic in order to determine its effect on the microbiome and the immune system.
Metabolic profiling during ex vivo machine perfusion of the human liver
STUDY_SUMMARY
As donor organ shortages persist, functional machine perfusion is under investigation to improve preservation of the donor liver. The transplantation of donation after circulatory death (DCD) livers is limited by poor outcomes, but its application may be expanded by ex vivo repair and assessment of the organ before transplantation. Here we employed subnormothermic (21āĀ°C) machine perfusion of discarded human livers combined with metabolomics to gain insight into metabolic recovery during machine perfusion. Improvements in energetic cofactors and redox shifts were observed, as well as reversal of ischemia-induced alterations in selected pathways, including lactate metabolism and increased TCA cycle intermediates. We next evaluated whether DCD livers with steatotic and severe ischemic injury could be discriminated from ātransplantableā DCD livers. Metabolomic profiling was able to cluster livers with similar metabolic patterns based on the degree of injury. Moreover, perfusion parameters combined with differences in metabolic factors suggest variable mechanisms that result in poor energy recovery in injured livers. We conclude that machine perfusion combined with metabolomics has significant potential as a clinical instrument for the assessment of preserved livers.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolic profiling during ex vivo machine perfusion of the human liver (part III)
STUDY_SUMMARY
As donor organ shortages persist, functional machine perfusion is under investigation to improve preservation of the donor liver. The transplantation of donation after circulatory death (DCD) livers is limited by poor outcomes, but its application may be expanded by ex vivo repair and assessment of the organ before transplantation. Here we employed subnormothermic (21āĀ°C) machine perfusion of discarded human livers combined with metabolomics to gain insight into metabolic recovery during machine perfusion. Improvements in energetic cofactors and redox shifts were observed, as well as reversal of ischemia-induced alterations in selected pathways, including lactate metabolism and increased TCA cycle intermediates. We next evaluated whether DCD livers with steatotic and severe ischemic injury could be discriminated from ātransplantableā DCD livers. Metabolomic profiling was able to cluster livers with similar metabolic patterns based on the degree of injury. Moreover, perfusion parameters combined with differences in metabolic factors suggest variable mechanisms that result in poor energy recovery in injured livers. We conclude that machine perfusion combined with metabolomics has significant potential as a clinical instrument for the assessment of preserved livers.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
The first 4 samples were a test run to see how efficient the analysis was and were run on a lipidomics platform. The next 12 samples were the used in the paper and were the same as the original 4 samples, but they were split into 3 biological replicates and run on the GC platform.
Toxicokinetics and Metabolomic Disrupting of the Flame Retardant Mixture Firemaster 550
STUDY_TYPE
Low Dose and High Dose Exposure to Firemaster (FM) 550
STUDY_SUMMARY
Pregnant lab rats (dams) were assigned to three groups: a control group, which was not exposed to Firemaster (FM) 550; a low-dose group, which ingested 100 Āµg of FM550 once daily throughout pregnancy; and a high-dose group, which ingested 1000 Āµg FM550 on the same schedule. The placentas were harvested immediately after birth, frozen on dry ice and pulverized in liquid nitrogen via mortar and pestle. The overall objective is to investigate the differences in the metabolic profiles of the placentas in each phenotypic group.
INSTITUTE
RTI International
DEPARTMENT
RCMRC
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E Cornwallis Road, Research Triangle Park, NC 27709
Controlled Human Exposure to Particulate Matter (PM) and Gaseous Co-Pollutants
STUDY_TYPE
Exposome Evaluation
STUDY_SUMMARY
This study is designed to provide the environmental aspects to support both the acquisition of study samples and the advancement of environmental chemical speciation information and data analyis needed. The aims of the study are as follows:1)Do the metabolomics profiles appear to be impacted by exposure to PM and NO2+PM. 2)are the metabolomic profiles related to the PM and NO2+PM distinct 3)which features (chemical or physical) of the PM and NO2+PM have the most significant impact on the metabolomic profiles.
INSTITUTE
RTI International
DEPARTMENT
RCMRC
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E Cornwallis Road, Research Triangle Park, NC, 27519, USA
The role of CFTR in the regulation of intrinsic defense mechanisms of exocrine secretions
STUDY_SUMMARY
This study was designed as a pilot to determine if the concentrations of selected ions in salivary gland secretions were influenced by the absence of CFTR on the apical surfaces of glandular cells and ductal epithelia. To this end, five cystic fibrosis children homozygous for ļPhe508 were to be recruited with their heterozygous non-CF mothers. For adaptation and development of the assays to a microtiter format, saliva samples were collected from non-CF control subjects initially to optimize the assays recognizing that the volumes for analyses would be small. The values obtained with these non-CF control samples were also compared to that of the homozygous and heterozygous CF subjects. As residual volume permits, these samples will be further analyzed for metabolic constituent profiles.
INSTITUTE
RTI International
DEPARTMENT
RCMRC
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E Cornwallis Road, Research Triangle Park, NC 27709
Impact Of High Sugar Diet On L-Arginine Metabolism In The Lung
STUDY_SUMMARY
Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. We need novel therapeutic agents that are affordable, can decrease the reliance on steroids, and can improve quality of life. This clinical and mechanistic study has the potential to impact treatment of a subset of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and NO biology in the airways of asthmatics. We will pursue a clinical trial in subjects not well controlled on standard drug therapy; this strategy will address whether L-arginine is efficacious in patients receiving standard of care medications. In studies using animal models, we and others have shown that interventions that augment NO levels, through either supplementation of L-arginine or inhibition of arginase, decrease allergic airway inflammation and hyperresponsiveness-the two hallmarks of asthma. Overall, we hypothesize that a responder subset of adult severe asthma patients will derive clinical benefit from supplemental L-arginine therapy and that these patients will have a lower exhaled NO concentrations (<20 ppb) and a higher NOS2/Arg1 mRNA and protein ratio in their airway epithelial cells than non-responders. We aim to: 1) test the hypothesis that uncontrolled, adult severe asthma patients with exhaled breath NO concentrations <20 ppb will have fewer asthma exacerbations over 3 months when treated with L-arginine compared to patients with FeNO > 25, 2) determine the mechanisms by which L-arginine affects the regulation of NOS and arginase enzymes in primary airway epithelial cell cultures from severe asthmatic subjects, and 3) test the hypothesis that inhaled nanoparticle carrier formulations of L-arginine will decrease airway inflammation, airway hyperresponsiveness, and airway fibrosis at lower doses than systemically administered L-arginine. The major impact of our study will be to identify the adult severe asthma cohort that will benefit from supplemental L-arginine therapy. Our ultimate goal is to develop novel therapeutic agents to treat adult severe asthma patients better. PUBLIC HEALTH RELEVANCE: Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. This clinical study has the potential to improve the care of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and nitric oxide biology in the lung. If we demonstrate that L-arginine supplementation can decrease asthma attacks in a subset of severe asthmatics, it will have great implications for future research as well as for the daily lives of patients with asthma.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
This study is a test of the untargeted metabolomics approach to studying samples from exposure studies. The samples are NIST standard SRM 3672 for pooled urine of smokers and SRM 3673 for pooled urine of non-smokers. A broad spectrum GCMS approach was used in this study.
ms3076 T1D poor glycemic control and control samples
STUDY_TYPE
Plasma metabolites in T1 diabetes
STUDY_SUMMARY
The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Type 1 Diabetes good glycemic control and controls samples
STUDY_TYPE
Plasma metabolites in T1 diabetes
STUDY_SUMMARY
The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Differences in mycoplasma growth due to different mediums
STUDY_SUMMARY
The object is to learn if there are variations in the lipid profiles of the three genomic variants (relative to one another) and if there are difference the lipid profiles due to growth in medium having different supplements. Mycoplasmas are eubacteria, but have only a single plasma membrane and no cell wall. They acquire FAs and cholesterol and other (perhaps many unknown) lipids from the medium which is complex and contains mammalian serum. Various mycoplasma species have been shown to contain a wide spectrum of bacterial lipids, but the composition is unknown for this mycoplasma species. We are particularly interested in ratios of membrane lipids among our strains, in part to gain clues about differences in metabolic pathways pertinent to membrane biogenesisĶ¾ and to predict any underlying features that could relate to the extremely different modes of cell propagation observed among these genomic constructs.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomics Approach to Allograft Assessment in Liver Transplantation
STUDY_TYPE
Retrospective analysis of biobanked liver tissue from organ donors
STUDY_SUMMARY
This pilot study is designed to apply several types of metabolomic analysis for liver allograft assessment with the aim of identifying candidate biomarkers for allograft function and to develop a methodology that could be applied to larger scale studies. The three main categories of metabolomic analysis are reflected in each of the specific aims, including a targeted profiling of central carbon metabolism, open lipidomic and metabolomic profiling for hypothesis generation and MALDI-IMS for tissue-based spatial analysis. Each of these approaches offers potential benefits that could be optimized in a protocol for larger scale studies depending the results of the pilot investigations.
Non targeted metabolomics of gastrocnemius tissue samples obtained from 20 month old (old) mice- Both Sham and after inducing lung injury
STUDY_TYPE
Non targeted metabolomic analysis
STUDY_SUMMARY
Introduction: Older patients are more likely to acquire and die from acute respiratory distress syndrome (ARDS) and muscle weakness may be more significant in older survivors. Recent data implicate muscle ring finger protein 1 (MuRF1) in lung injury-induced skeletal muscle atrophy in young mice and identify an alternative role for MuRF1 in cardiac metabolism regulation through inhibition of fatty acid oxidation. Objectives: To develop a model of lung injury-induced muscle wasting in old mice and to evaluate the skeletal muscle metabolomic profile of adult and old acute lung injury (ALI) mice. Methods: Young (2 month), adult (6 month) and old (20 month) male C57Bl6J mice underwent Sham (intratracheal H2O) or ALI [intratracheal E. coli lipopolysaccharide (i.t. LPS)] conditions and muscle functional testing. Metabolomic analysis on gastrocnemius muscle was performed using gas chromatography-mass spectrometry (GC-MS). Results: Old ALI mice had increased mortality and failed to recover skeletal muscle function compared to adult ALI mice. Muscle MuRF1 expression was increased in old ALI mice at day 3. Non-targeted muscle metabolomics revealed alterations in amino acid biosynthesis and fatty acid metabolism in old ALI mice. Targeted metabolomics of fatty acid intermediates (acyl-carnitines) and amino acids revealed a reduction in long chain acyl-carnitines in old ALI mice. Conclusion: This study demonstrates age-associated susceptibility to ALI-induced muscle wasting which parallels a metabolomic profile suggestive of altered muscle fatty acid metabolism. MuRF1 activation may contribute to both atrophy and impaired fatty acid oxidation, which may synergistically impair muscle function in old ALI mice.
INSTITUTE
University of North Carolina, Duke University
DEPARTMENT
UNC McAllister Heart Institute, Duke Molecular Physiology Institute
LABORATORY
Multiple Centers
LAST_NAME
Willis, Ilaiwy
FIRST_NAME
Monte, Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Non targeted metabolomics of gastrocnemius tissue samples obtained from 6 month old (adult) mice- Both Sham and after inducing lung injury
STUDY_TYPE
Non targeted metabolomic analysis
STUDY_SUMMARY
Introduction: Older patients are more likely to acquire and die from acute respiratory distress syndrome (ARDS) and muscle weakness may be more significant in older survivors. Recent data implicate muscle ring finger protein 1 (MuRF1) in lung injury-induced skeletal muscle atrophy in young mice and identify an alternative role for MuRF1 in cardiac metabolism regulation through inhibition of fatty acid oxidation. Objectives: To develop a model of lung injury-induced muscle wasting in old mice and to evaluate the skeletal muscle metabolomic profile of adult and old acute lung injury (ALI) mice. Methods: Young (2 month), adult (6 month) and old (20 month) male C57Bl6J mice underwent Sham (intratracheal H2O) or ALI [intratracheal E. coli lipopolysaccharide (i.t. LPS)] conditions and muscle functional testing. Metabolomic analysis on gastrocnemius muscle was performed using gas chromatography-mass spectrometry (GC-MS). Results: Old ALI mice had increased mortality and failed to recover skeletal muscle function compared to adult ALI mice. Muscle MuRF1 expression was increased in old ALI mice at day 3. Non-targeted muscle metabolomics revealed alterations in amino acid biosynthesis and fatty acid metabolism in old ALI mice. Targeted metabolomics of fatty acid intermediates (acyl-carnitines) and amino acids revealed a reduction in long chain acyl-carnitines in old ALI mice. Conclusion: This study demonstrates age-associated susceptibility to ALI-induced muscle wasting which parallels a metabolomic profile suggestive of altered muscle fatty acid metabolism. MuRF1 activation may contribute to both atrophy and impaired fatty acid oxidation, which may synergistically impair muscle function in old ALI mice.
INSTITUTE
University of North Carolina, Duke University
DEPARTMENT
UNC McAllister Heart Institute, Duke Molecular Physiology Institute
LABORATORY
Multiple Centers
LAST_NAME
Willis, Ilaiwy
FIRST_NAME
Monte, Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
targeted metabolomics of gastrocnemius tissue samples obtained from 6 month old (adult) mice- Both Sham and after inducing lung injury
STUDY_TYPE
targeted metabolomic analysis
STUDY_SUMMARY
Introduction: Older patients are more likely to acquire and die from acute respiratory distress syndrome (ARDS) and muscle weakness may be more significant in older survivors. Recent data implicate muscle ring finger protein 1 (MuRF1) in lung injury-induced skeletal muscle atrophy in young mice and identify an alternative role for MuRF1 in cardiac metabolism regulation through inhibition of fatty acid oxidation. Objectives: To develop a model of lung injury-induced muscle wasting in old mice and to evaluate the skeletal muscle metabolomic profile of adult and old acute lung injury (ALI) mice. Methods: Young (2 month), adult (6 month) and old (20 month) male C57Bl6J mice underwent Sham (intratracheal H2O) or ALI [intratracheal E. coli lipopolysaccharide (i.t. LPS)] conditions and muscle functional testing. Metabolomic analysis on gastrocnemius muscle was performed using gas chromatography-mass spectrometry (GC-MS). Results: Old ALI mice had increased mortality and failed to recover skeletal muscle function compared to adult ALI mice. Muscle MuRF1 expression was increased in old ALI mice at day 3. Non-targeted muscle metabolomics revealed alterations in amino acid biosynthesis and fatty acid metabolism in old ALI mice. Targeted metabolomics of fatty acid intermediates (acyl-carnitines) and amino acids revealed a reduction in long chain acyl-carnitines in old ALI mice. Conclusion: This study demonstrates age-associated susceptibility to ALI-induced muscle wasting which parallels a metabolomic profile suggestive of altered muscle fatty acid metabolism. MuRF1 activation may contribute to both atrophy and impaired fatty acid oxidation, which may synergistically impair muscle function in old ALI mice.
INSTITUTE
University of North Carolina, Duke University
DEPARTMENT
UNC McAllister Heart Institute, Duke Molecular Physiology Institute
LABORATORY
Multiple Centers
LAST_NAME
Willis, Ilaiwy
FIRST_NAME
Monte, Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Targeted metabolomics of gastrocnemius tissue samples obtained from 20 month old (old) mice- Both Sham and after inducing lung injury (part II)
STUDY_TYPE
Targeted metabolomic analysis
STUDY_SUMMARY
Introduction: Older patients are more likely to acquire and die from acute respiratory distress syndrome (ARDS) and muscle weakness may be more significant in older survivors. Recent data implicate muscle ring finger protein 1 (MuRF1) in lung injury-induced skeletal muscle atrophy in young mice and identify an alternative role for MuRF1 in cardiac metabolism regulation through inhibition of fatty acid oxidation. Objectives: To develop a model of lung injury-induced muscle wasting in old mice and to evaluate the skeletal muscle metabolomic profile of adult and old acute lung injury (ALI) mice. Methods: Young (2 month), adult (6 month) and old (20 month) male C57Bl6J mice underwent Sham (intratracheal H2O) or ALI [intratracheal E. coli lipopolysaccharide (i.t. LPS)] conditions and muscle functional testing. Metabolomic analysis on gastrocnemius muscle was performed using gas chromatography-mass spectrometry (GC-MS). Results: Old ALI mice had increased mortality and failed to recover skeletal muscle function compared to adult ALI mice. Muscle MuRF1 expression was increased in old ALI mice at day 3. Non-targeted muscle metabolomics revealed alterations in amino acid biosynthesis and fatty acid metabolism in old ALI mice. Targeted metabolomics of fatty acid intermediates (acyl-carnitines) and amino acids revealed a reduction in long chain acyl-carnitines in old ALI mice. Conclusion: This study demonstrates age-associated susceptibility to ALI-induced muscle wasting which parallels a metabolomic profile suggestive of altered muscle fatty acid metabolism. MuRF1 activation may contribute to both atrophy and impaired fatty acid oxidation, which may synergistically impair muscle function in old ALI mice.
INSTITUTE
University of North Carolina, Duke University
DEPARTMENT
UNC McAllister Heart Institute, Duke Molecular Physiology Institute
LABORATORY
Multiple Centers
LAST_NAME
Willis, Ilaiwy
FIRST_NAME
Monte, Amro
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Targeted metabolomics of MuRF1 overexpressing cardiomyocytes compared to their wildtype controls (part I)
STUDY_TYPE
Targeted metabolomic analysis
STUDY_SUMMARY
The transcriptional regulation of peroxisome proliferator-activated receptor (PPAR) a by post-translational modification, such as ubiquitin, has not been described. We report here for the first time an ubiquitin ligase (muscle ring finger-1/MuRF1) that inhibits fatty acid oxidation by inhibiting PPARa, but not PPARĆ/d or PPAR? in cardiomyocytes in vitro. Similarly, MuRF1 Tg+ hearts showed significant decreases in nuclear PPARa activity and acyl-carnitine intermediates, while MuRF1-/- hearts exhibited increased PPARa activity and acyl-carnitine intermediates. MuRF1 directly interacts with PPARa, mono-ubiquitinates it, and targets it for nuclear export to inhibit fatty acid oxidation in a proteasome independent manner. We then identified a previously undescribed nuclear export sequence in PPARa, along with three specific lysines (292, 310, 388) required for MuRF1's targeting of nuclear export. These studies identify the role of ubiquitination in regulating cardiac PPARa, including the ubiquitin ligase that may be responsible for this critical regulation of cardiac metabolism in heart failure.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister Heart Institute, Department of Internal Medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Targeted metabolomics of MuRF1 Knockdown cardiomyocytes compared to their wildtype controls (part 2)
STUDY_TYPE
Targeted metabolomic analysis
STUDY_SUMMARY
The transcriptional regulation of peroxisome proliferator-activated receptor (PPAR) Ī± by post-translational modification, such as ubiquitin, has not been described. We report here for the first time an ubiquitin ligase (muscle ring finger-1/MuRF1) that inhibits fatty acid oxidation by inhibiting PPARĪ±, but not PPARĪ²/Ī“ or PPARĪ³ in cardiomyocytes in vitro. Similarly, MuRF1 Tg+ hearts showed significant decreases in nuclear PPARĪ± activity and acyl-carnitine intermediates, while MuRF1ā/ā hearts exhibited increased PPARĪ± activity and acyl-carnitine intermediates. MuRF1 directly interacts with PPARĪ±, mono-ubiquitinates it, and targets it for nuclear export to inhibit fatty acid oxidation in a proteasome independent manner. We then identified a previously undescribed nuclear export sequence in PPARĪ±, along with three specific lysines (292, 310, 388) required for MuRF1s targeting of nuclear export. These studies identify the role of ubiquitination in regulating cardiac PPARĪ±, including the ubiquitin ligase that may be responsible for this critical regulation of cardiac metabolism in heart failure.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister Heart Institute, Department of Internal Medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
C2C12 stretch cessation models muscle atrophy and anaplerotic changes in metabolism
STUDY_TYPE
GC-MS non-targeted metabolomic profiling
STUDY_SUMMARY
Studies of skeletal muscle disuse either in patients on bed rest or experimentally in animals(immobilization) have demonstrated that decreased protein synthesis is common, with transient parallel increases in protein degradation. Muscle disuse atrophy involves a process of transition from slow to fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for fat oxidation, and substrate accumulation in atrophied muscles have been reported as has accommodation of the liver with an increased gluconeogenic capacity. Recent studies have modeled skeletal muscle disuse by using cyclic stretch of differentiated myotubes (C2C12), which mimics the loading pattern of mature skeletal muscle, followed by cessation of stretch.We utilized this model to determine the metabolic changes using non-targeted metabolomics analysis of the media. We identified increases in amino acids resulting from protein degradation (largely sarcomere) that occurs with muscle atrophy that are involved in feeding the Krebās cycle through anaplerosis. Specifically, we identified increased alanine/proline metabolism (significantly elevated proline, alanine, glutamine, and asparagine) and increased ļ”-ketoglutaric acid, the proposed Krebās cycle intermediate being fed by the alanine/proline metabolic anaplerotic mechanism. Additionally, several unique pathways not clearly delineated in previous studies of muscle unloading were seen, including: 1) elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and 2-keovaline) represent intermediates in the Ehlrich amino acid degradation pathway, which feeds into a metabolic pathway supplying acetyl-CoA and 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an intermediate of purine metabolism, was seen at 12 hours unloading. Given the interest in targeting different aspects of the ubiquitin proteasome system to inhibit protein degradation, this C2C12 system may allow the identification of direct and indirect alterations in metabolism due to anaplerosis or through other yet to be identified mechanisms using a non-targeted metabolomics approach.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister Heart Institute
LABORATORY
Mutliple Centers
LAST_NAME
Ilaiwy, WIllis
FIRST_NAME
Amro, Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Quantitative measurements of vitamin D in T1D poor control, good control, and controls.
STUDY_TYPE
Quantitative measurements of vitamin D
STUDY_SUMMARY
The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Plasma sphingolipid changes with autopsy-confirmed Lewy body or Alzheimer's pathology
STUDY_TYPE
targeted sphingolipid and fatty acid analyses
STUDY_SUMMARY
We identified four groups with available plasma 2 years before death: high (n = 12) and intermediate-likelihood DLB (n = 14) based on the third report of the DLB consortium; dementia with Alzheimer's pathology (.D n = 18); and cognitively normal with normal aging pathology (n = 21). Lipids were measured using ESI/MS/MS.
Quantitative measurements of free fatty acid in T1D poor control, good control, and controls.
STUDY_TYPE
Quantitative measurements of free fatty acid
STUDY_SUMMARY
The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Quantitative measurements of amino acids in T1D poor control, good control, and controls.
STUDY_TYPE
Quantitative measurements of amino acid
STUDY_SUMMARY
The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Metabolomic analysis of human plasma samples spiked with unknown chemicals
STUDY_TYPE
Untargeted metabolomics for sample classification
STUDY_SUMMARY
To conduct metabolomic studies comparing 20 males and 20 females as well as 10 pool controls and 5 spike-in samples using MS platform to perform both qualitative and quantitative metabolome analysis to classify the sample groups (male, female, pooled) and to identify the spiked-in compounds.
Characterizing commonalities and differences between the breast and prostate cancer metabotypes in African-American cohorts (part I)
STUDY_TYPE
Identify Breast Cancer Progression and Prostate Cancer correlative Metabolite Markers
STUDY_SUMMARY
Thus, we aimed to 1) understand mechanisms related to the onset and progression of BCa, 2) identify precursors and targets for prevention and therapy for each stage, grade and subtype which may contribute to the disparate impact of BCa in African American women, and 3) Identify common and different BCa metabolite markers versus PCa markers.
Characterizing commonalities and differences between the breast and prostate cancer metabotypes in African-American cohorts
STUDY_TYPE
Identify Breast Cancer Progression and Prostate Cancer correlative Metabolite Markers
STUDY_SUMMARY
Thus, we aimed to 1) understand mechanisms related to the onset and progression of BCa, 2) identify precursors and targets for prevention and therapy for each stage, grade and subtype which may contribute to the disparate impact of BCa in African American women, and 3) Identify common and different BCa metabolite markers versus PCa markers.
Metabotypes of Subjects with Adverse Reactions Following Vaccination
STUDY_TYPE
Metabolomic Profile of Human Serum
STUDY_SUMMARY
Metabolomics may help identify a particular metabolic signature āmetabotypeā in patients who are predisposed to developing AEFI such as a systemic reaction, or myocarditis that currently is difficult or impossible to identify prior to the development of the AEFI. This proposed pilot study looks at the metabolic profiles of a specific population of subjects who received the smallpox vaccine with or without other concomitantly administered vaccines to help determine if a unique metabotype can be identified in subjects who reported systemic reactions following immunization. In addition, this proposed study will look at the metabolic profile of several subjects with subclinical or clinically diagnosed myopericarditis to determine if these subjects have a unique metabotype. The ability to identify a unique metabotype would allow a clinician to potentially mitigate serious AEFI and ultimately improve the quality of immunization healthcare. If identified, these profiles might represent novel biomarkers of risk that can supplement existing clinical decision making for risk stratification or vaccine exemptions.
Metabolomic Profiling of the Malaria Box Reveals Antimalarial Target Pathways
STUDY_SUMMARY
Here we interrogated the in vitro metabolic effects of 189 drugs (including 169 of the drug-like compounds from the Malaria Box) using ultra-high performance liquid chromatography mass-spectrometry (UHPLC-MS). The resulting metabolic fingerprints provide information on the parasite biochemical pathways affected by pharmacologic intervention and offer a critical blueprint for selecting and advancing lead compounds as next-generation antimalarial drugs. Our results reveal several major classes of metabolic disruption, which allow us to predict the mode-of-action (MoA) for many of the Malaria Box compounds.
INSTITUTE
Penn State University
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millenium Science Complex, University Park, PA 16802
Metabolomics Analysis of Triple Negative Breast Cancer (BCa) Cell Lines
STUDY_TYPE
Metabolomics comparison of different breast cancer cell lines
STUDY_SUMMARY
We used untargeted metabolomic profiling to distinguish this form of BCa from estrogen receptor positive (ER+) subtypes (+/- HER2/neu) and determine that may explain why a commonly used chemotherapeutic, paclitaxel, is generally ineffective at eliciting long-term cytotoxic and/or cytostatic responses in cell line models of TNBC. This metabolomics study used broad spectrum 1H NMR to compare Luminal A (BT474, MCF-7) and triple-negative (MDA-MB-231, MDA-MB-468) BCa cell lines, to determine differences in the two subtypes as well as distinguish therapeutic treatment responses for identifying new targets for drug discovery.
INSTITUTE
RTI International
DEPARTMENT
Discovery Sciences
LABORATORY
STS/RCMRC
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040, East Cornwallis Road, Research Triangle Park, NC 27709
EMAIL
ssumner@rti.org
PHONE
919-541-7479
NUM_GROUPS
4
TOTAL_SUBJECTS
24
NUM_MALES
N/A
NUM_FEMALES
N/A
STUDY_COMMENTS
4 breast cancer cell lines, treated with paclitaxel compared to controls (3 replicates/line/condition)
PUBLICATIONS
J. Proteome Res., Article ASAP, DOI: 10.1021/acs.jproteome.6b00430
Quantitative measurements of TCA cycle metabolites in T1D poor control, good control, and controls.
STUDY_TYPE
Quantitative measurements of plasma levels of tricarboxylic acid (TCA) cycle metabolites
STUDY_SUMMARY
The objective of the study was to determine whether T1D with good glycemic control have persistent abnormalities of metabolites and pathways that exist in T1D with poor glycemic control.
Preconcentration of organic solutes in urine by bubble bursting
STUDY_TYPE
Sample preparation for MS analysis
STUDY_SUMMARY
The chemical sensitivity of urine metabolomics analysis is greatly compromised due to the large amounts of inorganic salts in urine (NaCl, KCl), which are detrimental to analytical instrumentation, e.g. chromatographic columns or mass spectrometers. Traditional desalting approaches applied to urine pretreatment suffer from the chemical losses, which reduce the information depth of analysis. We aimed to test a simple approach for the simultaneous preconcentration and desalting of organic solutes in urine based on the collection of induced bursting bubble aerosols above the surface of urine samples. Bursting bubbles were generated at ambient conditions by feeding gas through an air diffuser at the bottom of diluted (200 times in ultrapure water) urine solution (50-500 mL). Collected aerosols were analyzed by the direct-infusion electrospray ionization mass spectrometry (ESI-MS). The simultaneous preconcentration (ca. 6-12 fold) and desalting (ca. 6-10 fold) of organic solutes in urine was achieved by the bursting bubble sample pretreatment, which allowed ca. 3-times higher number of identified urine metabolites by high-resolution MS analysis. No notable chemical discrimination effects were observed. The increased degree of MS data clustering was demonstrated on the principal component analysis of data sets from the urine of healthy people and from the urine people with renal insufficiency. At least 10 times higher sensitivity of trace drug detection in urine was demonstrated for clenbuterol and salbutamol. Our results indicate the high versatility of bubble bursting as a simple pretreatment approach to enhance the chemical depth and sensitivity of urine analysis.
INSTITUTE
Research Center for Obstetrics, Gynecology and Perinatology
DEPARTMENT
Department of System Biology in Reproduction
LABORATORY
Laboratory for Proteomics and Metabolomics of Human Reproduction
Follicular fluid lipidomics reveals lipid alterations by LH addition during IVF cycles
STUDY_SUMMARY
Purpose Ovulation induction protocols are key components for performing assisted reproduction treatments successfully. The objective of the present study was to estimate how LH addition to controlled ovarian stimulation protocols may affect the follicular fluid lipid profile of women undergoing in vitro fertilization treatment. Methods We conducted the study using 28 self-paired samples, 14 per group. The patients received FSH during their first cycle of ovarian stimulation (FSH group). If treatment did not result in pregnancy, the same patients returned for a new cycle and received stimulus with the addition of LH to the previous protocol (Low-dose-LH group). Lipidomics analysis was performed by UPLC-MSE mass spectrometry. Potential lipid biomarkers were identified by the software Progenesis QI. Statistical analysis was performed using the SPSS 18.0 and MetaboAnalyst 2.0 software.
INSTITUTE
Universidade Federal de Sao Paulo
DEPARTMENT
Surgery
LABORATORY
Centro de Pesquisa em Urologia
LAST_NAME
Da Costa
FIRST_NAME
Livia
ADDRESS
Rua Embau 231 - Vila Clementino, Sao Paulo, Sao Paulo, 04039060, Brazil
EMAIL
liviadovale@hotmail.com
PHONE
551138074062
NUM_GROUPS
2
TOTAL_SUBJECTS
28
NUM_FEMALES
14
STUDY_COMMENTS
The groups consists of the same 14 women submitted to two different controlled ovarian stimulation protocols (FSH or LH group)
Heterologous expression and detection of Apratoxins in E. coli
STUDY_TYPE
Organic extraction of E coli cultures harboring apratoxin gene cluster
STUDY_SUMMARY
The apratoxin gene cluster was recovered from fosmid DNA library. The gene set responsible for the biosynthesis of the polyketide backbone was introduced in E. coli BAP strain and expressed at 30C for 24 hours. Two sets of control and experimental samples with culture broth pH adjusted at 8 and 12 respectively was extracted with ethyl acetate and dried.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Kallifidas
FIRST_NAME
Dimitris
ADDRESS
Medical Sciences Building, Rm P5-26, 1600 SW Archer Rd., FL32610
Emory University high-resolution metabolomic profiling of ring trial samples
STUDY_TYPE
Untargeted HRMS for cross-laboratory comparison of metabolomics platforms
STUDY_SUMMARY
Untargeted profiling of blinded plasma samples consisting of 20 females, 20 males, 10 pooled samples and 5 fortified pooled samples was completed using liquid chromatography with high-resolution mass spectrometry. All samples were analyzed using HILIC and C18 chromatography interfaced to a Thermo Scientific Orbitrap Fusion Tribrid high-resolution mass spectrometer operated in positive and negative ESI. Mass spectral data was extracted using adaptive processing algorithms at multiple settings and annotated using xMSannotator. Collected data were uploaded to the Metabolomics Workbench for evaluation of system performance.
INSTITUTE
Emory University School of Medicine
DEPARTMENT
Division of Pulmonary, Allergy and Critical Care Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Walker
FIRST_NAME
Douglas
ADDRESS
615 Michael St. Ste 225, Atlanta, GA, 30322, USA
EMAIL
douglas.walker@emory.edu
PHONE
4047275984
NUM_GROUPS
4
TOTAL_SUBJECTS
55
NUM_MALES
20
NUM_FEMALES
20
STUDY_COMMENTS
Contained 20 females, 20 males, 10 pooled samples and 5 fortified pooled samples
This targeted metabolomic analysis was performed on plasma samples from 39 normal controls (n=18 men and 21 women) and 45 subjects ((n = 22 men and 23 women) who met diagnostic criteria for ME/CFS by Institute of Medicine, Canadian, and Fukuda criteria.
INSTITUTE
University of California, San Diego
DEPARTMENT
The Mitochondrial and Metabolic Disease Center
LAST_NAME
Naviaux
FIRST_NAME
Robert
ADDRESS
214 Dikinson Street, CTF-C102, San Diego, CA, 92103
EMAIL
maviaux@ucsd.edu
PHONE
619-993-2904
NUM_GROUPS
2 groups for men (control and CFS) and 2 groups for women (control and CFS)
The alpha-1A adrenergic receptor agonist A61603 reduces cardiac polyunsaturated fatty acid-Heart raw data
STUDY_TYPE
GC-MS non-targeted metabolomic profiling
STUDY_SUMMARY
Studies of skeletal muscle disuse either in patients on bed rest or experimentally in animals(immobilization) have demonstrated that decreased protein synthesis is common, with transient parallel increases in protein degradation. Muscle disuse atrophy involves a process of transition from slow to fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for fat oxidation, and substrate accumulation in atrophied muscles have been reported as has accommodation of the liver with an increased gluconeogenic capacity. Recent studies have modeled skeletal muscle disuse by using cyclic stretch of differentiated myotubes (C2C12), which mimics the loading pattern of mature skeletal muscle, followed by cessation of stretch.We utilized this model to determine the metabolic changes using non-targeted metabolomics analysis of the media. We identified increases in amino acids resulting from protein degradation (largely sarcomere) that occurs with muscle atrophy that are involved in feeding the Krebās cycle through anaplerosis. Specifically, we identified increased alanine/proline metabolism (significantly elevated proline, alanine, glutamine, and asparagine) and increased ļ”-ketoglutaric acid, the proposed Krebās cycle intermediate being fed by the alanine/proline metabolic anaplerotic mechanism. Additionally, several unique pathways not clearly delineated in previous studies of muscle unloading were seen, including: 1) elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and 2-keovaline) represent intermediates in the Ehlrich amino acid degradation pathway, which feeds into a metabolic pathway supplying acetyl-CoA and 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an intermediate of purine metabolism, was seen at 12 hours unloading. Given the interest in targeting different aspects of the ubiquitin proteasome system to inhibit protein degradation, this C2C12 system may allow the identification of direct and indirect alterations in metabolism due to anaplerosis or through other yet to be identified mechanisms using a non-targeted metabolomics approach.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister Heart Institute
LABORATORY
Mutliple Centers
LAST_NAME
Ilaiwy, WIllis
FIRST_NAME
Amro, Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
The alpha-1A adrenergic receptor agonist A61603 reduces cardiac polyunsaturated fatty acid-Serum raw data
STUDY_TYPE
GC-MS non-targeted metabolomic profiling
STUDY_SUMMARY
Studies of skeletal muscle disuse either in patients on bed rest or experimentally in animals(immobilization) have demonstrated that decreased protein synthesis is common, with transient parallel increases in protein degradation. Muscle disuse atrophy involves a process of transition from slow to fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for fat oxidation, and substrate accumulation in atrophied muscles have been reported as has accommodation of the liver with an increased gluconeogenic capacity. Recent studies have modeled skeletal muscle disuse by using cyclic stretch of differentiated myotubes (C2C12), which mimics the loading pattern of mature skeletal muscle, followed by cessation of stretch.We utilized this model to determine the metabolic changes using non-targeted metabolomics analysis of the media. We identified increases in amino acids resulting from protein degradation (largely sarcomere) that occurs with muscle atrophy that are involved in feeding the Krebās cycle through anaplerosis. Specifically, we identified increased alanine/proline metabolism (significantly elevated proline, alanine, glutamine, and asparagine) and increased ļ”-ketoglutaric acid, the proposed Krebās cycle intermediate being fed by the alanine/proline metabolic anaplerotic mechanism. Additionally, several unique pathways not clearly delineated in previous studies of muscle unloading were seen, including: 1) elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and 2-keovaline) represent intermediates in the Ehlrich amino acid degradation pathway, which feeds into a metabolic pathway supplying acetyl-CoA and 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an intermediate of purine metabolism, was seen at 12 hours unloading. Given the interest in targeting different aspects of the ubiquitin proteasome system to inhibit protein degradation, this C2C12 system may allow the identification of direct and indirect alterations in metabolism due to anaplerosis or through other yet to be identified mechanisms using a non-targeted metabolomics approach.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister Heart Institute
LABORATORY
Mutliple Centers
LAST_NAME
Ilaiwy, WIllis
FIRST_NAME
Amro, Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Metabolic Adaptation of Staphylococcus aureus to Host Immunity
STUDY_TYPE
Metabolomics Analysis of Methicillin-Resistant Staphylococcus aureus (MRSA) to host immunity.
STUDY_SUMMARY
This project is intended to study the metabolic adaptation of Methicillin-Resistant Staphylococcus aureus (MRSA) to host immunity. Because of the nature of the samples RTI RCMRC worked with Dr. Anthony R. Richardson so that the samples would be extracted at the University of North Carolina at Chapel Hill under the condition that were optimized by RTI RCMRC for broad spectrum metabolomics analysis.
Utilizing Metabolomics to Understand Novel Anti-Desmoid Tumor Drugs
STUDY_SUMMARY
This pilot study will use broad spectrum metabolomics to study the tumorigenesis process of fibroblasts to desmoids by investigating paired desmoid and fibroblast cell lines, in addition to unaffected fibroblast cells. Additionally, this pilot study will explore the effects of two of the active drugs identified on the desmoid and fibroblast cells.
Utilizing Metabolomics to Understand Novel Anti-Desmoid Tumor Drugs
STUDY_SUMMARY
This pilot study will use broad spectrum metabolomics to study the tumorigenesis process of fibroblasts to desmoids by investigating paired desmoid and fibroblast cell lines, in addition to unaffected fibroblast cells. Additionally, this pilot study will explore the effects of two of the active drugs identified on the desmoid and fibroblast cells.
Metabolomic changes during active immunization with anti-METH vaccines.
STUDY_SUMMARY
A targeted metabolomics analysis was performed on serum samples from rats immunized with a methamphetamine-like hapten covalently bound to keyhole limpet hemocyanin (KLH) monomers, and controls. The subjects were approximately 300 g adult male Sprague Dawley rats. RTI International RCMRC received pre-immune serum from RI 11-03A (n=12) which is a matched control serum that was collected before starting immunizations, and week 17 bleeds from RI 11-03A (n=12), RI 11-03B (n=8), and RI 11-03C (n=12). The A-C nomenclature denotes A) complete antigen with adjuvant, B) complete antigen without adjuvant and C) only KLH carrier protein and adjuvant without a METH-like hapten. The ratsā serum samples were extracted and prepared for a combined flow injection (FIA) and LC-MS/MS assay (Biocrates AbsoluteIDQĀ® p180 kit). The Biocrates AbsoluteIDQĀ® p180 kit (Biocrates Life Sciences AG, Innsbruck, Austria) can quantitatively measure ~188 biologically relevant metabolites from five analyte groups (acylcarnitines, amino acids, biogenic amines, glycerophospho- and sphingolipids, and hexoses) on a triple quadrupole mass spectrometer.
Metabolomics of Mice Cohousing and Microbiota Transfer
STUDY_SUMMARY
The mice serum samples were extracted and prepared for a combined flow injection (FIA) and LC-MS/MS assay (Biocrates AbsoluteIDQĀ® p180 kit). The Biocrates AbsoluteIDQĀ® p180 kit (Biocrates Life Sciences AG, Innsbruck, Austria) can quantitatively measure ~188 biologically relevant metabolites from five analyte groups (acylcarnitines, amino acids, biogenic amines, glycerophospho- and sphingolipids, and hexoses) on a triple quadrupole mass spectrometer.
Cells are treated with vehicle or Doxycycline - to induce expression of GFP or ATG7
STUDY_SUMMARY
AGY571 cells engineered to express Dox-inducible GFP or ATG7 were treated with 500 ng/mL or 1 ug/mL Doxycycline for 48 hours. Assessing the effects of ATG7 reexpression in B cell lymphoma.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LABORATORY
John Cleveland
LAST_NAME
Fernandez
FIRST_NAME
Mario
ADDRESS
12902 Magnolia Drive, MRC 4-West
EMAIL
mario.fernandez@moffitt.org
PHONE
813-745-5140
NUM_GROUPS
5
TOTAL_SUBJECTS
15
STUDY_COMMENTS
Groups collected in triplicate per time point, either treated or untreated with Doxycycline.
CNS and peripheral metabolomics of calorie restriction in a mouse model of Alzheimerās disease (part I)
STUDY_SUMMARY
This pilot metabolomic study will evaluate brain specimens from an established mouse model of AD, the tq2576 mouse model of cerebral amyloid overexpression (APP), in comparison to their non-transgenic (NTG) littermates. These animals were either on a CR or ad libitum (AL) diet, and specimens were collected at two time points (5 and 15 months of age). Tissue from this cohorts of mice have already undergone microbiome analysis, and await coordinated brain and peripheral tissue assessments. Future analysis will include metabolomics, RNA-seq, and microarray data to assess the gut-brain microbiome system in neurodegenerative disorders.
CNS and peripheral metabolomics of calorie restriction in a mouse model of Alzheimerās disease (part II)
STUDY_SUMMARY
This pilot metabolomic study will evaluate cecal specimens from an established mouse model of AD, the tq2576 mouse model of cerebral amyloid overexpression, in comparison to their non-transgenic (ntg) littermates. These animals were either on a CR or ad libitum (AL) diet, and specimens were collected at two time points (5 and 15 months of age). Tissue from this cohorts of mice have already undergone microbiome analysis, and await coordinated brain and peripheral tissue assessments. Future analysis will include metabolomics, RNA-seq, and microarray data to assess the gut-brain microbiome system in neurodegenerative disorders.
Transpulmonary metabolomics in pulmonary arterial hypertension
STUDY_SUMMARY
We hypothesize that transpulmonary metabolomic profiling will demonstrate a PAH-specific metabolic signature. We will examine organ-specific metabolism by measuring blood flowing into (pulmonary artery) and out of (pulmonary artery wedge) the pulmonary circulation at the time of right heart catheterization (RHC). We will compare PAH to patients without PH and to a disease control cohort with PH due to left heart disease (pulmonary ventrical hypertension - PVH).
Uniquely Tumor-Selective Englerin A Profoundly Alters Lipid Metabolism in Renal Cell Carcinoma inducing ER-Stress and an Acute Inflammatory Response
STUDY_TYPE
Metabolomic effect of Englerin A on renal cell carcinoma
STUDY_SUMMARY
This targeted metabolomic analysis was performed on renal cell carcinoma A498 cells with or without anti-cancer drug Englerin treatment for 24 and 48 h.
INSTITUTE
University of California, San Diego
DEPARTMENT
Department of Pediatrics
LAST_NAME
Batova
FIRST_NAME
Ayse
ADDRESS
La Jolla, CA 92093
EMAIL
abatova@ucsd.edu
PHONE
619-543-1962
NUM_GROUPS
Two groups for 24 h treatment (control and Eglerin treatment) and two groups for 48 h treatment. Each has 4 replicates
Streptococcus mutans organic acids/amino acid profiles, effect of oxidative stress; pilot
STUDY_TYPE
Amino acid/organic acid profiles of wild-type and mutant strain in response to H2O2 treatment over time.
STUDY_SUMMARY
S. mutans UA159 and ĪspxA1 will be grown in BHI to OD600 = 0.4. Each culture will be split in to 4 samples. For the first sample, 3E10 cells will immediately be harvested by fast filtration, scraped off of membrane into ice-cold PBS, and pelleted by centrifugation. The remaining three samples will be treated with H2O2 to 0.5 mM, and the harvesting process repeated after 15, 30 and 60 minutes.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LABORATORY
Lemos-Abranches
LAST_NAME
Hardin
FIRST_NAME
Emily
ADDRESS
1395 Center Drive, PO Box 100424, Gainesville, FL 32610
Metabolomics of Saliva Samples Obtained from Subjects with Diabetes
STUDY_SUMMARY
In this research, were are investigating the metabolic profile changes associated with well- and poorly-controlled type 1 and 2 diabetes and if there are distinct metabolite compounds that may be associated with glycemic control. The PI of the study collected whole unstimulated saliva samples were from subjects with well- and poorly-controlled type 1 and type 2 diabetes. Subjects were selected based on the level of A1C (<7= well-controlled and >7 = poorly controlled). Saliva samples were shipped to RTI RCMRC for a broad spectrum reverse phase metabolomics analysis.
Enterococcus faecalis nucleotide profiles following mupirocin or decoyinine
STUDY_TYPE
Exposure to mupirocin or decoyinine
STUDY_SUMMARY
Triplicate samples of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split and exposed to DMSO, mupirocin (0.01mg/mL) or decoyinine (0.1mg/mL) for 15 minutes. 6E9 cells will be harvested by fast filtration, scraped off of membrane into ice-cold PBS, and pelleted by centrifugation, and stored at -80C until shipment.
INSTITUTE
Univeristy of Florida
DEPARTMENT
SECIM
LABORATORY
Lemos-Abranches
LAST_NAME
Kajfasz
FIRST_NAME
Jessica
ADDRESS
1395 Center Drive, PO Box 100424 Gainesville, FL 32610
Comparison between wild-type and mutant strain, as well as providing varying cell count input for WT
STUDY_SUMMARY
E. faecalis OG1RF and rel/relQ will be grown in FMC-AUG to OD600 = 0.25. 3E10 cells will be harvested by fast filtration, scraped off of membrane into ice-cold PBS, and pelleted by centrifugation. This process will be repeated with OG1RF only to harvest 3E9 and 3E8 cells. A second OG1RF sample of 3E10 cells will be scraped into formic acid; only acid-soluble extract willl be sent for this sample.
INSTITUTE
Univeristy of Florida
DEPARTMENT
SECIM
LABORATORY
Lemos-Abranches
LAST_NAME
Kajfasz
FIRST_NAME
Jessica
ADDRESS
1395 Center Drive, PO Box 100424 Gainesville, FL 32610
Exahustive degradation of nucleotide triphosphates
STUDY_TYPE
Comparison of degradation kinetics
STUDY_SUMMARY
The degradation kinetics of nucleotide triphosphates (ATP, GTP, UTP and CTP) were evaluated under boiling ethanol extraction conditions (95Ā°C) during 0 to 300 minutes.
INSTITUTE
University of Groningen
DEPARTMENT
Analytical Biochemistry
LAST_NAME
Bischoff
FIRST_NAME
Rainer
ADDRESS
Antonius Deusinglaan 1 (XB20), Building 3226, room 601, 9713 AV Groningen, The Netherlands
Effect of a complex matrix on the degradation of nucleotide triphosphates
STUDY_TYPE
Comparison of degradation kinetics
STUDY_SUMMARY
The effect of a complex cellular matrix extracted from yeast (S. cerevisiae, strain YSBN6 (MATa; genotype: FY3 ho::HphMX4 derived from the S288C parental strain)) on the degradation profiles of nucleotide triphosphates extracted under typical boiling ethanol conditions was evaluated.
INSTITUTE
University of Groningen
DEPARTMENT
Analytical Biochemistry
LAST_NAME
Bischoff
FIRST_NAME
Rainer
ADDRESS
Antonius Deusinglaan 1 (XB20), Building 3226, room 601, 9713 AV Groningen, The Netherlands
Effect of the culture medium on the degradation of nucleotide triphosphates
STUDY_TYPE
Endpoint measurement
STUDY_SUMMARY
The effect of the Verduyn culture medium on the degradation profiles of nucleotide triphosphates extracted under typical boiling ethanol conditions was evaluated.
INSTITUTE
University of Groningen
DEPARTMENT
Analytical Biochemistry
LAST_NAME
Bischoff
FIRST_NAME
Rainer
ADDRESS
Antonius Deusinglaan 1 (XB20), Building 3226, room 601, 9713 AV Groningen, The Netherlands
Effect of the chemical environment on the degradation of nucleotide triphosphates
STUDY_TYPE
Endpoint measurement
STUDY_SUMMARY
The influence of particular groups of compounds/metabolites, mainly comprising major central carbon metabolites including amino acids, organic acids, sugar phosphates, coenzymes, etc, on the degradation profiles of nucleotide triphosphates extracted under typical boiling ethanol conditions was evaluated.
INSTITUTE
University of Groningen
DEPARTMENT
Analytical Biochemistry
LAST_NAME
Bischoff
FIRST_NAME
Rainer
ADDRESS
Antonius Deusinglaan 1 (XB20), Building 3226, room 601, 9713 AV Groningen, The Netherlands
Amino Acid Quantifcation of obese patients on a 16 week caloric restriction from Plasma
STUDY_TYPE
timecourse, quantitative measurements of amino acid
STUDY_SUMMARY
Caloric restriction (CR) improves insulin sensitivity and reduces the incidence of diabetes in obese individuals. The underlying mechanisms whereby CR improves insulin sensitivity are not clear. We evaluated the effect of 16 weeks of CR on whole-body insulin sensitivity by pancreatic clamp before and after CR in 11 obese participants (BMI = 35 kg/m2) compared with 9 matched control subjects (BMI = 34 kg/m2). Compared with the control subjects, CR increased the glucose infusion rate needed to maintain euglycemia during hyperinsulinemia, indicating enhancement of peripheral insulin sensitivity. This improvement in insulin sensitivity was not accompanied by changes in skeletal muscle mitochondrial oxidative capacity or oxidant emissions, nor were there changes in skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels. However, CR lowered insulin-stimulated thioredoxin-interacting protein (TXNIP) levels and enhanced nonoxidative glucose disposal. These results support a role for TXNIP in mediating the improvement in peripheral insulin sensitivity after CR.
Amino Acid Quantifcation of obese patients on a 16 week caloric restriction from muscle biopsy (part II)
STUDY_TYPE
timecourse, quantitative measurements of amino acid
STUDY_SUMMARY
Caloric restriction (CR) improves insulin sensitivity and reduces the incidence of diabetes in obese individuals. The underlying mechanisms whereby CR improves insulin sensitivity are not clear. We evaluated the effect of 16 weeks of CR on whole-body insulin sensitivity by pancreatic clamp before and after CR in 11 obese participants (BMI = 35 kg/m2) compared with 9 matched control subjects (BMI = 34 kg/m2). Compared with the control subjects, CR increased the glucose infusion rate needed to maintain euglycemia during hyperinsulinemia, indicating enhancement of peripheral insulin sensitivity. This improvement in insulin sensitivity was not accompanied by changes in skeletal muscle mitochondrial oxidative capacity or oxidant emissions, nor were there changes in skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels. However, CR lowered insulin-stimulated thioredoxin-interacting protein (TXNIP) levels and enhanced nonoxidative glucose disposal. These results support a role for TXNIP in mediating the improvement in peripheral insulin sensitivity after CR.
D2 Glucose Quantifcation of obese patients on a 16 week caloric restriction from plasma
STUDY_TYPE
timecourse
STUDY_SUMMARY
Caloric restriction (CR) improves insulin sensitivity and reduces the incidence of diabetes in obese individuals. The underlying mechanisms whereby CR improves insulin sensitivity are not clear. We evaluated the effect of 16 weeks of CR on whole-body insulin sensitivity by pancreatic clamp before and after CR in 11 obese participants (BMI = 35 kg/m2) compared with 9 matched control subjects (BMI = 34 kg/m2). Compared with the control subjects, CR increased the glucose infusion rate needed to maintain euglycemia during hyperinsulinemia, indicating enhancement of peripheral insulin sensitivity. This improvement in insulin sensitivity was not accompanied by changes in skeletal muscle mitochondrial oxidative capacity or oxidant emissions, nor were there changes in skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels. However, CR lowered insulin-stimulated thioredoxin-interacting protein (TXNIP) levels and enhanced nonoxidative glucose disposal. These results support a role for TXNIP in mediating the improvement in peripheral insulin sensitivity after CR.
Impacts of high-fat diet (HFD), high-carbohydrate diet (HCD) and high-fat-high-carbohydrate diet (HFHCD) on metabolites in a farmed cyprinid fish Megalobrama amblycephala.
STUDY_TYPE
Dietary imbalance experiment
STUDY_SUMMARY
1H NMR-based metabolomic approach to measure the concentrations of metabolites in plasma and liver of four different diet groups: HFD, HCD, HFHCD and control. Multivariate statistical analyses were used to determine significantly changed metabolites between all group-pairs.
Metabolomic profiles along the gastrointestinal tract of the healthy dog
STUDY_SUMMARY
Introduction: The fecal microbiome is relevant to the health and disease of many species. The importance of the fecal metabolome has more recently been appreciated, but our knowledge of the microbiome and metabolome at other sites along the gastrointestinal tract remains deficient. Objective: To analyze the gastrointestinal microbiome and metabolome of healthy domestic dogs at four anatomical sites. Methods: Samples of the duodenal, ileal, colonic, and rectal contents were collected from six adult dogs after humane euthanasia for an unrelated study. The microbiota were characterized using Illumina sequencing of 16S rRNA genes. The metabolome was characterized by mass spectrometry-based methods. Results: Prevalent phyla throughout the samples were Proteobacteria, Firmicutes, Fusobacteria, and Bacteroidetes, consistent with previous findings in dogs and other species. A total of 530 unique metabolites were detected; 199 of these were identified as previously named compounds, but 141 of them had at least one significantly different site-pair comparison. Noteworthy examples include amino acids, which decreased from the small to large intestine; pyruvate, which was at peak concentrations in the ileum; and several phenol-containing carboxylic acid compounds that increased in the large intestine. Conclusion: The microbiome and metabolome vary significantly at different sites along the canine gastrointestinal tract.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Distinct signatures of dental plaque metabolic byproducts dictated by periodontal inflammatory status
STUDY_SUMMARY
Onset of chronic periodontitis is associated with an aberrant polymicrobial community, termed dysbiosis. Findings of a recent model of its etiology suggested that dysbiosis holds a conserved metabolic signature as an emergent property. The purpose of this study was to identify robust biomarkers for periodontal inflammation severity. Furthermore, we investigated disease-associated metabolic signatures of periodontal microbiota using a salivary metabolomics approach. Collection of whole saliva samples was performed before and after removal of supragingival plaque (debridement). Periodontal inflamed surface area (PISA) was employed as an indicator of periodontal inflammatory status. Based on multivariate analyses using pre-debridement salivary metabolomics data, we found that the metabolites associated with higher PISA included cadaverine and hydrocinnamate, while uric acid and ethanolamine were associated with lower PISA. Next, we focused on dental plaque metabolic byproducts by selecting significantly decreased salivary metabolites following debridement. Metabolite set enrichment analysis revealed that polyamine metabolism, arginine and proline metabolism, butyric acid metabolism, and lysine degradation were distinctive metabolic signatures of dental plaque in the high PISA group, which may have relevance to the metabolic signatures of disease-associated communities. Collectively, our findings identified potential biomarkers of periodontal inflammatory status, while they also provide insight into metabolic signatures of dysbiotic communities.
M-CMV-infected N. tabacum plants with six symptoms (vein clearing, mosaic, chlorosis, partial green recovery, complete green recovery and secondary mosaic) were analyzed by LC-MS & GC-MS. In addition, the pathogenesis biomarker might be found by this untargeted global metabolomic analysis.
The goal of this project is to analyze the metabolic pool distribution of cerebral metabolites in response to cocaine administration and withdrawal as compared to control
Weight loss and weight maintenance obtained with or without GLP-1 analogue treatment decrease branched chain amino acid levels.
STUDY_TYPE
RCT
STUDY_SUMMARY
RCT of the effect og liraglutide on weight maintenance. 58 individuals wer included and subjected to low calorie intake during 8 weeks. Individuals lost a mean of 12 kg during that period. Individuals were then randomized to receive 1.2 mg liraglutide daily for 1 year or serving as control. Metabolites were measured at inclusion (-8 weeks), randomization (0 weeks) and at 4 and 52 weeks during weight maintenance.
INSTITUTE
NNF Center for Metabolic Research
LAST_NAME
Engelbrechtsen
FIRST_NAME
Line
ADDRESS
Section of Metabolic Genetics, Universitetsparken 1, DIKU building 1st floor, 2100 Copenhagen
Understanding the response to endurance exercise using a systems biology approach: combining blood metabolomics, transcriptomics and miRNome in horses
STUDY_SUMMARY
Endurance exercise in horses implies adaptive processes involving affective, physiological, biochemical, and cognitive-behavioral response in an attempt to regain homeostasis. We hypothesized that the identification of the relationships between blood metabolome, transcriptome and miRNome during endurance exercise could provide significant insights into the molecular response to intense exercise or prediction of this response at basal status. In this perspective, the serum metabolome and whole-blood transcriptome and miRNome data were obtained from 10 horses before and after a 160 km endurance competition. Results: We obtained a global regulatory network based on 11 unique metabolites, 263 metabolic genes and 5 miRNAs whose expression was significantly altered at T1 (post- endurance competition) relative to T0 (baseline, pre- endurance competition). This network provided new insights into the cross talk between the distinct molecular pathways (e.g. energy and oxygen sensing, oxidative stress, and inflammation) that were not detectable when analyzing single metabolites or transcripts alone. This suggested that single metabolites and transcripts were carrying out multiple roles and thus sharing several biochemical pathways. Using a regulatory impact factor metric analysis, this regulatory network was further confirmed at the transcription factor and miRNA levels. In an extended cohort of 39 animals, multiple factor analysis confirmed the strong associations between lactate, methylene derivatives, miR-21-5p, miR-16-5p, and genes that coded proteins involved in metabolic reactions primarily related to energy, ubiquitin proteasome and lipopolysaccharide immune responses at T1. Multiple factorial analyses also identified potential biomarkers at T0 for an increased possibility of failure to finish an endurance competition.
In this study, two independent large cohorts of mature dates exhibiting substantial diversity in origin, varieties and fruit processing conditions were measured by metabolomics techniques in order to identify major determinants of the fruit metabotype. Additional samples reflecting different stages of date fruit ripening process has been included for 10 different fruit varieties.
Metabolic changes to maternal rat liver tissue during and post-pregnancy
STUDY_SUMMARY
Assessing metabolic changes to maternal rat liver tissue during and post-pregnancy. Liver tissue was harvested post-mortem from rats or mice without pregnancy (Nulliparous - NP), time course during pregnancy in days (P2-4, P11-13m P18-20), lactation day 10 (LD10), in during involution of the liver (I2, I4, I6, I8, I10) and 4-weeks regression after preganancy (R4).
Assessing metabolic changes to maternal rat liver tissue during and post-pregnancy (part II)
STUDY_SUMMARY
Assessing metabolic changes to maternal rat liver tissue during and post-pregnancy. Liver tissue was harvested post-mortem from rats or mice without pregnancy (Nulliparous - NP), time course during pregnancy in days (P2-4, P11-13m P18-20), lactation day 10 (LD10), in during involution of the liver (I2, I4, I6, I8, I10) and 4-weeks regression after preganancy (R4).
Determine how inhibition of autophagy/proteasome degradation or inhibition of protein synthesis in models of muscle insulin resistance affect amino acid metabolites
STUDY_SUMMARY
To determine which protein degradation pathways downstream of IR and IGF1R contribute to changes in amino acid and mitochondrial metabolite pools, we will treat control, M-IR-/-, MIGIRKO, and HFD obese mice with inhibitors of autophagy or proteasome. We will treat 5 animals each of control, M-IR-/-, MIGIRKO, and HFD mice with vehicle, colchicine to inhibit autophagy, or MG132 to inhibit proteasome activity, then measure amino acid in muscle tissue.
Investigating TCA concentrations in mice muscle tissue lacking insulin receptors and IGF-1 receptors
STUDY_SUMMARY
Quantitative measures of TCA cycle metabolites from control, M-IR-/-, M-IGF1R-/- , and MIGIRKO mice. Also compare mice on a chow diet to mice on a high fat diet (HFD).
Inhibition of autophagy/proteasome degradation or inhibition of protein synthesis in models of muscle insulin resistance affect TCA cycle
STUDY_SUMMARY
To determine which protein degradation pathways downstream of IR and IGF1R contribute to changes in amino acid and mitochondrial metabolite pools, we will treat control, M-IR-/-, MIGIRKO, and HFD obese mice with inhibitors of autophagy or proteasome. We will treat 5 animals each of control, M-IR-/-, MIGIRKO, and HFD mice with vehicle, colchicine to inhibit autophagy, or MG132 to inhibit proteasome activity, then measure TCA cycle in muscle tissue.
Inhibition of autophagy/proteasome degradation or inhibition of protein synthesis in models of muscle insulin resistance affect amino acids metabolites in serum
STUDY_SUMMARY
To determine which protein degradation pathways downstream of IR and IGF1R contribute to changes in amino acid and mitochondrial metabolite pools, we will treat control, M-IR-/-, MIGIRKO, and HFD obese mice with inhibitors of autophagy or proteasome. We will treat 5 animals each of control, M-IR-/-, MIGIRKO, and HFD mice with vehicle, colchicine to inhibit autophagy, or MG132 to inhibit proteasome activity, then measure amino acid metabolites in serum.
Metabolomics of longitudinal plasma samples from Macaca mulatta infected with Plasmodium cynomolgi B strain.
STUDY_TYPE
Longitudinal parasite infection and treatment of multiple individuals
STUDY_SUMMARY
Malaria-naive male rhesus macaques (Macaca mulatta), approximately three years of age, were inoculated intravenously with salivary gland sporozoites isolated at the Centers for Disease Control and Prevention from multiple Anopheles species (An. dirus, An. gambiae, and An. stephensi) and then profiled for parasitological, clinical, immunological, functional genomic, lipidomic, proteomic, and metabolomic measurements. The experiment was designed for 100 days, and pre- and post-100 day periods to prepare subjects and administer curative treatments respectively. The anti-malarial drug Artemether was subcuratively administered selectively to several subjects during the primary parasitemia to suppress clinical complications and to all animals for curative treatment of blood-stage infections to allow detection of relapses. One subject was euthanized during the 100-day experimental period due to clinical complications. The anti-malarial drugs Primaquine and Chloroquine were administered to all remaining subjects at the end of the study for curative treatment of the liver and blood-stage infections, respectively. Plasma samples were acquired every other day from capillary blood and during seven-time point collections of venous blood. Supplemental files, including a ReadMe with additional experimental details, all raw data, and analytical metadata, are provided as part of this submission.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine, Vaccine Center at Yerkes
LAST_NAME
Karpuzoglu
FIRST_NAME
Ebru
ADDRESS
Emory University, 954 Gatewood Rd, Room 208, Atlanta, GA 30329
EMAIL
mahpic@emory.edu
PHONE
N/A
TOTAL_SUBJECTS
5
STUDY_COMMENTS
Malaria Host Pathogen Interaction Center Experiment 04, 5 subjects 286 samples, "The experimental design and protocols for this study were approved by the Emory University Institutional Animal Care and Use Committee (IACUC). These results are a product of a consortium of researchers known as the Malaria Host Pathogen Interaction Center (MaHPIC). For more information on the MaHPIC, please visit http://www.systemsbiology.emory.edu/ . Within the MaHPIC, these data were collected as part of 'Experiment 04' (E04). To access other publicly available results from E04 and other MaHPIC Experiments, including clinical results (specifics on drugs administered, diet, and veterinary interventions), and other omics, visit http://plasmodb.org/plasmo/mahpic.jsp . This page will be updated as datasets are released to the public."
Measuring amino acid metabolites in insulin resistant and insulin deficient mouse tissue models
STUDY_SUMMARY
To compare models of insulin resistance to a model of loss of insulin signaling, we will also determine amino acid metabolites from muscle, using control and streptozotocin (STZ) treated mice as a model of insulin deficient diabetes.
Measuring acylcarnitine concentrations in insulin resistant and insulin deficient mouse tissue models
STUDY_SUMMARY
To compare models of insulin resistance to a model of loss of insulin signaling, we will also determine muscle acylcarnitine concentrations, using control and streptozotocin (STZ) treated mice as a model of insulin deficient diabetes. Changes in metabolite profiles will be correlated with activation of mTOR and FoxO pathways in muscle.
Measuring ceramide concentrations in insulin resistant and insulin deficient mouse tissue models
STUDY_SUMMARY
To compare models of insulin resistance to a model of loss of insulin signaling, we will also determine ceramide concentration in muscle, using control and streptozotocin (STZ) treated mice as a model of insulin deficient diabetes.
Investigating ceremide concentrations in mice muscle tissue lacking insulin receptors and IGF-1 receptors
STUDY_SUMMARY
Quantitative measures of ceramide metabolite concentrations in muscle from control, M-IR-/-, M-IGF1R-/- , and MIGIRKO mice. Also compare mice on a chow diet to mice on a high fat diet (HFD).
Measuring TCA cycle concentrations in insulin resistant and insulin deficient mouse tissue models
STUDY_SUMMARY
To compare models of insulin resistance to a model of loss of insulin signaling, we will also determine TCA cycle metabolites in muscle, using control and streptozotocin (STZ) treated mice as a model of insulin deficient diabetes.
Measuring NEFA concentrations in insulin resistant and insulin deficient mouse tissue models
STUDY_SUMMARY
To compare models of insulin resistance to a model of loss of insulin signaling, we will also determine nonesterified fatty acid metabolites in muscle, using control and streptozotocin (STZ) treated mice as a model of insulin deficient diabetes.
Plasma metabolomics profiling for fish maturation in blunt snout bream
STUDY_TYPE
Metabolomics experiment
STUDY_SUMMARY
We investigated the comprehensive metabolic profiles of plasma among immature females, mature females ready to spawn, as well as already spawned breeders of blunt snout bream (Megalobrama amblycephala). The purpose of this study was to screen out potential biomarkers for sexual mature female M. amblycephala compared to immature female individuals and already spawned breeders. The three groups were set up in this study, including one year old immature females, 2 years old sexually mature females ready to spawning and successfully spawning females of M. amblycephala. The plasma samples were collected to investigate the comprehensive metabolic profiles through UPLC-MS/MS based metabolomics analysis method. According to multivariate and univariate statistical analysis, plasma metabolite profiles of three groups were obviously separated, and the plasma metabolite profiles of immature female M. amblycephala were much more different from mature females ready to spawn as well as already spawned breeders. The differential plasma metabolites from three hormone related pathways including GnRH signaling pathway, steroid hormone biosynthesis and steroid biosynthesis, were further analyzed. A total of 29 metabolites were identified as differential biomarkers associated with the female maturation status
INSTITUTE
Huazhong Agricultural University, Wuhan
DEPARTMENT
College of Fisheries
LABORATORY
Key Lab of Freshwater Animal Breeding, Ministry of Agriculture
Prenatal exposures and Metabolite and Lipid Profiles
STUDY_SUMMARY
Our goal is to examine relationships between prenatal exposures (to metals, BPA, and phthalates) and metabolite and lipid profiles using untargeted approaches in mother-infant pairs from the Michigan Mother Infant Pairs (MMIP) cohort. Three plasma samples will be analyzed from each family: two maternal samples (from first trimester and time of admission for labor and delivery) and one child sample (from cord blood). We are interested in differences/correlations between family samples over time, and also the extent to which maternal exposures impact the lipidomes and metabolomes of both family members.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effects of Curcumin Supplementation on the Amino Acid Concentration of Older Adults: Relation to Vascular Function
STUDY_SUMMARY
Perform amino acid concentrations metabolite analysis related to nitric oxide biology, oxidative stress and inflammation in plasma before and after 12 weeks of oral curcumin (2000 mg/d) or placebo (double-blind, randomized) in men and women aged 45-79 years who are free from clinical cardiovascular disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Seals
FIRST_NAME
Douglas
ADDRESS
Department of Integrative Physiology University of Colorado Boulder, CO 80309
Effects of Curcumin Supplementation on the NonāEsterified Fatty Acids concentration of Older Adults: Relation to Vascular Function
STUDY_SUMMARY
Perform non-esterified fatty acid concentrations metabolite analysis related to nitric oxide biology, oxidative stress and inflammation in plasma before and after 12 weeks of oral curcumin (2000 mg/d) or placebo (double-blind, randomized) in men and women aged 45-79 years who are free from clinical cardiovascular disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Seals
FIRST_NAME
Douglas
ADDRESS
Department of Integrative Physiology University of Colorado Boulder, CO 80309
Effects of Curcumin Supplementation on the Ceramides Concentration of Older Adults: Relation to Vascular Function
STUDY_SUMMARY
Perform ceramides concentrations metabolite analysis related to nitric oxide biology, oxidative stress and inflammation in plasma before and after 12 weeks of oral curcumin (2000 mg/d) or placebo (double-blind, randomized) in men and women aged 45-79 years who are free from clinical cardiovascular disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Seals
FIRST_NAME
Douglas
ADDRESS
Department of Integrative Physiology University of Colorado Boulder, CO 80309
Effects of Curcumin Supplementation on the Acylcarnitine Concentration of Older Adults: Relation to Vascular Function
STUDY_SUMMARY
Perform acylcarnitine concentrations metabolite analysis related to nitric oxide biology, oxidative stress and inflammation in plasma before and after 12 weeks of oral curcumin (2000 mg/d) or placebo (double-blind, randomized) in men and women aged 45-79 years who are free from clinical cardiovascular disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Seals
FIRST_NAME
Douglas
ADDRESS
Department of Integrative Physiology University of Colorado Boulder, CO 80309
Our aim is to identify the LSR-driven metabolomics profile of breast cancer cells in lean and obesogenic environments. Breast cancer cell models with high or undetectable levels of LSR, including drug resistance models, were cultured in lean and obesogenic environments and comprehensive metabolomics profiling, including lipidomics-focused sub-analyses were performed. The metabolomics analyses using both approaches will help us determine if LSR enhances aggressive breast cancer phenotypes via modulation of cellular bioenergetic metabolism, ultimately contributing to poor patient outcome.
Broad spectrum, reverse phase LCMS metabolomics (Negative ion mode)
STUDY_SUMMARY
Our aim is to identify the LSR-driven metabolomics profile of breast cancer cells in lean and obesogenic environments. Breast cancer cell models with high or undetectable levels of LSR, including drug resistance models, were cultured in lean and obesogenic environments and comprehensive metabolomics profiling, including lipidomics-focused sub-analyses were performed. The metabolomics analyses using both approaches will help us determine if LSR enhances aggressive breast cancer phenotypes via modulation of cellular bioenergetic metabolism, ultimately contributing to poor patient outcome.
Effects of herb DG and KK01 on Type 2 Diabetes Mellitus (T2DM) through Lipidomics
STUDY_TYPE
LC-MS lipidomics
STUDY_SUMMARY
According to the results in animal test, KK01 is effective in controlling blood glucose increase with comparable effect as metformin and rosiglitazone. This study will conduct lipid profile comparison for serum samples generated from the animal tests. The comparison will be based on the following groups: 1) db/db mice + DG-high dose; 2) db/db mice +DG-low dose; 3) db/db mice + KK01-high dose; 4) db/db mice + KK01-low dose; 5) db/db mice + metformin; 6) db/db mice + rosiglitazone; 7) db/db mice + saline (disease model); and 8) wild type mice + saline (healthy model). The determined lipid marker(s) will be applied to elucidate the drug target(s) and mechanisms of DG and KK01. Furthermore, comparison of target(s) between KK01 and the first line drugs in diabetic treatment, e.g., metformin and rosiglitazone, will facilitate the finding of featured pathway(s) of KK01 differentiated from the established drugs. Comparison of drug target(s) between KK01 and DG can help to understand the synergistic effects of multiple constituents in the herb.
CHEAR Pooled Plasma Reference Material Proficiency Test
STUDY_TYPE
Cytokine Analysis
STUDY_SUMMARY
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed using the cytokine array workflow of the Eastern Regional Metabolomics Resource Core (protocols based on Ray Biotech's manufacturer recommendations). Six replicate aliquots of the pooled reference sample were anlyzed on 6 arrays.
INSTITUTE
RTI International
DEPARTMENT
Analytical Chemistry & Pharmaceuticals
LABORATORY
CHEAR CC - UARC
LAST_NAME
Fennell
FIRST_NAME
Timothy
ADDRESS
3040 E. Cornwallis Road, Research Triangle Park, NC 27709
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed using the GC-TOF MS workflow of the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Seven samples were injected of the sample reference material prepared in replicate.
CHEAR Plasma Dilution Reference Material Proficiency Test NMR (Part I)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed using the NMR metabolomics workflow of the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Data of six samples were acquired of the sample reference material prepared in replicate.
INSTITUTE
RTI International
DEPARTMENT
Discovery-Sciences-Technology (DST)
LABORATORY
RTI CHEAR ANALYTICAL HUB/UARC
LAST_NAME
Fennell
FIRST_NAME
Timothy
ADDRESS
3040 E Cornwallis Road, Research Triangle Park, NC 27709
CHEAR Plasma Extraction Reference Material Proficiency Test NMR (Part I)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed using the NMR metabolomics workflow of the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Data of six samples were acquired of the sample reference material prepared in replicate.
CHEAR Urine Reference Material Proficiency Test NMR (Part I)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Urine was provided by Emory University. The material was prepared and analyzed using the NMR metabolomics workflow of the Eastern Regional Metabolomics Resource Core. Data of six samples were acquired of the sample reference material prepared in replicate.
CHEAR Plasma Dilution Reference Material Proficiency Test NMR (Part II)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed using the NMR metabolomics workflow of the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Data of six samples were acquired of the sample reference material prepared in replicate.
CHEAR Plasma Extraction Reference Material Proficiency Test NMR (Part II)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed using the NMR metabolomics workflow of the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Data of six samples were acquired of the sample reference material prepared in replicate.
CHEAR Urine Reference Material Proficiency Test NMR (Part II)
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Urine was provided by Emory University. The material was prepared and analyzed using the NMR metabolomics workflow of the Eastern Regional Metabolomics Resource Core. Data of six samples were acquired of the sample reference material prepared in replicate.
Kidney tissue metabolomic profiling of diabetic nephropathy in the steptozotocin induced type-1 diabetes mouse model.
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
This metabolomics study evaluated kidney tissue from wild-type and meprin Ī² knockout mice after induction of diabetes with streptozotocin or treatment with sodium citrate control to understand how these factors influence the metabotype.
INSTITUTE
RTI International
DEPARTMENT
Discovery-Science-Technology
LABORATORY
NIH Eastern Regional Comphrehensive Metabolomics Resource Core at UNC Chapel Hill (ERCMRC)
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E Cornwallis Road, Research Triangle Park, NC 27709
Canine Diabetes - Preliminary Evaluation of Testing Methods
STUDY_TYPE
Single time point blood collection
STUDY_SUMMARY
Blood was collected from diabetic dogs and healthy control dogs. Diabetic dogs and control dogs were breed matched where possible. Timing of blood collection after a meal was matched as best as possible. Serum was frozen at -80C.
Triple Quadrupole Mass Spectrometer to measure low abundance isotope enrichment in individual muscle proteins
STUDY_TYPE
isotope encrichment and comparison of mass spectrometer platforms, timecourse
STUDY_SUMMARY
Stable isotope-labeled amino acids have long been used to measure the fractional synthesis rate of proteins, although the mass spectrometry platforms used for such analyses have changed throughout the years. More recently, tandem mass spectrometers such as triple quadrupoles have been accepted as the standard platform for enrichment measurement due to their sensitivity and the enhanced specificity offered by multiple reaction monitoring (MRM) experiments. The limit in the utility of such platforms for enrichment analysis occurs when measuring very low levels of enrichment from small amounts of sample, particularly proteins isolated from two-dimensional gel electrophoresis (2D-GE), where interference from contaminant ions impact the sensitivity of the measurement. We therefore applied a high resolution orbitrap mass spectrometer to the analysis of [ring-13C6]-phenylalanine enrichment in individual muscle proteins isolated with 2D-GE. Comparison of samples analyzed on both platforms revealed that the high resolution MS has significantly improved sensitivity relative to the triple quadrupole MS at very low-level enrichments due to its ability to resolve interferences in the m/z dimension.
High Resolution orbittrap Mass Spectrometer to measure low abundance isotope enrichment in individual muscle proteins
STUDY_TYPE
isotope encrichment and comparison of mass spectrometer platforms
STUDY_SUMMARY
Stable isotope-labeled amino acids have long been used to measure the fractional synthesis rate of proteins, although the mass spectrometry platforms used for such analyses have changed throughout the years. More recently, tandem mass spectrometers such as triple quadrupoles have been accepted as the standard platform for enrichment measurement due to their sensitivity and the enhanced specificity offered by multiple reaction monitoring (MRM) experiments. The limit in the utility of such platforms for enrichment analysis occurs when measuring very low levels of enrichment from small amounts of sample, particularly proteins isolated from two-dimensional gel electrophoresis (2D-GE), where interference from contaminant ions impact the sensitivity of the measurement. We therefore applied a high resolution orbitrap mass spectrometer to the analysis of [ring-13C6]-phenylalanine enrichment in individual muscle proteins isolated with 2D-GE. Comparison of samples analyzed on both platforms revealed that the high resolution MS has significantly improved sensitivity relative to the triple quadrupole MS at very low-level enrichments due to its ability to resolve interferences in the m/z dimension.
CHEAR Urine Reference Material Proficiency Test Biocrates
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Urine. The material was prepared and analyzed by way of LC-MS and Biocrates workflow employed by the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Six samples were injected of the sample reference material prepared in replicate.
CHEAR Plasma Reference Material Proficiency Test Biocrates
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed by way of LC-MS and Biocrates workflow employed by the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Six samples were injected of the sample reference material prepared in replicate.
Multi-omics based identification of specific biochemical changes associated with PfKelch13-mutant artemisinin resistant Plasmodium
STUDY_TYPE
Cell type comparison
STUDY_SUMMARY
Two clonaly artemisinin resistant parasitised red blood cells (trophozoite stage) were compared with artemsinin sensitive parasitised red blood cells by metabolomics analysis.
INSTITUTE
Monash Institute of Pharmaceutical Sciences, Monash University
DEPARTMENT
Drug Delivery, Disposition and Dynamics
LABORATORY
Creek lab
LAST_NAME
Creek
FIRST_NAME
Darren
ADDRESS
381 Royal Parade, Parkville, Melbourne, VIC3052, Australia
Inbred mice are used to investigate many aspects of human physiology, including susceptibility to disease and response to therapies. Despite increasing evidence that the composition and function of the murine intestinal microbiota can substantially influence a broad range of experimental outcomes, relatively little is known about microbiome dynamics within experimental mouse populations. We investigated changes in the intestinal microbiome between C57BL/6J mice spanning six generations (assessed at generations 1, 2, 3 and 6), following their introduction to a stringently controlled facility. Faecal microbiota composition and function were assessed by 16S rRNA gene amplicon sequencing and liquid chromatography mass spectrometry, respectively. Significant divergence of the intestinal microbiota between founder and second generation mice, as well as continuing inter-generational variance, was observed. Bacterial taxa whose relative abundance changed significantly included Akkermansia, Turicibacter and Bifidobacterium (p< 0.05), all of which are recognised as having the potential to substantially influence host physiology. Shifts in microbiota composition were mirrored by corresponding differences in the faecal metabolome (r=0.57, p=0.0001), with notable differences in levels of tryptophan pathway metabolites and amino acids, including glutamine, glutamate and aspartate. The magnitude of these changes in the intestinal microbiota and metabolome characteristics during acclimation were on a scale with those observed between populations housed in separate facilities, which differed in regards to husbandry, barrier conditions and dietary intake. The microbiome variance reported here has major implications for experimental reproducibility, and as a consequence, experimental design and the interpretation of research outcomes across as wide range of contexts.
INSTITUTE
South Australian Health and Medical Research Institute
DEPARTMENT
Infection and Immunity Theme
LAST_NAME
Rogers
FIRST_NAME
Geraint
ADDRESS
SAHMRI, North Terrace, Adelaide, SA 5000, Australia
Replication study: The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate
STUDY_SUMMARY
The Reproducibility Project: Cancer Biology seeks to address growing concerns about reproducibility in scientific research by conducting replications of selected experiments from a number of high-profile papers in the field of cancer biology. The papers, which were published between 2010 and 2012, were selected on the basis of citations and Altmetric scores (Errington et al., 2014). This Registered Report describes the proposed replication plan of key experiments from āThe common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarateā by Ward and colleagues, published in Cancer Cell in 2010 (Ward et al., 2010). The experiments that will be replicated are those reported in Figures 2, 3 and 5. Ward and colleagues demonstrate the mutations in isocitrate dehydrogenase 2 (IDH2), commonly found in acute myeloid leukemia (AML), abrogate the enzymeās wild-type activity and confer to the mutant neomorphic activity that produces the oncometabolite 2-hydroxyglutarate (2-HG) (Figures 2 and 3). They then show that elevated levels of 2-HG are correlated with mutations in IDH1 and IDH2in AML patient samples (Figure 5). The Reproducibility Project: Cancer Biology is a collaboration between the Center for Open Science and Science Exchange and the results of the replications will be published by eLife.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
Health Sciences Drive, Davis, California, 95616, USA
Investigating large scale metabolomics in mice serum lacking insulin receptors and IGF-1 receptors
STUDY_SUMMARY
Large scale metabolomics from control, M-IR-/-, M-IGF1R-/- , and MIGIRKO mice serum. Also compare mice on a chow diet to mice on a high fat diet (HFD).
Metabolomics marker of brown adipose tissue in men
STUDY_SUMMARY
Objective: We aimed to identify metabolites in serum that are associated with BAT volume and activity in men. Methods: We assessed 163 metabolites in fasted serum of a cohort of twenty two healthy lean men (age 24.1 (21.7 ā 26.6) years, BMI 22.1 (20.5 ā 23.4) kg/m2) who subsequently underwent a cold-induced [18F]FDG PET-CT scan to assess BAT volume and activity. In addition, we included three replication cohorts consisting of in total thirty-seven healthy lean men that were similar with respect to age and BMI compared to the discovery cohort.
Investigating large scale metabolomics in mice tissue lacking insulin receptors and IGF-1 receptors
STUDY_SUMMARY
Large scale metabolomics from control, M-IR-/-, M-IGF1R-/- , and MIGIRKO mice tissue. Also compare mice on a chow diet to mice on a high fat diet (HFD).
The goal of this project is to analyze the metabolic pool distribution of cerebral metabolites in response to cocaine administration and withdrawal as compared to control
IROA feasibility project; plasticizers as obesogens in zebrafish
STUDY_TYPE
(1) IROA label (2) Zebrafish exposed to DEHP
STUDY_SUMMARY
Zebrafish were fed IROA labelled nematodes (smaple 1-4); In a second experiment, zebrafish larvae were exposed to DEHP, a chemical that is a suspected obesogen.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Martyniuk
FIRST_NAME
Chris
ADDRESS
2187 Mowry Rd. Bldg 471
EMAIL
cmartyn@ufl.edu
PHONE
352-294-4636
NUM_GROUPS
2
TOTAL_SUBJECTS
20
STUDY_COMMENTS
Expt 2: Two groups (control) + DEHP-treated (n=10 larvae pools or biological replicates)
Plasma metabolomic profiling of diabetic nephropathy in the steptozotocin induced type-1 diabetes mouse model
STUDY_SUMMARY
This metabolomics study evaluated plasma from wild-type and meprin Ī² knockout mice after induction of diabetes with streptozotocin or treatment with sodium citrate control to understand how these factors influence the metabotype.
INSTITUTE
RTI International
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E. Cornwallis Road, Research Triangle Park, NC 27709
Urine metabolomic profiling of diabetic nephropathy in the steptozotocin induced type-1 diabetes mouse model.
STUDY_SUMMARY
This metabolomics study evaluated urine from wild-type and meprin Ī² knockout mice after induction of diabetes with streptozotocin or treatment with sodium citrate control to understand how these factors influence the metabotype.
INSTITUTE
RTI International
LABORATORY
NIH Eastern Regional Comphrehensive Metabolomics Resource Core at UNC Chapel Hill (ERCMRC)
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E. Cornwallis Road, Research Triangle Park, NC 27709
Metabolomics of immunoglobulin-producing cells in IgA nephropathy
STUDY_SUMMARY
IgA nephropathy (IgAN), the most common primary glomerulonephritis, is characterized by deposits of IgA-containing immune complexes in the kidney glomeruli, as first described by Berger and Hinglais in 1968. IgAN is a major cause of end-stage renal disease with its associated cardio-renal morbidity and mortality. Analyses of the IgA deposits revealed that the IgA is exclusively of the IgA1 subclass and that this IgA1 is aberrantly glycosylated, deficient in galactose in some O-glycans (Gd-IgA1). Patients with IgAN have elevated serum levels of Gd-IgA1 bound by anti-glycan autoantibodies in circulating immune complexes (CIC) that are fundamental in driving disease pathology in an autoimmune process. We have recently shown that elevated serum levels of Gd-IgA1 in patients with IgAN predict disease progression. Thus, understanding the mechanisms behind Gd-IgA1 production will improve future treatment options, as there is presently no disease-specific therapy. A total of 24 cell pellets (4 replicates from 6 cell lines) were analyzed by LCMS metabolomics. Immortalized immunoglobulin-producing cell lines were generated from peripheral-blood lymphocytes from patients with IgAN and healthy controls as described in Suzuki, H., Moldoveanu, Z., Hall, S., et al. IgA1-secreting cell lines from patients with IgA nephropathy produce aberrantly glycosylated IgA1. J Clin Invest. 2008, 118, 629-639.
INSTITUTE
RTI International
LABORATORY
NIH Eastern Regional Comphrehensive Metabolomics Resource Core at UNC Chapel Hill (ERCMRC)
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E. Cornwallis Road, Research Triangle Park, NC 27709
In this study the F8 population of 100 Recombinant Inbred Lines (RILs) obtained from a cross between Solanum lycopersicum X Solanum pimpinellifolium were intelligently allocated to two sub-populations optimized for the distribution of parental alleles using the R-procedure DesignGG (Li et al., 2009; Joosen et al., 2013); hence 50 RIL lines were used for dry seeds and 50 lines for 6h imbibed seeds
INSTITUTE
Wageningen UR
DEPARTMENT
Plant Physiology
LABORATORY
Wageningen Seed Lab, Lab
LAST_NAME
Ligterink
FIRST_NAME
Wilco
ADDRESS
Droevendaalsesteeg 1, NL-6708 PB Wageningen, The Netherlands
Maize plants were grown under three different temperature regimes: 1) normal day / normal night; 2) hot day / normal night; 3) hot day / hot night. Kernels from developing ears were taken 14, 16, 18, 22, 26 and 40 days after pollination.
Association of hemodialysis patient plasma trace metals with response to erythropoiesis stimulating agents
STUDY_TYPE
Metallomics
STUDY_SUMMARY
EDTA-Plasma from 110 hemodialysis patients participating in an NIDDK funded study were analyzed by ICP-MS for the concentration of As, Cd, Co, Cr, Cu, Mn, Mo, Ni, Pb, Sb, Se, Sn, V, and Zn. Associations were determined between trace metals and gender, race, hemodialysis status, hemoglobin at the time of draw (Hgb), total ESA dose for the month the sample was collected (EPO), and erythropoietin resistance index determined over the 6 months of treatment leading up to sample collection (ERI)
INSTITUTE
RTI International
LABORATORY
NIH Eastern Regional Comprehensive Metabolomics Resource Core at UNC Chapel Hill (ERCMRC)
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E. Cornwallis Road, Research Triangle Park, NC 27709, USA
Large Scale C18 Profiling of the Effects of Curcumin Supplementation of Older Adults: Relation to Vascular Function
STUDY_SUMMARY
Perform large scale profiling C18 metabolite analysis related to nitric oxide biology, oxidative stress and inflammation in plasma before and after 12 weeks of oral curcumin (2000 mg/d) or placebo (double-blind, randomized) in men and women aged 45-79 years who are free from clinical cardiovascular disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Seals
FIRST_NAME
Douglas
ADDRESS
Department of Integrative Physiology University of Colorado Boulder, CO 80309
Large Scale HILIC Profiling of the Effects of Curcumin Supplementation of Older Adults: Relation to Vascular Function
STUDY_SUMMARY
Perform large scale profiling HILIC metabolite analysis related to nitric oxide biology, oxidative stress and inflammation in plasma before and after 12 weeks of oral curcumin (2000 mg/d) or placebo (double-blind, randomized) in men and women aged 45-79 years who are free from clinical cardiovascular disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Seals
FIRST_NAME
Douglas
ADDRESS
Department of Integrative Physiology University of Colorado Boulder, CO 80309
Metabolomic study on a schizophrenia and type 2 diabetes susceptibility gene
STUDY_SUMMARY
a comprehensive serum metabolomic analysis in healthy subjects with different genotypes of rs12742393 (n=49 for AA, AC, and CC, respectively) using gas chromatographyātime-of-flight mass spectrometry and ultra-performance liquid chromatography quadruple time-of-flight mass spectrometry.
INSTITUTE
Shanghai Jiao Tong University Affiliated Sixth Peopleās Hospital
Metabolome analysis of the cecal contents of GF mice and GF mice colonized with dominant gut microbes present in the ceca of neonatal and adult mice
STUDY_SUMMARY
Metabolome profiles of GF or GF mice reconstituted with Esherichia coli (EC), Bacteroides acidifaciens (Bac), or Clostridia consortium (CL) were compared.
Exploratory research on first and second trimester urinary metabolic profiles and fetal growth restriction
STUDY_SUMMARY
From 2010 to 2012, women were recruited into The Infant Development and the Environment Study (TIDES) from obstetrical clinics affiliated with academic medical centers in four U.S. cities. Using previously collected first and second trimester urine samples from this prospective cohort of nearly 800 pregnancies, we designed a nested case control study aimed to determine whether maternal metabolic abnormalities/differences are associated with FGR. To find the patients with fetal growth restriction (FGR), we reviewed de-identified questionnaires and de-identified previously collected data, and performed a growth potential formula considering gestational age, infant gender, maternal and paternal height, and interaction of gestational age with maternal weight. Using a case:control ratio of 1:2, matched on study site, maternal age (Ā± 2 years), parity, and infant's sex, 53 cases were matched to 106 controls for a total of 159 patients, and 318 samples (one in each ttrimester). The samples were analyses by NMR spectroscopy, using Bruker IVDr platform. Identification of FGR cases and controls: FGR cases were determined using the AUDIPOG formula for the average predicted birthweight for an infant with specific characteristics of: maternal height, age, and prenatal weight, as well as infant sex, gestational age, and birth rank. AUDIPOG formula: avg_pred_bw = 10,228066774 - 0,646727171*GA + 0,0259713417*GAĀ² - 0,000291122*GAĀ³ - 0,045467351*sex + 0,0606013862*rank - 0,013592585*rankĀ² + 0,0009109473*rankĀ³ + 0,0003976103*MA + 0,0019992269*MH + 0,0169049061*MW - 0,000171266*MWĀ² + 5,8340462E-7*MWĀ³ The natural logarithm of this average predicted birthweight was used with the observed birthweight to determine a z-score and percentile for each infant. If an infant fell into the 10th percentile for his or her given characteristics, then they were considered as having FGR. Otherwise they were considered in the pool of controls.
INSTITUTE
Mayo Clinic
LAST_NAME
Vuckovic
FIRST_NAME
Ivan
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Effects of the Kinase Inhibitor Sorafenib on Serum Metabolism In Vivo using Non-targeted Metabolomics Analysis tissue).
STUDY_SUMMARY
The human kinome consists of ~500 kinases, including 150 proposed as therapeutic targets. progression, cell death, differentiation, and survival. It is not surprising, then, that new tyrosine kinase inhibitors (TKIs) developed to treat cancer also exhibit cardiotoxicity, including sorafenib. We hypothesized that sorafenib cardiotoxicity is related to its deleterious effects on specific cardiac metabolic pathways given the critical role of TKs in cardiac metabolism. FVB/N mice (10/group) were challenged with sorafenib or vehicle control daily for two weeks. Echocardiographic assessment of the heart identified systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Cardiac, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Compared to vehicle treated controls, sorafenib-treated hearts exhibited significant alterations in 11 metabolites, including markedly altered taurine/hypotaurine metabolism by pathway enrichment analysis (25-fold enrichment). These studies identify sorafenib-induced alterations in cardiac alanine and taurine/hypotaurine metabolic. Interventions to rescue or prevent sorafenib-related cardiotoxicity warrant consideration of therapies targeting the taurine/hypotaurine deficiency identified in the current study.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Effects of the Kinase Inhibitor Sorafenib on Muscle Metabolism In Vivo using Non-targeted Metabolomics Analysis tissue).
STUDY_SUMMARY
The human kinome consists of ~500 kinases, including 150 proposed as therapeutic targets. progression, cell death, differentiation, and survival. It is not surprising, then, that new tyrosine kinase inhibitors (TKIs) developed to treat cancer also exhibit cardiotoxicity, including sorafenib. We hypothesized that sorafenib cardiotoxicity is related to its deleterious effects on specific cardiac metabolic pathways given the critical role of TKs in cardiac metabolism. FVB/N mice (10/group) were challenged with sorafenib or vehicle control daily for two weeks. Echocardiographic assessment of the heart identified systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Cardiac, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Compared to vehicle treated controls, sorafenib-treated hearts exhibited significant alterations in 11 metabolites, including markedly altered taurine/hypotaurine metabolism by pathway enrichment analysis (25-fold enrichment). These studies identify sorafenib-induced alterations in cardiac alanine and taurine/hypotaurine metabolic. Interventions to rescue or prevent sorafenib-related cardiotoxicity warrant consideration of therapies targeting the taurine/hypotaurine deficiency identified in the current study.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Effects of the Kinase Inhibitor Sorafenib on Heart Metabolism In Vivo using Non-targeted Metabolomics Analysis tissue).
STUDY_SUMMARY
The human kinome consists of ~500 kinases, including 150 proposed as therapeutic targets. progression, cell death, differentiation, and survival. It is not surprising, then, that new tyrosine kinase inhibitors (TKIs) developed to treat cancer also exhibit cardiotoxicity, including sorafenib. We hypothesized that sorafenib cardiotoxicity is related to its deleterious effects on specific cardiac metabolic pathways given the critical role of TKs in cardiac metabolism. FVB/N mice (10/group) were challenged with sorafenib or vehicle control daily for two weeks. Echocardiographic assessment of the heart identified systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Cardiac, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Compared to vehicle treated controls, sorafenib-treated hearts exhibited significant alterations in 11 metabolites, including markedly altered taurine/hypotaurine metabolism by pathway enrichment analysis (25-fold enrichment). These studies identify sorafenib-induced alterations in cardiac alanine and taurine/hypotaurine metabolic. Interventions to rescue or prevent sorafenib-related cardiotoxicity warrant consideration of therapies targeting the taurine/hypotaurine deficiency identified in the current study.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Effects of the Kinase Inhibitor Sorafenib on Liver Metabolism In Vivo using Non-targeted Metabolomics Analysis tissue).
STUDY_SUMMARY
The human kinome consists of ~500 kinases, including 150 proposed as therapeutic targets. progression, cell death, differentiation, and survival. It is not surprising, then, that new tyrosine kinase inhibitors (TKIs) developed to treat cancer also exhibit cardiotoxicity, including sorafenib. We hypothesized that sorafenib cardiotoxicity is related to its deleterious effects on specific cardiac metabolic pathways given the critical role of TKs in cardiac metabolism. FVB/N mice (10/group) were challenged with sorafenib or vehicle control daily for two weeks. Echocardiographic assessment of the heart identified systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Cardiac, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Compared to vehicle treated controls, sorafenib-treated hearts exhibited significant alterations in 11 metabolites, including markedly altered taurine/hypotaurine metabolism by pathway enrichment analysis (25-fold enrichment). These studies identify sorafenib-induced alterations in cardiac alanine and taurine/hypotaurine metabolic. Interventions to rescue or prevent sorafenib-related cardiotoxicity warrant consideration of therapies targeting the taurine/hypotaurine deficiency identified in the current study.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Experiment HuA: Metabolomics of plasma samples from humans infected with Plasmodium vivax strain.
STUDY_TYPE
Longitudinal study and treatment of multiple individuals with Chloroquine
STUDY_SUMMARY
Patients with vivax malaria were enrolled in this study from June 2011 to December 2012 at the FundacĆ£o de Medicina Tropical Doutor Heitor Vieira Dourado (FMT-HVD), an infectious disease referral center located in Manaus, Western Brazilian Amazon. This study, which required a 42-day follow-up period, was approved by the FMT-HVD Institutional Review Board and the Brazilian National Ethics Committee (CONEP) (IRB approval #: CAAE: 12516713.8.0000.0005). All protocols and documentation were reviewed and sample shipments approved by the Emory IRB. Male and female patients were eligible for inclusion if aged 6 months to 60 years, bodyweight ?5 kg, presenting a blood parasite density from 250 to 100,000 parasites/microliter and axillary temperature ?37.5Ā°C or history of fever in the last 48 hours. Exclusion criteria were: use of antimalarials in the previous 30 days, refusal to be followed up for 42 days and any clinical complication. Patients received supervised treatment with 25 mg/kg of chloroquine (CQ) phosphate over a 3-day period (10 mg/kg on day 0 and 7.5 mg/kg on days 1 and 2). Primaquine (0.5 mg/kg per day for 7 days) was prescribed at the end of the 42-day follow-up period. Patients who vomited the first dose within 30 minutes after drug ingestion were re-treated with the same dose. Patients were evaluated on days 0, 1, 2, 3, 7, 14, 28 and 42 and, if they felt ill, at any time during the study period. Blood smear readings, complete blood counts, and diagnostic polymerase chain reaction (PCR) amplifications were performed at all time points. Three aliquots of 100 ĀµL of whole blood from the day of a recurrence were spotted onto filter paper for later analysis by high performance liquid chromatography (HPLC) to estimate the levels of CQ and desethylchloroquine (DCQ) as previously described. In this study, CQ-resistance with parasitological failure was defined as parasite recurrence in the presence of plasma concentrations of CQ and DSQ higher than 100 ng/mL and microsatellite analysis revealing the presence of the same clonal nature at diagnosis and recurrence. The CQ-sensitive control group consisted of patients with no parasitemia recurring during follow-up period. A group of 20 healthy individuals from Brazil was used as non-malarial control group. Within the MaHPIC, this project is known as āExperiment HuAā. This dataset was produced by Dean Jones at Emory University.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine, Vaccine Center at Yerkes
LAST_NAME
Galinksi
FIRST_NAME
Mary
ADDRESS
Emory University, Yerkes National Primate Research Center, 954 Gatewood Rd, Room 003, Atlanta, GA 30329
AH and TM samples were obtained from young-normotensive DBA/2J (3 and 7.5 months) and old-hypertensive DBA/2J mice (8-8.5, 10 and 12 month). Lipids were extracted using modified Bligh and Dyer method and subjected to mass spectrometric identification using appropriate class-specific lipid standards and ratiometric quantification. Corresponding aqueous phase (of extraction) protein concentrations were measured using Bradford method.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
AH and TM samples were obtained from young-normotensive DBA/2J (3 and 7.5 months) and old-hypertensive DBA/2J mice (8-8.5, 10 and 12 month). Lipids were extracted using modified Bligh and Dyer method and subjected to mass spectrometric identification using appropriate class-specific lipid standards and ratiometric quantification. Corresponding aqueous phase (of extraction) protein concentrations were measured using Bradford method.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
AH and TM samples were obtained from young-normotensive DBA/2J (3 and 7.5 months) and old-hypertensive DBA/2J mice (8-8.5, 10 and 12 month). Lipids were extracted using modified Bligh and Dyer method and subjected to mass spectrometric identification using appropriate class-specific lipid standards and ratiometric quantification. Corresponding aqueous phase (of extraction) protein concentrations were measured using Bradford method.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
AH and TM samples were obtained from young-normotensive DBA/2J (3 and 7.5 months) and old-hypertensive DBA/2J mice (8-8.5, 10 and 12 month). Lipids were extracted using modified Bligh and Dyer method and subjected to mass spectrometric identification using appropriate class-specific lipid standards and ratiometric quantification. Corresponding aqueous phase (of extraction) protein concentrations were measured using Bradford method.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
The Metabolomics of Oral Biofilms exposed to Arginine and Fluoride
STUDY_SUMMARY
The study aims to use global metabolomics to investigate: (1) the metabolic profile of supragingival dental plaque from adults with different caries-status and from specific healthy and carious tooth-sites; and (2) the metabolic changes occurring in response to the use of the arginine or fluoride toothpastes for 12 weeks.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LABORATORY
Nascimento/Garrett
LAST_NAME
Nascimento
FIRST_NAME
Marcelle
ADDRESS
1395 Center Drive, Room D9-6, PO Box 100415, Gainesville, FL 32610-0415
Fasting wildtype, tfeb -/- knockout, and lmna -/- knockout metabolite profiling of adult zebrafish
STUDY_SUMMARY
Inhibition of mechanistic target of rapamycin (mTOR) activity exerts cardioprotective functions. We propose to assess the metabolite profile in zebrafish cardiomyopathy models to test the cardioprotective role of mTOR-TFEB-autophagy and mTOR-lmna- autophagy signaling in heart, liver, muscle, brain, and kidney tissue. In addition mTOR signaling among zebrafish 2 hour post feeding, 24 hour post feeding, and 48 hour post feeding will be profiled. These studes will be used as a baseline and for protocol development before we assess changes in DOX-induced cardiomyopathy.
METABOLOMIC PROFILING OF FOLLICULAR FLUID FROM PATIENTS WITH INFERTILITY-RELATED DEEP ENDOMETRIOSIS.
STUDY_SUMMARY
the metaboloc qualitative profiling was performed by LC-MS in follicular fluid samples of controls and endometriosis patients undergoing in vitro fertilization treatment
Evaluation of specific concentrations for use in experimental protocol
STUDY_TYPE
GC-MS non-targeted metabolomic profiling
STUDY_SUMMARY
This study evaluated specific plasma concentrations and compared the optimal plasma extract volume established in the first study (Effects of dilution on analyte identification and quantification) with the volume previously used in the current institutional protocol. The findings of this study lead to recommendations for experimental design in GC-MS-based metabolomic profiling of human plasma.
Metabolomics of Exhaled Breath Condensate in Decompensated Heart Failure
STUDY_SUMMARY
The aim of the study was to test weather characteristic differences or changes in metabolic profile exist between exhaled breath condensate (EBC) and saliva of healthy individuals and heart failure patients. EBC NMR profiling was performed.
Metabolomics of Saliva in Decompensated Heart Failure
STUDY_SUMMARY
The aim of the study was to test weather characteristic differences or changes in metabolic profile exist between exhaled breath condensate (EBC) and saliva of healthy individuals and heart failure patients. Salival NMR profiling was performed.
Effects of dilution on analyte identification and quantification
STUDY_TYPE
GC-MS non-targeted metabolomic profiling
STUDY_SUMMARY
The limiting-dilution study evaluated the effects of sample dilution on the ability to identify and quantify analytes in plasma. The study was divided into 10 batches with identical experimental design spanning over a 16-day period. Each batch consisted of 33 aliquots with 11 different plasma extract volumes (0 ā 700 ĀµL) corresponding to 11 plasma concentrations repeated three times.
Multi-site, cross-sectional study, including subjects with Age-related Macular Degeneration (early, intermediate and late disease) and a control group of subjects without any macular diseases. Plasma metabolomic profiles were assessed using Nuclear Magnetic Resonance Spectroscopy (NMR). Multivariate statistics were performed to compare metabolomic profiles of AMD patients vs controls.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO_Aveiro Institute of Materials
LABORATORY
University of Aveiro
LAST_NAME
Gil
FIRST_NAME
Ana
ADDRESS
Campus de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Metablomic profiling in acc1-5 mutant and wild type arabidiopsis
STUDY_SUMMARY
This experiment tests the metabolic consequence of a mutation at the ACC1 gene (At1g36160). The allele of acc1-5 bearing an EMS mutation, which cause a single amino acid substitution from aspartic acid to asparagine. Seedlings from both the acc1-5 mutant and the wild type were harvested and analyzed via HILIC LC-MS. Of particular interest are metabolites which would be affected by depletion of malonyl-CoA pools (flavenoids) and primary metabolites.
Experiment 13: Uninfected Macaca mulatta exposed to pyrimethamine to produce and integrate clinical, hematological, and omics control measures.
STUDY_SUMMARY
Uninfected, malaria-naive, male rhesus macaques (Macaca mulatta), approximately two years of age, were inoculated intravenously with a preparation of salivary gland material derived from non-infected Anopheles dirus and profiled for clinical, hematological, functional genomic, lipidomic, proteomic, and metabolomic measurements. Samples were generated and analyzed to investigate the effects of the pharmacological intervention with the anti-malarial drug pyrimethamine on normal individuals. The experiment was designed for 100 days plus a follow-up period, with pyrimethamine administered at three different time points to coincide with the predicted treatment days of experimentally infected rhesus macaques. Capillary blood samples were collected daily for the measurement of CBCs and reticulocytes. Capillary blood samples were collected every other day to obtain plasma for metabolomic analysis. Venous blood samples and bone marrow aspirates were collected at seven time points before and after three rounds of drug administration for functional genomic, proteomic, and lipidomic analyses. Within the MaHPIC, this project is known as 'Experiment 13'. This dataset was produced by Dean Jones at Emory University. The following contributed to the creation of this dataset: The MaHPIC Consortium, John Barnwell, Monica Cabrera, Jeremy D. DeBarry, Mary Galinski, Trenton Hoffman, Jay Humphrey, Jianlin Jiang, Chet Joyner, Nicolas Lackman, Stacey Lapp, Esmeralda Meyer, Alberto Moreno, Mustafa Nural, and Suman Pakala. The experimental design and protocols for this study were approved by the Emory University Institutional Animal Care and Use Committee (IACUC).
INSTITUTE
Emory University
DEPARTMENT
School of Medicine, Vaccine Center at Yerkes
LAST_NAME
Galinksi
FIRST_NAME
Mary
ADDRESS
Emory University, Yerkes National Primate Research Center, 954 Gatewood Rd, Room 003, Atlanta, GA 30329
EMAIL
mahpic@emory.edu
PHONE
none
TOTAL_SUBJECTS
6
STUDY_COMMENTS
31 samples, "The experimental design and protocols for this study were approved by the Emory University Institutional Animal Care and Use Committee (IACUC). These results are a product of a consortium of researchers known as the Malaria Host Pathogen Interaction Center (MaHPIC). For more information on the MaHPIC, please visit http://www.systemsbiology.emory.edu/ . Within the MaHPIC, these data were collected as part of 'Experiment 13' (E13). To access other publicly available results from E13 and other MaHPIC Experiments, including clinical results (specifics on drugs administered, diet, and veterinary interventions), and other omics, visit http://plasmodb.org/plasmo/mahpic.jsp . This page will be updated as datasets are released to the public. See Pubmed ID:25453034 for the associated publication for this study."
PGD2 and other lipid mediator changes in mouse adipose associated with administration of an oral inhibitor of H-PGDS (HQL-79)
STUDY_SUMMARY
This is an additional experiment being added onto a previous mouse feeding study that aimed to identify changes in metabolites that occur in metabolic tissues in the obese state that are long-lasting and not reversed by weight loss. We observed in the previous mice feeding study that levels of PGD2 increased in HFD fed mice and stayed high after the diet switch. Other members of the Prostaglandin family followed a similar trend (15-deoxy PGJ2, PGJ2) and were specific to adipose tissue. Based on previously published data indicating that central injection of PGD2 stimulates food intake, we attempted to observe this effect using an oral PGD2 inhibitor of H-PGDS (HQL-79). In fact, the oral inhibitor of the H-PGDS (HQL-79) administered peripherally (oral gavage in mice at 30mg/kg dose) reduced daily food intake. Mice were divided into two groups termed Vehicle (Control) and HGL-79 (H-PGDS inhibitor). Each group was analyzed for lipid mediator changes (including PGD2) in adipose tissue by the Newman lab. Analytical results generally met quality control criterion with respect to surrogate recoveries and replicate precision. Surrogate recoveries were good for most oxylipins (58-76%), endocannabinoids (53-75%), and fatty acids (36%). Recovery precision was good for most analytes in these profiles, ranging from 6-28% RSD for most surrogates. The precision for the LTB4 surrogate was higher than most others (38%). Analytical precision was assessed by duplicate analysis of two separate study samples. Analytical precision was 62 - 69% of analytes having <30% RSD for all profiles and correlation analysis for the analytes within these samples ranged from 0.90-0.99 R2. The complete data set is in the associated excel file (Osborn HQL-79 ā Deliverable Data Newman Lab.xls). There were few statistically significant differences observed when comparing concentrations (pmol/gr) between the control and HGL-79 treatment groups. However, when we compared ratios we saw numerous differences between PGD2 and its metabolite d15-PGJ2 versus other prostaglandins. Specifically, ratios between PGD2 and other connected pathway metabolites indicate a shift toward PGE2 and PGF2a production instead of PGD2 (Figure 1) with HQL-79 treatment. The PGD2 and PGE2 metabolites ratio of d15-PGJ2/15-keto PGE2 was statistically significant (P<0.01) using a two-tailed t-test. The ratios of PGD2/PGE2 and PGD2/PGF2 had p values of P<0.09 and P=0.07), respectively. Considering that we were predicting changes that indicated less PGD2 production it may be justifiable to use one-tailed tests instead. In order to maintain consistency with the metabolomic data analysis in the previous study, I followed the same statistical protocol that Johannes preformed for the main Pilot study. Using R and Devium log transformed data. Since this was a two group comparision, if the data was normal a 2 tailed t-test was used and if not normal then Mann-Whitney was used. A far as the significance of a shift from PGD2 to PGE2 production, I found a nice review article that discusses in detail the role of prostaglandins in white adipose tissue (Flachs et al. 2013). In the review it cites articles that have shown PGE2 to induce UCP1, modulate lipolysis adipogenesis, and stimulate leptin release. On the other hand, PGD2 was shown to increase adipogenesis and weight gain. Its downstream product d15-PGJ2 has been shown to increase adipogenesis, adipocyte differentiation, and decrease leptin production. This is significant since I also observed that the ratio of d15-PGJ2 to 15-keto PGE2 (the downstream product of PGE2) was also decreased. Another prostaglandin whose ratio versus PGD2 was different in the inhibitor group was PGF2a which has been shown to increase glucose transport in adipose tissue.
INSTITUTE
U.S.D.A. Western Human Nutrition Research Center, University of California, Davis
PGD2 and other lipid mediator changes in mouse adipose associated with administration of an oral inhibitor of H-PGDS (HQL-79)
STUDY_SUMMARY
This is an additional experiment being added onto a previous mouse feeding study that aimed to identify changes in metabolites that occur in metabolic tissues in the obese state that are long-lasting and not reversed by weight loss. We observed in the previous mice feeding study that levels of PGD2 increased in HFD fed mice and stayed high after the diet switch. Other members of the Prostaglandin family followed a similar trend (15-deoxy PGJ2, PGJ2) and were specific to adipose tissue. Based on previously published data indicating that central injection of PGD2 stimulates food intake, we attempted to observe this effect using an oral PGD2 inhibitor of H-PGDS (HQL-79). In fact, the oral inhibitor of the H-PGDS (HQL-79) administered peripherally (oral gavage in mice at 30mg/kg dose) reduced daily food intake. Mice were divided into two groups termed Vehicle (Control) and HGL-79 (H-PGDS inhibitor). Each group was analyzed for lipid mediator changes (including PGD2) in adipose tissue by the Newman lab. Analytical results generally met quality control criterion with respect to surrogate recoveries and replicate precision. Surrogate recoveries were good for most oxylipins (58-76%), endocannabinoids (53-75%), and fatty acids (36%). Recovery precision was good for most analytes in these profiles, ranging from 6-28% RSD for most surrogates. The precision for the LTB4 surrogate was higher than most others (38%). Analytical precision was assessed by duplicate analysis of two separate study samples. Analytical precision was 62 - 69% of analytes having <30% RSD for all profiles and correlation analysis for the analytes within these samples ranged from 0.90-0.99 R2. The complete data set is in the associated excel file (Osborn HQL-79 ā Deliverable Data Newman Lab.xls). There were few statistically significant differences observed when comparing concentrations (pmol/gr) between the control and HGL-79 treatment groups. However, when we compared ratios we saw numerous differences between PGD2 and its metabolite d15-PGJ2 versus other prostaglandins. Specifically, ratios between PGD2 and other connected pathway metabolites indicate a shift toward PGE2 and PGF2a production instead of PGD2 (Figure 1) with HQL-79 treatment. The PGD2 and PGE2 metabolites ratio of d15-PGJ2/15-keto PGE2 was statistically significant (P<0.01) using a two-tailed t-test. The ratios of PGD2/PGE2 and PGD2/PGF2 had p values of P<0.09 and P=0.07), respectively. Considering that we were predicting changes that indicated less PGD2 production it may be justifiable to use one-tailed tests instead. In order to maintain consistency with the metabolomic data analysis in the previous study, I followed the same statistical protocol that Johannes preformed for the main Pilot study. Using R and Devium log transformed data. Since this was a two group comparision, if the data was normal a 2 tailed t-test was used and if not normal then Mann-Whitney was used. A far as the significance of a shift from PGD2 to PGE2 production, I found a nice review article that discusses in detail the role of prostaglandins in white adipose tissue (Flachs et al. 2013). In the review it cites articles that have shown PGE2 to induce UCP1, modulate lipolysis adipogenesis, and stimulate leptin release. On the other hand, PGD2 was shown to increase adipogenesis and weight gain. Its downstream product d15-PGJ2 has been shown to increase adipogenesis, adipocyte differentiation, and decrease leptin production. This is significant since I also observed that the ratio of d15-PGJ2 to 15-keto PGE2 (the downstream product of PGE2) was also decreased. Another prostaglandin whose ratio versus PGD2 was different in the inhibitor group was PGF2a which has been shown to increase glucose transport in adipose tissue.
INSTITUTE
U.S.D.A. Western Human Nutrition Research Center, University of California, Davis
Emory University high-resolution metabolomic profiling CHEAR pooled reference materials: Urine
STUDY_TYPE
Untargeted HRMS for cross-laboratory characterization of common reference material.
STUDY_SUMMARY
Proper QA/QC is an essential component of any analytical procedure. The variability in platforms and approaches used by the untargeted cores requires that each lab develop specific QA/QC protocols, making cross-laboratory assessments challenging. To address the difference in measures across the untargeted cores, the untargeted quality assurance working group (UQAWG) has identified the importance of using the same biological reference sample across the different laboratories. To reach this goal, Dr. Jones and Mr. Walker of Emory University volunteered to take the lead in identifying vendors, requesting quotes, purchasing the pooled material, delivering the specified volumes to each of the lab hubs and storing additional material on-site at Emory University. Based upon discussion with the UQAWG, it was decided that each participating laboratory (five total) will receive 500 mL of plasma and 2000 mL of urine, providing sufficient volume for daily analysis over the next five years. Initially, NIST standard reference materials were considered; however, limited supply and cost makes the use of NIST materials impractical. Vendors, including BioreclamationIVT, Lee Biosolutions Inc and Gemini Bio-Products were contacted to request quotes for preparing pooled plasma and urine meeting the following requirements: K2EDTA recovered whole blood (plasma), equal distribution male/female, equal distribution of Black/Hispanic/Caucasian donors and be collected from greater than 40 donors. In addition, we requested all blood samples be collected in FDA inspected facilities, provide collection details, provide general demographics for each donor and be delivered in 500 mL aliquots. BioreclamationIVT, who also provided the biological samples for the NIH metabolomics ring trial, was selected as the most appropriate vendor due to the ability to provide the requested materials, lead-time to deliver and price. To date, both pools have been purchased. The urine pool was received on July 21, 2016 and the plasma pool will be received July 26, 2016. Upon receipt, both pools will be stored at -80Ā°C. Summary demographics were provided by BioreclamationIVT, the urine pool was created by combining urine samples from 60 males and 60 females with an average age of 38 (range 18-60) and equal distribution of races. The plasma pool was created by combining recovered plasma obtained from 50 males and 50 females with an average age of 38.4 (range 19-72) and equal distribution of races. Both urine and plasma pools will be delivered to the lab hubs frozen in units of 500 mL. Upon receipt, plasma and urine pools should be aliquoted into volumes that provide sufficient material so that freeze-thaw of the pooled material is minimized. For unambiguous identification, the urine pool has been named UrineRef_20160721 and the plasma pool has been named PlasmaRef_20160726. The purpose of providing each untargeted lab hub with the pooled material is two-fold. Through analysis of UrineRef_20160721 (if study samples are urine) or PlasmaRef_20160726 (if study samples are plasma) in each batch of samples analyzed for CHEAR, it will be possible to have a common reference across the different lab hubs. Thus, the ability to compare analyses on different platforms will be greatly improved. The use of a consistent reference material in each batch will also facilitate reference standardization, a concept developed for retrospective quantification of high-resolution metabolomics (Go, Walker et al. 2015). Using reference standardization, quantification post-data acquisition is possible by referencing the pooled material analyzed with each series of samples. The pooled material can be characterized and analytes quantified using traditional analytical chemistry techniques, such as quantification by methods of addition or standardization with a certified reference material, such as NIST SRM 1950. Known concentrations within the pooled material can be used to determine an analyte response factor and calculate sample concentrations based on single-point calibration. The benefit of this approach is that targeted quantification is only required in the pooled sample (i.e. UrineRef_20160721 or PlasmaRef_20160726), chemicals selected for quantification do not need to be selected a priori, and population wide estimates of plasma chemical concentrations can be determined without having to re-analyze samples using a targeted approach. Current Clinical Biomarker Laboratory protocol requires three analytical replicates of pooled reference material per day (three technical replicates per analytical replicate per column configuration, or 18 injections/instrument-day), resulting in analyses of the reference material every 10 samples. Results from untargeted LC-HRMS analyses of pooled CHEAR reference materials. PlasmaRef_20160726 was analyzed with additional NIST reference materials: SRM 1950, SRM 1957 and SRM 1958. UrineRef_20160721 was analyzed with NIST reference materials SRM 3672 and 3673.
INSTITUTE
Emory University School of Medicine
DEPARTMENT
Division of Pulmonary, Allergy and Critical Care Medicine
Emory University high-resolution metabolomic profiling CHEAR pooled reference materials
STUDY_TYPE
Untargeted HRMS for cross-laboratory characterization of common reference material.
STUDY_SUMMARY
Proper QA/QC is an essential component of any analytical procedure. The variability in platforms and approaches used by the untargeted cores requires that each lab develop specific QA/QC protocols, making cross-laboratory assessments challenging. To address the difference in measures across the untargeted cores, the untargeted quality assurance working group (UQAWG) has identified the importance of using the same biological reference sample across the different laboratories. To reach this goal, Dr. Jones and Mr. Walker of Emory University volunteered to take the lead in identifying vendors, requesting quotes, purchasing the pooled material, delivering the specified volumes to each of the lab hubs and storing additional material on-site at Emory University. Based upon discussion with the UQAWG, it was decided that each participating laboratory (five total) will receive 500 mL of plasma and 2000 mL of urine, providing sufficient volume for daily analysis over the next five years. Initially, NIST standard reference materials were considered; however, limited supply and cost makes the use of NIST materials impractical. Vendors, including BioreclamationIVT, Lee Biosolutions Inc and Gemini Bio-Products were contacted to request quotes for preparing pooled plasma and urine meeting the following requirements: K2EDTA recovered whole blood (plasma), equal distribution male/female, equal distribution of Black/Hispanic/Caucasian donors and be collected from greater than 40 donors. In addition, we requested all blood samples be collected in FDA inspected facilities, provide collection details, provide general demographics for each donor and be delivered in 500 mL aliquots. BioreclamationIVT, who also provided the biological samples for the NIH metabolomics ring trial, was selected as the most appropriate vendor due to the ability to provide the requested materials, lead-time to deliver and price. To date, both pools have been purchased. The urine pool was received on July 21, 2016 and the plasma pool will be received July 26, 2016. Upon receipt, both pools will be stored at -80Ā°C. Summary demographics were provided by BioreclamationIVT, the urine pool was created by combining urine samples from 60 males and 60 females with an average age of 38 (range 18-60) and equal distribution of races. The plasma pool was created by combining recovered plasma obtained from 50 males and 50 females with an average age of 38.4 (range 19-72) and equal distribution of races. Both urine and plasma pools will be delivered to the lab hubs frozen in units of 500 mL. Upon receipt, plasma and urine pools should be aliquoted into volumes that provide sufficient material so that freeze-thaw of the pooled material is minimized. For unambiguous identification, the urine pool has been named UrineRef_20160721 and the plasma pool has been named PlasmaRef_20160726. The purpose of providing each untargeted lab hub with the pooled material is two-fold. Through analysis of UrineRef_20160721 (if study samples are urine) or PlasmaRef_20160726 (if study samples are plasma) in each batch of samples analyzed for CHEAR, it will be possible to have a common reference across the different lab hubs. Thus, the ability to compare analyses on different platforms will be greatly improved. The use of a consistent reference material in each batch will also facilitate reference standardization, a concept developed for retrospective quantification of high-resolution metabolomics (Go, Walker et al. 2015). Using reference standardization, quantification post-data acquisition is possible by referencing the pooled material analyzed with each series of samples. The pooled material can be characterized and analytes quantified using traditional analytical chemistry techniques, such as quantification by methods of addition or standardization with a certified reference material, such as NIST SRM 1950. Known concentrations within the pooled material can be used to determine an analyte response factor and calculate sample concentrations based on single-point calibration. The benefit of this approach is that targeted quantification is only required in the pooled sample (i.e. UrineRef_20160721 or PlasmaRef_20160726), chemicals selected for quantification do not need to be selected a priori, and population wide estimates of plasma chemical concentrations can be determined without having to re-analyze samples using a targeted approach. Current Clinical Biomarker Laboratory protocol requires three analytical replicates of pooled reference material per day (three technical replicates per analytical replicate per column configuration, or 18 injections/instrument-day), resulting in analyses of the reference material every 10 samples. Results from untargeted LC-HRMS analyses of pooled CHEAR reference materials. PlasmaRef_20160726 was analyzed with additional NIST reference materials: SRM 1950, SRM 1957 and SRM 1958. UrineRef_20160721 was analyzed with NIST reference materials SRM 3672 and 3673.
INSTITUTE
Emory University School of Medicine
DEPARTMENT
Division of Pulmonary, Allergy and Critical Care Medicine
Dysfunctional lipid metabolism underlies the effect of the perinatal DDT exposure on the development of metabolic syndrome
STUDY_SUMMARY
This study aims to identify changes in lipid mediators in the hypothalamus with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP (n=8 per group) or not treated (n=8 per group). Each group was analyzed for oxylipin, nitro lipids, endocannabinoid, and endocannabinoid-like monoacylglycerol and N-acylethanolamide changes to investigate alterations in lipid mediator signaling due to TPP exposure. Targeted metabolomic analysis of lipid mediators in rat hypothalamus samples was performed by the Newman lab.
INSTITUTE
U.S.D.A. Western Human Nutrition Research Center, University of California, Davis
Dysfunctional lipid metabolism underlies the effect of the perinatal DDT exposure on the development of metabolic syndrome
STUDY_SUMMARY
This study aims to identify changes in lipid mediators in the hypothalamus with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP (n=8 per group) or not treated (n=8 per group). Each group was analyzed for oxylipin, nitro lipids, endocannabinoid, and endocannabinoid-like monoacylglycerol and N-acylethanolamide changes to investigate alterations in lipid mediator signaling due to TPP exposure. Targeted metabolomic analysis of lipid mediators in rat hypothalamus samples was performed by the Newman lab.
INSTITUTE
U.S.D.A. Western Human Nutrition Research Center, University of California, Davis
Metabolomics measures of Macaca mulatta infected with Plasmodium coatneyi Hackeri strain to produce and integrate clinical, hematological, parasitological, and omics measures of acute, recrudescent, and chronic infections.
STUDY_TYPE
Longitudinal parasite infection and treatment of multiple individuals
STUDY_SUMMARY
Malaria-naive male rhesus macaques (Macaca mulatta), approximately four years of age, were inoculated intravenously with salivary gland sporozoites produced and isolated at the Centers for Disease Control and Prevention from multiple Anopheles species (An. dirus, An. gambiae, and An. stephensi) and then profiled for clinical, hematological, parasitological, immunological, functional genomic, lipidomic, proteomic, and metabolomic measurements. The experiment was designed for 100 days, and pre- and post-100 day periods to prepare subjects and administer curative treatments respectively. The anti-malarial drug artemether was subcuratively administered to all subjects at the initial peak of infection, one out of the five macaques received four additional subcurative treatments for subsequent recrudescence peaks. The experimental infection in one subject was ineffective but the macaque was followed-up for the same period of 100 days. The different clinical phases of the infection were clinically determined for each subject. Blood-stage curative doses of artemether were administered to all subjects at the end of the study. Capillary blood samples were collected daily for the measurement of CBCs, reticulocytes, and parasitemias. Capillary blood samples were collected every other day to obtain plasma for metabolomic analysis. Venous blood and bone marrow samples were collected at seven time points for functional genomic, proteomic, lipidomic, and immunological analyses. Within the MaHPIC, this project is known as āExperiment 03ā. This dataset was produced by Dean Jones at Emory University. The experimental design and protocols for this study were approved by the Emory University Institutional Animal Care and Use Committee (IACUC).
INSTITUTE
Emory University
DEPARTMENT
School of Medicine, Vaccine Center at Yerkes
LAST_NAME
Galinski
FIRST_NAME
Mary
ADDRESS
Emory University, Yerkes National Primate Research Center, 954 Gatewood Rd, Room 003, Atlanta, GA 30329
EMAIL
mahpic@emory.edu
PHONE
N/A
NUM_GROUPS
6
TOTAL_SUBJECTS
331
STUDY_COMMENTS
The experimental design and protocols for this study were approved by the Emory University Institutional Animal Care and Use Committee (IACUC). These results are a product of a consortium of researchers known as the Malaria Host Pathogen Interaction Center (MaHPIC). Contributors include: Monica Cabrera, Jeremy D. DeBarry, Mary G. Galinski, Jay Humphrey, Dean Jones, Ebru Karpuzoglu, Jessica C. Kissinger, Regina Joice, Esmeralda Meyer, Vishal Nayak, Mustafa Nural, Suman Pakala, ViLinh Tran, Karan Uppal, Loukia Williams. For more information on the MaHPIC, please visit http://www.systemsbiology.emory.edu/ . Within the MaHPIC, these data were collected as part of 'Experiment 03' (E03). To access other publicly available results from E03 and other MaHPIC Experiments, including clinical results (specifics on drugs administered, diet, and veterinary interventions), and other omics, visit http://plasmodb.org/plasmo/mahpic.jsp . This page will be updated as datasets are released to the public.
NEFA Profile Response to Triphenyl Phosphate Exposure
STUDY_SUMMARY
This study aims to identify changes in non-esterified fatty acid (NEFAs) in the plasma with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP or not treated. Each group was analyzed for non-esterified fatty acid (NEFA) changes to investigate alterations in NEFAs due to TPP exposure. Targeted analysis of NEFA in rat plasma samples was performed by the Newman lab.
INSTITUTE
U.S.D.A. Western Human Nutrition Research Center, University of California, Davis
DEPARTMENT
Nutrition
LAST_NAME
Newman
FIRST_NAME
John
ADDRESS
430 W. Health Sciences Dr., Davis, CA 95616
EMAIL
john.newman@ars.usda.gov
PHONE
+1-530-752-1009
STUDY_COMMENTS
The samples included a high degree of hemolysis exhibited in the plasma. One sample was lost during processing (Group E- Subject 78). Two samples were outliers for multiple analytes and were not included in the final data (E-117 & T-28). Of the samples reported in this data set, there were no missing values.
Untargeted LC-MS metabolomics analysis of human COPD plasma, HILIC & C18
STUDY_SUMMARY
Identify perturbed metabolites and pathways in human plasma collected from 131 COPD subjects. Subjects were either current or former smokers with various COPD phenotypes including emphysema, and exacerbations.
PIXiE: An Algorithm for Automated Ion Mobility Arrival Time Extraction and Collision Cross Section Calculation using Global Data Association
STUDY_SUMMARY
Motivation: Drift tube ion mobility spectrometry coupled with mass spectrometry (DTIMS-MS) is increasingly implemented in high throughput omics workflows, and new informatics approaches are necessary for processing the associated data. To automatically extract arrival times for molecules measured by DTIMS at multiple electric fields and compute their associated collisional cross sections (CCS), we created the PNNL Ion Mobility Cross Section Extractor (PIXiE). The primary application presented for this algorithm is the extraction of that can then be used to create a reference library of CCS valuesinformation necessary to create a reference library containing accurate masses, DTIMS arrival times and CCSs for use in high throughput omics analyses. Results: We demonstrate the utility of this approach by automatically extracting arrival times and calculating the associated CCSs for a set of endogenous metabolites and xenobiotics. The PIXiE-generated CCS values were within error of those calculated using commercially available instrument vendor software.
Urinary Volatile Compound, Associated with Chronic Inflammation In Interstitial Cystitis
STUDY_SUMMARY
Interstitial cystitis (IC)/bladder pain syndrome (BPS) is a clinical condition that manifests as a sensory hypersensitivity of unknown cause and is characterized by urinary frequency, bladder discomfort, and pelvic pain. In the present volatolomic study, we have analyzed the VOCs unique to urine specimens obtained from interstitial cystitis patients, in compassion to healthy controls.This is the novel finding from comprehensive and unbiased metabolomics analysis that urinary menthol is decreased in urine specimens from IC patients, and that the reduced menthol level in IC is potentially linked to the chronic inflammation, which is often observed in IC patients
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
Health Sciences Drive, Davis, California, 95616, USA
Impact Of High Sugar Diet On L-Arginine Metabolism In The Lung
STUDY_SUMMARY
Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. We need novel therapeutic agents that are affordable, can decrease the reliance on steroids, and can improve quality of life. This clinical and mechanistic study has the potential to impact treatment of a subset of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and NO biology in the airways of asthmatics. We will pursue a clinical trial in subjects not well controlled on standard drug therapy; this strategy will address whether L-arginine is efficacious in patients receiving standard of care medications. In studies using animal models, we and others have shown that interventions that augment NO levels, through either supplementation of L-arginine or inhibition of arginase, decrease allergic airway inflammation and hyperresponsiveness-the two hallmarks of asthma. Overall, we hypothesize that a responder subset of adult severe asthma patients will derive clinical benefit from supplemental L-arginine therapy and that these patients will have a lower exhaled NO concentrations (<20 ppb) and a higher NOS2/Arg1 mRNA and protein ratio in their airway epithelial cells than non-responders. We aim to: 1) test the hypothesis that uncontrolled, adult severe asthma patients with exhaled breath NO concentrations <20 ppb will have fewer asthma exacerbations over 3 months when treated with L-arginine compared to patients with FeNO > 25, 2) determine the mechanisms by which L-arginine affects the regulation of NOS and arginase enzymes in primary airway epithelial cell cultures from severe asthmatic subjects, and 3) test the hypothesis that inhaled nanoparticle carrier formulations of L-arginine will decrease airway inflammation, airway hyperresponsiveness, and airway fibrosis at lower doses than systemically administered L-arginine. The major impact of our study will be to identify the adult severe asthma cohort that will benefit from supplemental L-arginine therapy. Our ultimate goal is to develop novel therapeutic agents to treat adult severe asthma patients better. PUBLIC HEALTH RELEVANCE: Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. This clinical study has the potential to improve the care of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and nitric oxide biology in the lung. If we demonstrate that L-arginine supplementation can decrease asthma attacks in a subset of severe asthmatics, it will have great implications for future research as well as for the daily lives of patients with asthma.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Bioprospection of the aromatic potential of species from the Atlantic Rainforest in SĆ£o Paulo: occurrence, taxonomy and chemical, genetic and physiological characterization of plant populations
STUDY_TYPE
Chemcial profile of the volatiles from plants of the Atlantic Rainforest in SĆ£o Paulo
1D-1H-Nuclear Magnetic Resonance Metabolomics Reveals Age-related Changes in Metabolites Associated with Experimental Venous Thrombosis
STUDY_TYPE
Experimental venous thrombosis
STUDY_SUMMARY
Sodium heparin preserved whole blood samples were collected from young and old mice with and without induced VT. Samples were subjected to MeOH:CHCl3 extraction and the MeOH was assayed by 1H-NMR. Chenomx software was used for spectral analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Clinical Pharmacy
LABORATORY
The NMR Metabolomics Laboratory (Stringer)
LAST_NAME
Stringer
FIRST_NAME
Kathleen
ADDRESS
College of Pharmacy, University of Michigan, 428 Church Street, Ann Arbor, MI 48109
Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies
STUDY_SUMMARY
The use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as the smaller blood volume required, storage at room temperature, and ability to sample in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed the stability of polar metabolites and lipids in DBS samples collected in 2000-2001 and stored at room temperature. The identified and statistically significant molecules were then compared to matched serum samples stored at ā80Ā°C to determine if the DBS samples could be effectively used in a longitudinal study following metabolic disease. A total of 400 polar metabolites and lipids were identified in the serum and DBS samples using gas chromatograph/mass spectrometry (GC/MS), liquid chromatography (LC)/MS, and LC/ion mobility spectrometry-MS (LC/IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant metabolite was conserved in a case-control study of older diabetic males with low amounts of high-density lipoproteins and high body mass indices, triacylglycerides and glucose levels when compared to non-diabetic patients with normal levels, indicating that degradation in the DBS samples affects polar metabolite quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, 36 statistically significant lipids correlated in both sample types. The difference in the number of statistically significant polar metabolites and lipids indicated that the lipids did not degrade to as great of a degree as the polar metabolites in the DBS samples and lipid quantitation was still possible.
CHEAR PlasmaRef_20160726 was provided by Emory University. The material was prepared and analyzed using the Metabolomics Reverse Phase Broad Spectrum analysis on a Synapt G2-Si system. Six samples were injected of the sample reference material that were prepared in replicate.
INSTITUTE
RTI international
DEPARTMENT
ACP
LAST_NAME
Fennell
FIRST_NAME
Timothy
ADDRESS
3040 E Cornwallis Rd, Durham, North Carolina, 27709, USA
CHEAR UrineRef_20160721 was provided by Emory University. The material was prepared and analyzed using the Metabolomics Reverse Phase Broad Spectrum analysis on a Synapt G2-Si system. Six samples were injected of the sample reference material that were prepared in replicate.
INSTITUTE
RTI International
DEPARTMENT
ACP
LAST_NAME
Fennell
FIRST_NAME
Timothy
ADDRESS
3040 E Cornwallis Rd, Durham, North Carolina, 27709, USA
LC/MS-MS measurement of inosine to adenosine ratio as well as GC/MS measurement of sarcosine relative values in urine samples from prostate cancer and case control patients in a group of ancestry verified African American and European American.
Sphingolipid Analysis of Human Aqueous Humor in Glaucomatous and Control eyes
STUDY_SUMMARY
This study examined the profiles of sphinĀ¬golipids and ceramides present in the aqueous humor (AH) of human control and POAG donors. Furthermore, we quantitatively compared distinct differences between glaucomatous and age-matched control eyes, identifying potential molecules for further experimentation to determine their biological role in modulating cell behavior. Lipids were identified and ratiometrically quantified in a two-step process using precursor ion scan (PIS) or neutral loss scan (NLS) with appropriate class-specific lipid standards on a TSQ Quantum Access Max mass spectrometer following established procedures. We identified several species of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramides that were common between control and glaucomatous AQH. Some unique lipid species in these classes were also identified in controls but not in glaucoma and vice versa.
We determined the profiles of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramide, and their quantitative differences between control and glaucomatous trabecular meshwork (TM) derived from human donors.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
Smoking induced methylation plays a critical role in the accumulation of methylated metabolites, DNA adducts damage and altered metabolism in BCa. These deregulated metabolites can be detected non-invasively and can be used as causal biomarkers that can predict the risk of developing BCa in smokers
GC/MS measurement of sarcosine in urine samples from prostate cancer and case control patients in a group of ancestry verified African American and European Americans.
Human Aqueous Humor Phospholipids in Control and Glaucomatous eyes
STUDY_SUMMARY
An analysis of phospholipid (phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine and phosphatidylinositol) classes in human control and glaucomatous aqueous humor (AH). Lipid extraction was performed using a modification of the Bligh and Dyer method, protein concentrations were determined using the Bradfordās method, and select samples were confirmed with Densitometry of PHAST gels. Lipids were identified and subjected to ratiometric quantification using a TSQ Quantum Access Max triple quadrupole mass spectrometer utilizing precursor ion scan (PIS) or neutral ion loss scan (NLS) using appropriate class specific lipid standards in a two-step quantification process. Mass spectrometer data were analyzed using MzMine 2.23.
Validation of the application of targeted metabolomic appraoch in the diagnosis of CFS
STUDY_TYPE
Plasma metabolomic profiling
STUDY_SUMMARY
This study was to validate the utility of the developed targeted metabolomic method in the diagnosis of chronic fatigue syndrome (CFS). Clinical validation consisted of a cohort of 20 male CFS (53 Ā± 2.8 years old, mean Ā± SEM, range 21-67 y) and 18 male controls (53 Ā± 3.5 years old, mean Ā± SEM, range 23-69 y), who were enrolled in a previous study (Naviaux et al. 2016). These plasma samples were stored in -80 ĀŗC for about 1.5 years and reanalyzed on a different LC-MS/MS system by a different investigator.
INSTITUTE
University of California, San Diego
DEPARTMENT
Department of Medicine
LABORATORY
The Mitochondrial and Metabolic Disease Center
LAST_NAME
Naviaux
FIRST_NAME
Robert
ADDRESS
214 Dikinson Street, CTF-C102, San Diego, CA, 92103
EMAIL
rnaviaux@ucsd.edu
PHONE
619-993-2904
NUM_GROUPS
2
TOTAL_SUBJECTS
38
NUM_MALES
38
STUDY_COMMENTS
These plasma samples were stored in -80 ĀŗC for about 1.5 years and reanalyzed on a different LC-MS/MS system by a different investigator.
Metabolomic Profiling of Follicular Fluid from Patients with Polycystic Ovary Syndrome and Hyper Response to In Vitro Fertilization Treatment
STUDY_TYPE
MS quallitative analysis
STUDY_SUMMARY
The present study consisted in a metabolomic approach in follicular fluid samples from patients with polycystic ovary syndrome and hyper response to in vitro fertilization treatment.
INSTITUTE
Sao Paulo Federal University
DEPARTMENT
Department of Surgery, Division of Urology, Human Reproduction Section
A structural examination and collision cross section database of over 500 metabolities and xenobiotics using drift tube ion mobility
STUDY_TYPE
Library compilation of metabolomic standards
STUDY_SUMMARY
Standards of metabolomic pathways and external secondary metabolites and xenobiotics were analysed with Agilent 6560 Ion mobility QTOF MS platform to build a comprehensive library of Collision Cross Section values. All measurements were performed in triplicate in both positive and negative polarities with nitrogen gas and at seven different electric fields, so that values could be directly measured and random standard deviations (RSD) assessed for each molecule.
INSTITUTE
Pacific Northwest National Laboratory
DEPARTMENT
Integrative Omics
LABORATORY
Pacific Northwest National Laboratory
LAST_NAME
Baker
FIRST_NAME
Erin
ADDRESS
Pacific Northwest National Laboratory, 902 Battelle Boulevard, Richland, WA
Sphingolipid Analysis of hyper and normotensive DBA2J mice aqueous humor and trabecular meshwork
STUDY_SUMMARY
To determine the differential profiles of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramide and their quantitative differences between trabecular meshwork (TM) and aqueous humor (AH) derived from normotensive and hypertensive intraocular pressure states of DBA/2J mice.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
Untargeted metabolomic changes in Chlamydomonas reinhardtii treated with lipid inducing small molecules
STUDY_SUMMARY
A study to investigate the effect of small molecule lipid inducing compounds that leads to hyper accumulation of lipids in N replete cells of Chlamydomonas reinhardtii. These compounds were identified through a high throughput screening designed for that purpose. During that screening, we screened 43,783 compounds and identified 367 primary hits. These 367 hits were further retested using a 8-point dilution series (from 0.25 to 30 uM) and verified the activity of 250 compounds that induce the hyper lipid accumulating phenotype in algae. Once the hit compounds were identified and confirmed, we then performed extensive chemoinformatics analysis to look for common scaffolds and identified several common substructures. We then selected 15 top performing compounds from 5 diverse structural groups and tested biochemical parameters such as growth, lipid accumulating capacity, effect on photosynthetic rates, respiration rates, oxygen consumption rates, analysis of different lipid species to quantify and identify fatty acid species using GC-MS. To understand the global changes in the metabolome, 2 structurally different compounds were selected and compared with cells grown without compounds as control for untargeted metabolomics analysis.
Identification and metabolite profiling of chemical activators of lipid accumulation in green algae
STUDY_TYPE
GC-MS metabolite profiling of algal lipid activators
STUDY_SUMMARY
Microalgae are proposed as feedstock organisms useful for producing biofuels and co-products. However, several limitations must be overcome before algae-based production is economically feasible. Among these is the ability to induce lipid accumulation and storage without affecting biomass yield. To overcome this barrier, a chemical genetics approach was employed in which 43,783 compounds were screened against Chlamydomonas reinhardtii and 243 compounds were identified that increase triacylglyceride (TAG) accumulation without terminating growth. Identified compounds were classified by structural similarity and 15 selected for secondary analyses addressing impacts on growth fitness, photosynthetic pigments, and total cellular protein and starch concentrations. TAG accumulation was verified using GC-MS quantification of total fatty acids and targeted TAG and galactolipid (GL) measurements using LC-MRM/MS. These results demonstrated TAG accumulation does not necessarily proceed at the expense of GL. Untargeted metabolite profiling provided important insights into pathway shifts due to 5 different compound treatments and verified the anabolic state of the cells with regard to the oxidative pentose phosphate pathway, Calvin cycle, tricarboxylic acid cycle and amino acid biosynthetic pathways. Metabolite patterns were distinct from nitrogen starvation and other abiotic stresses commonly used to induce oil accumulation in algae. The efficacy of these compounds was also demonstrated in 3 other algal species. These lipid inducing compounds offer a valuable set of tools for delving into the biochemical mechanisms of lipid accumulation in algae and a direct means to improve algal oil content independent of the severe growth limitations associated with nutrient deprivation.
INSTITUTE
Univ of Nebraska-Lincoln
DEPARTMENT
Biochemistry
LABORATORY
FATTTLab
LAST_NAME
Wase
FIRST_NAME
Nishikant
ADDRESS
1901 Beadle Center, Vine Street, 1901 VINE STREET, Lincoln, NE, 68588-0664, USA
Non-targeted Metabolomics Analysis of Golden Retriever Muscular Dystrophy-Affected Muscles Reveals Alterations in Arginine and Proline Metabolism, and Elevations in Glutamic and Oleic Acid In Vivo
STUDY_SUMMARY
Like Duchenne muscular dystrophy (DMD), the Golden Retriever Muscular Dystrophy (GRMD) dog model of DMD exhibits is characterized by muscle necrosis, progressive paralysis, and pseudohypertrophy in specific skeletal muscles. This severe GRMD phenotype included atrophy of the biceps femoris (BF) and compared to unaffected normal dogs, while the long digital extensor (LDE) of the pelvic limb, serving as a hip flex and stifle extensor, is unaffected. A recent microarray analysis of GRMD identified alterations in genes associated with lipid metabolism and energy production. We, therefore, undertook a non-targeted metabolomics analysis of the GRMD BF (affected) and LDE (unaffected) using GC-MS to identify underlying metabolic defects specific for affected GRMD skeletal muscle. Of the 134 metabolites identified in BF, eight were significantly altered in GRMD BF compared to control BF (Glutamic Acid (2.48 fold vs. controls); Oleic Acid (1.76 fold vs. controls); Proline (1.73 fold vs. controls); Myoinositol-2- Phosphate (0.44 fold vs. controls); Fumaric Acid (0.40 fold vs. controls); Carnosine (0.40 fold vs. controls); Lactamide (0.33 fold vs. controls); and Stearamide (0.23 fold vs. controls). Pathway analysis of the T-test significant metabolites identified BF muscle metabolites significantly enriched for Arginine and proline metabolism (p=5.8E-4, FDR=0.04) and Alanine, aspartate, and glutamate metabolism (p=1.3E-3, FDR=0.05). The GRMD LDE previously reported to be unaffected, in contrast, had only one significantly altered metabolite (3-Phosphoglyceric Acid (0.35 Fold vs. controls)).The identification of elevated BF Oleic acid (a long-chain fatty acid) is consistent with recent microarray studies identifying altered lipid metabolism genes, while alterations in Arginine and Proline metabolism are consistent with recent studies identifying elevated L-arginine in DMD patient sera as a biomarker of disease (alterations in DMD or GRMD muscle itself have not previously been reported).Together, these studies demonstrate muscle-specific alterations in GRMD-affected muscle, which illustrate previously unidentified metabolic changes.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine, Department of Pathology & Laboratory, Department of Pharmacology
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm Road, MBRB 2336, Chapel Hill, North Carolina, 27599-7126, USA
EMAIL
monte_willis@med.unc.edu
PHONE
(984) 999-5431
STUDY_COMMENTS
Skeletal Muscle (Biceps femoris (BF), long digital extensor(LDE))
Human trabecular meshowrk phospholipid profiles of control and glaucomatous eyes
STUDY_SUMMARY
We compared phospholipid (phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, and phosphatidylinositol) profiles of control and glaucomatous trabecular meshwork (TM) derived from human donors.Lipid extraction was performed using a modification of the Bligh and Dyer method, protein concentrations were determined using the Bradford method, and for select samples confirmed with densitometry of PHAST gels. Lipids were identified and subjected to ratiometric quantification using a TSQ quantum Access Max triple quadrupole mass spectrometer with precursor ion scan (PIS) or neutral ion loss scan (NLS), using appropriate class specific lipid standards.
INSTITUTE
University of Miami
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
Non targeted metabolomic analysis of mouse and human bronchoalveolar lavage fluid, Aqueous(+) experiment (part I)
STUDY_TYPE
Comprehensive profile
STUDY_SUMMARY
Mouse BALF was collected from C57BL/6 mice (n = 10). Five mice were exposed to ambient air for one day (air control) and five mice were exposed to CS for nine months (smoking). Human BALF was collected from subjects from the COPDGene cohort (n = 5). COPD diagnosis was based on the ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC). Subjects were 45-70 years old, BMI 27-45, weight 76-125kg, and were categorized as follows: 1 male former smoker without COPD (FEV1/FVC = 0.82), 2 male current smokers without COPD (FEV1/FVC = 0.91 and 0.79), 1 female current smoker with moderate COPD (FEV1/FVC = 0.51), and 1 male current smoker with moderate COPD (FEV1/FVC = 0.64).
Non targeted metabolomic analysis of mouse and human bronchoalveolar lavage fluid, Lipid(-) experiment
STUDY_TYPE
Comprehensive profile
STUDY_SUMMARY
Mouse BALF was collected from C57BL/6 mice (n = 10). Five mice were exposed to ambient air for one day (air control) and five mice were exposed to CS for nine months (smoking). Human BALF was collected from subjects from the COPDGene cohort (n = 5). COPD diagnosis was based on the ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC). Subjects were 45-70 years old, BMI 27-45, weight 76-125kg, and were categorized as follows: 1 male former smoker without COPD (FEV1/FVC = 0.82), 2 male current smokers without COPD (FEV1/FVC = 0.91 and 0.79), 1 female current smoker with moderate COPD (FEV1/FVC = 0.51), and 1 male current smoker with moderate COPD (FEV1/FVC = 0.64)
Non targeted metabolomic analysis of mouse and human bronchoalveolar lavage fluid, Lipid(+) experiment (part III)
STUDY_TYPE
Comprehensive profile
STUDY_SUMMARY
Mouse BALF was collected from C57BL/6 mice (n = 10). Five mice were exposed to ambient air for one day (air control) and five mice were exposed to CS for nine months (smoking). Human BALF was collected from subjects from the COPDGene cohort (n = 5). COPD diagnosis was based on the ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC). Subjects were 45-70 years old, BMI 27-45, weight 76-125kg, and were categorized as follows: 1 male former smoker without COPD (FEV1/FVC = 0.82), 2 male current smokers without COPD (FEV1/FVC = 0.91 and 0.79), 1 female current smoker with moderate COPD (FEV1/FVC = 0.51), and 1 male current smoker with moderate COPD (FEV1/FVC = 0.64)
TCA Cycle Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine from 20 Participants of the DASH2 Clinical Trial
STUDY_SUMMARY
The objective of the study is to identify changes of urinary metabolite profiles associated with different responses to blood pressure to salt. Subjects are derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. 2001).We choose two groups subjects who meet the following conditions(the two groups are separately named A and B). We chose subjects on the Control diet .These subjects meet the blood pressure criteria described below:Group A subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic blood pressure from the low sodium visit is less than 140 and the diastolic blood pressure from low sodium visit is less than 90; 3) For group A: Either the systolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the systolic blood pressure from the low sodium visit, or the diastolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the diastolic blood pressure from the low sodium visit; 3) For group B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. +/- 5) from the systolic blood pressure from the low sodium visit, and the diastolic blood pressure from the high sodium visit is within 5 mmHg from the diastolic blood pressure from the low sodium visit. Use gas chromatography/mass spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry (LC/MS)analysis to find the differences of metabolic profiles between the high sodium level and the low sodium level, and compare the metabolic profiles of A with the metabolic profiles of B at the low and high sodium level. Note, this was used as a pilot, the rest of the samples are in another uploaded study.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Amino Acd Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine from 20 Participants of the DASH2 Clinical Trial (part II)
STUDY_SUMMARY
The objective of the study is to identify changes of urinary metabolite profiles associated with different responses to blood pressure to salt. Subjects are derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. 2001).We choose two groups subjects who meet the following conditions(the two groups are separately named A and B). We chose subjects on the Control diet .These subjects meet the blood pressure criteria described below:Group A subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic blood pressure from the low sodium visit is less than 140 and the diastolic blood pressure from low sodium visit is less than 90; 3) For group A: Either the systolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the systolic blood pressure from the low sodium visit, or the diastolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the diastolic blood pressure from the low sodium visit; 3) For group B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. +/- 5) from the systolic blood pressure from the low sodium visit, and the diastolic blood pressure from the high sodium visit is within 5 mmHg from the diastolic blood pressure from the low sodium visit. Use gas chromatography/mass spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry (LC/MS)analysis to find the differences of metabolic profiles between the high sodium level and the low sodium level, and compare the metabolic profiles of A with the metabolic profiles of B at the low and high sodium level. Note, this was used as a pilot, the rest of the samples are in another uploaded study.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Neuromodulator Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine from 20 Participants of the DASH2 Clinical Trial (part III)
STUDY_SUMMARY
The objective of the study is to identify changes of urinary metabolite profiles associated with different responses to blood pressure to salt. Subjects are derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. 2001).We choose two groups subjects who meet the following conditions(the two groups are separately named A and B). We chose subjects on the Control diet .These subjects meet the blood pressure criteria described below:Group A subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic blood pressure from the low sodium visit is less than 140 and the diastolic blood pressure from low sodium visit is less than 90; 3) For group A: Either the systolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the systolic blood pressure from the low sodium visit, or the diastolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the diastolic blood pressure from the low sodium visit; 3) For group B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. +/- 5) from the systolic blood pressure from the low sodium visit, and the diastolic blood pressure from the high sodium visit is within 5 mmHg from the diastolic blood pressure from the low sodium visit. Use gas chromatography/mass spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry (LC/MS)analysis to find the differences of metabolic profiles between the high sodium level and the low sodium level, and compare the metabolic profiles of A with the metabolic profiles of B at the low and high sodium level. Note, this was used as a pilot, the rest of the samples are in another uploaded study.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
TCA Cycle Metabolites of Dietary Salt Effects on Blood Pressure in Rat Urine (part IV)
STUDY_SUMMARY
We propose to analyze kidney tissue extract and urine samples from SS and SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the DASH2 trial. The analysis of the rat samples will be highly valuable for several reasons. First, it will to take the findings in human subjects back to animal models and prepare us for further mechanistic studies. We hypothesize at least some of the effects of dietary salt intake on metabolite profiles in human will be recapitulated or altered in the SS rat. If this is confirmed, we will have a highly informative animal model ready for mechanistic studies in which we can investigate the functional contribution of specific metabolites to hypertension and the mechanisms involved. Second, the rat study will allow us to take advantage of a new and unique transgenic SS.Fh1+ model that we recently developed that overexpresses fumarase (Fh1) on the genetic background of the SS rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced hypertension in SS rats.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Amino Acid Metabolites of Dietary Salt Effects on Blood Pressure in Rat Urine (part V)
STUDY_SUMMARY
We propose to analyze kidney tissue extract and urine samples from SS and SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the DASH2 trial. The analysis of the rat samples will be highly valuable for several reasons. First, it will to take the findings in human subjects back to animal models and prepare us for further mechanistic studies. We hypothesize at least some of the effects of dietary salt intake on metabolite profiles in human will be recapitulated or altered in the SS rat. If this is confirmed, we will have a highly informative animal model ready for mechanistic studies in which we can investigate the functional contribution of specific metabolites to hypertension and the mechanisms involved. Second, the rat study will allow us to take advantage of a new and unique transgenic SS.Fh1+ model that we recently developed that overexpresses fumarase (Fh1) on the genetic background of the SS rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced hypertension in SS rats.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Neuromodulator Metabolites of Dietary Salt Effects on Blood Pressure in Rat Urine (part VI)
STUDY_SUMMARY
We propose to analyze kidney tissue extract and urine samples from SS and SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the DASH2 trial. The analysis of the rat samples will be highly valuable for several reasons. First, it will to take the findings in human subjects back to animal models and prepare us for further mechanistic studies. We hypothesize at least some of the effects of dietary salt intake on metabolite profiles in human will be recapitulated or altered in the SS rat. If this is confirmed, we will have a highly informative animal model ready for mechanistic studies in which we can investigate the functional contribution of specific metabolites to hypertension and the mechanisms involved. Second, the rat study will allow us to take advantage of a new and unique transgenic SS.Fh1+ model that we recently developed that overexpresses fumarase (Fh1) on the genetic background of the SS rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced hypertension in SS rats.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
TCA Cycle Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine from DASH2 Clinical Trial (part VII)
STUDY_SUMMARY
The objective of the study is to identify changes of urinary metabolite profiles associated with different responses to blood pressure to salt. Subjects are derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. 2001).We choose two groups subjects who meet the following conditions(the two groups are separately named A and B). We chose subjects on the Control diet .These subjects meet the blood pressure criteria described below:Group A subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic blood pressure from the low sodium visit is less than 140 and the diastolic blood pressure from low sodium visit is less than 90; 3) For group A: Either the systolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the systolic blood pressure from the low sodium visit, or the diastolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the diastolic blood pressure from the low sodium visit; 3) For group B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. +/- 5) from the systolic blood pressure from the low sodium visit, and the diastolic blood pressure from the high sodium visit is within 5 mmHg from the diastolic blood pressure from the low sodium visit. Use gas chromatography/mass spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry (LC/MS)analysis to find the differences of metabolic profiles between the high sodium level and the low sodium level, and compare the metabolic profiles of A with the metabolic profiles of B at the low and high sodium level.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Amino Acid Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine from DASH2 Clinical Trial (part VIII)
STUDY_SUMMARY
The objective of the study is to identify changes of urinary metabolite profiles associated with different responses to blood pressure to salt. Subjects are derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. 2001).We choose two groups subjects who meet the following conditions(the two groups are separately named A and B). We chose subjects on the Control diet .These subjects meet the blood pressure criteria described below:Group A subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic blood pressure from the low sodium visit is less than 140 and the diastolic blood pressure from low sodium visit is less than 90; 3) For group A: Either the systolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the systolic blood pressure from the low sodium visit, or the diastolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the diastolic blood pressure from the low sodium visit; 3) For group B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. +/- 5) from the systolic blood pressure from the low sodium visit, and the diastolic blood pressure from the high sodium visit is within 5 mmHg from the diastolic blood pressure from the low sodium visit. Use gas chromatography/mass spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry (LC/MS)analysis to find the differences of metabolic profiles between the high sodium level and the low sodium level, and compare the metabolic profiles of A with the metabolic profiles of B at the low and high sodium level.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Neuromodulator Metabolites of Dietary Salt Effects on Blood Pressure in Human Urine from DASH2 Clinical Trial (part IX)
STUDY_SUMMARY
The objective of the study is to identify changes of urinary metabolite profiles associated with different responses to blood pressure to salt. Subjects are derived from The Dietary approaches to stop hypertension (DASH) diet, Sodium Intake and Blood Pressure Trial (Sacks FM et al PMID: 11136953,N Engl J Med. 2001).We choose two groups subjects who meet the following conditions(the two groups are separately named A and B). We chose subjects on the Control diet .These subjects meet the blood pressure criteria described below:Group A subjects conditions: 1) On Control diet. 2) Normotensive subjects: systolic blood pressure from the low sodium visit is less than 140 and the diastolic blood pressure from low sodium visit is less than 90; 3) For group A: Either the systolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the systolic blood pressure from the low sodium visit, or the diastolic blood pressure from the high sodium visit was greater than 10 mmHg higher than the diastolic blood pressure from the low sodium visit; 3) For group B: The systolic blood pressure from the high sodium visit is within 5 mmHg (i.e. +/- 5) from the systolic blood pressure from the low sodium visit, and the diastolic blood pressure from the high sodium visit is within 5 mmHg from the diastolic blood pressure from the low sodium visit. Use gas chromatography/mass spectrometry (GC/MS) analysis, and liquid chromatography/mass spectrometry (LC/MS)analysis to find the differences of metabolic profiles between the high sodium level and the low sodium level, and compare the metabolic profiles of A with the metabolic profiles of B at the low and high sodium level.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
TCA Cycle Metabolites of Dietary Salt Effects on Blood Pressure in Rat Urine and Kidney Tissue (part X)
STUDY_SUMMARY
We propose to analyze kidney tissue extract and urine samples from SS and SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the DASH2 trial. The analysis of the rat samples will be highly valuable for several reasons. First, it will to take the findings in human subjects back to animal models and prepare us for further mechanistic studies. We hypothesize at least some of the effects of dietary salt intake on metabolite profiles in human will be recapitulated or altered in the SS rat. If this is confirmed, we will have a highly informative animal model ready for mechanistic studies in which we can investigate the functional contribution of specific metabolites to hypertension and the mechanisms involved. Second, the rat study will allow us to take advantage of a new and unique transgenic SS.Fh1+ model that we recently developed that overexpresses fumarase (Fh1) on the genetic background of the SS rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced hypertension in SS rats.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Amino Acid Metabolites of Dietary Salt Effects on Blood Pressure in Rat Urine and Kidney Tissue (part I)
STUDY_SUMMARY
We propose to analyze kidney tissue extract and urine samples from SS and SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the DASH2 trial. The analysis of the rat samples will be highly valuable for several reasons. First, it will to take the findings in human subjects back to animal models and prepare us for further mechanistic studies. We hypothesize at least some of the effects of dietary salt intake on metabolite profiles in human will be recapitulated or altered in the SS rat. If this is confirmed, we will have a highly informative animal model ready for mechanistic studies in which we can investigate the functional contribution of specific metabolites to hypertension and the mechanisms involved. Second, the rat study will allow us to take advantage of a new and unique transgenic SS.Fh1+ model that we recently developed that overexpresses fumarase (Fh1) on the genetic background of the SS rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced hypertension in SS rats.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Neuromodulator Metabolites of Dietary Salt Effects on Blood Pressure in Rat Urine and Kidney Tissue (part XII)
STUDY_SUMMARY
We propose to analyze kidney tissue extract and urine samples from SS and SS.Fh1+ transgenic rats in addition to the analysis of urine samples from the DASH2 trial. The analysis of the rat samples will be highly valuable for several reasons. First, it will to take the findings in human subjects back to animal models and prepare us for further mechanistic studies. We hypothesize at least some of the effects of dietary salt intake on metabolite profiles in human will be recapitulated or altered in the SS rat. If this is confirmed, we will have a highly informative animal model ready for mechanistic studies in which we can investigate the functional contribution of specific metabolites to hypertension and the mechanisms involved. Second, the rat study will allow us to take advantage of a new and unique transgenic SS.Fh1+ model that we recently developed that overexpresses fumarase (Fh1) on the genetic background of the SS rat. Fumarase is a TCA cycle enzyme previously implicated in salt-induced hypertension in SS rats.
INSTITUTE
Mayo Clinic
LAST_NAME
Liang
FIRST_NAME
Mingyu
ADDRESS
Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226
Targeted NEFA in American Indian Adolescents (part I)
STUDY_SUMMARY
The goal of this study is to determine the effect of physical activity on the blood concentrations of several compounds that are proposed to be markers of diabetic risk. We will compare results from normal weight and obese American Indian adolescents, with high or low habitual physical activity. We will also compare the same blood markers in the obese group before and after 4 months of exercise to measure the benefit of the program.
INSTITUTE
Mayo Clinic
LAST_NAME
Short
FIRST_NAME
Kevin
ADDRESS
University of Oklahoma Health Sciences Center 1200 Childrenās Ave, Suite 4500 Oklahoma City, OK 73104
Targeted Amino Acids in American Indian Adolescents (part II)
STUDY_SUMMARY
The goal of this study is to determine the effect of physical activity on the blood concentrations of several compounds that are proposed to be markers of diabetic risk. We will compare results from normal weight and obese American Indian adolescents, with high or low habitual physical activity. We will also compare the same blood markers in the obese group before and after 4 months of exercise to measure the benefit of the program.
INSTITUTE
Mayo Clinic
LAST_NAME
Short
FIRST_NAME
Kevin
ADDRESS
University of Oklahoma Health Sciences Center 1200 Childrenās Ave, Suite 4500 Oklahoma City, OK 73104
Effects of Exercise on Dystrophic Mouse Muscle Amino Acids (part II)
STUDY_SUMMARY
We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to characterize the skeletal and cardiac muscle metabolomes. Half of the mice in each group will remain sedentary while the other half will be subjected to a single bout of treadmill exercise to exhaustion. Mice will be euthanized immediately postexercise and blood, hearts, and hindlimb muscles will be harvested and frozen as detailed in the General Methods. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of Exercise on Dystrophic Mouse Muscle TCA Cycle (part II)
STUDY_SUMMARY
We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to characterize the skeletal and cardiac muscle metabolomes. Half of the mice in each group will remain sedentary while the other half will be subjected to a single bout of treadmill exercise to exhaustion. Mice will be euthanized immediately postexercise and blood, hearts, and hindlimb muscles will be harvested and frozen as detailed in the General Methods. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of Exercise on Dystrophic Mouse Muscle Non-Esterified Fatty Acids (part III)
STUDY_SUMMARY
We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to characterize the skeletal and cardiac muscle metabolomes. Half of the mice in each group will remain sedentary while the other half will be subjected to a single bout of treadmill exercise to exhaustion. Mice will be euthanized immediately postexercise and blood, hearts, and hindlimb muscles will be harvested and frozen as detailed in the General Methods. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of NO Donor Therapy on the Dystrophic Mouse Muscle Amino Acids (part IV)
STUDY_SUMMARY
For this aim, we will only use male mdx mice. We will study three groups treated for 7 days with vehicle, naproxcinod (i.e., NO-naproxen), or naproxen (n = 10 each group). Two hours after the final treatment, half the mice in each group will be run to exhaustion on a treadmill. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of NO Donor Therapy on the Dystrophic Mouse Muscle Non-Esterified Fatty Acids (part V)
STUDY_SUMMARY
For this aim, we will only use male mdx mice. We will study three groups treated for 7 days with vehicle, naproxcinod (i.e., NO-naproxen), or naproxen (n = 10 each group). Two hours after the final treatment, half the mice in each group will be run to exhaustion on a treadmill. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of NO Donor Therapy on the Dystrophic Mouse Muscle TCA Cycle (part VI)
STUDY_SUMMARY
For this aim, we will only use male mdx mice. We will study three groups treated for 7 days with vehicle, naproxcinod (i.e., NO-naproxen), or naproxen (n = 10 each group). Two hours after the final treatment, half the mice in each group will be run to exhaustion on a treadmill. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of NO Donor Therapy on the Dystrophic Mouse Heart Metabolome (part VII)
STUDY_SUMMARY
For this aim, we will only use male mdx mice. We will study three groups treated for 7 days with vehicle, naproxcinod (i.e., NO-naproxen), or naproxen (n = 10 each group). Two hours after the final treatment, half the mice in each group will be run to exhaustion on a treadmill. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS).
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of NO Donor Therapy on the Dystrophic Mouse Quadricep Metabolome (part VIII)
STUDY_SUMMARY
For this aim, we will only use male mdx mice. We will study three groups treated for 7 days with vehicle, naproxcinod (i.e., NO-naproxen), or naproxen (n = 10 each group). Two hours after the final treatment, half the mice in each group will be run to exhaustion on a treadmill. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS).
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of Exercise on Dystrophic Mouse Heart Meabolome (part IX)
STUDY_SUMMARY
We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to characterize the skeletal and cardiac muscle metabolomes. Half of the mice in each group will remain sedentary while the other half will be subjected to a single bout of treadmill exercise to exhaustion. Mice will be euthanized immediately postexercise and blood, hearts, and hindlimb muscles will be harvested and frozen as detailed in the General Methods. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Effects of Exercise on Dystrophic Mouse Quadricep Meabolome (part X)
STUDY_SUMMARY
We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to characterize the skeletal and cardiac muscle metabolomes. Half of the mice in each group will remain sedentary while the other half will be subjected to a single bout of treadmill exercise to exhaustion. Mice will be euthanized immediately postexercise and blood, hearts, and hindlimb muscles will be harvested and frozen as detailed in the General Methods. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates.
INSTITUTE
Mayo Clinic
LAST_NAME
Thomas
FIRST_NAME
Gail
ADDRESS
Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033
Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice (part II)
STUDY_SUMMARY
In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (Ļ6) and omega-3 (Ļ3) fatty acids (linoleic and Ī±-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice (part III)
STUDY_SUMMARY
In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (Ļ6) and omega-3 (Ļ3) fatty acids (linoleic and Ī±-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
mTOR regulates metabolic adaptation of APCs in the lung microenvironment and controls the outcome of allergic inflammation
STUDY_SUMMARY
Antigen presenting cells (APCs) occupy diverse anatomical tissues, but their tissue-restricted homeostasis remains poorly understood. Here, working in mouse models of inflammation, we found that mTOR-dependent metabolic adaptation was required at discrete locations. mTOR was dispensable for DC homeostasis in secondary lymphoid tissues, but necessary to regulate cellular metabolism and accumulation of CD103+ DCs and alveolar macrophages in lung. Moreover, whilst numbers of mTOR-deficient lung CD11b+ DCs were not changed, they were metabolically reprogrammed to skew allergic inflammation from eosinophilic Th2 to neutrophilic Th17 polarity. The mechanism for this change was independent of translational control, but dependent on inflammatory DC which produced IL-23 and increased fatty acid oxidation. mTOR therefore mediates metabolic adaptation of APCs in distinct tissues, influencing the immunological character of allergic inflammation.
INSTITUTE
Emory University
LAST_NAME
Li; Gardinassi
FIRST_NAME
Shuzhao; Luiz
ADDRESS
Emory Woodruff Memorial Research Building, 1639 Pierce Dr NE, Atlanta, GA 30322
Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice (part IV)
STUDY_SUMMARY
In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (Ļ6) and omega-3 (Ļ3) fatty acids (linoleic and Ī±-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Changes in metabolites and lipid mediators associated with supervised exercise training for peripheral
STUDY_TYPE
Timecourse
STUDY_SUMMARY
Peripheral artery disease (PAD) is a leading cause of cardiovascular related morbidity and mortality, affecting over 8.5 million men and women in the United States and greater than 200 million individuals worldwide. The mainstay of treatment to improve lower limb symptoms is supervised walking therapy, which does not affect plaque morphology or alter conduit artery blood flow, but rather ameliorates endothelial dysfunction, enhances skeletal muscle metabolism and mitochondrial function, and suppresses inflammatory activation. In this pilot feasibility project we will employ metabolic and lipidomic techniques to measure the effects of supervised exercise therapy on primary metabolism, complex lipids, and lipid mediators, and correlate these effects with individual, subject-level measures of the response to exercise therapy among subjects with PAD. The overarching theme of this work is to identify metabolites, complex lipids, and lipid mediators that are associated with the inter-individual variability in the response of subjects with PAD to supervised exercise therapy. This knowledge will significantly enhance our understanding of the pathophysiology of lower extremity symptoms in PAD, as well as the manner in which supervised exercise therapy improves walking intolerance. It will identify novel therapeutic targets and pathways for pharmacologic manipulation in the treatment of PAD. Aside from having the potential to generate multiple high-impact publications, it will serve as the basis for a planned NIH R01 submission by the PI at the conclusion of the award period.
INSTITUTE
USDA
DEPARTMENT
Obesity and metabolism research unit
LAST_NAME
Newman
FIRST_NAME
John
ADDRESS
430 West Health Sciences Dr. Davis, Ca, 95616
EMAIL
john.newman@ars.usda.gov
PHONE
(530) 752-1009
STUDY_COMMENTS
Fatty acids measured but not against a calibration curve. Therefore values are expressed as a relative abundance of the entire samples set such that the sum of all subjects eguals 100%.
Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice (part I)
STUDY_SUMMARY
In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (Ļ6) and omega-3 (Ļ3) fatty acids (linoleic and Ī±-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Evaluation of plasma samples from the CHOICE clinical weight loss study in which obese post menopausal breast cancer survivors lost >15% initial body weight following either a low carbohydrate or a low fat dietary pattern. These samples will be interrogated for changes in metabolic intermediates and lipid metabolites that may be indicative of changes in prognosis for long term survival following treatment for breast cancer. Samples will be subjected to LCMS and shotgun lipidomics. In parallel, plasma and mammary gland fat pad from an obese rat model for breast cancer in which rats were subjected to a parallel level of weight loss will also be interrogated using the same procedures in an effort to inform the interpretation of the clinical data.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
The samples are frozen biopsies of whilte subcutaneous adipose tissues (50 mg). Samples were collected from 4 hormone-sensitive lipase (HSL) WT patients (adipose insulin sensitive), 3 HSL heterozygote patients and 1 HSL KO patient.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
These are fasted plasma samples collected from bipolar and control subjects following a 7-day diet journaling period. The purpose of the experiment is to evaluate potential differences in lipid concentrations/metabolism between bipolar and control subjects with the ability to correct for dietary intake, age, gender and medication use.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Rats were subjected to bilateral rotator cuff tears of the right and left supraspinatus muscle. Muscles were harvested from each shoulder at 0, 10, 30, or 60 days post surgery.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Determining lipid composition of diabetic microvascular complication-prone tissues and comparing tissues levels to plasma levels. Samples are in addition to plasma and kidney tissue samples ran (shotgun lipidomics) in June-July 2014.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Subjects were subjected to different dietary interventions. Indivduals (A1 to A19) were either randomized to a isocaloric diet with either 40-45% saturated fat diet or 40-45% monounsaturated fat diet for 4 weeks. Plasma was sampled at baseline and after 4 weeks on the diet in the fasting state. Multiple clinical values (Weight, blood pressure, pulse,total cholesterol, HDL, LDL and triglycerides) and hyperinsulinemic euglycemic clamps were assessed. Alternatively (XS-3 to XS 43) were provided a diet that was 2000 calories above the estimated isocaloric state for 2 weeks. Multiple clinical values (Weight, blood pressure, pulse,total cholesterol, HDL, LDL and triglycerides) were determined along with body composition, insulin and glucose values.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Characterization of Retinal Exudates in Coats Disease - lipid profile
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The aim of our study is to characterize the retinal lipid exudates through lipidomic assays. Lipidomic analysis will quantify and identify the retinal lipid exudates and provide a `lipid profile? of the exudates. The characterization of the retinal lipids will help in further understanding the pathogenesis of Coat?s disease. It may allow us to identify and therapeutically target a metabolic pathway to prevent retinal exudation abnormal in Coat?s disease.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Wild type (a/a) agouti mouse dams were randomized to one of three diets (control, mediterranean, western) two weeks prior to pairing with an agouti (Avy/a) sire. Diet exposure continued through pregnancy and lactation. All pups were weaned onto the control diet and then followed for metabolic phenotyping measures to 10 months of age. Comprehensive phenotyping (body composition, CLAMS, blood draw) was completed at 2, 4, and 8 months, with an OGTT at 8 months. Weekly weights were also recorded. The study examines whether prenatal dietary exposure to high fat diets (HFD) and bisphenol a (BPA, those groups will not be tested in this pilot) impacts metabolic programming in offspring as measured by hepatic steatosis, serum hormone levels, and epigenetic changes in hepatic lipid metabolism genes.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
8
TOTAL_SUBJECTS
6
STUDY_COMMENTS
Shotgun lipidomics will add insight into the potentially changing lipid composition of membranes in offspring following different prenatal diet exposures as a means of assessing risk of steatosis and metabolic changes.
LCR and HCR rats (from Nathan Qi's 2010 study) were dissected at rest or following 10 minutes of exercise. Mitochondria were isolated from frozen gastrocnemius skeletal muscle. Samples consist of these isolated mitochondria suspended in 50 uL of mitochondrial isolation buffer (IBM containing sucrose, salts, etc; see Katie Overmyer's notebook E, p~23).
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
4
TOTAL_SUBJECTS
12
STUDY_COMMENTS
Recommend extracting entire sample suspension, NOT pelleting and removing supernatant. Quantities are likely to be small (~1-3mg tissue each?) Protein assays were performed to estimate quantities; see Katie's notebook or contact Charles for these protein concentration estimates, but also RECOMMEND SAVING RESIDUAL PELLET to allow further normalization.
Plasma, kidney, sciatic nerve, and retina samples collected from control (db/m) and diabetic (db/db) mice. Samples snap frozen and stored at -80. Plasma volume measured and tissues weighed for lipid extraction.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
6
TOTAL_SUBJECTS
31
STUDY_COMMENTS
Plasma and kidney samples were originally prepped in June 2014 and analyzed on our first TripleTOF. George Michailidis has requested they be re-analyzed to compare results between platforms. N/R #91-111 are new samples that will be prepared for this experiment (n = 40). WHEN ANALYZING: need to separate by tissue (P, K, N, R) for imputation of missing values. Want raw data and normalized to IS.
The mice were injected via tail vein with either shBmal1 adenovirus or shLacz adenocirus as control (n=5 for each group), following 10 days of 5% ethanol diet feeding. Then the mice were dissected and liver tissues were flash freezed. 25mg liver tissues from each mouse of the same group were pooled.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effects of rosiglitazone treatment on lipid composition
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Reprograming of 'white' to 'brite' adipocytes with higher oxidative capacity and improved endocrine function represents a potentially important approach to address the dysfunctional adipocyte phenotypes in obesity. We find that chronic treatment with the PPAR? agonist (rosiglitazone, 1 uM for 7days in vitro) in white human adipose tissue induced metabolic changes. Our trancriptome analysis showed that higher mitochondrial and peroxisomal fatty acid oxidation pathways and other genes involved in lipid metabolism including (re)esterification are induced by rosiglitazone treatment. To understand the biochemical basis of brite vs. white human adipocytes, we will perform comprehensive metabolomic profiling of control and rosiglitazone treated tissues using unbiased lipidomics approach.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effect of high fat diet and streptozotocin treatment on neuropathy
STUDY_TYPE
MS analysis
STUDY_SUMMARY
C57BL6 male mice were fed either a control 10% fat (RD D12540-B) or a high fat 60% fat (RD 12492) diet from 5 wk age. STZ was given to some groups at 12 wk age. Chow was reversed in some groups (DR) from 60% HF to 10 % HF at 16 wk age. Mice were euthanized at either 16 wk or 24 wk age. Neuropathy phenotyping occured prior to all harvests.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Nonalcoholic steatohepatitis (NASH) changes with S.Q. Leptin administrations
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Samples are from individuals with documented fatty liver disease (NAFLD) from whom serum was drawn at baseline and after 1 and 12 months of S.Q. Leptin administrations. The effect of leptin on degree of steatosis and inflammation was assessed in liver biopsies at baseline and following 1 year of treatment.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Functional genomics study on Non Alcoholic fatty liver disease (NAFLD)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
In this experiment, vector , GCKR (WT), GCKR(P446L), PPP1R3B, TM6SF2 (WT and E167K) overexpression stable HuH-7_Lok cell lines as well as Non-target, GCKR shRNA (52621), PPP1R3B shRNA (2640), TM6SF2 shRNA (382426) knockdown HuH-7_Lok cell lines are used to do the lipidomics experiments 3 X 100 mm dishes of cells were seeded (1x106 cells/dish) for each cell line and incubated at 37? C for 24 hours, then replaced medium with delipidated serum DMEM medium. After 24 hours, treated with BSA-OA (200?M) for another 24 hours before the collection of cells Add 4mL/dish 50 mM ammonium acetate wash cells and then quench cells by adding liquid nitrogen covering all the surface of dish. Store the cell samples in -80?C freezer.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Functional genomics study on Non Alcoholic fatty liver disease (NAFLD) - part II
STUDY_TYPE
MS analysis
STUDY_SUMMARY
In this experiment, vector , GCKR (WT), GCKR(P446L), PPP1R3B, TM6SF2 (WT and E167K) overexpression stable HuH-7_Lok cell lines as well as Non-target, GCKR shRNA (52621), PPP1R3B shRNA (2640), TM6SF2 shRNA (382426) knockdown HuH-7_Lok cell lines are used to do the lipidomics experiments 3 X 100 mm dishes of cells were seeded (1x106 cells/dish) for each cell line and incubated at 37? C for 24 hours, then replaced medium with delipidated serum DMEM medium. After 24 hours, treated with BSA-OA (200?M) for another 24 hours before the collection of cells Add 4mL/dish 50 mM ammonium acetate wash cells and then quench cells by adding liquid nitrogen covering all the surface of dish. Store the cell samples in -80?C freezer.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Functional genomics study on Non Alcoholic fatty liver disease (NAFLD) - part III
STUDY_TYPE
MS analysis
STUDY_SUMMARY
In this experiment, vector , GCKR (WT), GCKR(P446L), PPP1R3B, TM6SF2 (WT and E167K) overexpression stable HuH-7_Lok cell lines as well as Non-target, GCKR shRNA (52621), PPP1R3B shRNA (2640), TM6SF2 shRNA (382426) knockdown HuH-7_Lok cell lines are used to do the lipidomics experiments 3 X 100 mm dishes of cells were seeded (1x106 cells/dish) for each cell line and incubated at 37? C for 24 hours, then replaced medium with delipidated serum DMEM medium. After 24 hours, treated with BSA-OA (200?M) for another 24 hours before the collection of cells Add 4mL/dish 50 mM ammonium acetate wash cells and then quench cells by adding liquid nitrogen covering all the surface of dish. Store the cell samples in -80?C freezer.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
ARetinal tissue knockout to study efflux of cholesterol (AGCA1/G1 double KO - TY and LysM cre)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
ABCA1 and AGCG1, which is important gene to efflux the cholesterol from the cell/tissue, are knocked out in specific retinal tissue. T:RPE specific, L:macrophage specific.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Retinal tissue knockout to study efflux of cholesterol (AGCA1/G1 double KO - TY and LysM cre) - part II
STUDY_TYPE
MS analysis
STUDY_SUMMARY
ABCA1 and AGCG1, which is important gene to efflux the cholesterol from the cell/tissue, are knocked out in specific retinal tissue. R:Rod specific, C:Cone specific.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Untargeted Lipidomics for hepatic lipid profile wild type versus knockout
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Use untargeted lipidomics to investigate differences in hepatic lipid profile between wild type and knockout mice. Mice were fed with high fat diet for 10 weeks and sacrificed under randomly fed condition. Liver were harvested freshly and frozen into liquid nitrogen immediately. Each sample was combined liver tissues from three individual mice in the same group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effect of fatty acids on macrophage (wild type versus knockout) lipid levels
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Macrophages from wild type or knockout mice with various treatments [none, BSA (control), palmitate + oleate, conditioned media from adipocytes treated with either BSA or palmitate + oleate). N=4/group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
On day 1, seed 1 million cells/dish Huh7_Lok cells (vector, GCKR(WT), GCKR(P446L), Non-Target, GCKR KD and PPP1R3B/OE) in 100 mm dishes with DMEM complete medium and incubate at 37C for 24 h. On day 2, replace DMEM medium with delipidated serum medium for these cells and incubate for another 24 h. On day 3, add heated Oleic Acid-Albumin into each dish at final concentration of 200?M for 24h. On day 4, collect cells with Trypsin/EDTA lysis and count cells with automated cell counter and calculate total cell numbers in each sample, then centrifuge cells down and take out the supernatant and keep cells in -80 C freezer.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Lipodomics and Gestational bisphenol A (BPA) exposure
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Gestational BPA exposure in sheep induces metabolic phenotype in sheep characterized by peripheral insulin resistance and increased adipocyte size. Because insulin sensitivity can be regulated by various agents including free fatty acids (FFA), we hypothesize that gestational BPA exposure alters circulating FFA inducing dyslipidemia, a marker of metabolic disorder. Because saturated FFA is associated with insulin resistance, determination of the FFA profile in these species aid in understanding the underlying mechanism. Additionally, because of the non-monotonic nature of responses to BPA exposure dose response studies are also needed. This study will therefore assess plasma lipid profile in sheep exposed to three different doses of BPA prenatally and compared with untreated control animals
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
4
TOTAL_SUBJECTS
28
STUDY_COMMENTS
Metabolic disorders such as obesity and diabetes are currently widespread with epidemic proportions. Recent studies have implicated that these disorders have developmental basis due to maternal exposure to adverse insults including endocrine disruptors such as Bisphenol A (BPA). As BPA is present in maternal serum, amniotic fluid, cord blood taken at birth, placenta, colostrum and breast milk suggests that BPA has developmental impacts. In fact developmental BPA exposure have been linked to intrauterine growth restriction and low birth weight offspring, risk factors for adult cardiometabolic abnormalities. Animal models are helpful to study the effect of developmental BPA exposure and determine associated mechanisms.
Biomarkers for different types of Multiple Scelerosis (MS) (BMS vs SPMS lipidomics)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Metabolomic Analysis in Secondary Progressive and Benign MS. Preliminary data: We have conducted whole blood cell microarray comparing gene expression profile of 20 SPMS and 13 BMS patients. The data revealed significant increase in pathways involving iron ion binding, oxygen transporter activity and hemoglobin functions. Specifically, the identified down regulated genes are involved in interacting selectively and non-covalently with iron (Fe) ions. Heme has been described as a potent pro-inflammatory molecule that can induce multiple innate immune responses, and cause excessive iron and heme-induced oxidative stress and cell death. In the brain, it causes neurodegeneration. In addition, our microarray preliminary results also show a statistically significant increase in arachidonate 12-lipoxygenase activity in SPMS compared to benign MS. Specific aim. To validate metabolites and lipid biomarkers in serum samples from patients with benign and secondary progressive MS. Our central hypothesis is that the gene expression in various types of multiple sclerosis likely create a systematic metabolomic/lipidomic environment that leads to progression in MS. We plan to compare and contrast metabolomics in SP-MS to BMS patients using the innovative technology at the Metabolomics Research Core at the University of Michigan. By monitoring lipid and metabolite changes in SP and BMS patients, we aim to identify molecular processes and biomarkers of axonal damage for MS. Then, we plan to translate such metabolomic biomarkers to correlate with our rich clinical, immunological and imaging readouts. The goal of this project is to develop a blood-based test to differentiate SP-MS from BMS patients. To accomplish this, we will first identify the gene expression signature associated with of SP-MS. Then, we will correlate these gene expression changes with lipids and metabolite changes in the blood by metabolomic network analysis. Because metabolism is the final fingerprint of functionality and has been implicated in neurodegeneration, metabolomic network analysis can be used for refining the relationships between specific gene expression and metabolites and axonal damage in progressive MS. Ultimately, we hope to develop blood, CSF and stool-based assays, and use this comprehensive profile to predict susceptibility and progression of MS. This is a new and innovative idea, which will hope to lead to future diagnostic and therapeutic development in MS.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Stable clones of Control lentiviral (CLV) or lentiviral Noxa knockdown cells (NshLV7) (as described in Lowman et al, 2010) were counted at T-18hr and 60E6 cells were resuspended in 100ml of fresh complete medium. At T0 cells were counted again and 20E6 were collected into triplicate tubes. Cells were washed 1X with ice cold PBS and resuspended in 300ul ice cold methanol. Cells were then snap frozen in liquid nitrogen and stored at -80C. An additional 10E6 cells were collected for protein concentration and Western blot analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Human oral keratinocytes growth and proliferation based on culture medium volume
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We seeded cells at density of 500k/T75 and cultured them at two different medium volume, one at 30ml, the other at 15ml. Culture medium is composed of 1x EDGS and 0.06mM calcium. We changed medium every day for 30ml culture, and every two days for 15ml culture. We collected spent medium when we changed medium. The collected spent medium was filtered through low protein binding 0.22um pore size filter and then stored at -80 degree C. For OA and SMT, spent medium was collected on the same day of D5 and D9 after cell seeding as marked on the tubes. For GK samples, 15ml and 30ml cultures were collected when cells reached at around 90% confluence, that is D5 for 15ml and D6 for 30 ml culture. The numbers 151, 152 were duplicated cell cultures in two T75s, same for 301 and 302. One major differences among GK, OA, and SMT is GK was from frozen cells; OA and SMT were fresh cells that were never frozen. Blank15 and Blank30 were complete medium without cells incubating in incubator for two days for Blank15 and one day for Blank30. Medium was collected in the same manner as others.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolites produced by strains associated with inflammation
STUDY_TYPE
MS analysis
STUDY_SUMMARY
EXPERIMENT 1: identify commonalities and differences between species at late log phase. EXPERIMENT 2: identify commonalities and differences within a species in function of the media used for the subset of bacteria cultured in more than one media. EXPERIMENT 3: identify commonalities and differences within a species in function of the growth phase for the subset of strains for which more than one time point was taken during the growth phase. Factor 1: growth phase for this slow growers as defined by early, mid and late log phase. Factor 2:rich media used for growth which are either NOS or OMIZ supplemented or not of TPP (thiamine pyrophosphate) a required supplement for some strains. Factor 3: cell pellet and supernatant of culture in which bacetria excrete small molecules. Factor 4. the bacterial species. All those bacteria come from the same genus and have been isolated in function of inflmmatory conditions in human.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Lanthanide-mineral induced alteration of bile acid metabolism in a murine model of steatohepatitis
STUDY_TYPE
MS analysis
STUDY_SUMMARY
120 mice (equal number of males and females) were randomized into 3 equal groups. One group is on a high-fat "Western-style" diet (HFWD) alone, a second group is on HFWD supplemented with lanthanides and calcium, and a third control group is supplemented with calcium alone. At study termination (18 months) we will harvest hepatocytes and colonic enterocytes for evaluation of epithelial gene expression patterns. We will also harvest cecal contents and feces for microbial profiling by 16S rRNA pyrosequencing. For this small pilot proposal, we wish to add an untargeted metabolomic analysis component. We will harvest liver (right medial lobe with gall bladder), serum, and feces. We will assay representative liver/gall bladder samples (8 from the HFWD group and 8 from the lanthanide/calcium supplemented group). Remaining liver samples and the serum and feces will be archived for future investigation. Liver is being targeted first since both steatohepatitis and hepatocellular carcinoma were seen in our previous study in mice on HFWD. Additionally, alterations of bile acid profiles and bile acid metabolism have been associated with both steatohepatitis and hepatic cancers.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Differences in bile acids composition between ASCL5 knockout and floxed mice.
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Understand the differences in Bile Acid composition between knockout and floxed mice at each intestinal segment. Also difference in metabolites between these two groups at each level of the intestine.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Pilot metabolomics study of aromatase inhibitor associated arthralgias
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The goal of the experiment is untargeted metabolomic and shotgun lipidomic profiling of serum samples collected from 50 patients before patients initiated therapy with an aromatase inhibitor and again after 3 months of aromatase inhibitor therapy. Half of the patients discontinued treatment within 6 months because of development of arthralgias during therapy and half remained on therapy for at least 24 months without development of significant arthralgias. We plan to analyse effects of AI therapy on metabolomic and lipidomic profiles, and to investigate associations between changes in profiles and development of symptoms.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Brain-Immune system-Gut Interaction in Chronic Mild Stress (CMS +/- Lacto)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Mice were devided into three groups (Naive untreated, Stressed untreated, Stressed + Lacto). The stressed groups were subjected to unpredictable chronic mild stress for seven weeks. Three weeks into the protocol, the +Lacto groups were administered probiotic Lactobacillus daily.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Cytokines correlation with metabolomic profiling of psoriatic and normal skin
STUDY_TYPE
MS analysis
STUDY_SUMMARY
These experiments are cytokine stimulations (TNF, IL-17, IFNg) of three keratinocyte lines (HAHA, PAK and BS4). Aim is to determine the metabolic changes induced by these cytokines and correlate with metabolomic profiling of psoriatic, uninvolved and normal skin.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Human vitreous was collected in the OR at time of indicated surgery. No treatments were involved. Diagnoses are: epiretinal membranes (ERM); proliferative diabetic retinopathy (PDR); retinal detachment (RD)
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Differences in metabolism in mouse embryonic fibroblasts (control vs SCF-beta-TrCP knockout)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Mouse embryonic fibroblasts were grown in DMED supplemented with 10%FBS. The same number of cells were plated in eight 6cm dishes. When the cells were ~70% confluent, four dishes received ethanol (carrier) and the other four received tamoxifen (1uM final) to induce knockout. The cells and media were harvested 4 days later. The media harvested from control and knockout dishes will be compared with the starting medium. The cells were washed with cold PBS twice while they are attached to the dishes. The whole dish with cells attached were immediately frozen in dry ice and wrapped with aluminum foil and stored at -80C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
NIH 3T3 fibroblast cells with Ceftriaxone Treatment
STUDY_TYPE
MS analysis
STUDY_SUMMARY
NIH 3T3 cells were plated at 1.5million cells per 10cM dish into 12 10cm dishes. 24hrs post plating media was changed on all plates. At the time of media change, 4 dishes were treated with a final concentration of 50uM ceftriaxone (24hrs ceftriaxone n=4). 23 hrs post media change, 4 separate dishes were treated with 50uM ceftriaxone for 1hr (1hr ceftriaxone n=4). 4 dishes were untreated (0hr, basal, n=4). 24 hrs post media change, media was quickly removed from all 12 plates, each plate was rapidly washed with MilliQ H2O and rapidly frozen with liquid nitrogen poured directly into the dishes. All plates were transferred to dry ice and stored at -80 until shipped.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Short chain Fatty Acid (SCFA) analysis in HIV patients
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We hypothesize that a microbiome enriched with anaerobes in HIV leads to SCFA production and immunomodulatory effects. The goal of the experiment is evaluating bacterial metabolic pathways leading to production of SCFA. DLCO stands for Diffusing capacity of the lungs for carbon monoxide.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
The effect of acetate on intestinal microorganisms
STUDY_TYPE
MS analysis
STUDY_SUMMARY
8 week old B6 Male: HFD+ Glucose, 9 day feeding; Blood collection from tail every 3 days; mice were fed glycerol (control) or GTA with increasing concentrations; Day 1-3 0.1g/kg; Day 3-6 1g/kg; Day 6-9 6g/kg
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Plasma from young adult mice, fasted 4 hr prior to bleeding. Key comparisons: Snell dwarf vs Snell normal. GHRKO KO vs WT. PAPPA KO vs Het. And two factor ANOVA: LivGHR KO, LivGHR WT vs GHRKO and GHRWT. Consistency between Snell, GHRKO, and PAPPA will be a major theme in the analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effect of DASH Diet on Gut Microbiome (SCFA from stool)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
This research will examine the effect of the currently recommended DASH diet versus a Vegetarian DASH diet on the gut microbiome and risk for cardiovascular disease in pre-hypertensive obese African American women. We will define the gut microbiota profile, short chain fatty acid production, breath hydrogen/methane response, plasma lipopolysaccharide production and other biomarkers of inflammation in response to diet type in obese African American pre-hypertensive females at baseline and following placement on the traditional DASH plan or DASH Vegetarian diet. We will also define these parameters in African American women adhering to a Vegetarian or Vegan Diet. Our goal is to determine how the gut microbiome modulates host physiology and immune function in response to diet type. By evaluating the effect of a recommended DASH dietary pattern versus a Vegetarian DASH plan on the gut microbiome and its fermentation products, we aim to identify novel information about how these diet types strategically reduce cardiovascular disease risk through gut:microbiota:host interaction.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
5
TOTAL_SUBJECTS
13
STUDY_COMMENTS
We will define the gut microbiota profile, short chain fatty acid production, breath hydrogen/methane response, plasma lipopolysaccharide production and other biomarkers of inflammation in response to diet type in obese African American pre-hypertensive females at baseline and following placement on the traditional DASH plan or DASH Vegetarian diet. We will also define these parameters in African American women adhering to a Vegetarian or Vegan Diet. Our goal is to determine how the gut microbiome modulates host physiology and immune function in response to diet type. By evaluating the effect of a recommended DASH dietary pattern versus a Vegetarian DASH plan on the gut microbiome and its fermentation products, we aim to identify novel information about how these diet types strategically reduce cardiovascular disease risk through gut:microbiota:host interaction.
Hamsters were treated with Clindamycin, or a Saline, or nothing at all. After four days animals were sacrificed and their cecal contents were collected to record change in their Microbiome. Along with the Microbiome analysis we are also performing metabolomics analysis to understand the influence of change in the microbiome with the metabolic changes (if any). The cecal contents collected were frozen immediately (using liquid nitrogen) and was frozen in -80C until analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic effects of weight reduction of very low energy diet vs dietary counseling
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The goal of the study is to assess the relative efficacy of a very low energy diet (VLED) using liquid meal replacement vs. standard of care dietary counseling and education (DCE) on the metabolic effects of weight reduction in the obese, subfertile population and assess ovulation and time to conception in these women. Subfertile women were recruited for this study. They completed an OGTT at baseline and again after completing a very low energy diet intervention.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Microbial ecology of nontuberculous mycobacteria (NTM) in cystic fibrosis (CF) sputum
STUDY_TYPE
MS analysis
STUDY_SUMMARY
These are sputum samples collected from 8 individuals with cystic fibrosis during the course of routine clinical care. They were initially stored at 4C for up to 24 hours then stored at -80C. No processing has been done to the sputum prior to freezing. For each individual there are sputum samples collected both before and after the individual had his or her first positive sputum culture for nontuberculous mycobacteria (NTM). The individuals experienced different clinical courses after their infection.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Diabetic heart adrenergic signaling and metabolism (STZ & Isoproterenol, High Fat)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We are examing the effect of both diabetes and adrenergic activation on the metabolomic profile. STZ treatment was initiated 5 months before hearts were harvested. High fat/low fat diets were given for 13 weeks. All hearts were excised in the morning. For the STZ/isoproterenol experiment, mouse anesthesia was initiated with 4% isoflurane, and maintained with 2% and a homeothermic controller. After 5 min of equilibration, control and fasted mice were injected intraperitoneally with either saline (S) or 25ug/kg isoproterenol (I). After an additional 5 min, the whole heart was quickly excised, washed with ice-cold distilled water, dried off with a chem whipe, and flash frozen in liquid nitrogen. Hearts were stored at -80, and shipped using dry ice. The hearts from the high fat diet experiement were prepared in the same way, but no injection was given and total anesthesia time was 5min instead of 10min.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic profiling of cyst fluid from patients with Intraductal Pancreatic Mucinous Neoplasm
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The pancreatic cyst fluids were collected using a syringe aspiration of the cyst fluid immediately after surgical resection of the lesion. The cyst fluid were then aliquoted and stored in -80C freezer until ready to use. We plan on performing untargeted metabolomics analysis on these pancreatic cyst fluid to find potential biomarkers that correlate with histopathological assessment and clinical behavior of these cystic lesions and thus to guide clinical management of patients.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effect of genetic lesions on glioblastoma (NP, NPA, NPAI, NS)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The cells in this experiment are primary cells derived from tumors with various genetic lesions. The purpose of this experiment is to determine the effect of the genetic lesion IDH1m (mutant isocitrate dehydrogenase) on the metabolic state of the cell. Four replicates of NPA versus four replicates of NPAI suspension cell neurospheres will be compared in an untargeted manner to identify all possible metabolic differences between the two types of genetic lesions.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics of lean males given water, antibiotic cocktail (ANMV), or neomycin then exposed to ozone
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Mice were given either water, an antibiotic cocktail (ANMV), or Neomycin in their drinking water for two weeks to reduce bacterial load, and then exposed to air or to ozone (2ppm for 3hrs). 24 hours later, pulmonary mechanics and airway responsiveness was assessed, and blood was collected via cardiac puncture. Serum was isolated and immediately stored at -80 C for further analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomic Profiles of Recovery from Traumatic Brain Injury
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We hypothesize that differences between the metabolome of TBI patients achieving full recovery at 3 months post-injury and those with protracted recovery will provide insights into the biology of recovery and yield novel biomarkers for predicting protracted recovery. To test our hypothesis, we will perform a case-control study examining the metabolomic profile of 128 TBI patients, 18 years and older, who either have complete functional recovery at 3 months post-injury or have functional decline at 3 months post-injury. Subjects were selected from an ongoing prospective cohort of traumatic brain injury (Head Injury Serum Markers for Assessing Response to Trauma, HeadSMART. Funding for HeadSMART was provided by ImmunArray.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Replicate populations of Aedes albopictus were reared under diapause-inducing short day photoperiod (8h light: 16h dark) and diapause-averting long day photoperiod (16h light:8h dark). Eggs were collected from each replicate and snap-frozen 11d post-oviposition. We hope to characterize the metabolomic and lipidomic profiles of diapausing eggs relative to non-diapause eggs.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Analysis of SCFA in the fecal matter of wt and Orai1kO mice
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The gut microbiome participates in numerous physiological functions and is believed to be shaped by the host intestine. We report here an unsuspected prominent role of Orai1-mediated pancreatic acinar cell exocytosis in shaping the microbiome. Deletion of Orai1 in pancreatic acini of mature mice maintained on solid diet resulted in 60-70% mortality. The mice have severe bacterial burden with dysbiosis resulting in systemic inflammation and death. This occurs despite prominent Paneth cell hyperplasia and activation of the intestinal innate immune response. Secretion of and bacterial killing by pancreatic defensins are markedly reduced, resulting in microbiota with decreased diversity and increased pathogenic bacteria. Survival and weight gain are not rescued by digestive enzyme supplements, but are by broad-spectrum antibiotics and purified liquid diet. These findings reveal an unexpected profound role of pancreatic secretion in shaping the microbiome
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics has recently been used to document age-related changes in key metabolic pathways in laboratory animals and as a biomarker to predict age. We study the ecology, evolution, behavior, and physiology of wild North American red squirrels where we are able to follow individual squirrels across their lifetime from birth until death. We are beginning to document aging in this natural population and are interested in understanding whether there is a signature of aging using metabolomics. We collected plasma samples from the oldest female and male squirrels in our study population and also from an equivalent number of younger squirrels. We will use an untargeted approach to provide an assessment of what metabolites differ among very old and young squirrels. The aim of these analyses is to allow us to identify if the same metabolites that have been identified as biomarkers of advanced age from laboratory mice are also biomarkers of advanced age in red squirrels. After we complete these untargeted analyses, we aim to develop a metabolomics panel for this species so that we can use a more targeted approach to assess how the metabolic profiles of specific pathways (glucose/fatty acid metabolism, redox homeostasis) change with age in offspring exposed to increased perinatal stress in our study species.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
6
TOTAL_SUBJECTS
33
STUDY_COMMENTS
Metabolomics has recently been used to document age-related changes in key metabolic pathways in laboratory animals and as a biomarker to predict age. We study the ecology, evolution, behavior, and physiology of wild North American red squirrels where we are able to follow individual squirrels across their lifetime from birth until death. We are beginning to document aging in this natural population and are interested in understanding whether there is a signature of aging using metabolomics.
Metabolome, body composition, and muscle performance in children
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The purpose of this study is to determine potential relationships between plasma metabolome patterns and body composition in addition to muscle quality and muscular performance in children. Children ages 8 to 10 will complete body composition testing including BMI and skin fold measurements. An ultrasound machine will be used to determine cross-sectional area and echo intensity in the vastus lateralis and rectus femoris muscles as a measure of muscle quality. Isokinetic leg extension strength, power, and fatigability will be determined from a series of maximal knee extensions using an isokinetic dynamometer. An untargeted metabolomics test will be used on a single plasma sample from each subject in order to examine plasma metabolome
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
We intend to analyze 89 samples for Untargeted Metabolomics on patients with and without Polycystic Ovarian Syndrome(PCOS). Blood is drawn from patients at four different time points; 1st) 0 Timepoint at clamp 2nd) 270 Timepoint at clamp 3rd) 0 Timepoint at OGTT and 4th) 120 Timepoint at OGTT (a couple of samples were missing 120, but we included a 30 min sample and a 90 min sample and for some subjects OGTT samples are not included but not listed here). All samples are drawn in EDTA tubes at the Children's Hospital of Colorado. Collected blood is processed and frozen immediately at -80 degrees celcius.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
This is a double blind study, to measure short chain fatty acids in colons of mouse that have gone through bariatric surgery and the mouse that have gone through sham surgery (as control group). We would like to get a ratio of different types of fatty acids in the colons of these animals. This project looks at the effects of bariatric surgery on short chain fatty acids produced by microbiota ultimately the effects on the growth and suppression of breast cancer.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effects of bariatric and antibiotic treatment surgery on mammary carcinogenesis
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The purpose of this study is to understand the effects of bariatric surgery on mammary carcinogenesis in C3 Tag (1) mouse model. My animals have gone through antibiotic treatments in drinking water vs. no abx in their water. Also animals have been though vertical sleeve gasteroctomy (VSG) vs sham surgery vs no surgery group. It is expected that these surgeries will affect the tumor proliferation and growth. One potential explanation can be the changes of SCFAs following the bariatric surgery.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics of rats after bariatric or sham surgery
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We would like to examine the levels of amino acids in rats who have underwent bariatric (VSG, vertical sleeve gastrectomy) or sham (control) surgery. Plasma was collected after an overnight fast and once again after refeeding the rats for 2 hours.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
The aim of this study is to determine whether changes in CoA degradation affect the composition of the acylcarntine pool in the liver. 7 wild-type and 7 Nudt7 knockout male mice (10-16 weeks of age) on a mixed background were fasted for 24h and allowed to refeed for 12-14hours before removing the liver. The liver samples were flash-frozen and stored at -80C until used.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Microbial changes following fecal microbiota transplantation in patients with recurrent C. difficile infection
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Fecal samples were collected from patients with recurrent Clostridium difficile infection before and after treatment with fecal microbiota transplantation. Fecal samples were frozen immediately after collections, and later put into ~50mg aliquots for later analysis (at -80 C).
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Measuring SCFA in mice gut following colonization by C. difficile
STUDY_TYPE
MS analysis
STUDY_SUMMARY
3 cages of 5-8 week old C57B/6 mice were placed on the antibiotic cefoperazone for 10 days followed by 2 days of reovery on plain water. One cage remained on plain water the entire time. Of the three antibiotic treated cages. One was mock infected (uninfected control), one was infected with C. difficile strain VPI 10463, while the other was infected with C. difficile strain 630. Twenty-four hours after the infection the mice were eutanized and the ceal content was collected and snap frozen in liquid nitrogen. Samples were stored in the -80 until analysis.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Murine bile acid profiles during CDI R20291 infection
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We are interested in understanding the bile acid profiles in cecal content from mice challeneged with R20291 C. difficle spores. We are also interested in the bile acid profiles of mice that are administered Ursodiol (UDCA) or vehcile (corn oil) during C. difficile infection. In vitro Ursodiol (UDCA) has significant impacts on germination and growth of C. difficile R20291.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic change in macrophages after efferocytosis
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Study metabolic pathway alteration in macrophages after efferocytosis (when macrophages eating apoptotic cells). Peritoneal macrophages were purified and plated in the cell-culture dishes. After culturing overnight. Apoptotic cells were overlayed to macrophages for 2 hours, then un-engulfed cells were removed through washing. Macrophages were left in the plate for another 4 hours before being lifted from the plate.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic change in macrophages after efferocytosis (part II)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The goal of the experiment is to study metabolic pathway alteration in macrophages after efferocytosis (when macrophages eating apoptotic cells). Peritoneal macrophages were purified and plated in the cell-culture dishes. After culturing overnight. Apoptotic cells were overlayed to macrophages for 2 hours, then un-engulfed cells were removed through washing. Macrophages were left in the plate for another 4 hours before being lifted from the plate.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolite-phenotype link in X-linked Adrenoleukodystrophy (fibroblast cell culture)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Untargeted metabolomics of control, AMN and ALD patient-derived fibroblasts. Fibroblasts cultured in DMEM with 15%FBS and antibiotic (pen/strep) in 100mm petridishes (1million/dish). After 24h cultures supplemented with fresh media (DMEM+15%FBS+antibiotic) for 3hr
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolite-phenotype link in X-linked Adrenoleukodystrophy (part II)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Untargeted metabolomics of control, AMN and ALD patient-derived fibroblasts. Human Postmortem brain tissue were obtained from NICHD Brain Bank. Brain regions were directed by the neuropathologist at Henry Ford Health System and sample tissue were immediately frozen at -80C until shipping to the UMICH Core.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Human patients were enrolled in a bariatric weight loss clinic and followed. Patients were male and female, normoglycemic,pre-diabetic and diabetic and with and without neuropathy. One 500 ul aliquot will be submitted for both untargeted metabolomics and shotgun lipidomics.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics of intensive weight management clinic (IWMC) weight loss
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Human patients were enrolled in a weight loss clinic and followed. Patients were male and female, normoglycemic,pre-diabetic and diabetic and with and without neuropathy.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics of mice inoculated with human microbiota
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Mice were inoculated with human microbiota. Peripheral blood was collected using superficial temporal phlebotomy into a tube containing EDTA-K2. Tubes were then inverted 5 times and placed on ice for no longer than one hour. Samples were centrifuged at 1500rcf for 15 minutes. Supernatant was transferred and quantified with a pipette. Tubes were placed on dry ice for between 10 minutes to an hour andl stored at -80C until shipped on dry ice. All steps during processing were performed in batches containing mulitple treatment groups.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Blood was collected from the facial vein of humanized mice immediately prior to euthansia via CO2 inhalation. Blood sat at room temperature for 20 minutes, and then was centrifuged at 12000g for 10 minutes at 4 C. Serum was removed to a new eppendorf tube and immediately placed on ice, and then samples were subsequently stored at - 20 C until the present.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics of mice inoculated with human stool (part III).
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Mice were inoculated with human stool samples from donors in a high gene copy number group or low gene copy number group. After several weeks, cecal contents were collected and frozen at -80. Aliquots were made on dry ice and then stored at -80 until shipment on dry ice. Analysis of short chain fatty acids (SCFA) of cecal content was performed.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
BeWo cells were plated in 100mm x 20mm cell culture plates and allowed to attach and acclimate for 24 hours. Cell culture media was then replaced with media containing mono(2-ethylhexyl)phthalate (MEHP) at either 90 or 180 micromolar concentrations or vehicle control (DMSO, 0.05% by volume). Cells were exposed for 24 hours followed by collection of cell media which was flash frozen in liquid nitrogen and then placed on dry ice. Cells were washed with sodium acetate and then immediately flash frozen by addition of ~8mL of liquid nitrogen to tissue culture plate. All samples were stored at -80 degrees C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Pregnant rats were treated for 5 days with 100mg/kg/day DEHP or vehicle control (corn oil) via a wafer which the rats consumed orally. After the fifth treatment, rats were euthanized and uterine horns were dissected. Amniotic fluid was collected from the gestational sac via syringe and collected into 1.5mL tubes. Tubes were then stored at -80 celsius
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Pregnant rats were treated for 5 days with 100mg/kg/day DEHP or vehicle control (corn oil) via a wafer which the rats consumed orally. After the fifth treatment, rats were euthanized and uterine horns were dissected. Amniotic fluid was collected from the gestational sac via syringe and collected into 1.5mL tubes. Tubes were then stored at -80 celsius.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Pregnant mice were treated for 14 days (from GD 0.5-13.5) with 125mg/kg/day DEHP or vehicle control (corn oil) via oral gavage. After the last treatment, mice were euthanized and placenta were removed and flash frozen in liquid nitrogen. Placentas were then stored at -80 celsius.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Thermal modalities are commonly used in sports medicine to affect tissue healing. Cold therapy is commonly used modalities, but the metabolic changes to muscle after cooling are not known. The objective of this study is to look at the effect of cooling on muscle metabolites and gene expression. There are a total of 8 subjects in the study. Each subject had an ice cup cryotherapy treatment ("cool" samples) to one leg for 15 minutes, and the other leg served as the control ("cntrl" samples). Two hours after the application of cryotherapy, a biopsy was taken from each thigh muscle. Muscle was minced with scissors and quickly snap frozen in liquid nitrogen.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
10
TOTAL_SUBJECTS
8
STUDY_COMMENTS
We would like to run the TCA Plus Analysis on each of the samples. For the statistical analysis, we would normalize the cool side to the ctrl side, and look at the relative change in metabolite abundance as a result of the cooling procedure. only used in sports medicine to affect tissue healing.
Comparison of the metabolome and lipidome of wild type and mdx/mTR mice
STUDY_TYPE
MS analysis
STUDY_SUMMARY
There are two groups of mice in the study, wild type mice (WT) and mdx/mTR mice (mdx or mdx/mTR) that are a model of musclar dystrophy. We collected the tibialis anterior muscles from these mice (N=6 in each group), and would like to perform TCA-plus analysis and shotgun lipidomics analysis on the mice, and compare across the two groups.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Rat Retinal Detachment Metabolomics Timecourse (part II)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Detachments were created in adult male Brown-Norway rats (300?400 g; Charles River Laboratories, Wilmington, MA). Briefly, rodents were anesthetized with a 50:50 mix of ketamine/xylazine, and pupils were dilated with topical phenylephrine (2.5%) and tropicamide (1%). A 20-gauge microvitreoretinal blade was used to create a sclerotomy 2 mm posterior to the limbus, carefully avoiding lens damage. A subretinal injector (Glaser, 32-gauge tip; BD Ophthalmic Systems, Franklin Lakes, NJ) was introduced through the sclerotomy into the vitreous cavity and then through a peripheral retinotomy into the subretinal space. Sodium hyaluronate (10 mg/mL, Healon O.D Abbott Medical Optics, Uppsala, Sweden) was slowly injected to detach the neurosensory retina from the underlying RPE. In all experiments, approximately one third to one half of the neurosensory retina was detached. Detachments were created in the left eye. The right eye served as the control, with all the steps of the procedure performed except for introduction of the subretinal injector and injection of the sodium hyaluronate. At varying intervals after creation of the detachment, the animals were euthanatized, and the eyes were enucleated.The retina was then dissected away from the retinal pigment epithelium taking just the detached portion in those eyes experimentally detached. All experiments were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and the guidelines established by the University Committee on Use and Care of Animals of the University of Michigan. Detachments were created in adult male Brown-Norway rats (300?400 g; Charles River Laboratories, Wilmington, MA).
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
10
TOTAL_SUBJECTS
27
STUDY_COMMENTS
All experiments were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and the guidelines established by the University Committee on Use and Care of Animals of the University of Michigan. Detachments were created in adult male Brown-Norway rats (300?400 g; Charles River Laboratories, Wilmington, MA).
Metabolomics of cilia and modulation of renal microcirculation
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Explore and explain the mechanism of regulation of metabolism biomarkers to modulate renal microcirculation. 1 - let rats rest for 5 days. 2 - test basal glucose and plasma. Store plasma in -20C for one day then transfer to -80. 3 - inject STZ 50mg/kg ip. 4 - after 3 days, check for increased glucose and extract plasma for early stage diabetic sample >300 mg/dl. Store in -20C and then -80C. 5 - one month after initial injection, extract plasma and store in -20C and then -80C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Tubuloglomerular feedback and salt-sensitive hypertension
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Collect whole blood in tubes containing EDTA, gently invert tubes 10 times and cool as soon as practical on ice. Centrifuge to separate plasma as soon as practical, transfer to appropriately labeled tubes, and freeze immediately. I test basal glucose and plasma store plasma in -20C for one day then transfer to -80C. The next day, inject STZ 60 mg/kg ip. After 3 days, check for increased glucose and extract plasma for early stage diabetic sample >300 mg/dl. Store in -20C and then -80C. 30 days after initial injection, extract plasma and store in -20C and then -80C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
One year caloric restriction (CR) compared to ad lib (AL) feeding on metabolic and clinical parameters to assess association with intrinsic oxidative capacity. High and low capacity running rats (HCR and LCR) were randomized to ad lib feeding vs. 40% caloric restriction starting at 8 weeks of age. Experiment was continued for 52 weeks when blood and tissue were collected. Exercise capacity, CLAMS assessment of fuel utilization and clinical parameters were determined at 8 weeks, 8 months and 12 months of age. Tissue was collected in the fed (2 hours after re-feeding) or fasted stated all groups.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
N/NIH rats were selected for either low (LRT) or high (HRT) response to training. Rats at 2 year of age were trained for 8 weeks with an improvement in oxidative capacity. Selection was also performed for high (HCR) or low (LCR) intrinsic running capacity. Young and old animals are compared to 12 week old animals.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Plasma Nucleotide/adenosine concentrations in patiens with scleroderma depending on exercise
STUDY_TYPE
MS analysis
STUDY_SUMMARY
This experiment was performed using the following cohorts: 1) healthy controls, 2) patients with scleroderma at low risk for pulmonary hypertension, 3) pateints with scleroderma at high risk for pulmonary hypertension ("SSc Cath"). Whole blood was drawn directly into stop soilution (1:1 ratio) at rest and again at peak exercise for each human subject. The samples were processed to plasma by MCRU and stored at -80 in the Biorepository.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Untargeted metabolomics and lipidomics of scleroderma PAH discovery cohort
STUDY_TYPE
MS analysis
STUDY_SUMMARY
This experiment was performed using the following cohorts: 1) healthy controls, 2) patients with scleroderma at low risk for pulmonary hypertension, 3) pateints with scleroderma at high risk for pulmonary hypertension who underwent a catheterization and were found to have normal pressures, borderline elevated pressures or pulmonary arterial hypertensino (PAH). Whole blood was drawn directly into EDTA tubes at rest and again at peak exercise for each human subject. The samples were processed to plasma by MCRU and stored at -80 in the Biorepository.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Differences in metabolite profiles and antioxidant activities between wild and cultivated black soybeans evaluated by a correlation analysis
STUDY_SUMMARY
Non-targeted metabolic profiling of 26 soybean varieties using liquid chromatography-mass spectrometry (LC-MS) including 15 wild black soybeans (WBS) and 11 cultivated black soybeans (CBS), combined with multivariate analysis, revealed significant differences in 25 differential metabolites
INSTITUTE
KIST
LAST_NAME
Xu
FIRST_NAME
Jiuliang
ADDRESS
saimdang-ro 679, Gangneung, Gangwon, 213510, Korea, South
Metabolite analysis of regional neural stem/progenitor cells
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The developing mammalian brain generates a variety of Neural Progenitor Cells (NPCs). Primary NPCs throughout the neuraxis are derived from the ventricular zone. Intermediate progenitor cells (IPCs) are produced uniquely in the telencephalon and contribute extensively to the neurons that comprise the cerebral cortex and basal ganglia. It is known that the fate of the diverse NPC populations is determined by the interplay of transcription factors and regulation by regional humoral cues. However, despite our recent appreciation that nutrient-regulated intracellular metabolic milieu (pO2, energy, and redox state) significantly influence cell fate, an unexplored area is whether NPCs have intrinsic metabolic identity, and if so, the mechanism by which molecular metabolism contributes to brain development.Little is known however, if intrinsic differences in cellular metabolism of regional NPCs make certain NPCs susceptible while others resistant to genetic and environmental insults. We conjectured that regional (fore-and hindbrain) NPCs are metabolically distinct.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolites of human neurons and stem cells (siNeuron metabolomics)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
There are a total of four samples each for this analysis. Each sample consists of the cells grown on three 10 cm culture plates. Each plate should have 2x106 cells for a total of 6x106 cells per sample when all three plates are combined. The first sample is undifferentiated human embryonic stem cells, the second sample is human glutamatergic neurons derived from those human embryonic stem cells, the third sample is undifferentiated human induced pluripotent stem cells and the fourth sample is human glutamatergic neurons derived from those human induced pluripotent stem cells.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
From Lawson et al., Obesity 2015 (PMID: 25865294). Methods: A randomized, placebo-controlled crossover study of single-dose intranasal oxytocin (24 IU) in 25 fasting healthy men was performed. After oxytocin/placebo, subjects selected breakfast from a menu and were given double portions. Caloric content of food consumed was measured. Visual analog scales were used to assess appetite, and blood was drawn for appetite-regulating hormones, insulin, and glucose before and after oxytocin/placebo. Indirect calorimetry assessed resting energy expenditure (REE) and substrate utilization. Here, T0 refers to immediately before OXT or Placebo was administered (fasting), and T55 refers to 55 minutes post-administration (fasting, immediately pre-meal).
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Looking for changes in lipid levels with knock out of ACADM (shACADM)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
GSC 8-11 cells (20 million) were transduced with lentivirus coding for shACADM 4, 6, or scr (control). ACADM is an enzyme responsible for the metabolism of medium chain fatty acids (C4-C12). A round of selection was performed using puromycin to ensure only cells that had been transduced survived. Knockdown of ACADM was confirmed via western blot. Pellets were collected by centrifuging cells for 10 min at max speed. The pellets were washed and each sample was split into quadruplicate.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic effects of novel aldehyde dehydrogenase inhibitors (ALDH) inhibitors
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Effect of novel aldehyde dehydrogenase inhibitors. Before an experiment medium was replaced to medium without glucose with addition of labeled glucose on C13. Next, cells were treated with ALDH inhibitors 673 or 773 for 1, 4, or 8 hr (673&773-Jan26)
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Michigan Biomarkers for Refractory Depression (Bluebird) metabolomics pilot study
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The goal of the project is discovery of clinical useful biomarkers for treatment-resistant depression. All participants were undergoing electroconvulsive therapy (ECT) for treatment-resistant depression (major depressive disorder or bipolar disorder) in 2012-2014. All subjects were Caucasian except BB060, who was Asian. Samples were collected from each subject at baseline just before the first ECT treatment (Time Point T0) and approximately 3 weeks later just before the tenth ECT treatment (Time Point T2). Whole blood was drawn in the morning in a fasted state (at least 7 hours) through an intravenous catheter or butterfly needle into 6-mL EDTA tubes (Lavender Top, Becton Dickinson Vacutainer K2EDTA additive blood collection tube, part #367863) and transported to the processing lab at the Michigan Clinical Research Unit within 30 minutes. Plasma was isolated by centrifugation for 10 minutes at 2000G at 4C. Plasma from multiple tubes was pooled and aliquotted in 200 uL volumes into 2-mL screw-cap cryovials and frozen at -80C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic Adaptations to Chronic and Acute Exercise in Overweight Adults (ATX-Study) preliminary data
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The primary purpose of this study is to examine the effects of chronic exercise training and an acute session of exercise on key risk factors associated with Metabolic Syndrome (e.g., glucose tolerance, blood lipid profile, and blood pressure) and alterations in subcutaneous adipose tissue structure and metabolic function in overweight adults. Human adipose tissue samples collected before, and one hour after a 1h exercise session at ~65% VO2max (maximal oxygen uptake)
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Lipidomics pilot project for mice exposure to bisphenol A and high fat diets
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Wild type (a/a) agouti mouse dams were randomized to one of three diets (control, Mediterranean, western) two weeks prior to pairing with an agouti (Avy/a) sire. Diet exposure continued through pregnancy and lactation. All pups were weaned onto the control diet and then followed for metabolic phenotyping measures to 10 months of age. Comprehensive phenotyping (body composition, CLAMS, blood draw) was completed at 2, 4, and 8 months, with an OGTT at 8 months. Weekly weights were also recorded. The study examines whether prenatal dietary exposure to high fat diets (HFD) and bisphenol a (BPA, those groups will not be tested in this pilot) impacts metabolic programming in offspring as measured by hepatic steatosis, serum hormone levels, and epigenetic changes in hepatic lipid metabolism genes. Shotgun lipidomics will add insight into the potentially changing lipid composition of membranes in offspring following different prenatal diet exposures as a means of assessing risk of steatosis and metabolic changes.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Six strain/genotype combinations (BKS wt, BKS db/+, ?) were fed control (5LOD) or high fat (54 % lard) chow from 5 to 36 weeks of age. Longitudinal changes in small and large nerve fiber function, glucose tolerance, fasting blood glucose, body weight etc. were assessed in all groups
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Nuclear magnetic resonance-based metabolomics approach to evaluate the prevention effect of Camellia nitidissima Chi on colitis-associated carcinogenesis
STUDY_TYPE
Original Research
STUDY_SUMMARY
Colorectal cancer (CRC) is one of the most common malignant tumors worldwide, occurring in the colon or rectum portion of large intestine. With marked antioxidant, anti-inflammation and anti- tumor activities, Camellia nitidissima Chi has been used as an effective treatment of cancer. The azoxymethane/dextran sodium sulfate (AOM/DSS) induced CRC mice model was established and the prevention effect of Camellia nitidissima Chi extracts on the evolving of CRC was evaluated by gross examination, histopathological inspection, serum biochemistry analysis, combined with nuclear magnetic resonance (NMR)-based metabolomics and correlation network analysis. The results showed that Camellia nitidissima Chi extracts could significantly inhibit AOM/DSS induced CRC, relieve the colonic pathology and ameliorate the serum biochemistry, and could signiļ¬cantly reverse the disturbed metabolism towards the normal state. Moreover, the butanol fraction showed a better efficiency than the water-soluble fraction of Camellia nitidissima Chi. The study pave the way for further development of Camellia nitidissima Chi extracts as a potent CRC inhibitor.
Absolute Quantification of 180 metabolites in serum from african american and european american in prostate cancer and case control samples
STUDY_SUMMARY
Metabolite concentrations were obtained in prostate cancer and case control plasma samples from AA and EA individuals using the AbsoluteIDQ kit p180 (Biocrates Life Science AG, Austria) according to the manufacturerās instructions. The analysis was performed using a QTRAP 6500 LC/MS/MS System (AB SCIEX, USA) equipped with an electrospray ionization source, Agilent G1367B autosampler and the Analyst 1.51 software (AB SCIEX, USA).
metabolome in a group of AA and EA matched pairs of prostate cancer (PCa) and benign adjacent tissues
STUDY_SUMMARY
10 ĀµL of suspended samples was injected and analyzed using a 6495 triple quadrupole mass spectrometer (Agilent Technologies, Santa Clara, CA) coupled to a HPLC system (Agilent Technologies, Santa Clara, CA) via Multiple reaction monitoring (MRM) of a total of 240 endogenous water soluble metabolites for steady-state analyses of samples. Those 240 compounds monitored were chosen due to their involvement in central pathways important in a number of malignancies. Source parameters were as follows: Gas temperature was 250 Ā°C; Gas flow was 14 l/min; Nebulizer was 20psi; Sheath gas temperature was 350 Ā°C; Sheath gas flow was 12 l/min; Capillary was 3000 V positive and 3000 V negative; Nozzle voltage was 1500 V positive and 1500 V negative. Approximately 8ā11 data points were acquired per detected metabolite. Samples were delivered to the MS via normal phase chromatography using either a 4.6 mm i.d Ć10 cm Amide XBridge HILIC column (Waters) or a Luna 3Āµ NH2 100A (Phenomenex) at 300 ĀµL/min
An untargeted metabolomics approach was utilized to determine urinary metabolites that could serve as small molecule biomarkers for treatment response to standard tuberculosis treatment. However, the majority of metabolites that most accurately distinguished patient samples at time of diagnosis from those at one month after the start of therapy lacked structural identification. The detection of unknown metabolite structures is a well-known limitation of untargeted metabolomics, and underscores a need for continued elucidation of novel metabolite structures. In this study, we sought to define the structure of a urine metabolite with an experimentally determined mass of 202.1326 Da, classified as molecular feature (MF) 202.1326. A hypothesized structure of N1-acetylisoputreanine was developed for MF 202.1326 using in silico tools and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In the absence of a commercial standard, synthetic N1-acetylisoputreanine was generated using enzymatic and chemical synthesis and LC-MS/MS was used to confirm the structure of MF 202.1326 as N1-acetylisoputreanine, a proposed terminal polyamine catabolite that had not been previously detected in biological samples. Further analysis demonstrated that N1-acetylisoputreanine and an alternative form of this metabolite, N1-acetylisoputreanine-Ī³-lactam, are both present in human urine and are likely end-products of polyamine metabolism.
Identifying metabolic adaptations characteristic of multiple myeloma cells via mass spectrometry-based metabolite profiling from Peripheral Plasma
STUDY_SUMMARY
Will be assessing the untargeted metabolomic profiles of high risk versus low risk smoldering myeloma patients based on peripheral blood plasma, bone marrow plasma and Cd138+ plasma cells. Peripheral Plasma ms5972
INSTITUTE
Mayo Clinic
LAST_NAME
Gonsalves
FIRST_NAME
Wilson
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Identifying metabolic adaptations characteristic of multiple myeloma cells via mass spectrometry-based metabolite profiling from Bone Marrow Plasma
STUDY_SUMMARY
Will be assessing the untargeted metabolomic profiles of high risk versus low risk smoldering myeloma patients based on peripheral blood plasma, bone marrow plasma and Cd138+ plasma cells. Bone Marrow Plasma ms5973
INSTITUTE
Mayo Clinic
LAST_NAME
Gonsalves
FIRST_NAME
Wilson
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Identifying metabolic adaptations characteristic of multiple myeloma cells via mass spectrometry-based metabolite profiling from (CD138+ Plasma Cells)
STUDY_SUMMARY
Will be assessing the untargeted metabolomic profiles of high risk versus low risk smoldering myeloma patients based on peripheral blood plasma, bone marrow plasma and Cd138+ plasma cells. Cd138+ plasma cells ms5974
INSTITUTE
Mayo Clinic
LAST_NAME
Gonsalves
FIRST_NAME
Wilson
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Identifying metabolic adaptations characteristic of multiple myeloma cells via targeted sphingolipids concentrations from bone marrow and peripheral plasma
STUDY_SUMMARY
Will be assessing the targeted sphingolipids concentrations of high risk versus low risk smoldering myeloma patients based on peripheral blood plasma and bone marrow plasma.
INSTITUTE
Mayo Clinic
LAST_NAME
Gonsalves
FIRST_NAME
Wilson
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Identifying metabolic adaptations characteristic of multiple myeloma cells via amino acids concentrations from bone marrow plasma
STUDY_SUMMARY
Will be assessing the targeted amino acids concentrations of high risk versus low risk smoldering myeloma patients based on peripheral blood plasma and bone marrow plasma.
INSTITUTE
Mayo Clinic
LAST_NAME
Gonsalves
FIRST_NAME
Wilson
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Large Untargeted Profiling of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted FFA Composition of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted NEFA Panel of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted Cholesterol Profiling of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted Sphingolipids Panel of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted Galactosyl Sphingolipids Concentrations of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted Sphinomyelin Panel of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeted TCA Panel of Myelin to Enhance Recovery of Function after SCI
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Insights into myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) phenotypes through comprehensive metabolomics
STUDY_TYPE
Observational
STUDY_SUMMARY
The pathogenesis of ME/CFS, a disease characterized by unexplained debilitating fatigue, cognitive dysfunction, sleep disturbances, orthostatic intolerance, fever, lymphadenopathy and irritable bowel syndrome (IBS), is poorly understood. There are no validated diagnostic tests or interventions to mitigate disease. Here we report association modeling, biomarker discovery, biochemical enrichment analysis and topological network visualization of plasma metabolomic, fecal bacterial metagenomic and clinical data from 50 ME/CFS patients and 50 healthy controls. Through targeted and untargeted metabolomics analyses we confirm earlier reports of specific alterations in plasma levels of choline, carnitine and complex lipid metabolism in ME/CFS. We also demonstrate that patients with ME/CFS and IBS have a unique metabolomic profile that includes increased plasma levels of ceramide, a waxy lipid implicated in suppression of electron transport, insulin and leptin resistance and apoptosis. Integration of fecal metagenomic and plasma metabolomic data resulted in a stronger predictive model of ME/CFS (cross-validated AUC=0.836) than either metagenomic (cross-validated AUC=0.745) or metabolomic (cross-validated AUC=0.820) analysis alone. Our findings may provide insights into the pathogenesis of ME/CFS and ME/CFS subtypes, and suggest pathways for the development of diagnostic and therapeutic strategies.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
Key: mecfs : 1 in this column indicates case, while 0 indicates control Ibs: 1 in this column indicates the patient does have disease, 0 indicates free of ibs
Insights into myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) phenotypes through comprehensive metabolomics
STUDY_TYPE
Observational
STUDY_SUMMARY
The pathogenesis of ME/CFS, a disease characterized by unexplained debilitating fatigue, cognitive dysfunction, sleep disturbances, orthostatic intolerance, fever, lymphadenopathy and irritable bowel syndrome (IBS), is poorly understood. There are no validated diagnostic tests or interventions to mitigate disease. Here we report association modeling, biomarker discovery, biochemical enrichment analysis and topological network visualization of plasma metabolomic, fecal bacterial metagenomic and clinical data from 50 ME/CFS patients and 50 healthy controls.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
Key: mecfs : 1 in this column indicates case, while 0 indicates control Ibs: 1 in this column indicates the patient does have disease, 0 indicates free of ibs
Insights into myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) phenotypes through comprehensive metabolomics (part III))
STUDY_TYPE
Observational
STUDY_SUMMARY
The pathogenesis of ME/CFS, a disease characterized by unexplained debilitating fatigue, cognitive dysfunction, sleep disturbances, orthostatic intolerance, fever, lymphadenopathy and irritable bowel syndrome (IBS), is poorly understood. There are no validated diagnostic tests or interventions to mitigate disease. Here we report association modeling, biomarker discovery, biochemical enrichment analysis and topological network visualization of plasma metabolomic, fecal bacterial metagenomic and clinical data from 50 ME/CFS patients and 50 healthy controls. Through targeted and untargeted metabolomics analyses we confirm earlier reports of specific alterations in plasma levels of choline, carnitine and complex lipid metabolism in ME/CFS. We also demonstrate that patients with ME/CFS and IBS have a unique metabolomic profile that includes increased plasma levels of ceramide, a waxy lipid implicated in suppression of electron transport, insulin and leptin resistance and apoptosis. Integration of fecal metagenomic and plasma metabolomic data resulted in a stronger predictive model of ME/CFS (cross-validated AUC=0.836) than either metagenomic (cross-validated AUC=0.745) or metabolomic (cross-validated AUC=0.820) analysis alone. Our findings may provide insights into the pathogenesis of ME/CFS and ME/CFS subtypes, and suggest pathways for the development of diagnostic and therapeutic strategies.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
Key: MECFS: 1 in this column indicates case, while 0 indicates control IBS: 1 in this column indicates the patient does have disease, 0 indicates free of IBS
Insights into myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) phenotypes through comprehensive metabolomics (part IV)
STUDY_TYPE
Observational
STUDY_SUMMARY
The pathogenesis of ME/CFS, a disease characterized by unexplained debilitating fatigue, cognitive dysfunction, sleep disturbances, orthostatic intolerance, fever, lymphadenopathy and irritable bowel syndrome (IBS), is poorly understood. There are no validated diagnostic tests or interventions to mitigate disease. Here we report association modeling, biomarker discovery, biochemical enrichment analysis and topological network visualization of plasma metabolomic, fecal bacterial metagenomic and clinical data from 50 ME/CFS patients and 50 healthy controls. Through targeted and untargeted metabolomics analyses we confirm earlier reports of specific alterations in plasma levels of choline, carnitine and complex lipid metabolism in ME/CFS. We also demonstrate that patients with ME/CFS and IBS have a unique metabolomic profile that includes increased plasma levels of ceramide, a waxy lipid implicated in suppression of electron transport, insulin and leptin resistance and apoptosis. Integration of fecal metagenomic and plasma metabolomic data resulted in a stronger predictive model of ME/CFS (cross-validated AUC=0.836) than either metagenomic (cross-validated AUC=0.745) or metabolomic (cross-validated AUC=0.820) analysis alone. Our findings may provide insights into the pathogenesis of ME/CFS and ME/CFS subtypes, and suggest pathways for the development of diagnostic and therapeutic strategies.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
Key: MECFS: 1 in this column indicates case, while 0 indicates control IBS: 1 in this column indicates the patient does have disease, 0 indicates free of IBS
The goal is to look at measures of overall metabolic state in relationship to hormonal environment and measures of mood/depression or cognitive function (behavioral and functional MRI images). There are some relationships in this data set between glucose/insulin levels and cognitive function, and we would like to use the untargeted assays to further investigate this relationship. Factors related to visual/spatial memory are included in the design. Factors 1 and 2 (Left Hippocampus, Right Hippocampus) are levels of activation in the hippocampus during a visual memory task, (data missing for some of the samples). Factors 3-5 (BVMT % Retained, BVMT Learning T, BVMT Delay T) are results from a cognitive test of spatial memory function.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
The goal of this study was to characterize the metabolic impact manifested in mouse lung following acute ozone exposure, comparing differential effects experienced by lean and obese model mice.
Analysis of blood plasma lipid composition in patients with diabetic kidney disease In this untargeted lipidomics study we are comparing the lipidomics profile of progressors versus nonprogressors. A subgroup of patients have a follow up samples and therefore their baseline data are comared to the follow up visit data.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Dynamics of metabolism, proliferation and differentiation in Toxoplasma gondii mutants deficient in glycolysis and gluconeogenesis
STUDY_TYPE
Time course 13C labeling of Toxoplasma gondii using glucose and glutamine
STUDY_SUMMARY
The focus of this study was to profile the metabolic fates of glucose and glutamine in wild type, glycolytic mutant (hexokinase KO) and gluconeogenesis mutant (PEPCK1 KO) tachyzoite stage T. gondii parasites. 13C6-glucose and 13C5-15N1-glutamine were used as metabolic tracers. Metabolites from glycolysis, pentosephosphate pathway and Kerbs cycle were identified and their isotopomer abundance was quantified in order to visualize metabolic flux across the indicated pathways. A time course (from 0 minutes to 120 minutes) analysis was carried out in the labeling experiments. Here, host cell free extracellular tachyzoite stage T. gondii parasites were used.
INSTITUTE
CSIR-National Chemical Laboratory
DEPARTMENT
Biochemical Sciences Division
LABORATORY
Molecular Parasitology Laboratory
LAST_NAME
Shanmugam
FIRST_NAME
Dhanasekaran
ADDRESS
Dr. Homi Bhabha Road, Pune, 411008, Maharashtra, India
Integrated nutrigenomic and metabolomic analysis of Africans with variable diet
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Untargeted metabolic profiling and lipidomics profiling will be used to characterize a broad array of metabolites from plasma in 450 ethnically diverse individuals from Ethiopia, Tanzania, and Botswana with diverse diets. Mass spectrometry will be used to quantify metabolites previously found to be associated with cardiometabolic risk as well as the most informative metabolites from the untargeted screen. We will test for association of selected biomarkers with diet, geography, ancestry, and phenotypic variation. Metabolites obtained will be correlated with diet as well as clinical and anthropometric phenotypes. Using existing tools, we will assemble metabolites that associate with the various phenotypes into pathways and larger networks to provide insights into factors that relate to cardiometabolic health and disease. Finally, we will integrate metabolic and phenotypic data with genetic data from the SNP array and with high coverage whole genome sequence data. We will identify loci that play a role in local adaptation to diverse diets and will identify genetic variants associated with metabolite levels (mQTLs) and with the phenotypic traits listed above.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
43
TOTAL_SUBJECTS
450
STUDY_COMMENTS
Targeted mass spectrometry will be used to quantify metabolites previously found to be associated with cardiometabolic risk as well as the most informative metabolites from the untargeted screen. We will test for association of selected biomarkers with diet, geography, ancestry, and phenotypic variation. Metabolites obtained will be correlated with diet as well as clinical and anthropometric phenotypes. Using existing tools, we will assemble metabolites that associate with the various phenotypes into pathways and larger networks to provide insights into factors that relate to cardiometabolic health and disease. Finally, we will integrate metabolic and phenotypic data with genetic data from the SNP array and with high coverage whole genome sequence data. We will identify loci that play a role in local adaptation to diverse diets and will identify genetic variants associated with metabolite levels (mQTLs) and with the phenotypic traits listed above.
Looking for changes in lipid levels with knock out of ACADM (shACADM)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
GSC 8-11 cells (20 million) were transduced with lentivirus coding for shACADM 4, 6, or scr (control). ACADM is an enzyme responsible for the metabolism of medium chain fatty acids (C4-C12). A round of selection was performed using puromycin to ensure only cells that had been transduced survived. Knockdown of ACADM was confirmed via western blot. Pellets were collected by centrifuging cells for 10 min at max speed. The pellets were washed and each sample was split into quadruplicate.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
2 hydroxyglutarate prodution in neurospheres from IDH1 mouse glioma model #2
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Evaluation of 2HG and a-ketoglutarate in tumor neurospheres (NS) genertated from the mice brain tumors haboring specific genetic lesions: NRAS overexpression, p53 knockdown plus or minus IDH1 mutated and ATRX knockdown.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
We are interested in using both spontaneous and stimulated Raman microscopy to visualize these metabolomic changes as spectral alterations. We have two isogneic cell lines of normal human astrocytes differing only by a point mutation in the IDH-1 gene. We will work with the metabolomics core to elucidate the changes in central metabolism and lipid synthesys in an effort to determine the precise biochemical alterations underlying observed spectral differences. We wil then use a selective inhibitor of the IDH1 R132H to demonstrate to attempt to return TCA metabolome and lipidome to WT phenotype. Lastly, we will use a cell-permeablized variant of 2HG (2R-octyl-alpha-hydroxyglutarate) to recaptitulate the R132H mutant phenotype in wild-type cells, providing strong evidence that 2HG accumulation uderlies the metabolomic (and thus, spectral) changes observed.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
2
TOTAL_SUBJECTS
3
STUDY_COMMENTS
The IDH-1 R132H mutation in low grade gliomas preceptitates oncogenesis through a neomorphic enzyme function: the reduction of alpha-ketoglutarate (aKG) to 2-hydroxygutarate (2HG). 2HG accumukates to extremely high (~100mM) concentrations in IDH1 mutant cells and has substantial, predictable metabolic effects, including inhibition of a-KG dependent dioxygenases and hypermethylation of histones and chromatin.
We are interested in using both spontaneous and stimulated Raman microscopy to visualize these metabolomic changes as spectral alterations. We have two isogneic cell lines of normal human astrocytes differing only by a point mutation in the IDH-1 gene. We will work with the metabolomics core to elucidate the changes in central metabolism and lipid synthesys in an effort to determine the precise biochemical alterations underlying observed spectral differences. We wil then use a selective inhibitor of the IDH1 R132H to demonstrate to attempt to return TCA metabolome and lipidome to WT phenotype. Lastly, we will use a cell-permeablized variant of 2HG (2R-octyl-alpha-hydroxyglutarate) to recaptitulate the R132H mutant phenotype in wild-type cells, providing strong evidence that 2HG accumulation uderlies the metabolomic (and thus, spectral) changes observed.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolic profiling of cyst fluid from patients with Intraductal Pancreatic Mucinous Neoplasm
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The pancreatic cyst fluids were collected using a syringe aspiration of the cyst fluid immediately after surgical resection of the lesion. The cyst fluid were then aliquoted and stored in -80C freezer until ready to use. We plan on performing untargeted metabolomics analysis and unknown lipid analysis and identification on these pancreatic cyst fluid to find potential biomarkers that correlate with histopathological assessment and clinical behavior of these cystic lesions and thus to guide clinical management of patients. Abbreviations: IPMN - Intraductal Pancreatic Mucinous Neoplasm; MCN - mucinous cystic neoplasm; SCA - serous cystadenoma.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
SJGBM2 and MGG8 glioma cells were stable transfected with IDH1-R132H mutated. The expression of IDH1-R132H was confirmed by Western Blot assay. The aim of this project is analyze the production of 2HG in the stable transfected cells (SJGBM-IDH1m and MGG8-IDH1m) compared with the control group WT.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Human cord blood serum samples (200) were provided by David Christiani. The 200 study samples, total pool samples, and external CHEAR Plasma Reference Material samples were prepared and analyzed using the Biocrates AbsoluteIDQ p180 Kit.
Human cord blood serum samples (200) were provided by David Christiani. The 200 study samples, total pool samples, and external CHEAR Plasma Reference Material samples were prepared for NMR data collection and signals were library matched to metabolites to determine semi-quantitative concentration data using Chenomx NMR Suite 8.1.
INSTITUTE
RTI International
LAST_NAME
Fennell
FIRST_NAME
Timothy
ADDRESS
3040 E Cornwallis Rd, Durham, NC 27709
EMAIL
fennell@rti.org
PHONE
9194852781
TOTAL_SUBJECTS
200
STUDY_COMMENTS
Due to sample volume limitations: study samples, study pools, and CHEAR reference samples were prepared using 150 uL of sample and diluted with 100 ul of 0.9% Saline buffer. This is a deviation from the CHEAR Proficiency testing in which 400 ul of CHEAR reference sample was used and diluted with 300 uL of 0.9% Saline buffer. Samples were then transfered into a 3 mm NMR tube, in this study, as opposed to a 5 mm NMR tube for the Proficiency testing. Data for this study were acquired on a 600 MHz Bruker NMR spectrometer. Data for the procficiency testing were acquired on a 700 MHz Bruker NMR spectrometer.
Metabolic analysis of Normal Mouse Lung Fiboblasts with/without TGFbeta treatment
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Glycolysis/TCA/Nucleotide analysis (especially interested in alpha-ketoglutarate) and NAD+ and related metabolite analysis for parp1 ko and parp1 wild type lung tissue after saline or bleomycin treatment
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
We are interested in understanding the bile acid profiles from the feces of fecal microbiota transplant FMT patients that successfully recover from recurrent C. difficile infection. We have are submitting fecal samples from patients prior to their FMT and post FMT. We are also interested in the bile acid profiles of the donor stool that is used in successful transplants. Bile acids are important for C. difficile spore germination and outgrowth.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Detachments were created in adult male Brown-Norway rats (300?400 g; Charles River Laboratories, Wilmington, MA). Briefly, rodents were anesthetized with a 50:50 mix of ketamine/xylazine, and pupils were dilated with topical phenylephrine (2.5%) and tropicamide (1%). A 20-gauge microvitreoretinal blade was used to create a sclerotomy 2 mm posterior to the limbus, carefully avoiding lens damage. A subretinal injector (Glaser, 32-gauge tip; BD Ophthalmic Systems, Franklin Lakes, NJ) was introduced through the sclerotomy into the vitreous cavity and then through a peripheral retinotomy into the subretinal space. Sodium hyaluronate (10 mg/mL, Healon O.D Abbott Medical Optics, Uppsala, Sweden) was slowly injected to detach the neurosensory retina from the underlying RPE. In all experiments, approximately one third to one half of the neurosensory retina was detached. Detachments were created in the left eye. The right eye served as the control, with all the steps of the procedure performed except for introduction of the subretinal injector and injection of the sodium hyaluronate. At varying intervals after creation of the detachment, the animals were euthanatized, and the eyes were enucleated.The retina was then dissected away from the retinal pigment epithelium taking just the detached portion in those eyes experimentally detached. All experiments were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and the guidelines established by the University Committee on Use and Care of Animals of the University of Michigan.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
6.5 X 10^6 cells were plated in 10 cm Tissue Culture dishes and allowed to recover overnight at 37 degrees and 5% CO2. In the am, supernatant was removed, and 12 mL of medium containing either Murine Norovirus-1 (MNV) at an MOI=5 or medium containing a v/v match of mock lysate was added to the cells. Plates were rocked on ice for 1 hour, then cells were washed 3X with cold DPBS++, and plain medium as added to cells. Plates were then incubated for 7.5 hours at 37 degrees and 5% CO2. Cells were washed with 12 mL of 150 mM Ammonium Acetate, swirled 8 times, and immediately quenched with liquid nitrogen. Cells were then frozen at -80 degrees.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
2
TOTAL_SUBJECTS
1
STUDY_COMMENTS
BIOHAZARD - MURINE NOROVIRUS - SHOULD BE ENTIRELY NON-INFECTIOUS DUE TO THE LIQUID NITROGEN, BUT PLEASE HANDLE AND DISPOSE OF IN BIOHAZARD WASTE. IMPORTANT NOTE: I request that the metabolites of the TCA+ assay that are to be assessed quantitatively please be sure to include ATP, ADP, AMP, NAD, NADH, NADP, NADPH, E4P, S7P, 6PG, G3P H6P and Lactate. Also, I would request that both reduced and oxidized Glutathione as well as Fructose 1,6-bisphosphate be included in the non-quantitative data.
Purine and TCA measurements in salt sensitive (SS) hypertension
STUDY_TYPE
MS analysis
STUDY_SUMMARY
SS rats were surgically implanted with a chronic servo-control cuff, the purpose of which is to maintain normal pressure to the left kidney while the right kidney is exposed to high blood pressure. After 7 days of high salt treatment, which induces high blood pressure in the SS rat, rats were anesthetized with pentobarbital, kidneys were flushed and removed. The renal medulla was separated from the cortex using scissors, and the renal medullas were frozen in liquid nitrogen and stored at -80 C.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Deubiquitinase inhibitor compound C6 effect on cellular metabolism
STUDY_TYPE
MS analysis
STUDY_SUMMARY
6.0 X 10^6 cells were plated in 10 cm Tissue Culture dishes and allowed to recover overnight at 37 degrees and 5% CO2. In the am, supernatant was removed, and 12 mL of fresh medium with either 2.5 micromolar Compound C6 or a v/v match of DMSO (vehicle control) was added to the cells. Cells were incubated at 37 degrees in 5% CO2 for 30 minutes. Cells were washed 3X with cold DPBS++ and 10 mL of medium was added to the cells with either 1.0 micromolar Compound C6 or v/v match of DMSO. Cells were incubated at 37 degrees and 5% CO2 for 7.0 hours. Cells were washed with 10 mL of 150 mM Ammonium Acetate, swirled, washed again with 5 mL, and immediately quenched with liquid nitrogen. Cells were then frozen at -80 degrees.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
7
TOTAL_SUBJECTS
1
STUDY_COMMENTS
I request that the metabolites of the TCA+ assay that are to be assessed quantitatively please be sure to include ATP, ADP, AMP, NAD, NADH, NADP, NADPH, E4P, S7P, 6PG, G3P, H6P, and Lactate in particular. Also, I would request that both reduced and oxidized Glutathione as well as Fructose 1,6-bisphosphate be included in the non-quantitative data.
Pilot Human Muscle oral glucose tolerance test (OGTT)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Obesity is associated with metabolic inflexibility, the inability to switch from lipid to glucose oxidation during physiological conditions favoring excess glucose and insulin, which could contribute to the increased incidence of type 2 diabetes. Despite this knowledge, the role of skeletal muscle metabolites during these physiological conditions remain unclear and warrant further investigation. The purpose of this pilot study is to determine if: 1) there are differences in skeletal muscle glycolysis and TCA metabolites and amino acids between lean and obese individuals in the fasted state, and 2) if these metabolites change after 50 min of glucose ingestion, and 3) if there are differences in these metabolites between lean and obese individuals after 50 min of glucose ingestion.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
In the first group of samples, we are looking into the different metabolites and their concentrations within our human embryonic stem cells over the course of differentiation. Our goal is to study how the metabolism of stem cells are changing as they undergo this conversion to become hepatocyte-like cells. In the second group of samples labeled HyCell, we would like to understand the concentrations of metabolites within our CHO cells in order to update the parameters for our metabolic model. Included are two samples from different days of a suspension batch culture which may have different lactate production qualities.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Obesity and marrow and marrow adipose tissue (MAT) metabolomics
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Male C57Bl6j mice were fed HFD for 16-20 weeks and marrow adipose tissue (MAT) and bone marrow collected to evaluate lipid content/lipid type. In addition, marrow evaluations for glycolysis/TCA cycle and acylcarnitines will be performed.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Effect of cooling on muscle tissue metabolism (part II)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Thermal modalities are commonly used in sports medicine to affect tissue healing. Cold therapy is commonly used modalities, but the metabolic changes to muscle after cooling are not known. The objective of this study is to look at the effect of cooling on muscle metabolites and gene expression. There are a total of 8 subjects in the study. Each subject had an ice cup cryotherapy treatment ("cool" samples) to one leg for 15 minutes, and the other leg served as the control ("cntrl" samples). Two hours after the application of cryotherapy, a biopsy was taken from each thigh muscle. Muscle was minced with scissors and quickly snap frozen in liquid nitrogen.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
10
TOTAL_SUBJECTS
8
STUDY_COMMENTS
We would like to run the TCA Plus Analysis on each of the samples. For the statistical analysis, we would normalize the cool side to the ctrl side, and look at the relative change in metabolite abundance as a result of the cooling procedure. only used in sports medicine to affect tissue healing.
Diabetic heart adrenergic signaling and metabolism (STZ & Isoproterenol, High Fat)
STUDY_TYPE
MS analysis
STUDY_SUMMARY
We are examing the effect of both diabetes and adrenergic activation on the metabolomic profile. STZ treatment was initiated 5 months before hearts were harvested. High fat/low fat diets were given for 13 weeks. All hearts were excised in the morning. For the STZ/isoproterenol experiment, mouse anesthesia was initiated with 4% isoflurane, and maintained with 2% and a homeothermic controller. After 5 min of equilibration, control and fasted mice were injected intraperitoneally with either saline (S) or 25ug/kg isoproterenol (I). After an additional 5 min, the whole heart was quickly excised, washed with ice-cold distilled water, dried off with a chem whipe, and flash frozen in liquid nitrogen. Hearts were stored at -80, and shipped using dry ice. The hearts from the high fat diet experiement were prepared in the same way, but no injection was given and total anesthesia time was 5min instead of 10min.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Metabolomics of muscle in insulin sensitive and resistant obese indivduals.
STUDY_TYPE
MS analysis
STUDY_SUMMARY
"OC" samples: muscle biopsy samples collected from insulin sensitive / insulin resistant obese indivduals before undergoing a hyperinsulinemic-euglycemic clamp. Subjects were previously categorized for the rate of fatty acid release from adipose tissue into the bloodstream as a measure of insuline resistance. Low-FA, Med-FA, and High-FA stand for low, medium, and high rate of release.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
"PA" samples: matched plasma and muscle samples from an obese subject mild-intensity exercise training study, V1 is the first visit (untrained), V4 is post 3 months training, 1 day after exercise and V5 is post 3 months training, 3 days after last exercise.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statinsā varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naĆÆve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.
Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statinsā varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naĆÆve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.
Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statinsā varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naĆÆve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.
Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statinsā varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naĆÆve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.
Innovative and novel therapies are urgently needed for the treatment of patients with severe asthma, especially those who are refractory to standard-of-care bronchodilators and inhaled corticosteroids. The Zeki lab is investigating the role of the mevalonate (MA) pathway, in the pathogenesis of airway inflammation and remodeling. Although statins all inhibit HMGCR in the same manner in terms of enzyme binding site, the statinsā varied physiochemical properties with respect to their polarity (i.e. lipophilicity) result in very different immune and lipid effects. The major significance of this work is to advance a new class of inhaler therapies for asthma; the statins which work by an entirely different mechanism than current ICS/LABA mainstays. Evidence suggests that statins may have an additive benefit to corticosteroids in asthma, thereby confirming a unique mechanism, namely via MVA pathway inhibition. This becomes particularly important in the severe asthma population which is highly corticosteroid-resistant, is poorly controlled with high exacerbation rates and hospitalizations, and has the highest healthcare costs of all asthma phenotypes. In essence, the potential public health impact of even an incremental improvement in asthma symptom control cannot be underestimated. Even the prevention of 1 asthma attack preserves lung function and reduces the adverse personal and financial impact. This study aimed to determine if statin polarity affects airway drug concentration and systemic drug absorption and to determine the effect of inhaled statins on naĆÆve airway immune cell populations and alveolar-capillary membrane and epithelial barrier integrity in healthy rhesus monkeys. In this particular component of the study, we investigated the metabolic effects resulting from the use of statins in these healthy rhesus monkeys. Specifically, the Newman lab analyzed for lipid mediator (oxylipin, endocannabinoid, fatty acid, and nitro lipid) in lung and trachea tissue, plasma, and BAL and bile acid changes in the lung and trachea tissue and plasma.
Targeting Myelin NEFA of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
STUDY_SUMMARY
Targeting mouse myelin NEFA of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeting Myelin FFA Compostion of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
STUDY_SUMMARY
Targeting mouse myelin FFA composition of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeting Myelin Cholesterol of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
STUDY_SUMMARY
Targeting mouse myelin Cholesterol of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeting Myelin Sphingolipids of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
STUDY_SUMMARY
Targeting Myelin Sphingolipids of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeting Myelin Galactosyl Sphingolipids of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
STUDY_SUMMARY
Targeting mouse myelin Galactosyl Shingolipids of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
INSTITUTE
Mayo Clinic
LAST_NAME
Scarisbrick
FIRST_NAME
Isobel
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Targeting Myelin Sphinomyelin concentrations of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
STUDY_SUMMARY
Targeting Mouse Myelin Sphingomyelin concentrations of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Targeted FFA Composition in Kallikrein 6 Mice after SCI
STUDY_SUMMARY
Targeted FFA Composition in Kallikrein 6 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Urine metabolic profiles of pregnant women in relation to environmental exposures
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The purpose of this study is to broaden our understanding of how trace metals alone and in combination with other chemical exposures contribute to these adverse pregnancy outcomes that can have significant repercussive effects in the life of the child. Additionally, our aim is to use cutting-edge metabolomics techniques to better understand mechanisms underlying these associations in order to identify opportunities for prevention when reducing exposure is difficult or impossible. Utilizing the CHEAR resources, we plan to measure trace metal concentrations in urine from the third trimester of pregnancy along with metabolomics profiles from the same time point. Doing so will allow us to address the following specific aims: SEE NOTES
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
1159
TOTAL_SUBJECTS
390
STUDY_COMMENTS
SPECIFIC AIM 1: Assess the impact of urinary trace metal concentrations from the 3rd trimester of pregnancy on birth outcomes, including spontaneous and placentally-mediated preterm delivery as well as fetal growth as characterized by repeated ultrasound measures. SPECIFIC AIM 2: Investigate the cumulative association between trace metal and phthalate exposures during pregnancy on birth outcomes. SPECIFIC AIM 3: Explore metabolomics profiles in urine from the third trimester of pregnancy in relation to: environmental exposures individually and in mixtures; birth outcomes; and previously measured oxidative stress biomarkers.
Targeted NEFA in Kallikrein 6 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Targeted Cholesterol Profiling of Myelin to Enhance Recovery of Function after SCI. Second Set of Samples
STUDY_SUMMARY
Tissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
Targeted Sphingolipids in Kallikrein 6 Mice after SCI
STUDY_SUMMARY
Targeted Sphingolipids in Kallikrein 6 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Targeted Galactosyl Sphingolipid Concentration in Kallikrein 6 Mice after SCI
STUDY_SUMMARY
Targeted Galactosyl Sphingolipids in Kallikrein 6 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Targeted Sphingomyelin Concentrations in Kallikrein 6 Mice after SCI
STUDY_SUMMARY
Targeted Sphingomyelin in Kallikrein 6 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Targeting Myelin NEFA in PAR1 and PAR2 Mice after SCI
STUDY_SUMMARY
Targeting Myelin NEFA in PAR1 and PAR2 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21 or 60 mouse spinal cord (SC). There are 30 total sampes, n=5 for each group. wild type control, PAR1 KO and PAR2 KO at P21 and P60 days.
Targeting Myelin FFA Compostion in PAR1 and PAR2 Mice after SCI
STUDY_SUMMARY
Targeting Myelin FFA composition in PAR1 and PAR2 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21 or 60 mouse spinal cord (SC). There are 30 total sampes, n=5 for each group. wild type control, PAR1 KO and PAR2 KO at P21 and P60 days.
Targeting Myelin Cholesterol in PAR1 and PAR2 Mice after SCI
STUDY_SUMMARY
Targeting Myelin Cholesterol in PAR1 and PAR2 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21 or 60 mouse spinal cord (SC). There are 30 total sampes, n=5 for each group. wild type control, PAR1 KO and PAR2 KO at P21 and P60 days.
Targeting Myelin Ceramides in PAR1 and PAR2 Mice after SCI
STUDY_SUMMARY
Targeting Myelin ceramides in PAR1 and PAR2 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21 or 60 mouse spinal cord (SC). There are 30 total sampes, n=5 for each group. wild type control, PAR1 KO and PAR2 KO at P21 and P60 days.
Identification of Race-Associated Metabolite Biomarkers for Hepatocellular Carcinoma
STUDY_SUMMARY
Introduction: Metabolomics provides simultaneous assessment of a broad range of metabolites that can potentially serve as indicators of the overall physiology status as well as the response to host and environmental stimuli. It has been broadly used for biomarker discovery and characterization of complex diseases such as cancer. The evaluation of the changes in the levels of metabolites in samples stratified by race could lead to the identification of more reliable biomarkers than those obtained through whole-population-based approaches. In this study, we used plasma samples collected from patients recruited at MedStar Georgetown University Hospital to investigate metabolites that may be associated with hepatocellular carcinoma (HCC) in a race-specific manner. Methods: Plasma samples were protein depleted using a solution composed of acetonitrile:isopropanol:water (3:3:2) containing a mixture of isotope-labeled internal standards. The extracted metabolites were trimethylsilyl derivatized prior to analysis by GC-MS. A quality control (QC) sample derived by pooling plasma from multiple subjects was run in between samples to assess reproducibility. A mixture of fatty acids methyl esters (FAMEs) and alkane standards was run for retention index calibration. The mixture of isotope-labeled internal standards was used to evaluate the reproducibility of the GC-MS data across multiple runs. Preliminary Data: Plasma samples collected from 40 HCC cases and 44 patients with liver cirrhosis were analyzed. The cirrhotic controls were frequency matched with the HCC cases by demographic variables. The participants included 19 African Americans (AA) and 50 European Americans (EA). The analysis targeted 46 metabolites for quantitative analysis by Agilent GC-qMS in selected ion monitoring (SIM) mode. The data were pre-processed by the Automated Mass Spectral Deconvolution and Identification System (AMDIS) for peak detection, deconvolution, and identification. The resulting peaks were aligned using Mass Profiler Professional (MPP) from Agilent Technologies. A LASSO regression model was applied to select metabolites with significant changes in HCC vs. cirrhosis in three groups: (1) AA and EA combined; (2) AA only; and (3) EA only. Also, metabolites that distinguish HCC cases from cirrhosis in the three groups were selected by considering only those subjects with Hepatitis C virus (HCV) infection. The performances of the metabolites selected by LASSO in each group were evaluated through a leave-one-out cross-validation. We identified race-specific metabolites that differentiated HCC cases from cirrhotic controls, yielding better area under the ROC curve compared to alpha-fetoprotein (AFP) - the serological marker widely used for the diagnosis of HCC. Novel Aspect: We identified race-associated metabolites that are significantly altered in HCC vs. cirrhosis, suggesting the potential role of race in HCC.
INSTITUTE
Georgetown University
DEPARTMENT
Oncology, Georgetown University Medical Center
LABORATORY
Ressom Lab (PI: Habtom W. Ressom, email address hwr@georgetown.edu)
LAST_NAME
Cristina
FIRST_NAME
Di Poto
ADDRESS
3970 Reservoir Rd., NW, Research Bldg, Room W325, Washington, DC, 20007, USA
Large Untargeted Profiling in PAR1 and PAR2 Mice after SCI
STUDY_SUMMARY
Large Untargeted Profiling in NEFA in PAR1 and PAR2 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21 or 60 mouse spinal cord (SC). There are 30 total sampes, n=5 for each group. wild type control, PAR1 KO and PAR2 KO at P21 and P60 days.
In this study, two independent large cohorts of mature dates exhibiting substantial diversity in origin, varieties and fruit processing conditions were measured by metabolomics techniques in order to identify major determinants of the fruit metabotype. Additional samples reflecting different stages of date fruit ripening process has been included for 10 different fruit varieties. This study includes updated date photographs and combined results data (GCMS/LCMS) and technical validation data, see downloadable files section to access this information.
Untargeted metabolomic profile of oak and wine yeast strains
STUDY_SUMMARY
Metabolomic profiles were compiled from oak and wine yeast parents over three extraction times (batch). Included in this study are extraction controls.
INSTITUTE
Washington University in St. Louis
LAST_NAME
Swain Lenz
FIRST_NAME
Devjanee
ADDRESS
4515 McKinley Avenue, Saint Louis, Missouri, 63110, USA
Large Untargeted Profiling in Kallikrein 6 Mice after SCI
STUDY_SUMMARY
Large untargeted profiling mouse myelin of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Metabolomic profiles were compiled from reciprocal hemizygotes (oak/win hybrid, genes AUA1, ARG82) (batch). Included in this study are extraction controls.
INSTITUTE
Washington University in St. Louis
LAST_NAME
Swain Lenz
FIRST_NAME
Devjanee
ADDRESS
4515 McKinley Avenue, Saint Louis, Missouri, 63110, USA
Maternal Hypoxemia and oxidative stress in the fetus, newborn, and adult. exercise training for peripheral artery disease
STUDY_TYPE
Disease model
STUDY_SUMMARY
Gestational hypoxia presents a significant stress to an unborn fetus that can lead to significant complications related to fetal growth restriction and resulting in diseases in the newborn as well as those manifesting later in life. Recent evidence indicates that inflammation and oxidative stress are contributing factors to hypoxia-related diseases. The Center for Perinatal Biology at Loma Linda University has studied gestational chronic hypoxia in a sheep model for over 20 years to study dysfunction of vascular and nonvascular tissues derived from mothers, fetuses and offspring. In this project we are attempting to use metabolomics to assess metabolic dysregulation in vascular tissues along with markers of oxidative stress and inflammation in the mother and offspring to determine the extent of dysregulation due to chronic hypoxia. Untargeted metabolomics analysis focused on sheep plasma and arteries from the lung, resistance arteries in the brain, uterine arteries, and cultured human myocytes will be used to explore markers of glucose and lipid metabolism disruption. Targeted analyses of oxylipins and endocannabinoids will be used on the same samples to explore markers of oxidative stress and inflammation, which should be increased during hypoxia. This study should delineate pathways and biomarkers that help explain how hypoxia leads to the development of neonatal as well as adult-onset diseases associated with chronic hypoxia that are inter-related with fetal growth restriction.
Maternal Hypoxemia and oxidative stress in the fetus, newborn, and adult. exercise training for peripheral artery disease (part II)
STUDY_TYPE
Disease model
STUDY_SUMMARY
Gestational hypoxia presents a significant stress to an unborn fetus that can lead to significant complications related to fetal growth restriction and resulting in diseases in the newborn as well as those manifesting later in life. Recent evidence indicates that inflammation and oxidative stress are contributing factors to hypoxia-related diseases. The Center for Perinatal Biology at Loma Linda University has studied gestational chronic hypoxia in a sheep model for over 20 years to study dysfunction of vascular and nonvascular tissues derived from mothers, fetuses and offspring. In this project we are attempting to use metabolomics to assess metabolic dysregulation in vascular tissues along with markers of oxidative stress and inflammation in the mother and offspring to determine the extent of dysregulation due to chronic hypoxia. Untargeted metabolomics analysis focused on sheep plasma and arteries from the lung, resistance arteries in the brain, uterine arteries, and cultured human myocytes will be used to explore markers of glucose and lipid metabolism disruption. Targeted analyses of oxylipins and endocannabinoids will be used on the same samples to explore markers of oxidative stress and inflammation, which should be increased during hypoxia. This study should delineate pathways and biomarkers that help explain how hypoxia leads to the development of neonatal as well as adult-onset diseases associated with chronic hypoxia that are inter-related with fetal growth restriction.
Metabolomic investigations on Nesterenkonia flava from different origins revealed significant intraspecies differences between marine and terrestrial actinomycetes
STUDY_SUMMARY
Marine is one of the most important resources of microorganisms, including bacteria, actinomycetes, and fungi. As marine and terrestrial environments differ a lot in many aspects it is not surprising that the species and characteristics of microorganisms living there are very different. Interestingly, many marine microorganisms can find their congeners of the same species from terrestrial resources. The aim of this work is to evaluate the intraspecies differences between marine and terrestrial actinomycetes on metabolic level and to uncover the mechanism responsible for the differences. To address this, we carried out comparative metabolomics study on Nesterenkonia flava strains isolated from marine and terrestrial environments. The results showed that marine strains were clearly distinguished from their terrestrial congeners on the principal components analysis (PCA) scores plot of intracellular metabolites. The markers responsible for the discrimination of marine and terrestrial strains were figured out using loading plot from partial least squares discrimination analysis (PLS-DA). Pathway analysis based on PLS-DA, univariate analysis, and correlation analysis of metabolites showed that the major differential metabolites between the terrestrial N. flava and the marine ones were involved in osmotic regulation, redox balancing, and energy metabolism. Together, these insights provide clues as to how the previous living environment of microbes affect their current metabolic performances under laboratory cultivation conditions.
INSTITUTE
Third Institute of Oceanography, State Oceanic Administration
Micronutrient deficiencies, environmental exposures and severe malaria: Risk factors for adverse neurodevelopmental outcomes in Ugandan children
STUDY_TYPE
Untargeted high-resolution mass spectrometry profiling
STUDY_SUMMARY
Micronutrient deficiencies and environmental exposures have been to known to adversely impact brain and nervous system functions in adults and children worldwide. However, few studies have examined the short and long-term impact of these risk factors on neurodevelopmental outcomes in children in low-income countries, where the effects are likely to be more pronounced due to limited resources for monitoring and insufficient regulations. Biological risk factors of relevance include micronutrient deficiencies such as zinc and exposure to heavy metals such as lead and mercury. Studies have suggested an association between neurodevelopmental impairment and micronutrient deficiency as well as exposure to a number of heavy metals and environmental toxins. Moreover, findings also suggest that risk factors for adverse developmental outcomes that are independently significant may have the potential for causing cumulative increases in adverse effects. In Sub-Saharan Africa, severe malaria is a leading risk factor for long-term neurocognitive impairment in children. Zinc deficiency or exposure to heavy metals could influence risk of severe malaria, modify the risk of neurocognitive impairment in children with severe malaria, or independently affect risk of neurocognitive impairment. Untargeted analyses for potential environmental exposures or metabolomic changes in children with cerebral malaria vs. without cognitive impairment or in children with higher vs. lower cognitive scores, could also identify new risk factors for neurodevelopmental impairment in Ugandan children with cerebral malaria.In our completed study in Kampala, we assessed neurologic and developmental impairment in children with cerebral malaria [CM] or severe malarial anemia [SMA], as compared to health community children from the same extended household as the children with CM or SMA. As an extension of this study, we are interested in determining levels of micronutrients such as zinc in the population, and in addition, determining exposure levels of heavy metals (lead, mercury, copper, manganese etc.) in samples collected from children with severe malaria and community controls. The primary hypotheses of this study is that nutrient deficiencies or exposure to heavy metals influence short and long term neurocognitive outcomes in healthy community children and in children with severe malaria, and that children with cerebral malaria have specific metabolomic changes that relate to long-term neurocognitive impairment. The specific aims of our study are:Aim 1: To determine levels of zinc, heavy metals, and biomarkers associated with inflammation in children presenting with different forms of severe malaria (SM) and in healthy community children (CC). The working hypothesis of this aim is that 1) children with SM will have lower zinc levels compared to CC; 2) children with SM will present with higher toxic metal exposure and higher levels of biomarkers associated with inflammation than CC.Aim 2: To investigate how micronutrient deficiency, toxic metal exposure and inflammatory biomarkers affect short and long term neurodevelopmental outcomes and growth in children with severe malaria and community children (CC).The working hypothesis of this aim is that the lower levels of zinc, and presence of toxic metals in high concentrations will independently contribute to worsening neurodevelopmental outcomes and worsening growth over time in children with severe malaria and in community children. An alternate hypothesis is that micronutrient deficiency, toxic metal exposure and inflammatory states may interact with each other and with severe malaria to produce greater neurodevelopmental impairment, i.e., that the contribution is not independent but interactive.Aim 3: To determine whether the CSF metabolome differs according to level of neurodevelopmental impairment in children with cerebral malaria. The working hypothesis of this aim is that neurodevelopmental impairment in children with cerebral malaria is associated with changes in the CSF metabolome.
INSTITUTE
Emory University School of Medicine
DEPARTMENT
Division of Pulmonary, Allergy and Critical Care Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Walker
FIRST_NAME
Douglas
ADDRESS
615 Michael St. Ste 225, Atlanta, GA, 30322, USA
EMAIL
douglas.walker@emory.edu
PHONE
(404) 727 5984
TOTAL_SUBJECTS
141
STUDY_COMMENTS
CSF pools from elderly individuals included for QA/QC. Study specific pools were not created due to limited sample volumes provided (<100uL).
Fatty Acid Oxidation is Impaired in An Orthologous Mouse Model of Autosomal Dominant Polycystic Kidney Disease
STUDY_SUMMARY
Autosomal Dominant Polycystic Kidney Disease (ADP.D MIM ID's 173900, 601313, 613095) is estimated to affect almost 1/1000 and is the most common genetic cause of end stage renal disease (Torres et al., 2007). While advances have been made in slowing the progression of some other forms of chronic kidney disease, standard treatments have not reduced the need for renal replacement therapy in ADPKD (Spithoven et al., 2014). Unfortunately, several experimental interventions also have recently failed to show significant benefit in slowing the rate of functional decline (Serra et al., 2010; Walz et al., 2010; Schrier et al., 2014; Torres et al., 2014), and the only positive study reported very modest effects (Torres et al., 2012). These findings suggest new treatment strategies are required. A central dogma of molecular genetics is that discovery of the causative genes will lead to identification of key pathways and potential targets for intervention. In the case of ADPKD, the two genes mutated in the disorder, PKD1 and PKD2, were identified almost 20 years ago and yet their functions remain poorly understood. The PKD1 gene product, polycystin-1 (PC1), encodes a large membrane protein that requires the PKD2 gene product, polycystin-2 (PC2), for its trafficking to the primary cilium where the two are thought to form a receptor channel complex (Kim et al., 2014; Cai et al., 2014). What the complex senses and what it signals remains controversial. The primary cilium has emerged as a key player in the pathogenesis of PKD as mutations in dozens of different genes that encode either essential ciliary components or factors in ciliary signaling pathways result in PKD. A recent report suggests that the relationship between the polycystin complex and ciliary signaling is complicated, however.While ablation of primary cilia by mutation of core ciliary components results in cysts, these same perturbations done in the setting of Pkd1 or Pkd2 inactivation results in significant attenuation of cystic disease (Ma et al., 2013). These data suggest that the polycystin complex provides a suppressive signal for a novel, cilia-dependent growth-promoting pathway that is independent of MAPK/ERK, mTOR, or cAMP pathways, three effector pathways previously implicated as major drivers of cyst growth. The identities of the growth-promoting and growth-inhibiting pathways remain unknown. We have taken a systems-based approach to study Pkd1 gene function. Building on our previous work identifying markedly different outcomes in animals with induced Pkd1 inactivation before or after P12 and correlating this susceptibility with metabolic status (Piontek et al., 2007; Menezes et al., 2012), we now show that female sex is partially protective in adult-induced Pkd1 inactivation, that sex differences in metabolic status may account for this effect, and that cells lacking Pkd1 have abnormal fatty acid oxidation. Finally, manipulating diet in Pkd1 mouse models, we demonstrate a positive correlation between lipid content in mouse chow and cystic kidney disease severity. Our results therefore suggest that abnormal lipid metabolism is an intrinsic component of PKD and an important modifier of disease progression.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Diet, genetics and gut microbiome drive dynamic changes in plasma metabolites [plasma]
STUDY_TYPE
Plasma data
STUDY_SUMMARY
C57BL/6J mice (B6) and 129S1 mice (129J) were purchased from Jackson Laboratory (Bar Harbor, ME) and 129S6 mice (129T) were purchased from Taconic Farms (Germantown, NY). Mice were maintained on normal chow containing 22% calories from fat, 23% from protein and 55% from carbohydrates (Mouse Diet 9F 5020, PharmaServ, Framingham, MA) or a high fat diet (Open Source Diet, D12492, Research Diets, New Brunswick, NJ) containing 60% calories from fat, 20% from protein and 20% from carbohydrates. For antibiotic treatment, 6-week old mice were treated with either placebo, vancomycin (1g/L) or metronidazole (1g/L) (Sigma-Aldrich, St. Louis, MO) in drinking water then started on HFD from age 7 to 11 weeks. The mice were fasted for 2 hours and anesthetized with isoflurane before collecting cecum and plasma.
Diet, genetics and gut microbiome drive dynamic changes in plasma metabolites [cecal]
STUDY_TYPE
Cecal data
STUDY_SUMMARY
C57BL/6J mice (B6) and 129S1 mice (129J) were purchased from Jackson Laboratory (Bar Harbor, ME) and 129S6 mice (129T) were purchased from Taconic Farms (Germantown, NY). Mice were maintained on normal chow containing 22% calories from fat, 23% from protein and 55% from carbohydrates (Mouse Diet 9F 5020, PharmaServ, Framingham, MA) or a high fat diet (Open Source Diet, D12492, Research Diets, New Brunswick, NJ) containing 60% calories from fat, 20% from protein and 20% from carbohydrates. For antibiotic treatment, 6-week old mice were treated with either placebo, vancomycin (1g/L) or metronidazole (1g/L) (Sigma-Aldrich, St. Louis, MO) in drinking water then started on HFD from age 7 to 11 weeks. The mice were fasted for 2 hours and anesthetized with isoflurane before collecting cecum and plasma.
Breathprinting Reveals Malaria-Associated Biomarkers and Mosquito Attractants
STUDY_SUMMARY
Current evidence suggests that malaria infection could alter patient breath metabolites, a phenomenon that could be exploited to create a breath-based diagnostic test. Indications include the preferential attraction of the Anopheles mosquito vector upon infection and a distinct breath profile with the progression of experimental, sub-microscopic malaria. However, these observations have yet to be extended to the clinic. To investigate whether natural human malaria infection leads to a characteristic breath profile, we performed a field study in Malawi. Breath volatiles from pediatric patients with and without uncomplicated falciparum malaria were analyzed by thermal desorption-gas chromatography/mass spectrometry. Using an unbiased, correlation-based analysis, we find that children with malaria have a distinct shift in overall breath composition. Leveraging these differences, highly accurate classification of infection status was achieved with a suite of six compounds. In addition, we find that malaria-infected children have significantly higher breath levels of two mosquito-attractant terpenes, Ī±-pinene and 3-carene. Thus, our work attests to the viability of breath analysis for malaria diagnosis, identifies candidate compounds for follow-up studies, and identifies biologically plausible chemical mediators for increased mosquito attraction to malaria-infected patients.
INSTITUTE
Washington University School of Medicine
LAST_NAME
Schaber
FIRST_NAME
Chad
ADDRESS
4938 Parkview Place, MPRB/FLoor 6, Entry 5, St. Louis, MO, 63110, USA
Evidence that COG0325 proteins are involved in PLP homeostasis
STUDY_SUMMARY
Pyridoxal 5'-phosphate (PLP) is an essential cofactor for nearly 60 Escherichia coli enzymes but is a highly reactive molecule that is toxic in its free form. How PLP levels are regulated and how PLP is delivered to target enzymes are still open questions. The COG0325 protein family belongs to the fold-type III class of PLP enzymes and binds PLP but has no known biochemical activity although it occurs in all kingdoms of life. Various pleiotropic phenotypes of the E. coli COG0325 (yggS) mutant have been reported, some of which were reproduced and extended in this study. Comparative genomic, genetic and metabolic analyses suggest that these phenotypes reflect an imbalance in PLP homeostasis. The E. coli yggS mutant accumulates the PLP precursor pyridoxine 5'-phosphate (PNP) and is sensitive to an excess of pyridoxine but not of pyridoxal. The pyridoxine toxicity phenotype is complemented by the expression of eukaryotic yggS orthologs. It is also suppressed by the presence of amino acids, specifically isoleucine, threonine and leucine, suggesting the PLP-dependent enzyme transaminase B (IlvE) is affected. These genetic results lay a foundation for future biochemical studies of the role of COG0325 proteins in PLP homeostasis.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
STUDY_COMMENTS
LB stands for Luria Bertani medium. MG stands for minimal growth For strains grown on MG OD 0.5 is the mid log growth phase and OD 1.0 is the late log growth phase For strains grown on LB OD 1 is the mid log growth phase and OD 2.0 is the late log growth phase
Gut Microbiota Modulate Brain Insulin Sensitivity and Neurobehavior
STUDY_TYPE
Metabolite profiling of cecal contents and brains of mice under diet-induced obesity (DIO) with and without antibiotic treatments.
STUDY_SUMMARY
C57BL/6J mice were purchased from Jackson Laboratory and maintained on either a normal chow containing 22% of calories from fat, 23% from protein, and 55% from carbohydrates (Mouse diet 9F 5020; PharmaServ) or a high-fat diet (Open Source Diet, D12492; Research Diets) containing 60% of calories from fat, 20% from protein, and 20% from carbohydrates for 6 weeks. During the last 2 weeks, some of the HFD mice were treated with vancomycin or metronidazole (1 g/L in the drinking water). All mice were housed at 22Ā°C on a 12 h light/dark cycle. All animal studies were approved by the IACUC of Joslin Diabetes Center (# 97-05) and Harvard Medical School (# 05131) and were in accordance with NIH guidelines. The metabolite profiling was conducted on plasma, hypothalamus and nucleus accumbens.
Mechanism Behind Stay Green Trait in Bread Wheat (Triticum aestivum L.)
STUDY_TYPE
Comparison
STUDY_SUMMARY
Two different wheat genotypes were treated with the high temperature and control conditions under full irrigated condition. Leaf tissues were collected for all 2-different treatments with six replicates after 7 and 10 days of high temperature treatment.
Retrospective serum samples from patients with early Lyme disease, other diseases, and healthy controls were analyzed for small molecule metabolites by liquid chromatography-mass spectrometry (LC-MS). A metabolomics data workflow was applied to select a biosignature for classifying early Lyme disease and non-Lyme disease patients. A statistical model of the biosignature was trained using the patients' LC-MS data, and subsequently applied as an experimental diagnostic tool with LC-MS data from additional patient sera.
INSTITUTE
Colorado State University
DEPARTMENT
MIP
LABORATORY
Belisle
LAST_NAME
Belisle
FIRST_NAME
John
ADDRESS
200 West Lake, Campus Delivery 0922, Colorado State University, Fort Collins, CO, 80523, USA
EMAIL
john.belisle@colostate.edu
PHONE
970-491-5384
NUM_GROUPS
9
STUDY_COMMENTS
All samples and data used in training are provided. All samples and data used in testing are provided, except healthy controls that were experimentally heat treated.
GC-MS analysis of Quercus ilex organs (leaves, roots and acorns)
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Qualitative metabolomics study on leaves, roots and acorns from Quercus ilex plantlets. We analyzed polar(metanol:water) and apolar (chloroform) fractions.
INSTITUTE
Universidad de CĆ³rdoba
DEPARTMENT
Department Biochemistry and Molecular Biology
LABORATORY
Agroforestry and Plant Biochemistry, Proteomics and Systems Biology
Free fatty acids in mouse plasma were identified and quantified by LC-MS. Through differential feeding and PHZ (phnylhydrazine) dosing, coupled with mass spectrometry, we identified the long chain fatty acid C24:5 as a natural RXRA ligand, which was dynamically increased in concentration in response to hematopoietic stress. Collectively, these data demonstrate that natural RXRA ligands are present and are dynamically regulated in vivo in mouse hematopoietic cells.
NMR comparison of urine samples by 1D NOESY presat and PURGE
STUDY_TYPE
NMR development for metabolomics
STUDY_SUMMARY
Different water suppression pulse sequences, namely 1D NOESY presat and optimized PURGE were compared for urine samples. The original PURGE sequence was also tested to show the postential for lineshape distortions with the pulse sequence.
Comparison of T2 filters for quantification of small molecules in plasma by 1D NMR
STUDY_TYPE
NMR development for metabolomics
STUDY_SUMMARY
A pooled plasma sample obtained from Red Cross has been seperated into 20 samples. 10 of them were simply mixed with a D2O phosphate buffer, while the other 10 were centrifuged first, then dried and resuspended in a 90/10 H2O/D2O phosphate buffer. The 20 NMR samples were analysed by NMR (4 spectra per sample, but different pulse sequences and parameters were used for each group). Standard addition was used in order to measure the concentration of 5 metabolites: valine, lactate, glucose, phenylalanine and formate.
Analysis of the effects of Tyrosine Kinase Inhibitors Sunitinib and Erlotinib on Heart, Metabolism In Vivo
STUDY_SUMMARY
Background.More than 90 tyrosine kinases have been implicated in the pathogenesis of malignant transformation and tumor angiogenesis. Tyrosine kinase inhibitors (TKIs) have emerged as effective therapies in treating cancer by exploiting this kinase dependency. The tyrosine kinase inhibitor erlotinib targets epidermal growth factor receptor (EGFR), whereas sunitinib targets primarily vascular endothelial growth factor receptor (VEGRF) and platelet-derived growth factor receptor (PDGFR). TKIs impact the function of non-malignant cells had have on- and off-target toxicities, including cardiotoxicities. Most of the reports of sunitinib have identified cardiotoxic effects, whereas erlotinib was less often found to have these effects. We hypothesized that the deleterious effects of TKIs were related to their impact on cardiac metabolism. Methods. C57BL/6 mice (10/group) were treated with therapeutic doses of sunitinib (40 mg/kg), erlotinib (50 mg/kg), or vehicle daily for 2 weeks. Echocardiographic assessment of the heart in vivo identified significant systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Heart, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Results. Compared to vehicle treated controls, sunitinib treated mice had significant decreases insystolic function, whereas erlotinib treated mice did not. Non-Targeted metabolomics analysis of heart identified identified significant decreases in Docosahexaenoic acid (DHA), Arachidonic Acid/Eicosapentaenoic acid (EPA), O-Phosphocolamine, and 6-Hydroxynicotinic acid after sunitinib treatment. DHA was significantly decreased in skeletal muscle (quadriceps femoris), with elevations in cholesterol were identified in liver and elevated ethanol amine in serum. In contrast, erlotinib affected only one metabolite elevated (spermidine significantly increased).Conclusions.Mice treated with sunitinib had exhibited systolic dysfunction within two weeks, with significantly lower heart and skeletal muscle levels of long chain omega-3 fatty acids docosahexaenoic acid (DHA), arachidonic acid (AA)/eicosapentaenoic acid (EPA) and increased serum O-Phosphocholine phospholipid. This is the first link between sunitinib-induced cardiotoxicity and depletion in the polyunsaturated fatty acids (PUFAs) and inflammatory mediators DHA and AA/EPA in the heart, possibly by affecting mitochondria function where they have vital roles on calcium channels.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
EMAIL
monte_willis@med.unc.edu
PHONE
919-360-7599
STUDY_COMMENTS
Cardiac tissue, Gastrocnemius Muscle, Liver, and Serum
Analysis of the effects of Tyrosine Kinase Inhibitors Sunitinib and Erlotinib on Liver Metabolism In Vivo
STUDY_SUMMARY
Background.More than 90 tyrosine kinases have been implicated in the pathogenesis of malignant transformation and tumor angiogenesis. Tyrosine kinase inhibitors (TKIs) have emerged as effective therapies in treating cancer by exploiting this kinase dependency. The tyrosine kinase inhibitor erlotinib targets epidermal growth factor receptor (EGFR), whereas sunitinib targets primarily vascular endothelial growth factor receptor (VEGRF) and platelet-derived growth factor receptor (PDGFR). TKIs impact the function of non-malignant cells had have on- and off-target toxicities, including cardiotoxicities. Most of the reports of sunitinib have identified cardiotoxic effects, whereas erlotinib was less often found to have these effects. We hypothesized that the deleterious effects of TKIs were related to their impact on cardiac metabolism. Methods. C57BL/6 mice (10/group) were treated with therapeutic doses of sunitinib (40 mg/kg), erlotinib (50 mg/kg), or vehicle daily for 2 weeks. Echocardiographic assessment of the heart in vivo identified significant systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Heart, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Results. Compared to vehicle treated controls, sunitinib treated mice had significant decreases insystolic function, whereas erlotinib treated mice did not. Non-Targeted metabolomics analysis of heart identified identified significant decreases in Docosahexaenoic acid (DHA), Arachidonic Acid/Eicosapentaenoic acid (EPA), O-Phosphocolamine, and 6-Hydroxynicotinic acid after sunitinib treatment. DHA was significantly decreased in skeletal muscle (quadriceps femoris), with elevations in cholesterol were identified in liver and elevated ethanol amine in serum. In contrast, erlotinib affected only one metabolite elevated (spermidine significantly increased).Conclusions.Mice treated with sunitinib had exhibited systolic dysfunction within two weeks, with significantly lower heart and skeletal muscle levels of long chain omega-3 fatty acids docosahexaenoic acid (DHA), arachidonic acid (AA)/eicosapentaenoic acid (EPA) and increased serum O-Phosphocholine phospholipid. This is the first link between sunitinib-induced cardiotoxicity and depletion in the polyunsaturated fatty acids (PUFAs) and inflammatory mediators DHA and AA/EPA in the heart, possibly by affecting mitochondria function where they have vital roles on calcium channels.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
EMAIL
monte_willis@med.unc.edu
PHONE
919-360-7599
STUDY_COMMENTS
Cardiac tissue, Gastrocnemius Muscle, Liver, and Serum
Analysis of the effects of Tyrosine Kinase Inhibitors Sunitinib and Erlotinib on Serum Metabolism In Vivo using Non-Targeted Metabolomics Analysis
STUDY_SUMMARY
Background.More than 90 tyrosine kinases have been implicated in the pathogenesis of malignant transformation and tumor angiogenesis. Tyrosine kinase inhibitors (TKIs) have emerged as effective therapies in treating cancer by exploiting this kinase dependency. The tyrosine kinase inhibitor erlotinib targets epidermal growth factor receptor (EGFR), whereas sunitinib targets primarily vascular endothelial growth factor receptor (VEGRF) and platelet-derived growth factor receptor (PDGFR). TKIs impact the function of non-malignant cells had have on- and off-target toxicities, including cardiotoxicities. Most of the reports of sunitinib have identified cardiotoxic effects, whereas erlotinib was less often found to have these effects. We hypothesized that the deleterious effects of TKIs were related to their impact on cardiac metabolism. Methods. C57BL/6 mice (10/group) were treated with therapeutic doses of sunitinib (40 mg/kg), erlotinib (50 mg/kg), or vehicle daily for 2 weeks. Echocardiographic assessment of the heart in vivo identified significant systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Heart, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Results. Compared to vehicle treated controls, sunitinib treated mice had significant decreases insystolic function, whereas erlotinib treated mice did not. Non-Targeted metabolomics analysis of heart identified identified significant decreases in Docosahexaenoic acid (DHA), Arachidonic Acid/Eicosapentaenoic acid (EPA), O-Phosphocolamine, and 6-Hydroxynicotinic acid after sunitinib treatment. DHA was significantly decreased in skeletal muscle (quadriceps femoris), with elevations in cholesterol were identified in liver and elevated ethanol amine in serum. In contrast, erlotinib affected only one metabolite elevated (spermidine significantly increased).Conclusions.Mice treated with sunitinib had exhibited systolic dysfunction within two weeks, with significantly lower heart and skeletal muscle levels of long chain omega-3 fatty acids docosahexaenoic acid (DHA), arachidonic acid (AA)/eicosapentaenoic acid (EPA) and increased serum O-Phosphocholine phospholipid. This is the first link between sunitinib-induced cardiotoxicity and depletion in the polyunsaturated fatty acids (PUFAs) and inflammatory mediators DHA and AA/EPA in the heart, possibly by affecting mitochondria function where they have vital roles on calcium channels.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
EMAIL
monte_willis@med.unc.edu
PHONE
919-360-7599
STUDY_COMMENTS
Cardiac tissue, Gastrocnemius Muscle, Liver, and Serum
Analysis of the effects of Tyrosine Kinase Inhibitors Sunitinib and Erlotinib on Muscle Metabolism In Vivo
STUDY_SUMMARY
Background.More than 90 tyrosine kinases have been implicated in the pathogenesis of malignant transformation and tumor angiogenesis. Tyrosine kinase inhibitors (TKIs) have emerged as effective therapies in treating cancer by exploiting this kinase dependency. The tyrosine kinase inhibitor erlotinib targets epidermal growth factor receptor (EGFR), whereas sunitinib targets primarily vascular endothelial growth factor receptor (VEGRF) and platelet-derived growth factor receptor (PDGFR). TKIs impact the function of non-malignant cells had have on- and off-target toxicities, including cardiotoxicities. Most of the reports of sunitinib have identified cardiotoxic effects, whereas erlotinib was less often found to have these effects. We hypothesized that the deleterious effects of TKIs were related to their impact on cardiac metabolism. Methods. C57BL/6 mice (10/group) were treated with therapeutic doses of sunitinib (40 mg/kg), erlotinib (50 mg/kg), or vehicle daily for 2 weeks. Echocardiographic assessment of the heart in vivo identified significant systolic dysfunction consistent with cardiotoxicity compared to vehicle treated controls. Heart, skeletal muscle, liver, and serum were flash frozen and prepped for non-targeted GC-MS metabolomics analysis. Results. Compared to vehicle treated controls, sunitinib treated mice had significant decreases insystolic function, whereas erlotinib treated mice did not. Non-Targeted metabolomics analysis of heart identified identified significant decreases in Docosahexaenoic acid (DHA), Arachidonic Acid/Eicosapentaenoic acid (EPA), O-Phosphocolamine, and 6-Hydroxynicotinic acid after sunitinib treatment. DHA was significantly decreased in skeletal muscle (quadriceps femoris), with elevations in cholesterol were identified in liver and elevated ethanol amine in serum. In contrast, erlotinib affected only one metabolite elevated (spermidine significantly increased).Conclusions.Mice treated with sunitinib had exhibited systolic dysfunction within two weeks, with significantly lower heart and skeletal muscle levels of long chain omega-3 fatty acids docosahexaenoic acid (DHA), arachidonic acid (AA)/eicosapentaenoic acid (EPA) and increased serum O-Phosphocholine phospholipid. This is the first link between sunitinib-induced cardiotoxicity and depletion in the polyunsaturated fatty acids (PUFAs) and inflammatory mediators DHA and AA/EPA in the heart, possibly by affecting mitochondria function where they have vital roles on calcium channels.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
EMAIL
monte_willis@med.unc.edu
PHONE
919-360-7599
STUDY_COMMENTS
Cardiac tissue, Gastrocnemius Muscle, Liver, and Serum
Untargeted metabolomics analysis of ischemia-reperfusion injured hearts ex vivo from sedentary and exercise trained rats.
STUDY_SUMMARY
The effects of exercise on the heart and its resistance to disease are well-documented. Recent studies have identified exercise-induced resistance to arrhythmia is due to the preservation of mitochondrial membrane potential. To identify novel metabolic changes that occurred parallel to these mitochondrial alterations, we performed non-targeted metabolomics analysis on hearts from sedentary (Sed) and exercise- trained (Ex) rats challenged with isolated heart ischemia-reperfusion injury (I/R). Eight weeks old Sprague- Dawley rats were treadmill trained five days/week for six weeks (exercise duration and intensity progressively increased to 1 hour at 30 m/min up to 10.5% incline, 75-80% VO2mx). The recovery of pre-ischemic function for sedentary rat hearts was 28.8+/-5.4% (N=12) compared to exercise trained hearts which recovered 51.9%+/-5.7 (N=14)(p<0.001). Non-targeted GC-MS metabolomics analysis of 1) Sedentary rat hearts; 2) Exercise-trained rat hearts; 3) Sedentary rat hearts challenged with global ischemia-reperfusion (I/R) injury; and 4) Exercise-trained rat hearts challeged with global I/R (10/group) revealed 20 statistically significant metabolites between groups by ANOVA using Metaboanalyst (p<0.001). Enrichment analysis of these metabolites for pathway-associated metabolic sets indicated a >10 fold enrichment for ammonia recycling and protein biosynthesis (L-Glutamic acid; L-Proline; L-Histidine; L-Serine; L-Aspartic acid; L-Glutamine)(p<=4.05E-05, FDR=0.0024). Subsequent comparison of the sedentary hearts post-I/R and exercise-trained hearts post-I/R further identified significant differences in metabolites related to Aminoacyl-tRNA biosynthesis and nitrogen metabolism (4) (p<=1.24E-05, FDR<=5.07E-4). These studies shed light on novel mechanisms in which exercise-induced cardioprotection occurs in I/R which complement both the mitochondrial stabilization and antioxidant mechanisms recently described. These findings also link protein synthesis and protein degradation (protein quality control mechanisms) with exercise-linked cardioprotection and mitochondrial susceptibility for the first time in cardiac I/R.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
TAp73 is a marker of glutamine addiction in medulloblastoma
STUDY_TYPE
siRNA constructs targeting p73 (ID: 2671-AMBION) and a non-targeting control siRNA (scramble) were transfected with 10āpM siRNA with lipofectamine 3000 according to the supplierās protocol for 48 hours
STUDY_SUMMARY
Metabolically-targeted therapies hold the promise of offering an effective and less toxic treatment for tumours including medulloblastoma, the most common malignant brain tumour of childhood. Current treatment relies on the sensitivity of these tumours to DNA damage that was discovered more than 50 years ago. Finding new tumour-specific susceptibilities to complement sensitivity to DNA damage is key to developing new more effective adjuvant therapies. The specific metabolic program of tumours is an attractive vulnerability, as restriction diet are low cost and easy to implement. Here, we present compelling pre-clinical evidence that glutamine restriction diet can be used as an adjuvant treatment for p73-expressing medulloblastoma.
INSTITUTE
Queen Mary University of London
DEPARTMENT
Blizard Institute
LABORATORY
Centre for Genomics and Child Health
LAST_NAME
Marino
FIRST_NAME
Silvia
ADDRESS
4 Newark Street, E1 2AT, London
EMAIL
s.marino@qmul.ac.uk
PHONE
+44 20 7882 2360
NUM_GROUPS
2
TOTAL_SUBJECTS
18
STUDY_COMMENTS
We include 3 biological replicate with 3 technical replicates for each condition.
Alterations in Lipid, Amino Acid, and Energy Metabolism Distinguish Crohn Disease from Ulcerative Colitis and Control Subjects by Serum Metabolomic Profiling
STUDY_SUMMARY
Non-invasive biomarkers are needed in inflammatory bowel disease (IBD) to help define disease activity and identify underlying pathogenic mechanisms. We hypothesized that serum metabolomics, which produces unique metabolite profiles, can aid in this search. The aim of this study was to characterize serum metabolomic profiles in patients with IBD, and to assess for differences between patients with ulcerative colitis (UC), Crohn disease (CD), and non-IBD subjects. Serum samples from 20 UC, 20 CD, and 20 non-IBD control subjects were obtained along with patient characteristics, including medication use and clinical disease activity. Non-targeted metabolomic profiling was performed using ultra-high performance liquid chromatography/mass spectrometry (UPLC-MS/MS) optimized for basic or acidic species and hydrophilic interaction liquid chromatography (HILIC/UPLC-MS/MS).
INSTITUTE
Vanderbilt University Medical Center
LAST_NAME
Elizabeth
FIRST_NAME
Scoville
ADDRESS
1030C MRB IV, 2215 Garland Avenue, Nashville, TN, 37232, USA
Evidence that the metabolite repair enzyme NAD(P)HX epimerase has a moonlighting function
STUDY_SUMMARY
NAD(P)H-hydrate epimerase (EC 5.1.99.6) is known to help repair NAD(P)H hydrates (NAD(P)HX), which are damage products existing as R and S epimers. The S epimer is reconverted to NAD(P)H by a dehydratase; the epimerase facilitates epimer interconversion. Epimerase deficiency in humans causes a lethal disorder attributed to NADHX accumulation. However, bioinformatic evidence suggests caution about this attribution by predicting that the epimerase has a second function connected to vitamin B6 (pyridoxal 5'-phosphate and related compounds). Specifically, (i) the epimerase is fused to a B6 salvage enzyme in plants, (ii) epimerase genes cluster on the chromosome with B6-related genes in bacteria, and (iii) epimerase and B6-related genes are coexpressed in yeast and Arabidopsis. The predicted second function was explored in Escherichia coli, whose epimerase and dehydratase are fused and encoded by the yjeF gene. The putative NAD(P)HX epimerase active site has a conserved lysine residue (K192 in E. coli YjeF). Changing this residue to alanine cut in-vitro epimerase activity by ā„95% but did not affect dehydratase activity. Mutant cells carrying the K192A mutation had essentially normal NAD(P)HX levels, showing that the mutation had very little or no effect on NAD(P)HX repair in vivo. However, these cells showed metabolome changes, particularly in amino acids, that exceeded those in cells lacking the entire yjeF gene. The K192A mutant cells also had lower levels of free pyridoxal 5'-phosphate than wild-type cells. These results provide strong circumstantial evidence that the epimerase has a metabolic function beyond NAD(P)HX repair and that this function involves vitamin B6.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Pharmacometabolomic analysis of rat hippocampus and plasma in response to chronic stress and albiflorin treament
STUDY_TYPE
Animal study
STUDY_SUMMARY
Male Sprague-Dawley (SD) rats were exposed to one random stressor per day for 35 d. Following four weeks of stress exposure, rats with stress-induced depression-like behaviors were treated daily with either saline, fluoxetine or albiflorin for 7 days via intragastric gavage. Rat heparinized plasma was collected after drug treatment. After behavior tests, rats were sacrificed and the hippocampus was collected. Pharmacometabolomic analysis was conducted for both plasma and hippocampal samples
INSTITUTE
Beijing Woner Biotech Co. Ltd
LAST_NAME
Zhang
FIRST_NAME
Zuoguang
ADDRESS
No.9, Kexing Road, Fengtai, Beijing, Beijing, 101111, China
Murine vitamin A deficiency results in a hypermetabolic state and alterations in bacterial community structure and metabolism. (Liver)
STUDY_SUMMARY
Vitamin A deficiency (A-) is a significant public health problem. To better understand how vitamin A status influences gut microbiota and host metabolism, we systematically analyzed urine, cecum, serum, and liver samples from vitamin A sufficient (A+) and A- mice using 1H NMR-based metabolomics, quantitative (q)PCR, and 16S rRNA gene sequencing coupled with multivariate data analysis. The microbiota in the cecum of A- mice showed compositional as well as functional shifts compared to the microbiota from A+ mice. Targeted 1H NMR analyses revealed significant changes in microbial metabolite concentrations including higher butyrate and hippurate and decreased acetate and 4-hydroxyphenylacetate in A+ relative to A- mice. Bacterial butyrate-producing genes including butyryl-CoA:acetate CoA-transferase and butyrate kinase were significantly higher in bacteria from A+ versus bacteria from A- mice. A - mice had disturbances in multiple metabolic pathways including alterations in energy metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein biosynthesis) and the A- host showed metabolites indicative of a hypermetabolic state (higher levels of amino acids and nucleic acids). A- mice had hyperglycemia, liver dysfunction, changes in bacterial metabolism, and altered gut microbial communities. Moreover, integrative analyses indicated a strong correlation between gut microbiota and host energy metabolism pathways in the liver. Vitamin A regulates the microbiota, bacterial metabolism and the effects of vitamin A on the microbiota results in alterations to host metabolism.
INSTITUTE
Pennsylvania State University
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
101 Life science building, University park, PA, 16803
Murine vitamin A deficiency results in a hypermetabolic state and alterations in bacterial community structure and metabolism. (Serum)
STUDY_SUMMARY
Vitamin A deficiency (A-) is a significant public health problem. To better understand how vitamin A status influences gut microbiota and host metabolism, we systematically analyzed urine, cecum, serum, and liver samples from vitamin A sufficient (A+) and A- mice using 1H NMR-based metabolomics, quantitative (q)PCR, and 16S rRNA gene sequencing coupled with multivariate data analysis. The microbiota in the cecum of A- mice showed compositional as well as functional shifts compared to the microbiota from A+ mice. Targeted 1H NMR analyses revealed significant changes in microbial metabolite concentrations including higher butyrate and hippurate and decreased acetate and 4-hydroxyphenylacetate in A+ relative to A- mice. Bacterial butyrate-producing genes including butyryl-CoA:acetate CoA-transferase and butyrate kinase were significantly higher in bacteria from A+ versus bacteria from A- mice. A - mice had disturbances in multiple metabolic pathways including alterations in energy metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein biosynthesis) and the A- host showed metabolites indicative of a hypermetabolic state (higher levels of amino acids and nucleic acids). A- mice had hyperglycemia, liver dysfunction, changes in bacterial metabolism, and altered gut microbial communities. Moreover, integrative analyses indicated a strong correlation between gut microbiota and host energy metabolism pathways in the liver. Vitamin A regulates the microbiota, bacterial metabolism and the effects of vitamin A on the microbiota results in alterations to host metabolism.
INSTITUTE
Pennsylvania State University
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
101 Life science building, University park, PA, 16803
Murine vitamin A deficiency results in a hypermetabolic state and alterations in bacterial community structure and metabolism.(Urine)
STUDY_SUMMARY
Vitamin A deficiency (A-) is a significant public health problem. To better understand how vitamin A status influences gut microbiota and host metabolism, we systematically analyzed urine, cecum, serum, and liver samples from vitamin A sufficient (A+) and A- mice using 1H NMR-based metabolomics, quantitative (q)PCR, and 16S rRNA gene sequencing coupled with multivariate data analysis. The microbiota in the cecum of A- mice showed compositional as well as functional shifts compared to the microbiota from A+ mice. Targeted 1H NMR analyses revealed significant changes in microbial metabolite concentrations including higher butyrate and hippurate and decreased acetate and 4-hydroxyphenylacetate in A+ relative to A- mice. Bacterial butyrate-producing genes including butyryl-CoA:acetate CoA-transferase and butyrate kinase were significantly higher in bacteria from A+ versus bacteria from A- mice. A - mice had disturbances in multiple metabolic pathways including alterations in energy metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein biosynthesis) and the A- host showed metabolites indicative of a hypermetabolic state (higher levels of amino acids and nucleic acids). A- mice had hyperglycemia, liver dysfunction, changes in bacterial metabolism, and altered gut microbial communities. Moreover, integrative analyses indicated a strong correlation between gut microbiota and host energy metabolism pathways in the liver. Vitamin A regulates the microbiota, bacterial metabolism and the effects of vitamin A on the microbiota results in alterations to host metabolism.
INSTITUTE
Pennsylvania State University
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
101 Life science building, University park, PA, 16803
Murine vitamin A deficiency results in a hypermetabolic state and alterations in bacterial community structure and metabolism. (Cecal contents)
STUDY_SUMMARY
Vitamin A deficiency (A-) is a significant public health problem. To better understand how vitamin A status influences gut microbiota and host metabolism, we systematically analyzed urine, cecum, serum, and liver samples from vitamin A sufficient (A+) and A- mice using 1H NMR-based metabolomics, quantitative (q)PCR, and 16S rRNA gene sequencing coupled with multivariate data analysis. The microbiota in the cecum of A- mice showed compositional as well as functional shifts compared to the microbiota from A+ mice. Targeted 1H NMR analyses revealed significant changes in microbial metabolite concentrations including higher butyrate and hippurate and decreased acetate and 4-hydroxyphenylacetate in A+ relative to A- mice. Bacterial butyrate-producing genes including butyryl-CoA:acetate CoA-transferase and butyrate kinase were significantly higher in bacteria from A+ versus bacteria from A- mice. A - mice had disturbances in multiple metabolic pathways including alterations in energy metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein biosynthesis) and the A- host showed metabolites indicative of a hypermetabolic state (higher levels of amino acids and nucleic acids). A- mice had hyperglycemia, liver dysfunction, changes in bacterial metabolism, and altered gut microbial communities. Moreover, integrative analyses indicated a strong correlation between gut microbiota and host energy metabolism pathways in the liver. Vitamin A regulates the microbiota, bacterial metabolism and the effects of vitamin A on the microbiota results in alterations to host metabolism.
INSTITUTE
Pennsylvania State University
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
101 Life science building, University park, PA, 16803
Metabolomics Linking Air Pollution, Obesity and Type 2 Diabetes
STUDY_TYPE
Untargeted high-resolution mass spectrometry profiling
STUDY_SUMMARY
The overall goal of this proposal is to use blood non-targeted high resolution metabolomics (HRM) to investigate the hypothesis that regional air pollution (NO2, PM2.5 and O3) and traffic-related air pollution exposures (traffic-related particulate matter components including EC2.5 and PM2.5 transition metals, and CALINE model-predicted NOx) alter key metabolic pathway(s) and that these alterations are associated with obesity and type 2 diabetes-related traits during the important developmental period of adolesence in the ongoing prospective Chidlren's Health study (CHS). Specific Aim 1 will examine the adverse impact of environmental chemicals in fasting blood samples measured by HRM on obesity (i.e., total body fat and body mass index (BMI)), metabolic dysfunction (e.g., fasting glucose and insulin concentrations and insulin resistance), and obesity-induced inflammation (i.e., leptin) among 104 Southern California adolescents enrolled in the CHS. Specific Aim 2 will examine associations of childhood exposures to PM2.5 and traffic-related air pollutants (i.e., CALINE model-predicted NOx) with biological metabolites identified in fasting blood samples using HRM among 104 adolescents in the CHS. Specific Aim 3 will investigate the metabolic pathways linking air pollution exposures and obesity and type 2 diabetes-related traits using pathway analysis under bayesian hierarchical model among 104 adolescents in the CHS.
INSTITUTE
Emory University School of Medicine
DEPARTMENT
Division of Pulmonary, Allergy and Critical Care Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Walker
FIRST_NAME
Douglas
ADDRESS
615 Michael St. Ste 225, Atlanta, GA, 30322, USA
EMAIL
douglas.walker@emory.edu
PHONE
(404) 727 5984
TOTAL_SUBJECTS
104
STUDY_COMMENTS
Both CHEAR and Clinical Biomarker Laboratory pooled plasma samples were used for quality control. Study specific sample pools were not created
Insights into the pathogenesis of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) through metabolomic profiling of cerebrospinal fluid (part I)
STUDY_SUMMARY
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a disabling illness characterized by six months or more of unexplained profound fatigue with post-exertional malaise, sleep abnormalities, cognitive dysfunction and autonomic disturbances. Focusing on the pathogenesis of central nervous system abnormalities in ME/CFS, we pursued metabolomics analysis of cerebrospinal fluid (CSF) in 32 ME/CFS cases, 40 subjects with multiple sclerosis (MS), another fatiguing illness, and 19 healthy subjects with no neurological disease (ND). MS/ND subjects were frequency matched for age and sex to ME/CFS subjects. Three untargeted metabolomic assays for primary metabolites, biogenic amines and complex lipids were performed with gas chromatography time-of-flight (GC-TOF) and liquid chromatographyātandem mass spectrometry (LC-MS/MS) yielding profiles for 525 known metabolites. Mannose was a cardinal biomarker in ME/CFS subjects with reduced levels in ME/CFS compared to both MS and ND subjects. Levels of acetylcarnitine were reduced in ME/CFS vs. MS subjects. The predictive power of metabolomic analysis for diagnosis of ME/CFS vs. ND was higher (cross-validated AUC 0.875; 95% CI: 0.726~0.949) than with cytokine analysis alone (cross-validated AUC 0.865; 95% CI: 0.673~0.952) and improved with integration of both metabolomics and cytokine analyses (cross-validated AUC 0.916; 95% CI: 0.791~0.969). Our findings confirm the biological basis of ME/CFS, and may enable new methods for diagnosis and insight into cognitive and autonomic disturbances in this syndrome.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Insights into the pathogenesis of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) through metabolomic profiling of cerebrospinal fluid (part II)
STUDY_SUMMARY
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a disabling illness characterized by six months or more of unexplained profound fatigue with post-exertional malaise, sleep abnormalities, cognitive dysfunction and autonomic disturbances. Focusing on the pathogenesis of central nervous system abnormalities in ME/CFS, we pursued metabolomics analysis of cerebrospinal fluid (CSF) in 32 ME/CFS cases, 40 subjects with multiple sclerosis (MS), another fatiguing illness, and 19 healthy subjects with no neurological disease (ND). MS/ND subjects were frequency matched for age and sex to ME/CFS subjects. Three untargeted metabolomic assays for primary metabolites, biogenic amines and complex lipids were performed with gas chromatography time-of-flight (GC-TOF) and liquid chromatographyātandem mass spectrometry (LC-MS/MS) yielding profiles for 525 known metabolites. Mannose was a cardinal biomarker in ME/CFS subjects with reduced levels in ME/CFS compared to both MS and ND subjects. Levels of acetylcarnitine were reduced in ME/CFS vs. MS subjects. The predictive power of metabolomic analysis for diagnosis of ME/CFS vs. ND was higher (cross-validated AUC 0.875; 95% CI: 0.726~0.949) than with cytokine analysis alone (cross-validated AUC 0.865; 95% CI: 0.673~0.952) and improved with integration of both metabolomics and cytokine analyses (cross-validated AUC 0.916; 95% CI: 0.791~0.969). Our findings confirm the biological basis of ME/CFS, and may enable new methods for diagnosis and insight into cognitive and autonomic disturbances in this syndrome.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Insights into the pathogenesis of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) through metabolomic profiling of cerebrospinal fluid
STUDY_SUMMARY
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a disabling illness characterized by six months or more of unexplained profound fatigue with post-exertional malaise, sleep abnormalities, cognitive dysfunction and autonomic disturbances. Focusing on the pathogenesis of central nervous system abnormalities in ME/CFS, we pursued metabolomics analysis of cerebrospinal fluid (CSF) in 32 ME/CFS cases, 40 subjects with multiple sclerosis (MS), another fatiguing illness, and 19 healthy subjects with no neurological disease (ND). MS/ND subjects were frequency matched for age and sex to ME/CFS subjects. Three untargeted metabolomic assays for primary metabolites, biogenic amines and complex lipids were performed with gas chromatography time-of-flight (GC-TOF) and liquid chromatographyātandem mass spectrometry (LC-MS/MS) yielding profiles for 525 known metabolites. Mannose was a cardinal biomarker in ME/CFS subjects with reduced levels in ME/CFS compared to both MS and ND subjects. Levels of acetylcarnitine were reduced in ME/CFS vs. MS subjects. The predictive power of metabolomic analysis for diagnosis of ME/CFS vs. ND was higher (cross-validated AUC 0.875; 95% CI: 0.726~0.949) than with cytokine analysis alone (cross-validated AUC 0.865; 95% CI: 0.673~0.952) and improved with integration of both metabolomics and cytokine analyses (cross-validated AUC 0.916; 95% CI: 0.791~0.969). Our findings confirm the biological basis of ME/CFS, and may enable new methods for diagnosis and insight into cognitive and autonomic disturbances in this syndrome.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Insights into the pathogenesis of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) through metabolomic profiling of cerebrospinal fluid
STUDY_SUMMARY
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a disabling illness characterized by six months or more of unexplained profound fatigue with post-exertional malaise, sleep abnormalities, cognitive dysfunction and autonomic disturbances. Focusing on the pathogenesis of central nervous system abnormalities in ME/CFS, we pursued metabolomics analysis of cerebrospinal fluid (CSF) in 32 ME/CFS cases, 40 subjects with multiple sclerosis (MS), another fatiguing illness, and 19 healthy subjects with no neurological disease (ND). MS/ND subjects were frequency matched for age and sex to ME/CFS subjects. Three untargeted metabolomic assays for primary metabolites, biogenic amines and complex lipids were performed with gas chromatography time-of-flight (GC-TOF) and liquid chromatographyātandem mass spectrometry (LC-MS/MS) yielding profiles for 525 known metabolites. Mannose was a cardinal biomarker in ME/CFS subjects with reduced levels in ME/CFS compared to both MS and ND subjects. Levels of acetylcarnitine were reduced in ME/CFS vs. MS subjects. The predictive power of metabolomic analysis for diagnosis of ME/CFS vs. ND was higher (cross-validated AUC 0.875; 95% CI: 0.726~0.949) than with cytokine analysis alone (cross-validated AUC 0.865; 95% CI: 0.673~0.952) and improved with integration of both metabolomics and cytokine analyses (cross-validated AUC 0.916; 95% CI: 0.791~0.969). Our findings confirm the biological basis of ME/CFS, and may enable new methods for diagnosis and insight into cognitive and autonomic disturbances in this syndrome.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic adaptation of a deep sea Microbacterium sediminis to prolonged low temperature under high pressure
STUDY_SUMMARY
Low temperature is the most wide-spread āhostileā environmental factor on earth while at the same time the most common condition for marine organisms. However, the unique adaptive mechanisms that enable the survival of marine microorganisms under low temperature are unclear. Since low temperature is always accompanied by high pressure and other adverse conditions in marine environment, here we studied the metabolic adaptation of a marine strain to prolonged low temperature under high pressure. The strain studied is a psychrotolerant Microbacterium sediminis isolated from deep sea sediment. By using 1H nuclear magnetic resonance (NMR)-based metabolomics approach, we detected the spectral data of polar extracts of the strain M. sediminis, and applied multivariate statistical analysis methods together with univariate analysis to analyze metabolic profiles associated to different conditions. The metabolic profiles of the M. sediminis strain cultivated under high pressure at low temperature were distinctly different from those cultivated under high pressure at normal temperature. We identified the differential metabolites which were responsible for distinguishing the metabolic profiles and compared their relative intensities between groups. We also compared the different adaptive responses of the strain at different growth stages to the prolonged low temperature under high pressure. We proposed that the low-temperature adapting process of the M. sediminis strain involves, 1) the regulation of osmotic pressure using amino acids as possible alternative osmolytes, and, 2) the strengthen of glycolysis and the maintenance of TCA cycle via amino acids anaplerotic reaction. We put forward that the main difference of adaptation to low temperature for the strain at different growth stages was related to energy metabolism. Our findings improved the understanding of the low-temperature adaptive mechanisms for marine microorganisms under high pressure on the metabolic level.
INSTITUTE
Third Institute of Oceanography, State Oceanic Administration
Biomarkers of NAFLD progression: a lipidomics approach to an epidemic. Part 1:Liver
STUDY_TYPE
Lipidomics Study
STUDY_SUMMARY
The spectrum of nonalcoholic fatty liver disease (NAFLD) includes steatosis, nonalcoholic steatohepatitis (NASH), and cirrhosis. Recognition and timely diagnosis of these different stages, particularly NASH, is important for both potential reversibility and limitation of complications. Liver biopsy remains the clinical standard for definitive diagnosis. Diagnostic tools minimizing the need for invasive procedures or that add information to histologic data are important in novel management strategies for the growing epidemic of NAFLD. We describe an 'omics' approach to detecting a reproducible signature of lipid metabolites, aqueous intracellular metabolites, SNPs, and mRNA transcripts in a double-blinded study of patients with different stages of NAFLD that involves profiling liver biopsies, plasma, and urine samples. Using linear discriminant analysis, a panel of 20 plasma metabolites that includes glycerophospholipids, sphingolipids, sterols, and various aqueous small molecular weight components involved in cellular metabolic pathways, can be used to differentiate between NASH and steatosis. This identification of differential biomolecular signatures has the potential to improve clinical diagnosis and facilitate therapeutic intervention of NAFLD.
Biomarkers of NAFLD progression: a lipidomics approach to an epidemic. Part 1:Plasma
STUDY_TYPE
Lipidomics Study
STUDY_SUMMARY
The spectrum of nonalcoholic fatty liver disease (NAFLD) includes steatosis, nonalcoholic steatohepatitis (NASH), and cirrhosis. Recognition and timely diagnosis of these different stages, particularly NASH, is important for both potential reversibility and limitation of complications. Liver biopsy remains the clinical standard for definitive diagnosis. Diagnostic tools minimizing the need for invasive procedures or that add information to histologic data are important in novel management strategies for the growing epidemic of NAFLD. We describe an 'omics' approach to detecting a reproducible signature of lipid metabolites, aqueous intracellular metabolites, SNPs, and mRNA transcripts in a double-blinded study of patients with different stages of NAFLD that involves profiling liver biopsies, plasma, and urine samples. Using linear discriminant analysis, a panel of 20 plasma metabolites that includes glycerophospholipids, sphingolipids, sterols, and various aqueous small molecular weight components involved in cellular metabolic pathways, can be used to differentiate between NASH and steatosis. This identification of differential biomolecular signatures has the potential to improve clinical diagnosis and facilitate therapeutic intervention of NAFLD.
Biomarkers of NAFLD progression: a lipidomics approach to an epidemic. Part 3:Urine
STUDY_TYPE
Lipidomics Study
STUDY_SUMMARY
The spectrum of nonalcoholic fatty liver disease (NAFLD) includes steatosis, nonalcoholic steatohepatitis (NASH), and cirrhosis. Recognition and timely diagnosis of these different stages, particularly NASH, is important for both potential reversibility and limitation of complications. Liver biopsy remains the clinical standard for definitive diagnosis. Diagnostic tools minimizing the need for invasive procedures or that add information to histologic data are important in novel management strategies for the growing epidemic of NAFLD. We describe an 'omics' approach to detecting a reproducible signature of lipid metabolites, aqueous intracellular metabolites, SNPs, and mRNA transcripts in a double-blinded study of patients with different stages of NAFLD that involves profiling liver biopsies, plasma, and urine samples. Using linear discriminant analysis, a panel of 20 plasma metabolites that includes glycerophospholipids, sphingolipids, sterols, and various aqueous small molecular weight components involved in cellular metabolic pathways, can be used to differentiate between NASH and steatosis. This identification of differential biomolecular signatures has the potential to improve clinical diagnosis and facilitate therapeutic intervention of NAFLD.
Plasma metabolic fingerprint for breast cancer (MS) - part I
STUDY_TYPE
control - case
STUDY_SUMMARY
Breast cancer is a highly heterogeneous disease associated with metabolic reprogramming. The shifts in the metabolome caused by breast cancer still lack data from Latin populations of Hispanic origin. In this pilot study, metabolomic and lipidomic approaches were performed to establish a plasma metabolic fingerprint of Colombian Hispanic women with breast cancer. NMR, GC-MS and LC-MS datasets were combined and compared. Statistics showed discrimination between breast cancer and healthy subjects on all analytical platforms.
Discovery of Lipidome Alterations Following Traumatic Brain Injury via High-Resolution Metabolomics
STUDY_TYPE
Untargeted lipidomics
STUDY_SUMMARY
Traumatic brain injury (TBI) can occur across wide segments of the population, presenting in a heterogeneous manner that makes diagnosis inconsistent and management challenging. Biomarkers offer the potential to objectively identify injury status, severity, and phenotype by measuring the relative concentrations of endogenous molecules in readily accessible biofluids. Through a data-driven, discovery approach, novel biomarker candidates for TBI were identified in the serum lipidome of adult male Sprague-Dawley rats in the first week following moderate controlled cortical impact (CCI). Serum samples were analyzed in positive and negative ion modes by Ultra Performance Liquid Chromatography Mass Spectrometry (UPLC-MS). A predictive panel for the classification of injured and uninjured sera samples, consisting of 26 dysregulated species belonging to a variety of lipid classes, was developed with a cross-validated accuracy of 85.3% using omniClassifier software to optimize feature selection,. Polyunsaturated fatty acids (PUFAs) and PUFA-containing diacylglycerols were found to be upregulated in sera from injured rats, while changes in sphingolipids and other membrane phospholipids were also observed, many of which map to known secondary injury pathways. Overall, the identified biomarker panel offers viable molecular candidates representing lipids that may readily cross the blood brain barrier (BBB) and aid in the understanding of TBI pathophysiology.
Karenia brevis allelopathy compromises the lipidome, membrane integrity, and photosynthetic efficiency of competitors
STUDY_TYPE
Untargeted lipidomics
STUDY_SUMMARY
Allelopathy, or the release of compounds that inhibit competitors, is a form of interference competition that is common among bloom-forming phytoplankton. Allelopathy is hypothesized to play a role in bloom propagation and maintenance and is well established in the red tide dinoflagellate Karenia brevis. K. brevis typically suppresses competitor growth through unknown mechanisms over the course of many days. When we investigated the effects of allelopathy on the lipidomes of two competing phytoplankton, Asterionellopsis glacialis and Thalassiosira pseudonana using nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS)- based metabolomics, we found that the lipidomes of both species were significantly altered, however A. glacialis maintained a more robust response whereas T. pseudonana saw significant alterations in fatty acid synthesis, cell membrane integrity, and a decrease in photosynthetic efficiency. Membrane- associated lipids were significantly suppressed for T. pseudonana exposed to allelopathy to the point of permeabilizing the cell membrane of living cells. The dominant mechanisms of K. brevis allelopathy appear to target lipid biosynthesis affecting multiple physiological pathways suggesting that exuded compounds have the ability to significantly alter competitor physiology and give K. brevis a competitive edge over sensitive species.
Longitudinal Metabolomics of the Human Microbiome in Inflammatory Bowel Disease
STUDY_SUMMARY
A number of factors contribute to the complex array of small molecules that occur in stool; including diet, gut flora, and gut function. Comprehensive profiling of the stool metabolome therefore can provide detailed phenotypic information on health status, metabolic interactions between the host and the microbiome, and interactions among gut microbes. Here, we applied metabolomics to characterize stool samples collected longitudinally from inflammatory bowel disease (IBD) patients and non-IBD controls who participated in the Integrative Human Microbiome Project (iHMP). A total of 546 stool samples were analyzed using a platform comprised of four complementary liquid chromatography tandem mass spectrometry (LC-MS) methods designed to measure polar metabolites and lipids. Each method used high resolution/accurate mass (HRAM) profiling to measure both metabolites of confirmed identity and yet to be identified metabolite peaks. 81,867 de-isotoped LC-MS peaks were measured, out of which 597 were annotated based on confirmation with authentic reference standards. Pooled stool extracts inserted and analyzed throughout the analysis queues to evaluate analytical reproducibility showed a median coefficient of variation of 5.1% among known metabolites and 24.2% across all 81,867 features. Owing to differences in water content and heterogeneity among stool samples, total median scaling was used to standardize the metabolomics data. In addition to being accessible at the Metabolomics Workbench repository, these metabolomics data will be incorporated into a multiāomic database (https://www.hmpdacc.org/ihmp/) that will enable the study of associations between the gut microbiome and IBD.
MuRF1-Related Metabolic Alterations in HL-1 Cardiomyocyte Induced by Cyclic Stretch
STUDY_TYPE
Cardiomyocyte cell culture
STUDY_SUMMARY
We collected cell media and performed GC-MS non-targeted metabolomics to identify the role of MuRF1 in the dynamic metabolic changes in cardiomyocytes.
MuRF1-Related Metabolomic Changes in Stretched and Unloaded HL-1 Cells
STUDY_SUMMARY
Introduction: Left ventricular assist devices (LVADs) provide significant pressure and volume unloading, which reverse key structural features of heart failure, including hypertrophy, fibrosis, and altered sympathetic innervation. This has led to LVADs increasing utilization as both a bridge or destination therapy for heart failure. While distinct metabolic changes occur in the human myocardium with LVAD placement, the specific molecular mechanisms underlying these changes have not been identified. Objectives: To identify the role of MuRF1 in the metabolic changes in cardiomyocytes unloading in vitro. Methods: HL-1 atrial cardiomyocyte cells were plated on silastic membranes coated with gelatin/fibronectin and transduced with AdshRNA MuRF1 (or AdshRNA Scramble control) to knock-down MuRF1 protein to <25% of controls and subjected to 15% biaxial stretch at 1 Hz using the Flexcell FX-5000ā¢ Compression System in serum free DMEM (or no-stretch). After 6 hours stretch, media was collected in parallel with time-matched no-stretch controls, followed by serial collections at 1, 3, 6, and 12 hours unloading (i.e. after termination of stretch). Media was analyzed by untargeted metabolomics using GC-MS. Results: After 6 hours stretch, control HL-1 cell media (AdshRNA Scramble) had 19 significantly altered metabolites compared to non-stretched control cell media (AdshRNA Scramble) by t-test, involved in: 1) glyoxylate and dicarboxylate metabolism (p=0.00087043, FDR=0.071375), 2) methane metabolism (p=0.0041113, FDR=0.089824), 3) glycine, serine and threonine metabolism (p=0.0043816, FDR=0.089824), and 4) aminoacyl-tRNA biosynthesis(p=0.0060141, FDR=0.09863). To determine the role of MuRF1 in stretch, we additionally compared the AdshRNA Scramble groups to AdshRNA MuRF1 +/- stretch at 6 hours and identified 41 significant metabolites (of 79 identified) by ANOVA, involved in: 1) phenylalanine, tyrosine, and tryptophan biosynthesis (p=0.0036482, FDR=0.05983), 2) aminoacyl-tRNA biosynthesis (p=3.74E-07, FDR=3.06E-05), and 3) valine, leucine, and isoleucine biosynthesis (p=0.0021624, FDR=0.053255). We next compared the 6 hour stretch time point of the four groups to the 1, 3, 6, and 12 hours unloading conditions to identify MuRF1-associated metabolites during the unloading period. At 12 hours of unloading (representative 1, 3, and 6 hours unloading), MuRF1 knock-down cell media had 35 (of 82 named metabolites) significantly different metabolites by ANOVA involved in 1) phenylalanine, tyrosine and tryptophan biosynthesis (p=0.0023668, FDR=0.048519), 2) aminoacyl-tRNA biosynthesis (p=0.0011403, FDR=0.0032631), 3) phenylalanine metabolism (P= 0.0011403, FDR 0.042673),and arginine and proline metabolism (p=0.0015612, FDR 0.042673). A final analysis comparing all four groups across all five time points identified 21 significant metabolites. When MuRF1 was knocked down (no stretch), these 21 metabolites were significantly increased without stretch. In response to stretch and unloading, these metabolites were further decreased in AdshRNA Scramble (control) cell media decreased. In contrast, these stretch and unloading induced decreases were attenuated in AdshRNA MuRF1 cell media. These metabolites included nine (9) amino acids (citrulline, phenylalanine, tyrosine, asparagine, 2-ketovaline, and -ketoleucine/ketoisoleucine, threonine, isoleucine, leucine), three (3) metabolic co-factors (Pantothenic acid, Myoinositol, 5,6-Dihydrouracil), and one fatty acid (stearic acid). Conclusion: These studies identify for the first time a role for MuRF1 in generating key amino acids recently reported in LVAD unloading in human myocardium using an in vitro model of atrial cardiomyocyte unloading.
INSTITUTE
University of North Carolina
DEPARTMENT
McAllister heart Institute, Department of Internal medicine
LABORATORY
Multiple Centers
LAST_NAME
Willis
FIRST_NAME
Monte
ADDRESS
111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Probing the metabolic phenotype of breast cancer cells by multiple tracer stable isotope resolved metabolomics (part I)
STUDY_SUMMARY
Breast cancers vary by their origin and specific set of genetic lesions, which gives rise to distinct phenotypes and differential response to targeted and untargeted chemotherapies. To explore the functional differences of different breast cell types, we performed Stable Isotope Resolved Metabolomics (SIRM) studies of one primary breast (HMEC) and three breast cancer cells (MCF-7, MDAMB-231, and ZR75-1) having distinct genotypes and growth characteristics, using 13C6-glucose, 13C-1+2-glucose, 13C5,15N2-Gln, 13C3-glycerol, and 13C8-octanoate as tracers. These tracers were designed to probe the central energy producing and anabolic pathways (glycolysis, pentose phosphate pathway, Krebs Cycle, glutaminolysis, nucleotide synthesis and lipid turnover). We found that glycolysis was not associated with the rate of breast cancer cell proliferation, glutaminolysis did not support lipid synthesis in primary breast or breast cancer cells, but was a major contributor to pyrimidine ring synthesis in all cell types; anaplerotic pyruvate carboxylation was activated in breast cancer versus primary cells. We also found that glucose metabolism in individual breast cancer cell lines differed between in vitro cultures and tumor xenografts, but not the metabolic distinctions between cell lines, which may reflect the influence of tumor architecture/microenvironment.
INSTITUTE
University of Kentucky
LAST_NAME
Lane
FIRST_NAME
Andrew
ADDRESS
Rm 516 Biopharm Complex, 789 S. Limestone St.,Univ. of Kentucky, Lexington, KY 40536
Probing the metabolic phenotype of breast cancer cells by multiple tracer stable isotope resolved metabolomics (part II)
STUDY_SUMMARY
Breast cancers vary by their origin and specific set of genetic lesions, which gives rise to distinct phenotypes and differential response to targeted and untargeted chemotherapies. To explore the functional differences of different breast cell types, we performed Stable Isotope Resolved Metabolomics (SIRM) studies of one primary breast (HMEC) and three breast cancer cells (MCF-7, MDAMB-231, and ZR75-1) having distinct genotypes and growth characteristics, using 13C6-glucose, 13C-1+2-glucose, 13C5,15N2-Gln, 13C3-glycerol, and 13C8-octanoate as tracers. These tracers were designed to probe the central energy producing and anabolic pathways (glycolysis, pentose phosphate pathway, Krebs Cycle, glutaminolysis, nucleotide synthesis and lipid turnover). We found that glycolysis was not associated with the rate of breast cancer cell proliferation, glutaminolysis did not support lipid synthesis in primary breast or breast cancer cells, but was a major contributor to pyrimidine ring synthesis in all cell types; anaplerotic pyruvate carboxylation was activated in breast cancer versus primary cells. We also found that glucose metabolism in individual breast cancer cell lines differed between in vitro cultures and tumor xenografts, but not the metabolic distinctions between cell lines, which may reflect the influence of tumor architecture/microenvironment.
INSTITUTE
University of Kentucky
LAST_NAME
Lane
FIRST_NAME
Andrew
ADDRESS
Rm 516 Biopharm Complex, 789 S. Limestone St.,Univ. of Kentucky, Lexington, KY 40536
Probing the metabolic phenotype of breast cancer cells by multiple tracer stable isotope resolved metabolomics (part III)
STUDY_SUMMARY
Breast cancers vary by their origin and specific set of genetic lesions, which gives rise to distinct phenotypes and differential response to targeted and untargeted chemotherapies. To explore the functional differences of different breast cell types, we performed Stable Isotope Resolved Metabolomics (SIRM) studies of one primary breast (HMEC) and three breast cancer cells (MCF-7, MDAMB-231, and ZR75-1) having distinct genotypes and growth characteristics, using 13C6-glucose, 13C-1+2-glucose, 13C5,15N2-Gln, 13C3-glycerol, and 13C8-octanoate as tracers. These tracers were designed to probe the central energy producing and anabolic pathways (glycolysis, pentose phosphate pathway, Krebs Cycle, glutaminolysis, nucleotide synthesis and lipid turnover). We found that glycolysis was not associated with the rate of breast cancer cell proliferation, glutaminolysis did not support lipid synthesis in primary breast or breast cancer cells, but was a major contributor to pyrimidine ring synthesis in all cell types; anaplerotic pyruvate carboxylation was activated in breast cancer versus primary cells. We also found that glucose metabolism in individual breast cancer cell lines differed between in vitro cultures and tumor xenografts, but not the metabolic distinctions between cell lines, which may reflect the influence of tumor architecture/microenvironment.
INSTITUTE
University of Kentucky
LAST_NAME
Lane
FIRST_NAME
Andrew
ADDRESS
Rm 516 Biopharm Complex, 789 S. Limestone St.,Univ. of Kentucky, Lexington, KY 40536
Metabolomics Analysis of Aqueous Humour in the Presence of Glaucoma Drainage Devices
STUDY_TYPE
Metabolomics Evaluation of Aqueous Humor Samples from Left (treated) and Right (Control) Eyes of Rabbits.
STUDY_SUMMARY
Metabolomic experiments were carried out on aqueous humor samples provided by Ramesh Ayyala. Through this collaboration, we propose to study aqueous humor from control and experimental rabbits to better understand metabolic differences in the aqueous humor samples.
Define alterations in the gut metabolome of mice infected with C. difficile
STUDY_TYPE
Time course experiment
STUDY_SUMMARY
C57BL/6 mice were treated with antibiotics to make them susceptible to C. difficile. Mice were challenged with C. difficile and the infection was monitored for 30 hours post challenge. Mice were sacrificed at 0 hr, 12 hr, 24 hr, and 30 hr post challenge with C. difficile and gut content was collected for untargeted metabolomic analysis. The aim of this project was to define the gut metabolome throughout C. difficile infection.
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3915 (CwlM)
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv0100
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Metabolome profiles in urogenital schistosomiasis and associated pathologies
STUDY_SUMMARY
Metabolic fingerprint aid discovery of new biomarkers. Blood and urine were collected from volunteers in Eggua community, Nigeria, after ethical approval. LC-ESI-MS were used to analyse samples. Bioinformatics and statistical tools were used to detect important metabolites
INSTITUTE
University of Ibadan, Nigeria
DEPARTMENT
Zoology (Cell biology and Genetics unit)
LABORATORY
Centre for Materials Characterization, National Chemical Laboratory, Pune, India
LAST_NAME
Adebayo
FIRST_NAME
Adewale
ADDRESS
Cell Biology and Genetics unit, Department of zoology, University of ibadan, Nigeria/ National Centre for Cell Science, Pune, India
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv2553c
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv2247
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3208
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
"Utilizing Ion Mobility Spectrometry and Mass Spectrometry for the Analysis of Polycyclic Aromatic Hydrocarbons, Polychlorinated Biphenyls, Polybrominated Diphenyl Ethers and Their Metabolites mass spectrometry"
STUDY_TYPE
Structural analysis of xenobiotics using ion mobility
STUDY_SUMMARY
Standards of xenobiotics were analyzed with Agilent 6560 Ion mobility QTOF MS platform to find Collision Cross Section values. All measurements were performed in triplicate in both positive and negative polarities with nitrogen gas and at seven different electric fields, so that values could be directly measured and random standard deviations (RSD) assessed for each molecule.
INSTITUTE
Pacific Northwest National Laboratory
DEPARTMENT
Integrative Omics
LABORATORY
Pacific Northwest National Laboratory
LAST_NAME
Baker
FIRST_NAME
Erin
ADDRESS
Pacific Northwest National Laboratory 902 Battelle Boulevard Richland, WA
Predicting and Defining Steroid Resistance in Pediatric Nephrotic Syndrome using Plasma Metabolomics
STUDY_TYPE
Discovery Metabolomics
STUDY_SUMMARY
Paired citrate plasma samples were collected from 27 steroid-sensitive and 14 steroid-resistant nephrotic syndrome participants prior to beginning treatment and after an average of 7 weeks (range 3-19 wks) of treatment with a corticosteroid.
INSTITUTE
RTI International
DEPARTMENT
Discovery-Sciences-Technology
LABORATORY
NIH Eastern Regional Comprehensive Metabolomics Resource Core at UNC Chapel Hill (ERCMRC)
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
3040 E. Cornwallis Road, Research Triangle Park, NC 27709, USA
EMAIL
susan_sumner@unc.edu
PHONE
704-250-5000
TOTAL_SUBJECTS
86
STUDY_COMMENTS
Factor Description 1. Treatment Response R = Steroid Resistant; S = Steroid Sensitive; Steroid resistance was defined as failure to enter complete remission after 6-8 weeks of daily oral steroids; Steroid sensitive was defined as clinical remission after 6-8 weeks of daily oral steroids. 2. Draw Number Draw 1 sample, āPreā, was collected at the time of disease presentation before even a single dose of glucocorticoids, and Draw 2 sample, āPostā, was collected after 6-10 weeks of Glucocorticoid therapy. 3. Age Age in years at the time of Draw 1, or 'Pre', sample collection Other sample data Weeks difference Time in weeks between draw 1, 'pre', and draw 2, 'post', treatment sample collection Gender M = male; F = female Race self-reported (Native American or Alaskan Native/Asian/Black or African American/Native Hawaiian or Other Pacific Islander/White/More than one race) Ethnicity self-reported (Hispanic or Latino/Not Hispanic or Latino/Unknown)
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3285
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3651
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3799c
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Activity-Based Metabolic Profiling of M. tuberculosis H37Rv Rv3916c
STUDY_TYPE
timecourse
STUDY_SUMMARY
These experiments involve incubating various proteins of unknown function in the M. tuberculosis genome with whole-cell lysate and assaying changes in metabolite levels over time (e.g. 0, 15, 30min) to try to infer what reaction they might catalyze.
INSTITUTE
Weill Cornell Medical College, NY
LAST_NAME
Rhee
FIRST_NAME
Kyu
ADDRESS
Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY
Specific Aims: (a) Identify sex differences in serum untargeted metabolomics profiles during late adolescence, (b) examine changes in metabolomics profiles between early and late adolescence in the same children, and (c) evaluate associations of exposure to prenatal and concurrent exposure to PAHs, metals, and EDCs with serum metabolite profiles among boys and girls, separately, during late adolescence.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
EMAIL
mkachman@med.umich.edu
PHONE
(734) 232-8175
NUM_GROUPS
2
TOTAL_SUBJECTS
207
STUDY_COMMENTS
The overall objective of this project is to expand upon available data from Early Life Exposure in Mexico to ENvironental Toxicants (ELEMENT) Project to examine effects of toxicant exposures during three sensitive developmental timeframes ā the prenatal period, early adolescence, and late adolescence ā on adiposity and metabolic risk in adolescence with an emphasis on sex differences and the related molecular underpinnings. In addition to existing exposure data on BPA, metals and phthalates, we propose to add measures of polycyclic aromatic hydrocarbon (PAH) metabolites in archived urine samples from mothers and children. We also propose to assess the epigenome at three developmental periods (prenatal aka in utero, early and late adolescence) and the circulating metabolome during late adolescence. We will incorporate data on toxicant exposures and āomics, including untargeted metabolomics of 404 teenagers from the ELEMENT cohorts, to addressing several specific aims.
Mechanisms for Insulin Resistance in Polycystic Ovary Syndrome: aminoadipic acid, lysine concentrations, and enrichment (part I)
STUDY_SUMMARY
"To determine whether altered lysine and Ī±-aminoadipic acid (AAA) kinetics explain previous observations of increased lysine and AAA concentrations in PCOS compared to controls, as measured by baseline lysine and AAA flux in PCOS versus healthy controls using [Ī±-15N]-lysine and [13C]-AAA stable isotope tracers as well as by comparing the conversion of [Ī±-15N]-lysine to [15N]-AAA. To evaluate how hyperinsulinemia affects lysine and AAA kinetics. Changes in lysine and AAA flux during a hyperinsulinemic-euglycemic clamp will be evaluated in healthy controls and compared to the baseline changes in lysine and AAA flux in PCOS."
INSTITUTE
Mayo Clinic
LAST_NAME
Chang
FIRST_NAME
Al
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Mechanisms for Insulin Resistance in Polycystic Ovary Syndrome: aminoadipic acid, lysine concentrations, and enrichment (part II)
STUDY_SUMMARY
"To determine how metformin therapy changes lysine and AAA kinetics in PCOS and whether this is associated with improvements in insulin sensitivity. Changes in lysine and AAA flux before and after three months of metformin therapy will be compared to women with PCOS randomized to no treatment for three months.
INSTITUTE
Mayo Clinic
LAST_NAME
Chang
FIRST_NAME
Alice
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Using Metabolomics and Lipidomics Analysis to Explore Metabolites and Pathways Associated Increased Airway Hyperresponsiveness in Patients with Asthma who are Identified by Race (African American) and Genotype (ADRB2 Arg16/Arg)
STUDY_TYPE
Open-label, prospective cohort study
STUDY_SUMMARY
Asthma is a heterogeneous disease largely defined by chronic airway inflammation with similar symptomatology in patients that includes wheezing, shortness of breath, chest tightness and cough. However, underlying these common symptoms are varying endotypes with distinct pathophysiological processes. Metabolomic studies in patients with asthma are emerging and suggest that metabolomics can characterize distinct asthma phenotypes. In a completed study, we identified a population of patients with asthma who have increased airway hyperresponsiveness (airway hyperresponsiveness is a marker for asthma disease severity) who are characterized by race (African American) and genotype (ADRB2 Arg16/Arg) compared with patients who have less airway hyperresponsiveness (African Americans and whites with differing ADRB2 genotypes). This group may represent a distinct endotype of asthma with unique metabolomic and lipidomic characteristics. The aims of this project are to (1) use metabolomic and lipidomic analysis to identify metabolites present in plasma in this population of patients with asthma who have increased airway hyperresponsiveness (African Americans who carry the ADRB2 Arg16/Arg genotype) and patients with asthma who have less airway hyperresponsiveness (African Americans and whites with differing ADRB2 genotypes); and (2) identify pathways that will improve the understanding of increased airway hyperresponsiveness in this population. We hypothesize that there will be unique metabolic pathways in the population with increased airway hyperresponsiveness that will be distinct from pathways in patients with lower airway hyperresponsiveness. In this project will use data and samples that were previously collected as part of the NIH funded project āPharmacogenetics of Ī²2-Agonists in Asthmaā (Blake, PI K23 HL081245). Blood was collected in 55 African Americans and whites after receiving 2-weeks treatment with inhaled fluticasone. Samples were stored on ice until processed and plasma frozen at -80Ā°C. If our findings indicate distinct metabolic pathways are present using global metabolomic and lipodomic analysis, we will seek to replicate our findings using samples and data from phenotypically well characterized participants who participated in trials conducted through the American Lung Association Airways Clinical Research Centers network, of which Nemours has been a highly productive site since 1999. Future controlled trials would be conducted to evaluate treatments based upon molecular pathways identified through metabolomic and lipidomic analysis.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Beecher
FIRST_NAME
Chris
ADDRESS
PO Box 100219 Gainesville FL 32610-0219 , Southeast Center for Integrated Metabolomics
EMAIL
chris@iroatech.com
PHONE
(352) 294-4385
NUM_GROUPS
4
TOTAL_SUBJECTS
55
STUDY_COMMENTS
Nubmer of groups : 4 (race x diplotype); SECIM pilot and feasibility, NIH U24 DK097209
Isolated T cells were activated for 48-72 hours in the presence and absence of 5mM DFMO, an ODC inhibitor
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Hesterberg
FIRST_NAME
Rebecca
ADDRESS
12902 Magnolia Dr
EMAIL
rebecca.hesterberg@moffitt.org
PHONE
813-270-9181
NUM_GROUPS
4
TOTAL_SUBJECTS
16
STUDY_COMMENTS
OT-1 mice express a transgenic T cell receptor in all CD8+ T cells that binds with high affinity to a peptide derived from the protein ovalbumin (SIINFEKL).
Determine metabolomics signatures important for the ability of Streptococus gallolyticus to promote colon cancer cell proliferation
STUDY_SUMMARY
HT29 cells were treated with Sg TX20005 stationary phase, Sg TX20005 exponential phase (negative control), and Sg TX20008 stationary phase (negative control), and media only (baseline control). The cells were washed and aliquoted. One aliquot was pelleted and stored at -80C for metabolomics analysis. DNA was also extracted from the other aliquot for DNA methylation studies.
Pediatric obesity has more than doubled in children and tripled in adolescents over the past 30 years. Recent findings demonstrate that differences in energy harvesting bacteria promote obesity in the host and appear to be influenced by early life factors such as mode of delivery, maternal obesity, and breastfeeding. The goal of this proposal is to investigate how human milk impacts the infant gut microbiome during the first 12-months of life and identify the microbe-host interactions that mediates the protective role of breastfeeding on infant adiposity. The results of this exploratory study will characterize factors that influence microbial transmission between mothers and offspring and identify human milk compounds that stabilize a healthy infant microbiome with potential to reduce pediatric obesity.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LABORATORY
R1-187
LAST_NAME
Carney
FIRST_NAME
Olivia
ADDRESS
Clinical and Translational Research Building, University of Florida College of Medicine, 2004 Mowry Road, Gainesville, FL 32608
EMAIL
ocarney1@ufl.edu
PHONE
352-294-8361
NUM_GROUPS
3
TOTAL_SUBJECTS
12
STUDY_COMMENTS
12 samples (4 mothers with 3 human milk fractions: fat, skim and whole)
The Influence of Sugar, Artificial Sweeteners, and the Microbiome on Rodent TCA Concentrations (part I)
STUDY_SUMMARY
Targeted TCA concentrations of rodents treated with diets rich in commonly used artifically sweeteners will be assessed in this study. We hypothesized that a specific subset of plasma metabolites are generated as a result from a diet rich in commonly used artificial sweeteners and their subsequent processing by the gut microbiome, which could ultimately lead to impaired glycemic control and negative physiological health outcomes. To test this hypothesis we administered normal, high glucose, fructose, aspartame, and acesulfame potassium diets to rats for 3 weeks, followed by a plasma collection through cardiac puncture and metabolic analysis. We also treated the gut microbiota with in rats with the same diets plus bacitracin/streptomycin to observe how alterations of the microbiome influence the plasma metabolic profile in these animals. The resulting data will give us insights into the influence of high sugar and artificial sweetener diets on homeostatic metabolic processes and dive into the symbiotic relationship of the gut microbiome with this process.
The Influence of Sugar, Artificial Sweeteners, and the Microbiome on Rodent Diacylglycerols (part II)
STUDY_SUMMARY
Targeted diacylglycerols panel of rodents treated with diets rich in commonly used artifically sweeteners will be assessed in this study. We hypothesized that a specific subset of plasma metabolites are generated as a result from a diet rich in commonly used artificial sweeteners and their subsequent processing by the gut microbiome, which could ultimately lead to impaired glycemic control and negative physiological health outcomes. To test this hypothesis we administered normal, high glucose, fructose, aspartame, and acesulfame potassium diets to rats for 3 weeks, followed by a plasma collection through cardiac puncture and metabolic analysis. We also treated the gut microbiota with in rats with the same diets plus bacitracin/streptomycin to observe how alterations of the microbiome influence the plasma metabolic profile in these animals. The resulting data will give us insights into the influence of high sugar and artificial sweetener diets on homeostatic metabolic processes and dive into the symbiotic relationship of the gut microbiome with this process.
The Influence of Sugar, Artificial Sweeteners, and the Microbiome on Rodent Triglyceride Composition (part III)
STUDY_SUMMARY
Targeted triglyceride concentration panel of rodents treated with diets rich in commonly used artifically sweeteners will be assessed in this study. We hypothesized that a specific subset of plasma metabolites are generated as a result from a diet rich in commonly used artificial sweeteners and their subsequent processing by the gut microbiome, which could ultimately lead to impaired glycemic control and negative physiological health outcomes. To test this hypothesis we administered normal, high glucose, fructose, aspartame, and acesulfame potassium diets to rats for 3 weeks, followed by a plasma collection through cardiac puncture and metabolic analysis. We also treated the gut microbiota with in rats with the same diets plus bacitracin/streptomycin to observe how alterations of the microbiome influence the plasma metabolic profile in these animals. The resulting data will give us insights into the influence of high sugar and artificial sweetener diets on homeostatic metabolic processes and dive into the symbiotic relationship of the gut microbiome with this process.
The Influence of Sugar, Artificial Sweeteners, and the Microbiome on Rodent Amino Acids (part IV)
STUDY_SUMMARY
Targeted Amino Acid panel of rodents treated with diets rich in commonly used artifically sweeteners will be assessed in this study. We hypothesized that a specific subset of plasma metabolites are generated as a result from a diet rich in commonly used artificial sweeteners and their subsequent processing by the gut microbiome, which could ultimately lead to impaired glycemic control and negative physiological health outcomes. To test this hypothesis we administered normal, high glucose, fructose, aspartame, and acesulfame potassium diets to rats for 3 weeks, followed by a plasma collection through cardiac puncture and metabolic analysis. We also treated the gut microbiota with in rats with the same diets plus bacitracin/streptomycin to observe how alterations of the microbiome influence the plasma metabolic profile in these animals. The resulting data will give us insights into the influence of high sugar and artificial sweetener diets on homeostatic metabolic processes and dive into the symbiotic relationship of the gut microbiome with this process.
Lipidomics of inflammation-induced optic nerve regeneration
STUDY_TYPE
untargeted LC-MS/MS profiling
STUDY_SUMMARY
In adult mammals, retinal ganglion cells (RGCs) fail to regenerate their axons when damaged. As a result, RGCs die after acute injury and in progressive degenerative diseases such as glaucoma; such damage can lead to permanent vision loss and blindness. Little is known about the roles of lipids in axon injury and repair despite their fundamental importance in composition of cell membranes, myelin sheaths and mediation of signaling pathways. Study of the lipidome in the biology of optic nerve (ON) regeneration has been largely neglected. A better understanding of the roles that lipids play in RGC biology may enhance understanding of RGC-related diseases and point to novel treatments. Established experimental models of ON regeneration allow exploration of molecular determinants of RGC axon regenerative success and failure. In this study, we used high-resolution liquid chromatography-tandem mass spectrometry to analyze lipidomic profiles of the ON and retina in an ON crush model with and without intravitreal Zymosan injections to enhance regeneration. Our results reveal profound remodeling of retina and ON lipidomes that occur after injury. In the retina, Zymosan treatment largely abrogates widespread lipidome alterations. In the ON, Zymosan induces lipid profiles that are distinct from those observed in naĆÆve and vehicle-injected crush controls. We have identified a number of lipid species, classes and fatty acids that may be involved in the mechanisms of axon damage and repair. Lipids upregulated during RGC regeneration may be interesting candidates for further functional studies.
U13C Glucose Tracing of Young and Old Pancreatic Islets
STUDY_SUMMARY
Pancreatic islets from young and old mice were traced with either 2.8mM or 16.8mM U13C glucose to determine the effect of age upon glucose metabolism in basal and stimulated states.
Timecourse of U13C glucose labeling of pancreatic islets in young mice
STUDY_SUMMARY
Pancreatic islets from young mice were traced with 16.8mM U13C glucose over a 90 minute timecourse to determine the rate of metabolite labeling during glucose stimulation.
Metabolite and gene expression profiles suggest a putative mechanism through which high dietary carbohydrates reduce the content of hepatic betaine in Megalobrama amblycephala
STUDY_SUMMARY
plasma of Megalobrama amblycephala fed with control diet, high carbohydrates diet,betaine diet with 16 weeks and betaine diet with 4 weeks.
INSTITUTE
College of Fisheries Huazhong Agricultural University
Impact of thiamine metabolites and spent medium from Chlorella sorokiniana on metabolism in the green algae Auxenochlorella prototheciodes (part I)
STUDY_SUMMARY
The purpose of this study is to determine how thiamine metabolites impact central metabolism in Auxenochlorella protothecoides when grown in the presence of glucose. We hypothesize that thiamine metabolites alleviate bottlenecks in the TCA cycle and gluconeogensis, thus allowing for greater starch production when they are present. Cells were grown in bioreactors: 3 control cultures with no thiamine metabolites, 3 cultures received thiamine, 3 recieved HMP, and 3 were grown on residual medium from another algae species - Chlorella sorokiniana. We suspect that this residual medium also contains thiamine metabolites. Samples were taken daily from each of these 12 cultures over a 5 day time course so that we can observe build-up of metabolites over time.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Impact of thiamine metabolites and spent medium from Chlorella sorokiniana on metabolism in the green algae Auxenochlorella prototheciodes (part II)
STUDY_SUMMARY
The purpose of this study is to determine how thiamine metabolites impact central metabolism in Auxenochlorella protothecoides when grown in the presence of glucose. We hypothesize that thiamine metabolites alleviate bottlenecks in the TCA cycle and gluconeogensis, thus allowing for greater starch production when they are present. Cells were grown in bioreactors: 3 control cultures with no thiamine metabolites, 3 cultures received thiamine, 3 recieved HMP, and 3 were grown on residual medium from another algae species - Chlorella sorokiniana. We suspect that this residual medium also contains thiamine metabolites. Samples were taken daily from each of these 12 cultures over a 5 day time course so that we can observe build-up of metabolites over time.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
The Influence of Sugar, Artificial Sweeteners, and the Microbiome on Rodent Large Scale Profiling (part V)
STUDY_SUMMARY
Large scale profiling of rodents treated with diets rich in commonly used artifically sweeteners will be assessed in this study. We hypothesized that a specific subset of plasma metabolites are generated as a result from a diet rich in commonly used artificial sweeteners and their subsequent processing by the gut microbiome, which could ultimately lead to impaired glycemic control and negative physiological health outcomes. To test this hypothesis we administered normal, high glucose, fructose, aspartame, and acesulfame potassium diets to rats for 3 weeks, followed by a plasma collection through cardiac puncture and metabolic analysis. We also treated the gut microbiota with in rats with the same diets plus bacitracin/streptomycin to observe how alterations of the microbiome influence the plasma metabolic profile in these animals. The resulting data will give us insights into the influence of high sugar and artificial sweetener diets on homeostatic metabolic processes and dive into the symbiotic relationship of the gut microbiome with this process.
High Resolution GC-MS Metabolomics of Non-Human Primate Serum
STUDY_TYPE
Non-human Primate Serum
STUDY_SUMMARY
Rationale: Metabolomics analyses using gas chromatography mass spectrometry (GC-MS) - based metabolomics are heavily impeded by the lack of high-resolution mass spectrometers and limited spectral libraries to complement the excellent chromatography that GC platforms offer, a challenge that is being addressed with the implementation of high resolution (HR) platforms such as GC-Orbitrap-MS. Methods: We used serum samples from a non-human primate (NHP), a baboon (Papio hamadryas), with suitable quality controls to quantify the chemical space using an advanced HR MS platform for confident metabolite identification and robust quantification to assess the suitability of the platform for routine clinical metabolomics research. In a comparative approach, we also analyzed the same serum samples using a two-dimensional gas chromatography time-of-flight mass-spectrometer (2D GC-ToF-MS) for metabolite identification and quantification following established standard protocols. Results: Overall, the 2D GC-ToF-MS and GC-Orbitrap-MS analyses enabled identification and quantification of 555 total metabolites from the NHP serum with a spectral similarity score Rsim ā„ 900 and S/R ratio of > 25. A common set of 30 metabolites with HMDB and KEGG IDs were quantified in the serum samples by both platforms where the 2D GC-ToF-MS enabled quantification of a total 384 metabolites (118 HMDB IDs) and the GC-Orbitrap-MS analysis quantification of a total 200 metabolites (47 HMDB IDs). Conclusions: Our study provides insights into the benefits and limitations of the use of a higher mass accuracy instrument for untargeted GC-MS-based metabolomics with multi-dimensional chromatography in future studies addressing clinical conditions or exposome studies.
Metabolome profiles in urogenital schistosomiasis and associated pathologies (part II)
STUDY_SUMMARY
Metabolic fingerprint aid discovery of new biomarkers. Blood and urine were collected from volunteers in Eggua community, Nigeria, after ethical approval. LC-ESI-MS were used to analyse samples. Bioinformatics and statistical tools were used to detect important metabolites
INSTITUTE
University of Ibadan, Nigeria
DEPARTMENT
Zoology (Cell biology and Genetics unit)
LABORATORY
Centre for Materials Characterization, National Chemical Laboratory, Pune, India
LAST_NAME
Adebayo
FIRST_NAME
Adewale
ADDRESS
Cell Biology and Genetics unit, Department of zoology, University of ibadan, Nigeria/ National Centre for Cell Science, Pune, India
Metabolomic analysis of TB vs heathy including time to TB.
STUDY_SUMMARY
Within the GC6-74 cohorts, 4,462 HIV-negative healthy household contacts of 1,098 index TB cases were recruited from 2006 to 2012 at four African sites included in this study i.e. SUN (Stellenbosch University, South Africa), MRC (Medical Research Council Unit, The Gambia), AHRI (Armauer Hansen Research Institute, Ethiopia) and MAK (Makerere University, Uganda). The study had an exclusion period of 3 months, such that participants, who were diagnosed with active TB within 3 months of enrolment, were excluded from analysis to prevent inclusion of participants who had incipient or asymptomatic clinical TB disease at enrolment. Additional exclusion criteria were positive HIV rapid test, current or previous anti-retroviral treatment, history of TB, pregnancy, participation in drug and/or vaccine clinical trials and chronic disease diagnosis or immunosuppressive therapy within the past 6 months, and living in the study area for less than 3 months. A total of 97 individuals who developed active TB within the two year follow-up period were included in this study and matched at a ratio of 1:4 with participants who remained healthy during the 2-year follow-up period (controls); matching, by site, age class, sex, and wherever possible year of recruitment. Initial samples collected upon enrolment were termed baseline (BL) samples. Further samples were taken 6 and 18 months post-exposure, provided that the participant had remained TB free at the time of sample collection. Metabolic profiling was carried out for each study site, using either serum or plasma samples. For a small number of samples, an insufficient amount of plasma was available, so the sample was diluted using RPMI buffer.
GC6-74 metabolomics of TB vs healthy (Part 2: Serum)
STUDY_TYPE
Metabolomic analysis of TB vs healthy including time to TB.
STUDY_SUMMARY
Within the GC6-74 cohorts, 4,462 HIV-negative healthy household contacts of 1,098 index TB cases were recruited from 2006 to 2012 at four African sites included in this study i.e. SUN (Stellenbosch University, South Africa), MRC (Medical Research Council Unit, The Gambia), AHRI (Armauer Hansen Research Institute, Ethiopia) and MAK (Makerere University, Uganda). The study had an exclusion period of 3 months, such that participants, who were diagnosed with active TB within 3 months of enrolment, were excluded from analysis to prevent inclusion of participants who had incipient or asymptomatic clinical TB disease at enrolment. Additional exclusion criteria were positive HIV rapid test, current or previous anti-retroviral treatment, history of TB, pregnancy, participation in drug and/or vaccine clinical trials and chronic disease diagnosis or immunosuppressive therapy within the past 6 months, and living in the study area for less than 3 months. A total of 97 individuals who developed active TB within the two year follow-up period were included in this study and matched at a ratio of 1:4 with participants who remained healthy during the 2-year follow-up period (controls); matching, by site, age class, sex, and wherever possible year of recruitment. Initial samples collected upon enrolment were termed baseline (BL) samples. Further samples were taken 6 and 18 months post-exposure, provided that the participant had remained TB free at the time of sample collection. Metabolic profiling was carried out for each study site, using either serum or plasma samples. For a small number of samples, an insufficient amount of plasma was available, so the sample was diluted using RPMI buffer.
Metabolomic analysis of TB vs heathy including time to TB.
STUDY_SUMMARY
Within the GC6-74 cohorts, 4,462 HIV-negative healthy household contacts of 1,098 index TB cases were recruited from 2006 to 2012 at four African sites included in this study i.e. SUN (Stellenbosch University, South Africa), MRC (Medical Research Council Unit, The Gambia), AHRI (Armauer Hansen Research Institute, Ethiopia) and MAK (Makerere University, Uganda). The study had an exclusion period of 3 months, such that participants, who were diagnosed with active TB within 3 months of enrolment, were excluded from analysis to prevent inclusion of participants who had incipient or asymptomatic clinical TB disease at enrolment. Additional exclusion criteria were positive HIV rapid test, current or previous anti-retroviral treatment, history of TB, pregnancy, participation in drug and/or vaccine clinical trials and chronic disease diagnosis or immunosuppressive therapy within the past 6 months, and living in the study area for less than 3 months. A total of 97 individuals who developed active TB within the two year follow-up period were included in this study and matched at a ratio of 1:4 with participants who remained healthy during the 2-year follow-up period (controls); matching, by site, age class, sex, and wherever possible year of recruitment. Initial samples collected upon enrolment were termed baseline (BL) samples. Further samples were taken 6 and 18 months post-exposure, provided that the participant had remained TB free at the time of sample collection. Metabolic profiling was carried out for each study site, using either serum or plasma samples. For a small number of samples, an insufficient amount of plasma was available, so the sample was diluted using RPMI buffer.
Identification of unique metabolite networks between Latino and Caucasian patients with nonalcoholic fatty liver disease (NAFLD)
STUDY_TYPE
Populations comparison
STUDY_SUMMARY
Nonalcoholic fatty liver disease (NAFLD) is a spectrum of liver pathology ranging from simple steatosis to nonalcoholic steatohepatitis (NASH); the latter is characterized by inflammation and fibrosis. Risk factors for NALFD include obesity, diabetes, hyperlipidemia, and hypertensionāall of which are features of metabolic syndrome. NAFLD is a very heterogeneous disease, as it presents in different patterns in males and females and in patients from different ethnicities, with unclear predictors for development and severity of disease. Previous studies have shown that NAFLD is 1.4 times more frequent in Hispanics than in Caucasians. One of the major challenges in NAFLD is the lack of accurate, noninvasive biomarkers for the detection of the most aggressive presentation, NASH. The gold standard for the diagnosis is liver biopsy, which is an invasive procedure associated with possible complications. Noninvasive diagnosis of NASH is a major unmet medical need and there are no ethnicity-specific biomarkers that can diagnose this condition and predict its progression. Therefore, the main gap in knowledge that this proposal and line of research will address is the characterizing the different plasma and liver metabolomics profile of patients with fatty liver from two ethnicities (Latinos vs. Caucasians) and of both sexes. The overall hypothesis of the present study is that the higher incidence of nonalcoholic fatty liver (NAFL) in Latino patients is reflected in a different plasma and liver metabolomics profile compared to Caucasian patients with further sex-related differences. Characterization of metabolite networks can aid in identifying the mechanistic underpinnings of sex and ethnic driven differences in NAFL which could help diagnose and establish a prognosis of this condition, especially in the critical transition from NAFL to the more aggressive nonalcoholic steatohepatitis (NASH).To address this hypothesis, plasma metabolomics profile of samples from male and female Latino and Caucasian bariatric surgery patients with NAFL and from healthy subjects will be compared. Metabolomics findings will be related with liver pathology and liver transcriptome profiles from intraoperatively obtained liver biopsies using correlation, network, and pathway analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomics biomarkers and the risk of overall mortality and ESRD in CKD: results from the Progredir Cohort.
STUDY_TYPE
ongoing cohort study
STUDY_SUMMARY
454 CKD participants were originally recruited from the outpatient services from Hospital das Clinicas, Sao Paulo. Extensive baseline data and biobank were collected between 2012-2013 in the Clinical Research Center, Universitary Hospital, Sao Paulo. Participants are being actively followed for events of mortality, renal replacement therapy and CVD. Death certificates are obtained from State and National Registries. For the this uploaded data, events were determined up to May 2017 (median follow-up time of 3 y). Censoring data was considered as the last day of contact.
INSTITUTE
Sao Paulo University
DEPARTMENT
Clinical Research Center, Universitary Hospital, Sao Paulo University
LABORATORY
Laboratory of Genetics and Molecular Cardiology, Heart Institute
Events are overall mortality, RRT, and a composite outcome of mortality and RRT. In addition, we also provided the variable for competitive data analysis (RRT considering the competing effect of death). Please see below for description of Study Design. Cox_mortality: death events (for Cox proportional hazard models) Cox_RRT: renal replacement therapy events (for Cox proportional hazard models) Cox_composite_deathRRT: death events AND renal replacement therapy events (for Cox proportional hazard models) Competitive_RRTvs death: death or renal replacement therapy events (for competitive risk analysis) Time_death: follow-up time for death events and censored-participants (for Cox models) Time_Cox_RRT: follow-up time for renal replacement therapy events and censored-participants (for Cox models) Time_composite_deathRRT: follow-up time for death AND renal replacement therapy and censored-participants (for Cox models) Time_competitive_TRSvsDeath: follow-up time for renal replacement therapy, death or censored-participants (for competitive analysis)
Metabolomic profiles in healthy research cats receiving clindamycin with a synbiotic or a placebo: a randomized, controlled trial
STUDY_TYPE
Double-blind randomized controlled trial
STUDY_SUMMARY
Antibiotic-associated gastrointestinal signs (AAGS) occur commonly in cats. Co-administration of synbiotics is associated with decreased AAGS in people, potentially due to stabilization of the fecal microbiome and metabolome. The purpose of this double-blinded randomized-controlled trial was to compare AAGS and the fecal microbiome and metabolome between healthy cats that received clindamycin with a placebo or synbiotic. Methods. 16 healthy domestic shorthair cats from a research colony were randomized to receive 150 mg clindamycin with either a placebo (8 cats) or commercially-available synbiotic (8 cats) once daily for 21 days with reevaluation 603 days thereafter. All cats ate the same diet. Food consumption, vomiting, and fecal score were recorded. Fecal samples were collected daily on the last 3 days of baseline (days 5-7), treatment (26-28), and recovery (631-633). Sequencing of 16S rRNA genes and gas chromatography time-of-flight mass spectrometry was performed. Clinical signs, alpha and beta diversity metrics, dysbiosis indices, proportions of bacteria groups, and metabolite profiles were compared between treatment groups using repeated measures ANOVAs. Fecal metabolite pathway analysis was performed. P<0.05 was considered significant. The Benjamini & Hochbergās False Discovery Rate was used to adjust for multiple comparisons. Results. Median age was 6 and 5 years, respectively, for cats in the placebo and synbiotic groups. Hyporexia, vomiting, diarrhea, or some combination therein were induced in all cats. Though vomiting was less in cats receiving a synbiotic, the difference was not statistically significant. Bacterial diversity decreased significantly on days 26-28 in both treatment groups. Decreases in Actinobacteria (Bifidobacterium, Collinsella, Slackia), Bacteriodetes (Bacteroides), Lachnospiraceae (Blautia, Coprococcus, Roseburia), Ruminococcaceae (Faecilobacterium, Ruminococcus), and Erysipelotrichaceae (Bulleidia, [Eubacterium]) and increases in Clostridiaceae (Clostridium) and Proteobacteria (Aeromonadales, Enterobacteriaceae) occurred in both treatment groups, with incomplete normalization by days 631-633. Derangements in short-chain fatty acid, bile acid, indole, sphingolipid, benzoic acid, cinnaminic acid, and polyamine profiles also occurred, some of which persisted through the terminal sampling timepoint and differed between treatment groups. Discussion. Cats administered clindamycin commonly develop AAGS, as well as short- and long-term dysbiosis and alterations in fecal metabolites. Despite a lack of differences in clinical signs between treatment groups, significant differences in their fecal metabolomic profiles were identified. Further investigation is warranted to determine whether antibiotic-induced dysbiosis is associated with an increased risk of future AAGS or metabolic diseases in cats and whether synbiotic administration ameliorates this risk.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
NUM_GROUPS
2
TOTAL_SUBJECTS
16 subjects/3 timepoints/47 samples
STUDY_COMMENTS
Samples from 1 subject lacking for one timepoint, resulting in a total of 47 samples. Genomic DNA was extracted from 100 mg of feces from each pooled sample using a commercially available kit (PowerSoilĀ®, Mo Bio, Carlsbad, CA USA) according to manufacturerās protocol for a total of 11 pooled samples.
Metabolomic Analysis of plasma samples from Non-Allergic Subjects and Profilin-Allergic Patients overexposed to Grass Pollen
STUDY_TYPE
Allergy severity comparison
STUDY_SUMMARY
Prevalence and severity of allergic diseases have increased worldwide. To date, respiratory allergy phenotypes are not fully characterized and, along with inflammation progression, treatment is increasingly complex and expensive. Profilin sensitization constitutes a good model to study the progression of allergic inflammation. Our aim was to identify the underlying mechanisms and the associated biomarkers of this progression, focusing on severe phenotypes, using transcriptomics and metabolomics. Methods: 25 subjects were included in the study. Plasma samples were analyzed using Gas and Liquid Chromatography coupled to Mass Spectrometry (GC-MS and LC-MS, respectively). Individuals were classified in 4 groups ā ānon-allergicā, āmildā, āmoderateā and āsevereā ā based on their clinical history, their response to an oral challenge test with profilin, and after a refinement using a mathematical metabolomic model. PBMCs were used for microarray analysis.
INSTITUTE
The Centre of Metabolomics and Bioanalysis
DEPARTMENT
Analytical chemistry
LAST_NAME
Obeso Montero
FIRST_NAME
David
ADDRESS
Avda. Monteprincipe s/n 28668 Boadilla del Monte, Madrid, EspaƱa
Metabolomic profiles in healthy research cats receiving clindamycin with a synbiotic or a placebo: a randomized, controlled trial
STUDY_TYPE
Double-blind randomized controlled trial
STUDY_SUMMARY
Antibiotic-associated gastrointestinal signs (AAGS) occur commonly in cats. Co-administration of synbiotics is associated with decreased AAGS in people, potentially due to stabilization of the fecal microbiome and metabolome. The purpose of this double-blinded randomized-controlled trial was to compare AAGS and the fecal microbiome and metabolome between healthy cats that received clindamycin with a placebo or synbiotic. Methods. 16 healthy domestic shorthair cats from a research colony were randomized to receive 150 mg clindamycin with either a placebo (8 cats) or commercially-available synbiotic (8 cats) once daily for 21 days with reevaluation 603 days thereafter. All cats ate the same diet. Food consumption, vomiting, and fecal score were recorded. Fecal samples were collected daily on the last 3 days of baseline (days 5-7), treatment (26-28), and recovery (631-633). Sequencing of 16S rRNA genes and gas chromatography time-of-flight mass spectrometry was performed. Clinical signs, alpha and beta diversity metrics, dysbiosis indices, proportions of bacteria groups, and metabolite profiles were compared between treatment groups using repeated measures ANOVAs. Fecal metabolite pathway analysis was performed. P<0.05 was considered significant. The Benjamini & Hochbergās False Discovery Rate was used to adjust for multiple comparisons. Results. Median age was 6 and 5 years, respectively, for cats in the placebo and synbiotic groups. Hyporexia, vomiting, diarrhea, or some combination therein were induced in all cats. Though vomiting was less in cats receiving a synbiotic, the difference was not statistically significant. Bacterial diversity decreased significantly on days 26-28 in both treatment groups. Decreases in Actinobacteria (Bifidobacterium, Collinsella, Slackia), Bacteriodetes (Bacteroides), Lachnospiraceae (Blautia, Coprococcus, Roseburia), Ruminococcaceae (Faecilobacterium, Ruminococcus), and Erysipelotrichaceae (Bulleidia, [Eubacterium]) and increases in Clostridiaceae (Clostridium) and Proteobacteria (Aeromonadales, Enterobacteriaceae) occurred in both treatment groups, with incomplete normalization by days 631-633. Derangements in short-chain fatty acid, bile acid, indole, sphingolipid, benzoic acid, cinnaminic acid, and polyamine profiles also occurred, some of which persisted through the terminal sampling timepoint and differed between treatment groups. Discussion. Cats administered clindamycin commonly develop AAGS, as well as short- and long-term dysbiosis and alterations in fecal metabolites. Despite a lack of differences in clinical signs between treatment groups, significant differences in their fecal metabolomic profiles were identified. Further investigation is warranted to determine whether antibiotic-induced dysbiosis is associated with an increased risk of future AAGS or metabolic diseases in cats and whether synbiotic administration ameliorates this risk.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
EMAIL
ofiehn@ucdavis.edu
PHONE
(530) 754-8258
NUM_GROUPS
2
TOTAL_SUBJECTS
16 subjects/3 timepoints/43 samples
STUDY_COMMENTS
Samples from 5 cats (4 placebo, 1 synbiotic) were not available at the second timepoint due to withdrawal from treatment due to severity of gastrointestinal signs. Genomic DNA was extracted from 100 mg of feces from each pooled sample using a commercially available kit (PowerSoilĀ®, Mo Bio, Carlsbad, CA USA) according to manufacturerās protocol for a total of 11 pooled samples.
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (Part I)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part II)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part III)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part IV)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part V)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part VI)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part VII)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part VIII)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Validating Quantitative Untargeted Lipidomics Across Nine Liquid ChromatographyāHigh-Resolution Mass Spectrometry Platforms (part IX)
STUDY_SUMMARY
Liquid chromatographyāmass spectrometry (LCāMS) methods are most often used for untargeted metabolomics and lipidomics. However, methods have not been standardized as accepted ābest practiceā documents, and reports lack harmonization with respect to quantitative data that enable interstudy comparisons. Researchers use a wide variety of high-resolution mass spectrometers under different operating conditions, and it is unclear if results would yield different biological conclusions depending on the instrument performance. To this end, we used 126 identical human plasma samples and 29 quality control samples from a nutritional intervention study. We investigated lipidomic data acquisitions across nine different MS instruments (1 single TOF, 1 Q/orbital ion trap, and 7 QTOF instruments). Sample preparations, chromatography conditions, and data processing methods were kept identical. Single-point internal standard calibrations were used to estimate absolute concentrations for 307 unique lipids identified by accurate mass, MS/MS spectral match, and retention times. Quantitative results were highly comparable between the LCāMS platforms tested. Using partial least-squares discriminant analysis (PLS-DA) to compare results between platforms, a 92% overlap for the most discriminating lipids based on variable importance in projection (VIP) scores was achieved for all lipids that were detected by at least two instrument platforms. Importantly, even the relative positions of individual samples on the PLS-DA projections were identical. The key for success in harmonizing results was to avoid ion saturation by carefully evaluating linear dynamic ranges using serial dilutions and adjusting the resuspension volume and/or injection volume before running actual study samples.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Metabolomic Markers of Dietary Patterns in the Costa Rica Study
STUDY_TYPE
MS analysis
STUDY_SUMMARY
The study will examine associations between dietary patterns and the metabolome on a random subset (n=80) of control subjects in a population-based case-control study of myocardial infarction in Costa Rican adults. Each sample will undergo complementary LC-MS-based approaches, specifically untargeted metabolite profiling and targeted lipidomic profiling. Subsequently we will test for associations between 1) previously derived dietary patterns and plasma molecular features and 2) the top plasma correlates of dietary patterns and metabolic syndrome.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Amino Acid Concentrations of Primary Sclerosing Cholangitis (part I)
STUDY_SUMMARY
To qualitatively and quantitatively analyze enterohepatically-circulated molecules using targeted amino acid concentrations of peripheral blood collected from primary sclerosing cholangitis (PSC) patients compared to normal and diseased controls. There are three groups of patients. (1) Normal donor controls (ND), (2) Patients with Primary Sclerosing Cholangitis (PSC), and (3) Disease Controls (DC) which are patients with liver disease other than PSC.
INSTITUTE
Mayo Clinic
LAST_NAME
O'Hara
FIRST_NAME
Steven
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Non-Esterified Fatty Acids of Primary Sclerosing Cholangitis (part II)
STUDY_SUMMARY
To qualitatively and quantitatively analyze enterohepatically-circulated molecules using targeted free fatty acid concentrations of peripheral blood collected from primary sclerosing cholangitis (PSC) patients compared to normal and diseased controls. There are three groups of patients. (1) Normal donor controls (ND), (2) Patients with Primary Sclerosing Cholangitis (PSC), and (3) Disease Controls (DC) which are patients with liver disease other than PSC.
INSTITUTE
Mayo Clinic
LAST_NAME
O'Hara
FIRST_NAME
Steven
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Acyl Carnitines Concentrations of Primary Sclerosing Cholangitis (part III)
STUDY_SUMMARY
To qualitatively and quantitatively analyze enterohepatically-circulated molecules using targeted acyl carnitines concentrations of peripheral blood collected from primary sclerosing cholangitis (PSC) patients compared to normal and diseased controls. There are three groups of patients. (1) Normal donor controls (ND), (2) Patients with Primary Sclerosing Cholangitis (PSC), and (3) Disease Controls (DC) which are patients with liver disease other than PSC.
INSTITUTE
Mayo Clinic
LAST_NAME
O'Hara
FIRST_NAME
Steven
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Bile Acid of Primary Sclerosing Cholangitis (part IV)
STUDY_SUMMARY
To qualitatively and quantitatively analyze enterohepatically-circulated molecules using targeted bile acid concentrations of peripheral blood collected from primary sclerosing cholangitis (PSC) patients compared to normal and diseased controls. There are three groups of patients. (1) Normal donor controls (ND), (2) Patients with Primary Sclerosing Cholangitis (PSC), and (3) Disease Controls (DC) which are patients with liver disease other than PSC.
INSTITUTE
Mayo Clinic
LAST_NAME
O'Hara
FIRST_NAME
Steven
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Natural genetic variation in C. elegans identified genomic loci controlling metabolite levels
STUDY_TYPE
Metabolomics in C. elegans
STUDY_SUMMARY
Metabolic homeostasis is sustained by complex biological networks that respond to nutrient availability. Genetic and environmental factors may disrupt this equilibrium leading to metabolic disorders, including obesity and type 2 diabetes. To identify the genetic factors controlling metabolism, we performed quantitative genetic analysis using a population of 199 recombinant inbred lines (RILs) in the nematode Caenorhabditis elegans. We focused on the genomic regions that control metabolite levels by measuring fatty acid (FA) and amino acid (AA) composition in the RILs using targeted metabolomics. The genetically diverse RILs showed a large variation in their FA and AA levels with a heritability ranging from 32-82%. We detected strongly co-correlated metabolite clusters and 36 significant metabolite QTL (mQTL). We focused on mQTL displaying highly significant linkage and heritability, including an mQTL for the FA C14:1 on Chromosome I, and another mQTL for the FA C18:2 on Chromosome IV. Using introgression lines (ILs) we were able to narrow down both mQTL to a 1.4 Mbp and a 3.6 Mbp region, respectively. RNAi-based screening focusing on the Chromosome I mQTL identified several candidate genes for the C14:1 mQTL, including lagr-1, Y87G2A.2, nhr-265, nhr-276, and nhr-81. Overall, this systems approach provides us with a powerful platform to study the genetic basis of C. elegans metabolism. Furthermore, it allows us to investigate interventions, such as nutrients and stresses that maintain or disturb the regulatory network controlling metabolic homeostasis, and identify gene-by-environment interactions.
INSTITUTE
Academic Medical Center of Amsterdam
LAST_NAME
Gao
FIRST_NAME
Arwen
ADDRESS
Meibergdreef 9, Amsterdam, North-Holland, 1105 AZ, Netherlands
Gut microbiome structure and metabolic activity in inflammatory bowel disease
STUDY_SUMMARY
The inflammatory bowel diseases (IBD), which include Crohnās disease (CD) and ulcerative colitis (UC), are multifactorial, chronic conditions of the gastrointestinal tract. While IBD has been associated with dramatic changes in the gut microbiota, changes in the gut metabolome -- the molecular interface between host and microbiota -- are less-well understood. To address this gap, we performed untargeted LC-MS metabolomic and shotgun metagenomic profiling of cross-sectional stool samples from discovery (n=155) and validation (n=65) cohorts of CD, UC, and non-IBD control subjects. Metabolomic and metagenomic profiles were broadly correlated with fecal calprotectin levels (a measure of gut inflammation). Across >8,000 measured metabolite features, we identified chemicals and chemical classes that were differentially abundant (DA) in IBD, including enrichments for sphingolipids and bile acids, and depletions for triacylglycerols and tetrapyrroles. While >50% of DA metabolite features were uncharacterized, many could be assigned putative roles through metabolomic āguilt-by-associationā (covariation with known metabolites). DA species and functions from the metagenomic profiles reflected adaptation to oxidative stress in the IBD gut, and were individually consistent with previous findings. Integrating these data, however, we identified 122 robust associations between DA species and well-characterized DA metabolites, indicating possible mechanistic relationships that are perturbed in IBD. Finally, we found that metabolome- and metagenome-based classifiers of IBD status were highly accurate and, like the vast majority of individual trends, generalized well to the independent validation cohort. Our findings thus provide an improved understanding of perturbations of the microbiome-metabolome interface in IBD, including identification of many potential diagnostic and therapeutic targets.
A pilot study of urine metabolomics in two female and one male subject towards an outpatient estimate of circadian phase using urine storage as a factor (part II)
STUDY_SUMMARY
Serial urine samples were collected at each void (approximately every 3 hours) from subjects during a 6-day inpatient protocol. The total volume of each sample was measured, and then 5 mL was aliquoted into a 7 mL tube and elivered on ice to the processing lab, where the samples were then stored at either -20 degrees or -80 degrees (see details below). At the end of the study, samples were transported (~2 blocks) from the processing lab to our -20 or -80 freezer for storage. The samples being sent represent samples from two female subjects (3634A and 3635A) and one male subject (3624A). These subjects all spent 6 days in the lab: 3 baseline days where the subjects slept for 8 hours at night (at habitual times as determined during the screening period) and 16 hours of ambulatory wake in ambient light, followed by 50 hours of continuous wakefulness in which the subject was kept in a semi-recumbent position in bed under dim light and fed hourly isocaloric snacks (called a "constant routine"). We are requesting untargeted profiling of samples from these subjects (plus 6-sulphatoxymelatonin profile) to determine how the concentrations of different metabolites vary across the 24-hour period, and specifically to compare this circadian variation in each metabolite during a 48-hour ambulatory period versus a 48-hour constant routine period.
INSTITUTE
Mayo Clinic
LAST_NAME
Hilaire
FIRST_NAME
Melissa
ADDRESS
221 Longwood Avenue, Suite BL438 Boston, Massachusetts 02115
A pilot study of urine metabolomics in a female subject towards an outpatient estimate of circadian phase
STUDY_SUMMARY
Serial urine samples were collected at each void (approximately every 3 hours) from subjects during a 6-day inpatient protocol. The total volume of each sample was measured, and then 5 mL was aliquoted into a 7 mL tube and delivered on ice to the processing lab, where the samples were then stored at -80 degrees. At the end of the study, samples were transported (~2 blocks) from the processing lab to our -80 freezer for storage. The samples being sent represent samples from one female subject. This subject spent 6 days in the lab: 3 baseline days where the subject slept for 8 hours at night (at habitual times as determined during the screening period) and 16 hours of ambulatory wake in ambient light, followed by 50 hours of continuous wakefulness in which the subject was kept in a semi-recumbent position in bed under dim light and fed hourly isocaloric snacks (called a "constant routine"). We are requesting untargeted profiling of 25 samples (sample #: 52919-52943) to determine how the concentrations of different metabolites vary across the 24-hour period, and specifically to compare this circadian variation in each metabolite during a 48-hour ambulatory period versus a 48-hour constant routine period.
INSTITUTE
Mayo Clinic
LAST_NAME
Hilaire
FIRST_NAME
Melissa
ADDRESS
221 Longwood Avenue, Suite BL438 Boston, Massachusetts 02115
Untargeted high-resolution mass spectrometry profiling
STUDY_SUMMARY
Urban environments remain a poorly understood toxic environment for children with asthma, where improved exposure characterization and estimation of exposurehealth outcome relationships are clearly needed. The goal of this project is to investigate the interactions between relevant environmental exposures and asthma severity in a year-long longitudinal study of urban children with asthma. Environmental and clinical samples are being collected at 3 seasonal visits. Using these samples, we will measure the effects of multiple relevant exposures (environmental tobacco smoke (ETS), polycyclic aromatic hydrocarbons (PAHs), phthalates, and volatile organic compounds (VOCs)) on biological responses (metabolomics, oxidative stress, inflammatory markers, and endocannabinoids) and asthma outcomes. Our overall hypothesis is that relevant environmental exposures and their interactions drive disease severity in urban children with asthma. We will test this hypothesis by investigating the following aims: Aim 1: To investigate how environmental exposures (ETS, PAHs, phthalates, and VOCs) and their interactions contribute to asthma severity in urban children. Aim 2: To determine if environmental exposures in children with asthma are associated with changes in in biological responses (metabolomics, oxidative stress, inflammatory markers, and endocannabinoids). Aim 3: To determine which biological responses mediate the relationships between environmental exposures and asthma severity. Aim 4: To compare environmental exposures and biological responses in asthmatic and non-asthmatic children
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Uppal
FIRST_NAME
Karan
ADDRESS
615 Michael St. Ste 225, Atlanta, GA, 30322, USA
EMAIL
kuppal2@emory.edu
PHONE
(404) 727 5027
TOTAL_SUBJECTS
66
STUDY_COMMENTS
Both CHEAR and Clinical Biomarker Laboratory pooled plasma samples were used for quality control. Study specific sample pools were not created
Amino Acid Concentrations in Muscle Tissue of Muscle Wasting in Cancer Cachexia (part I)
STUDY_SUMMARY
Amino Acid Concentrations of Muscle Wasting in Cancer Cachexia. Muscle samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
TCA Concentrations in Muscle Tissue of Muscle Wasting in Cancer Cachexia (part II)
STUDY_SUMMARY
TCA Concentrations of Muscle Wasting in Cancer Cachexia. Muscle samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Multi-Platform Physiologic and Metabolic Phenotyping Reveals Microbial Toxicity
STUDY_SUMMARY
The gut microbiota are susceptible to modulation by environmental stimuli and therefore can serve as biological sensors. Recent evidence suggests that xenobiotics can disrupt the interaction between the microbiota and host. Here, we describe an approach that combines in vitro microbial incubation (isolated cecal contents from mice), flow cytometry, and mass spectrometry- and 1H NMR-based metabolomics to evaluate xenobiotic-induced microbial toxicity. Tempol, a stabilized free radical scavenger known to remodel the microbial community structure and function in vivo, was studied to assess its direct effects on the gut microbiota. Microbiota were isolated from mouse cecum and were exposed to tempol for 4 h under strict anaerobic condition. The flow cytometry data suggested short term exposure of the microbiota to tempol is associated with disrupted membrane physiology as well as compromised metabolic activity. Mass spectrometry and NMR metabolomics revealed that tempol exposure significantly disrupted microbial metabolic activity, specifically indicated by changes in short chain fatty acids, branched chain amino acids, amino acids, nucleotides, glucose, and oligosaccharides. In addition, a mouse study with tempol (5 days gavage) showed similar microbial physiologic and metabolic changes, indicating the in vitro approach reflected in vivo conditions. Our results, through evaluation of microbial viability, physiology and metabolism, and comparison of in vitro and in vivo exposures with tempol, suggests that physiologic and metabolic phenotyping provides unique insight into gut microbiota toxicity.
Multi-Platform Physiologic and Metabolic Phenotyping Reveals Microbial Toxicity (part II)
STUDY_SUMMARY
The gut microbiota are susceptible to modulation by environmental stimuli and therefore can serve as biological sensors. Recent evidence suggests that xenobiotics can disrupt the interaction between the microbiota and host. Here, we describe an approach that combines in vitro microbial incubation (isolated cecal contents from mice), flow cytometry, and mass spectrometry- and 1H NMR-based metabolomics to evaluate xenobiotic-induced microbial toxicity. Tempol, a stabilized free radical scavenger known to remodel the microbial community structure and function in vivo, was studied to assess its direct effects on the gut microbiota. Microbiota were isolated from mouse cecum and were exposed to tempol for 4 h under strict anaerobic condition. The flow cytometry data suggested short term exposure of the microbiota to tempol is associated with disrupted membrane physiology as well as compromised metabolic activity. Mass spectrometry and NMR metabolomics revealed that tempol exposure significantly disrupted microbial metabolic activity, specifically indicated by changes in short chain fatty acids, branched chain amino acids, amino acids, nucleotides, glucose, and oligosaccharides. In addition, a mouse study with tempol (5 days gavage) showed similar microbial physiologic and metabolic changes, indicating the in vitro approach reflected in vivo conditions. Our results, through evaluation of microbial viability, physiology and metabolism, and comparison of in vitro and in vivo exposures with tempol, suggests that physiologic and metabolic phenotyping provides unique insight into gut microbiota toxicity.
INSTITUTE
The Pennsylvania State University (Penn State)
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
650 toftrees ave Apt #108, State College, Pa 16802
Acyl Carnitines Concentrations in Muscle Tissue of Muscle Wasting in Cancer Cachexia (part-IV)
STUDY_SUMMARY
Acyl Carnitines Concentrations of Muscle Wasting in Cancer Cachexia. Muscle samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
NEFA Panel in Muscle Tissue of Muscle Wasting in Cancer Cachexia (part-V)
STUDY_SUMMARY
NEFA Panel of Muscle Wasting in Cancer Cachexia. Muscle samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
NEFA Panel in Serum for Muscle Wasting in Cancer Cachexia (part-VI)
STUDY_SUMMARY
NEFA Panel of Muscle Wasting in Cancer Cachexia. Serum samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Amino Acid Concentrations in Serum for Muscle Wasting in Cancer Cachexia (part-VII)
STUDY_SUMMARY
Amino Acid Concentrations of Muscle Wasting in Cancer Cachexia. Serum samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
TCA Concentrations in Serum for Muscle Wasting in Cancer Cachexia (part-VIII)
STUDY_SUMMARY
TCA Concentrations of Muscle Wasting in Cancer Cachexia. Serum samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Sphingolipid Concentrations in Serum for Muscle Wasting in Cancer Cachexia (part-IX)
STUDY_SUMMARY
Sphingolipid Concentrations of Muscle Wasting in Cancer Cachexia. Serum samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Acyl Carnitines Concentration in Serum for Muscle Wasting in Cancer Cachexia (part-X)
STUDY_SUMMARY
Acyl Carnitines Concentrations of Muscle Wasting in Cancer Cachexia. Serum samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Sphingolipid Concentrations in Muscle Tissue of Muscle Wasting in Cancer Cachexia (part-III)
STUDY_SUMMARY
Sphingolipid Concentrations of Muscle Wasting in Cancer Cachexia. Muscle samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Large Scale Profling in Muscle Tissue for Muscle Wasting in Cancer Cachexia (part-XI)
STUDY_SUMMARY
Large Scale Profiling of muscle wasting in Cancer Cachexia. Muscle samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Large Scale Profiling in Serum for Muscle Wasting in Cancer Cachexia (part-XII)
STUDY_SUMMARY
Large Scale Profiling of muscle wasting in Cancer Cachexia. Serum samples from 10 control patients, 10 weight stable pancreatic cancer patients, and 10 pancreatic cancer patients with significant weight loss. Samples are divided evenly between males and females.
INSTITUTE
Mayo Clinic
LAST_NAME
Guttridge
FIRST_NAME
Denis
ADDRESS
520 Biomedical Research Tower 460 W. 12th Avenue Columbus, OH 43210
Lipidomic profiling of heart and plasma of mice following swim training versus pressure overload
STUDY_SUMMARY
Lipid profiling was performed on hearts and plasma from mice subjected to a physiological stimulus (4 weeks of swim exercise training) or pathological stimulus (4 weeks of pressure overload ā transverse aortic constriction; TAC)
Characterization of metabolomics profile changes during development of post-traumatic epilepsy in Rat Plasma (part-I)
STUDY_SUMMARY
Characterize the metabolomics profile changes during progression of the transition from traumatic brain injury (TBI) to post-traumatic epilepsy (PTE). To do so, three experiments will be performed. PTE animal model will be developed using ferrous chloride injections. Metabolomics profile changes will be obtained before TBI, after TBI, and after PTE development These temporal changes in metabolomics profile during the course of PTE development will be collected. We will also collect cerebrospinal fluid (CSF) at each time point. In addition, we will collect the brain tissue from the center of injury, around the injury, and from the non-injured area for mass spectrometry. In this study, the rat plasma at each time point is analyzed.
Characterization of metabolomics profile changes during development of post-traumatic epilepsy in Rat Cerebrospinal Fluid (part-II)
STUDY_SUMMARY
Characterize the metabolomics profile changes during progression of the transition from traumatic brain injury (TBI) to post-traumatic epilepsy (PTE). To do so, three experiments will be performed. PTE animal model will be developed using ferrous chloride injections. Metabolomics profile changes will be obtained before TBI, after TBI, and after PTE development These temporal changes in metabolomics profile during the course of PTE development will be collected. We will also collect cerebrospinal fluid (CSF) at each time point. In addition, we will collect the brain tissue from the center of injury, around the injury, and from the non-injured area for mass spectrometry. In this study, Rat CSF is analyzed at end of study.
Characterization of metabolomics profile changes during development of post-traumatic epilepsy in Rat Brain Tissue (part-III)
STUDY_SUMMARY
Characterize the metabolomics profile changes during progression of the transition from traumatic brain injury (TBI) to post-traumatic epilepsy (PTE). To do so, three experiments will be performed. PTE animal model will be developed using ferrous chloride injections. Metabolomics profile changes will be obtained before TBI, after TBI, and after PTE development These temporal changes in metabolomics profile during the course of PTE development will be collected. We will also collect cerebrospinal fluid (CSF) at each time point. In addition, we will collect the brain tissue from the center of injury, around the injury, and from the non-injured area for mass spectrometry. In this study, the rat brain tissue at end of study is analyzed.
H3K27M cells and glutamine metabolomics 1 million cell test (part-I)
STUDY_SUMMARY
Testing TCA concentrations of Diffuse Intrinsic Pontine Gliomas (DIPG) cellines with H3K27M mutations. One million cells are tested with a TCA concentrations panel. We are a high volume center for treating malignant gliomas, which gives us an advantage in obtaining tissue for these relatively rare tumors. We have developed several DIPG patient derived cell lines and xenografts that bear all the key molecular features of this disease including the H3K27M mutation and global H3K27 hypomethylation. These cells are low in passage and we think these lines more closely resemble the patients tumor pathology then established cell lines that have been in culture/mice for numerous years.
TCA cycle metabolomics of H3K27M cells grown in regular glutamine media, glutamine free media, and glutamine free media with alpha-ketoglutarate (Part-II)
STUDY_SUMMARY
Testing TCA concentrations of Diffuse Intrinsic Pontine Gliomas (DIPG) cellines with H3K27M mutations. Preliminary studies show H3K27M tumor cells are addicted to Gln for survival. Removal of Gln from media resulted in tumor cell death which was rescued by the addition of Ī±-KG. These data show that Gln is taken up and metabolized by H3K27M tumor cells and that Gln derived Ī±-KG is critical for the survival of these tumors. Interestingly, tumor cell death with Gln deprivation was similar to the effect of the JMJD3 inhibitor GSKJ4. Therefore, Gln derived Ī±-KG may be required for both anaplerosis and to drive JMJD3 demethylation. We hypothesize that H3K27M tumors are reliant on Ī±-KG that is derived from Gln to drive the TCA cycle and further decrease H3K27 methylation levels. Furthermore, inhibition of Gln metabolism may represent a novel therapeutic approach for tumors with this mutation. In this study, TCA cycle metabolomics are analyzed of H3K27M cells grown in regular glutamine media, glutamine free media, and glutamine free media with alpha-ketoglutarate.
TCA Isotopmer metabolomics of H3K27M Cells grown in regular media, glutamine enriched regular media, and glucose encriched regular media (Part-III)
STUDY_SUMMARY
TCA Isotopmer metabolomics of H3K27M Cells grown in regular media, glutamine enriched regular media, and glucose encriched regular media. TCA isotopomers traced for 0, 12, or 24 hours were measured.
TCA cycle metabolomics of H3K27M Cell Nucleus Fraction and Cell Mitonchonrdial Fraction (Part-IV)
STUDY_SUMMARY
Testing TCA concentrations of Diffuse Intrinsic Pontine Gliomas (DIPG) cellines with H3K27M mutations. Preliminary studies show H3K27M tumor cells are addicted to Gln for survival. Removal of Gln from media resulted in tumor cell death which was rescued by the addition of Ī±-KG. These data show that Gln is taken up and metabolized by H3K27M tumor cells and that Gln derived Ī±-KG is critical for the survival of these tumors. Interestingly, tumor cell death with Gln deprivation was similar to the effect of the JMJD3 inhibitor GSKJ4. Therefore, Gln derived Ī±-KG may be required for both anaplerosis and to drive JMJD3 demethylation. We hypothesize that H3K27M tumors are reliant on Ī±-KG that is derived from Gln to drive the TCA cycle and further decrease H3K27 methylation levels. Furthermore, inhibition of Gln metabolism may represent a novel therapeutic approach for tumors with this mutation. In this study, TCA cycle metabolomics are analyzed of H3K27M cells grown in regular glutamine media, glutamine free media, and glutamine free media with alpha-ketoglutarate. Additionally, cell nucleus and cell mitochrondial fractions are run separately.
Influence of Data-Processing Strategies on Normalized Lipid Levels using an Open-Source LC-HRMS/MS Lipidomics Workflow
STUDY_SUMMARY
Lipidomics is an emerging field with significant potential for improving clinical diagnosis and our understanding of health and disease. While the diverse biological roles of lipids contribute to their clinical utility, the unavailability of lipid internal standards representing each species, make lipid quantitation analytically challenging. The common approach is to employ one or more internal standards for each lipid class examined and use a single point calibration for normalization (relative quantitation). To aid in standardizing and automating this relative quantitation process, we developed LipidMatch Normalizer (LMN) http://secim.ufl.edu/secim-tools/ which can be used in most open source lipidomics workflows. While the effect of lipid structure on relative quantitation has been investigated, applying LMN we show that data-processing can significantly affect lipid semi-quantitative amounts. Polarity and adduct choice had the greatest effect on normalized levels; when calculated using positive versus negative ion mode data, one fourth of lipids had greater than 50 % difference in normalized levels. Based on our study, sodium adducts should not be used for statistics when sodium is not added intentionally to the system, as lipid levels calculated using sodium adducts did not correlate with lipid levels calculated using any other adduct. Relative quantification using smoothing versus not smoothing, and peak area versus peak height, showed minimal differences, except when using peak area for overlapping isomers which were difficult to deconvolute. By characterizing sources or variation introduced during data-processing and introducing automated tools, this work helps increase through-put and improve data-quality for determining relative changes across groups.
INSTITUTE
University of Florida
DEPARTMENT
Chemistry
LABORATORY
Richard Yost Laboratory
LAST_NAME
Levy
FIRST_NAME
Allison
ADDRESS
214 Leigh Hall, PO Box 117200, Gainesville, Florida, 32611, USA
Metabolic profiling of identified single cells in Xenopus laevis embryos
STUDY_TYPE
Metabolic profiling of single cells
STUDY_SUMMARY
Single D11 cells were identified in 16-cell embryos of Xenopus laevis. Metabolites were extracted, and the extracts were analyzed using a custom-built capillary electrophoresis electrospray ionization platform coupled to a quadrupole time-of-flight mass spectrometer. The resulting metadata was analyzed by Trace, a custom-design software, designed to extract molecular feautres from trace-sensitive metabolomics experiments. The results were validated against molecular features that were extracted by manual curation of the same raw mass spectrometer files.
INSTITUTE
University of Maryland
DEPARTMENT
Department of Chemistry & Biochemistry
LABORATORY
Nemes Laboratory
LAST_NAME
Nemes
FIRST_NAME
Peter
ADDRESS
0107 Chemistry Building, 8051 Regents Dr, College Park, MD 20742
EMAIL
nemes@umd.edu
PHONE
3014050373
NUM_GROUPS
5 biological replicates (different cells from different embryos) + 1-to-3 technical replicates (same extract analyzed multiple times)
TOTAL_SUBJECTS
5 different D11 cells were analyzed, each from a different embryo
PUBLICATIONS
Trace: Machine Learning of Signal Images for Trace-Sensitive Mass Spectrometry ā A Case Study from Single-Cell Metabolomics
An integrated, high-throughput strategy for multi-omic systems level analysis
STUDY_SUMMARY
This report details the automation, benchmarking, and application of a strategy for transcriptomic, proteomic, and metabolomic analyses from a common sample. The approach, Sample Preparation for multi-Omics Technologies (SPOT), provides equivalent performance to typical individual omic preparation methods, but it greatly enhances throughput and minimizes the resources required for multi-omic experiments. SPOT was applied to a multi-omics time course experiment for zinc-treated HL60 cells.
Untargeted Discovery in Primary Metabolism: Collision Cross Section as a Molecular Descriptor in Ion Mobility Spectrometry
STUDY_TYPE
Database Library
STUDY_SUMMARY
In this work we have established a collision cross section (CCS) library of primary metabolites based on analytical standards in the Mass Spectrometry Metabolite Library of Standards (MSMLS). Using a commercially available ion mobility-mass spectrometer (IM-MS, Agilent 6560), from the 554 unique compounds in the MSMLS plate library we obtained a total of 1246 CCS measurements over a wide range of biochemical classes and adduct types. Resulting data analysis showed that the curated CCS library provides broad molecular coverage of metabolic pathways and highlights intrinsic mass/mobility relationships for specific metabolite superclasses. The separation and characterization of isomeric metabolites were assessed as well as an investigation to determine the analytical separation efficiency in both the mass (m/z) and mobility (CCS/ĆāCCS) dimension required for untargeted metabolomic analyses. To further demonstrate the analytical utility of CCS as an additional molecular descriptor, a well characterized biological sample of human plasma serum (NIST 1950) was examined by LC-IM-MS and a detailed analysis of carbohydrate isomers by ion mobility is presented.
INSTITUTE
Vanderbilt University
DEPARTMENT
Chemistry
LABORATORY
McLean Laboratory
LAST_NAME
Dodds
FIRST_NAME
James
ADDRESS
7330 Stevenson Ct., Nashville, Tennessee, 37235, USA
Metabolome analysis on multi-connected biparental chromosome segment substitution line populations in rice
STUDY_SUMMARY
rice flag leaves at heading stage from three chromosome substitution line populations, which were respectively constructed by introducing genomic segments from japonica cultivar Niponbare, indica cultivar Minghui 63 and wild accession ACC10, to an indica cultivar Zhenshan 97, were collected. Metabolomics profile was conducted to generate quantitative trait loci that may affect contents of metabolites, and candidate genes were assigned.
Single-cell Profiling of Cationic and Anionic Metabolites in Live Frog (Xenopus) Embryos using Microprobe Capillary Electrophoresis Mass Spectrometry
STUDY_TYPE
Metabolic profiling of single cells
STUDY_SUMMARY
The goal of this study was to enable the analysis of anionic and cationic metabolites from the same identified single cell in Xenopus laevis embryos. A 10 nL portion of identified animal-ventral (V1) cells was aspirated from 8-cell embryos, and metabolites were extracted from the aspirate, before characterization of cationic and anionic compounds using a custom-built capillary electrophoresis (CE) electrospray ionization (ESI) mass spectrometry platform. A total of ~250 cationic molecular features and ~150 anionic molecular features were detected, including 76 metabolites that were identified in this study. Pathway analysis of the identified metabolites highlighted arginine-proline metabolism of significance.
INSTITUTE
University of Maryland
DEPARTMENT
Department of Chemistry & Biochemistry
LABORATORY
Nemes Laboratory
LAST_NAME
Nemes
FIRST_NAME
Peter
ADDRESS
0107 Chemistry Building 8051 Regents Drive
EMAIL
nemes@umd.edu
PHONE
3014050373
NUM_GROUPS
4 biological replicates (each different cell from a different embryo) + 1-to-2 technical replicates (same extract analyzed multiple times)
TOTAL_SUBJECTS
4 different V1 cells were analyzed, each from a different embryo
Determination of mode of action of anti-malalrial drugs using untargeted metabolomics
STUDY_SUMMARY
The mode of action of a representative active compound was investigated using an unbiased metabolomics approach, which has previously been shown to reveal both novel and established modes of action of antimalarials (Creek et al 2016, DOI: 10.1128/AAC.01226-16). The active antimalarial OSM-S-313, and the inactive analogue OSM-S-291, were incubated with trophozoite stage P. falciparum parasites for five hours alongside reference compounds including atovaquone (ATV), chloroquine (CQ), dihydroartemisisin (DHA) and three PfATP4 inhibitors, MMV00073, MMV397264 and MMV390482. Metabolomics analysis of cell pellets and spent media allowed reproducible detection of diverse metabolites from a range of metabolic pathways, with the most significant OSM-S-313-induced perturbations observed within peptide, lipid and energy metabolism, suggesting a specific impact on parasite metabolism.
INSTITUTE
Monash University
LAST_NAME
Creek
FIRST_NAME
Darren
ADDRESS
Monash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
PAMP-triggered changes in the exometabolome of Arabidopsis suspension cells
STUDY_SUMMARY
The goal of this study was to determine how the exometabolome of defense-elicited Arabidopsis suspension cells inhibits virulence gene expression and growth of a plant pathogenic bacterium Pseudomonas syringae. Arabidopsis T87 suspension cells were treated with the pathogen-associated molecular pattern elf26 or a DMSO-control treatment for six hours, then incubated in fresh media for one hour. The conditioned medium (exudate) was collected from each culture by centrifugation and 0.22 um filter to remove plant cells. These samples are designated T=6 mock and T=6 elf26 in our experimental design. We also prepared samples in the same manner from control-treated cells except without any pre-treatment time prior to one hour exudate production. These samples are labeled T=0 mock. A total of seven biological replicates of each treatment condition were analyzed, with each replicate prepared from cells grown in separate flasks. The exudates were prepared in four independent experiments performed on separate days (1 biological replicate from first experiment, 2 biological replicates from each of the 3 remaining experiments). Four samples of the culture medium, one from each of the four independent experiments, are included.
TCA Isotopomers in Neuromyelitis Optica Patients (part - I)
STUDY_SUMMARY
Patients with an inflammatory disease of the central nervous system known as neuromyelitis optica (NMO) experience increased levels of depression. These patients have an antibody that recognizes a type of cell in the brain called astrocytes ā binding of this antibody to astrocytes triggers a stress response in the cell that results in the development of brain lesions that cause disability and cognitive disturbances. We recently observed a change in the level of glutamate in a part of the brain involved in depression in patients with NMO. Glutamate is a chemical that is used in the brain for communication between neurons ā reduced levels of glutamate are thought to trigger depression by reducing neuronal activity in specific circuits. Based on this observation and the known role of astrocytes in maintaining glutamate levels in the brain, we hypothesize that the NMO antibody disturbs metabolic activity in astrocytes and thereby reduces glutamate and triggers depression. We intend to trace the metabolic response induced in astrocytes by the NMO antibody using TCA isotopomers. It is our hope that we will not only learn something about the mechanisms of astrocyte dysregulation in neuromyelitis optica, but that we will learn something about the mechanisms of depression in general that may lead to new therapies for this disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Howe
FIRST_NAME
Charles
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
TCA Concentrations in Neuromyelitis Optica Patients (part - II)
STUDY_SUMMARY
Patients with an inflammatory disease of the central nervous system known as neuromyelitis optica (NMO) experience increased levels of depression. These patients have an antibody that recognizes a type of cell in the brain called astrocytes ā binding of this antibody to astrocytes triggers a stress response in the cell that results in the development of brain lesions that cause disability and cognitive disturbances. We recently observed a change in the level of glutamate in a part of the brain involved in depression in patients with NMO. Glutamate is a chemical that is used in the brain for communication between neurons ā reduced levels of glutamate are thought to trigger depression by reducing neuronal activity in specific circuits. Based on this observation and the known role of astrocytes in maintaining glutamate levels in the brain, we hypothesize that the NMO antibody disturbs metabolic activity in astrocytes and thereby reduces glutamate and triggers depression. We intend to measure TCA concentration in NMO astrocytes. It is our hope that we will not only learn something about the mechanisms of astrocyte dysregulation in neuromyelitis optica, but that we will learn something about the mechanisms of depression in general that may lead to new therapies for this disease.
INSTITUTE
Mayo Clinic
LAST_NAME
Howe
FIRST_NAME
Charles
ADDRESS
200 First St. SW, Rochester, Minnesota, 55905, USA
Global Metabolomics of the Placenta Reveals Distinct Metabolic Profiles between Maternal and Fetal Placental Tissues Following Delivery in Non-Labored Women
STUDY_TYPE
Timecourse
STUDY_SUMMARY
We evaluated the metabolic alterations in maternal and fetal placental tissues from non-labored women undergoing cesarean section using samples collected from 5 min to 24 h following delivery. Using 1H-NMR, we identified 14 metabolites that significantly differed between maternal and fetal placental tissues (FDR-corrected p-value < 0.05), with 12 metabolites elevated in the maternal tissue, reflecting the flux of these metabolites from mother to fetus. In the maternal tissue, 4 metabolites were significantly altered at 15 min, 10 metabolites at 30 min, and 16 metabolites at 1 h postdelivery, while 11 metabolites remained stable over 24 h. In contrast, in the fetal placenta tissue, 1 metabolite was significantly altered at 15 min, 2 metabolites at 30 min, and 4 metabolites at 1 h postdelivery, while 22 metabolites remained stable over 24 h. Our study provides information on the metabolic profiles of maternal and fetal placental tissues delivered by cesarean section and reveals that there are different metabolic alterations in the maternal and fetal tissues of the placenta following delivery.
INSTITUTE
University of Florida
DEPARTMENT
Biochemistry & Molecular Biology
LAST_NAME
Jacquelyn
FIRST_NAME
Walejko
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry and Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245
Denver Asthma Panel Study-CHEAR Ancillary Study (part II)
STUDY_SUMMARY
Urban environments remain a poorly understood toxic environment for children with asthma, where improved exposure characterization and estimation of exposurehealth outcome relationships are clearly needed. The goal of this project is to investigate the interactions between relevant environmental exposures and asthma severity in a year-long longitudinal study of urban children with asthma. Environmental and clinical samples are being collected at 3 seasonal visits. Using these samples, we will measure the effects of multiple relevant exposures (environmental tobacco smoke (ETS), polycyclic aromatic hydrocarbons (PAHs), phthalates, and volatile organic compounds (VOCs)) on biological responses (metabolomics, oxidative stress, inflammatory markers, and endocannabinoids) and asthma outcomes. Our overall hypothesis is that relevant environmental exposures and their interactions drive disease severity in urban children with asthma. We will test this hypothesis by investigating the following aims: Aim 1: To investigate how environmental exposures (ETS, PAHs, phthalates, and VOCs) and their interactions contribute to asthma severity in urban children. Aim 2: To determine if environmental exposures in children with asthma are associated with changes in in biological responses (metabolomics, oxidative stress, inflammatory markers, and endocannabinoids). Aim 3: To determine which biological responses mediate the relationships between environmental exposures and asthma severity. Aim 4: To compare environmental exposures and biological responses in asthmatic and non-asthmatic children
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Uppal
FIRST_NAME
Karan
ADDRESS
615 Michael St. Ste 225, Atlanta, GA, 30322, USA
EMAIL
kuppal2@emory.edu
PHONE
(404) 727 5027
TOTAL_SUBJECTS
169
STUDY_COMMENTS
Both CHEAR pooled urine samples and Clinical Biomarker Laboratory pooled plasma samples were used
Maternal gut microbiome, pregnancy exposure to environmental contaminants, and child health outcomes: A pilot study exploring potential effect modifications by diet
STUDY_TYPE
CHEAR Pilot Study
STUDY_SUMMARY
Using specimens collected as part of a pilot RCT of a diet intervention in pregnancy, we propose to ship our time 2 blood and urine samples (collected post-diet intervention or usual care at 36 weeks gestation) to: 1) Assess biomarker confirmation of the diet intervention effect via: (a) carotenoids and/or other antioxidants (which have been shown to be a good marker of fruit and vegetable intake); and (b) untargeted metabolomics. 2) Assess pregnancy exposure to metals and/or other environmental contaminants. 3) Combine the biomarker data proposed here in aims 1 and 2 with the data being analyzed at MSU (diet survey data and butyrate composition of stool) to explore potential bi-directional effects on gut microbiome, environmental contaminants, and dietary factors.
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Department of Environmental Medicine and Public Health
Development of a weaned pig model of enterotoxigenic E.coli-induced environmental enteropathy
STUDY_TYPE
Animal Model
STUDY_SUMMARY
Environmental Enteropathy (EE) is a subclinical condition primarily effecting developing countries believed to be caused by chronic fecal-oral contamination. The condition is characterized by chronic gut inflammation, malabsorption, stunting of growth, stunted villi, and reduced efficacy of oral vaccines. Due to the similarity of the gastrointestinal tract and immune response of swine and humans, the piglet is an attractive model for studying this condition. In this present study, piglets were challenged with either a chronic or acute dose of pathogenic E.coli in an attempt to mimic the symptoms of EE over a 7 day trial period. Throughout the study a number of biological samples were collected for analysis and are presented here, including feces for untargeted metabolomics analysis.
1H-NMR urinary metabolomic profiling for diagnosis of gastric cancer (part II)
STUDY_SUMMARY
Background: Metabolomics has shown promise in gastric cancer (GC) detection. This research sought to identify whether GC has a unique urinary metabolomic profile compared with benign gastric disease (BN) and healthy (HE) patients. Methods: Urine from 43 GC, 40 BN, and 40 matched HE patients was analysed using 1H nuclear magnetic resonance (1H-NMR) spectroscopy, generating 77 reproducible metabolites (QC-RSD <25%). Univariate and multivariate (MVA) statistics were employed. A parsimonious biomarker profile of GC vs HE was investigated using LASSO regularised logistic regression (LASSO-LR). Model performance was assessed using Receiver Operating Characteristic (ROC) curves. Results: GC displayed a clear discriminatory biomarker profile; the BN profile overlapped with GC and HE. LASSO-LR identified three discriminatory metabolites: 2-hydroxyisobutyrate, 3-indoxylsulfate, and alanine, which produced a discriminatory model with an area under the ROC of 0.95. Conclusions: GC patients have a distinct urinary metabolite profile. This study shows clinical potential for metabolic profiling for early GC diagnosis.
INSTITUTE
University of Alberta
LAST_NAME
Broadhurst
FIRST_NAME
David
ADDRESS
Department of Medicine, 4126A Katz Group Centre for Pharmacy & Health, University of Alberta, Edmonton, AB T6G 2E1, Canada.
EMAIL
d.broadhurst@ecu.edu.au
PHONE
+61 8 6304 2705
PUBLICATIONS
Chan, A. W., Mercier, P., Schiller, D., Bailey, R., Robbins, S., Eurich, D. T., Sawyer, M. B., Broadhurst, D. (2016). 1H-NMR urinary metabolomic profiling for diagnosis of gastric cancer. British Journal of Cancer, 114(1), 59-62. doi:10.1038/bjc.2015.414
Pediatric Inner-City Environmental Exposures at School and Home and Asthma Study
STUDY_TYPE
CHEAR Study
STUDY_SUMMARY
SICAS 1 and SICAS 2 have extraordinary opportunity to evaluate the role of diet, environmental exposures and asthma in the context of school and home specific exposures and capitalize on all the data we are already collecting. Asthma affects 25 million Americans, particularly urban minority children. Children spend nearly all day in school, yet little is known about the role of a childās exposure to widely disseminated industrial chemicals on asthma morbidity. Early animal models and population studies have begun to identify an association between phenolic chemical exposure and asthma development through proposed increased regulation of an individualās allergic immune response. This study, nested within a school-based environmental intervention trial, (School Inner-City Asthma Intervention Study, SICAS2) , will enable urinary biomarker analyses during a school-based academic year-long environmental intervention trial to analyze the source and impact of exposures on urinary environmental exposure biomarker levels as well as the relationship between these biomarkers levels and asthma morbidity. We are poised to leverage the clinical and exposure data being collected in the clinical trial and generate cross-sectional urinary phenol biomarker data (at baseline) within the resources of CHEAR. If successful, our study will assess the impact of exposures on these biomarker levels and the impact that these exposures have on asthma morbidity, controlling comprehensively for other personal, home, and school environmental factors associated with asthma outcomes. We hypothesize that exposure to environmental exposures (e.g. phenols, phthalates, environmental tobacco smoke) in urban school children and higher urinary biomarkers will preliminarily be associated with higher asthma morbidity. Specific aims are: Aim 1. To determine the source of exposure to environmental exposures (e.g. phenols, phthalates, environmental tobacco smoke) in inner-city school children as assessed by questionnaire, product use assessment and comprehensive school and home environmental assessment of children with physician-diagnosed asthma. Aim 2. To determine whether urinary phenol/phathalate/cotinine biomarkers are associated with asthma control (e.g. asthma symptoms, such as asthma-related symptom days (primary outcome), and other phenotypes of asthma/allergic symptoms and inflammation such as allergic sensitization, health care utilization and pulmonary lung function
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Department of Environmental Medicine and Public Health
Two groups of rats were fed either a high salt diet or a low salt diet. This study aims to look at salt intake in correlation to altering other metabolites and the onset of hypertension
Lipidomics for wildlife disease etiology and biomarker discovery: a case study of pansteatitis outbreak in South Africa (part-I)
STUDY_TYPE
lipidomics
STUDY_SUMMARY
Lipidomics is a promising tool to determine biomarkers and elucidate mechanisms associated with anthropogenic-induced stress in wildlife. Therefore, we examine the application of lipidomics for in situ studies on Mozambique tilapia (Oreochromis mossambicus) in Loskop Dam, South Africa. Mortality events of aquatic life associated with an environmentally-derived inflammatory disease, pansteatitis, have occurred in this area. The lipidome of adipose tissue (n = 31) and plasma (n = 51) from tilapia collected from at Loskop Dam were characterized using state of the art liquid chromatography coupled to high-resolution tandem mass spectrometry. Lipid profiles reflected pansteatitis severity and were significantly different between diseased and healthy individuals. Over 13 classes of lipids associated with inflammation, cell death, and/or oxidative damage were upregulated in pansteatitis-affected adipose tissue, including ether-lipids, short-chained triglyceride oxidation products, sphingolipids, and acylcarnitines. Ceramides showed a 1000-fold increase in the most affected adipose tissues, illustrating its potential as sensitive and novel indicators of disease severity. In plasma, triglycerides were found to be downregulated in pansteatitis-affected tilapia. As comprehensive coverage of the lipidome aids in the elucidation of possible disease mechanisms, application of lipidomics could be applied to the understanding of other environmentally-derived inflammatory conditions, such as those caused by obesogens.
INSTITUTE
South East Center for Integrated Metabolomics
DEPARTMENT
Department of Pathology, Immunology and Laboratory Medicine
LABORATORY
SECIM
LAST_NAME
Koelmel
FIRST_NAME
Jeremy
ADDRESS
Department of Pathology, Immunology and Laboratory Medicine, University of Florida, 1395 Center Dr, Room M641c
EMAIL
jeremykoelmel@gmail.com
PHONE
7187300454
NUM_GROUPS
8
TOTAL_SUBJECTS
51
NUM_MALES
28
NUM_FEMALES
23
STUDY_COMMENTS
Adipose and plasma do not overlap exactly in subjects ran
1H-NMR Analysis of Skin and Blubber of Nose Dolphin Metabolome Reveal the Functional Metabolomic Dichotomy of The Organs
STUDY_TYPE
Dolphin Skin and Blubber
STUDY_SUMMARY
The common bottlenose dolphin (Tursiops truncatus) is carnivorous cetacean thriving in marine environment are one of the most common apex predators found in coastal and estuarine ecosystems. Although recent studies have focused on capturing the circulating metabolomes of these organisms, with respect to pollutants and exposures of the marine environment, the skin and blubber are important protective organs that have not been probed. Using 1HNMR based untargeted metabolomics we quantified 51 metabolites belonging to 74 different metabolic pathways in the skin and blubber of bottle nose dolphins (n=5) samples collected in 2017 from the coast of Mexico. Results indicate that the skin and blubber metabolism are quantitatively different. These metabolite abundances could help discriminate the tissue-types using supervised and unsupervised PCA and PLSDA analysis. Heat maps and random forest analysis point to unique metabolites that are important classifiers of the tissue-type. The altered metabolic patterns, mainly linking fatty acid metabolism and ketogenic amino acids, seem to constitute a characteristic of blubber, while the skin showed diverse metabolites involved in gluceoneogenic pathways. 1H NMR spectra allowed the identification of metabolites associated with these organ types, such as pyruvic acid, arginine, ornithine, 2-hydroxybutyric acid, 3-hydroxyisobutyric acid, and acetic acid, as discriminatory and classifying metabolites. These results would lead to further understanding of dolphin skin and blubber metabolism for better efforts in their conservation as well as a measure of marine pollution and ecotoxicology.
INSTITUTE
Wake Forest Baptist Medical Center
DEPARTMENT
Department of Internal Medicine
LAST_NAME
Misra
FIRST_NAME
Biswapriya
ADDRESS
Medical Center Boulevard NRC Building, G#43, Medical Center Boulevard, Winston Salem, NC, 27157, USA
Fingerprinting gastrointestinal diseases by 1H NMR
STUDY_SUMMARY
We studied 64 patients admitted to the Florence main hospital emergency room with severe abdominal pain. A blood sample was drawn from each patient at admission, and the corresponding sera underwent 1H NMR metabolomics fingerprinting. Unsupervised PCA analysis showed a significant discrimination between a group of patients with symptoms of upper abdominal pain and a second group consisting of patients with diffuse abdominal/intestinal pain. Prompted by this observation, supervised statistical analysis (OPLS-DA) showed a very good discrimination (> 90 %) between the two groups of symptoms. Actually in the present study, upper abdominal pain may result from either symptomatic gallstones, cholecystitis or pancreatitis, while diffuse abdominal/intestinal pain may result from either intestinal ischemia, strangulated obstruction or mechanical obstruction. Although limited by the small number of samples from each of these six conditions, discrimination of these diseases was attempted. In the first symptom group, > 70% discrimination accuracy was obtained among symptomatic gallstones, pancreatitis and cholecystitis, while for the second symptom group > 85% classification accuracy was obtained for intestinal ischemia, strangulated obstruction and mechanical obstruction. No single metabolite stands up as a possible biomarker for any of these diseases, while the contribution of the whole 1H NMR serum fingerprint seems to be a promising candidate, to be confirmed on larger cohorts, as a first-line discriminator for these diseases.
Evaluation of the short-term effects of the allelochemical umbelliferone on Triticum durum L. metabolism through GC-MS based untargeted metabolomics
STUDY_SUMMARY
Allelopathy is a plant defense mechanism by which they protect themselves from competitive species using specialized biochemicals in the form of secretion or volatiles released to the environment. Though, umbelliferone is a well-known allelochemical, its mechanism of action in a short-term treatment is far from established. We used ā 10ādays old wheat seedlings treated with104 ĀµM umbelliferone over a time course experiment covering 6 times points, i.e., 0h, 6h, 12h, 24h, 48h, and 96h and compared the metabolomic changes to control (mock-treated) plants. Using gas chromatography mass-spectrometry (GC-MS) based metabolomics efforts, we collectively obtained quantitative data on 177 metabolites that were derivatized (either derivatized singly or multiple times) or not representing 139 non-redundant (unique) metabolites. Out of these 139 metabolites, 118 were associated with a unique HMDB identifier, while 113 were associated with a KEGG identifier. Relative quantification of these metabolites across the time-course of umbelliferone treatment, revealed 22 compounds (sugars, fatty acids, secondary metabolites, organic acids, and amino acids) that changed significantly (repeated measures ANOVA, P-value < 0.05) with time. Using multivariate partial least squares discriminant analysis (PLS-DA) we showed the grouping of samples based on time-course across the control and umbelliferone treated plants, whereas the metabolite-metabolite Pearson correlation revealed tightly formed clusters of umbelliferone-derived metabolites, fatty acids, amino acids, and carbohydrates. Also, time-course of umbelliferone treatment revealed, that phospho-L-serine, maltose, and dehydroquinic acid were the top three metabolites showing highest importance in discrimination among the time-points. The above indicate a system-wide changes induced by umbelliferone, through dysregulation of primary as well as specialized metabolism.
INSTITUTE
UniversitĆ Mediterranea di Reggio Calabria
DEPARTMENT
Dipartimento AGRARIA
LAST_NAME
Araniti
FIRST_NAME
Fabrizio
ADDRESS
Department AGRARIA, University Mediterranea of Reggio Calabria, LocalitĆ Feo di Vito, SNC I-89124, Reggio Calabria RC, Italy
Metabolite Extractions data from the Strains Synechococcus sp. PCC 7002, Synechococcus elongatus PCC 11801 (New strain, Manuscript submitted) and Synechococcus elongatus PCC 7942
Lipidomics for wildlife disease etiology and biomarker discovery: a case study of pansteatitis outbreak in South Africa (part-II)
STUDY_TYPE
lipidomics
STUDY_SUMMARY
Lipidomics is a promising tool to determine biomarkers and elucidate mechanisms associated with anthropogenic-induced stress in wildlife. Therefore, we examine the application of lipidomics for in situ studies on Mozambique tilapia (Oreochromis mossambicus) in Loskop Dam, South Africa. Mortality events of aquatic life associated with an environmentally-derived inflammatory disease, pansteatitis, have occurred in this area. The lipidome of adipose tissue (n = 31) and plasma (n = 51) from tilapia collected from at Loskop Dam were characterized using state of the art liquid chromatography coupled to high-resolution tandem mass spectrometry. Lipid profiles reflected pansteatitis severity and were significantly different between diseased and healthy individuals. Over 13 classes of lipids associated with inflammation, cell death, and/or oxidative damage were upregulated in pansteatitis-affected adipose tissue, including ether-lipids, short-chained triglyceride oxidation products, sphingolipids, and acylcarnitines. Ceramides showed a 1000-fold increase in the most affected adipose tissues, illustrating its potential as sensitive and novel indicators of disease severity. In plasma, triglycerides were found to be downregulated in pansteatitis-affected tilapia. As comprehensive coverage of the lipidome aids in the elucidation of possible disease mechanisms, application of lipidomics could be applied to the understanding of other environmentally-derived inflammatory conditions, such as those caused by obesogens.
INSTITUTE
South East Center for Integrated Metabolomics
DEPARTMENT
Department of Pathology, Immunology and Laboratory Medicine
LABORATORY
SECIM
LAST_NAME
Koelmel
FIRST_NAME
Jeremy
ADDRESS
Department of Pathology, Immunology and Laboratory Medicine, University of Florida, 1395 Center Dr, Room M641c
Glutathione (GSH) is a tripeptide that is implicated in various crucial physiological processes including redox buffering and protection against heavy metal toxicity. GSH is abundant in plants, with reported intracellular concentrations typically in the 1-10 millimolar range. Various aminotransferases can inadvertently transaminate the amino group of the Ī³-glutamyl moiety of GSH to produce deaminated glutathione (dGSH), a metabolite damage product. It was recently reported that an amidase known as Nit1 participates in dGSH breakdown in mammals and yeast. Plants have a hitherto uncharacterized homolog of the Nit1 amidase. We show that recombinant Arabidopsis Nit1 (At4g08790) has efficient amidase activity towards dGSH. Ablating the Arabidopsis Nit1 gene causes a massive accumulation of dGSH and other marked changes to the metabolome. All plant Nit1 sequences examined had predicted plastidial targeting peptides with a potential second start codon whose use would eliminate the targeting peptide. In vitro transcription/translation assays show that both potential translation start codons were used and subcellular localization of GFP fusions confirmed both cytosolic and plastidial localization. Further, we show that Arabidopsis enzymes convert GSH to dGSH at a rate of 2.8 pmol min-1 mg-1 in vitro. Our data demonstrate that plants have a dGSH repair system that is directed to at least two subcellular compartments via the use of alternative translation start sites.
Nude mice orthotopicly implanted with human glioma cell lines
STUDY_SUMMARY
Malignant gliomas are considered to be one of the deadliest human cancers, accounting for about 60% of all primary brain tumors. Cachexia is a complex metabolic derangement and muscle atrophy syndrome, which causes high mortalities in patients with advanced cancers including brain tumors. However, cachexia symptoms induced by gliomas and mechanisms underlying muscle atrophy are unclear. Herein, we developed a glioma cachexia model using nude mice orthotopicly implanted with two glioma cell lines (WHO II CHG5 and WHO IV U87). U87 mice developed more severe cachexia symptoms than CHG5 mice, including more evident anorexia, greater body weight loss and mortality. Unlike non-central nervous system cancer cachexia, glioma cachexia did not induce remarkable systemic inflammation but massive multi-organ atrophy. It also caused significantly decreased skeletal muscle mass and strength, which were associated with down-regulated myosin and AKT, and up-regulated AMPK, FOXO and Atrogin1. Interestingly, expressions of MuRF1, MyoD1, eIF3f, desmin and vimentin were not significantly changed. Consistently, NMR-based metabolomic analyses revealed pronounced metabolic derangements in cachectic gastrocnemius relative to controls. Glucose, glycerol, 3-hydroxybutyrate and glycine were remarkably down-regulated, whereas largely released amino acids due to proteolysis including glutamate, arginine, leucine and isoleucine were up-regulated in cachectic gastrocnemius. Moreover, glucose and lipid metabolism, protein biosynthesis and amino acid metabolism were disturbed dramatically in both glioma-bearing mice. U87 mice showed more changed metabolite levels and altered metabolic pathways. This work uncovers malignant grade-dependent glioma cachexia symptoms and metabolic derangements of skeletal muscle for the first time, and provides hints for new therapeutic approaches.
The proteomic and metabolomic characterization of exercise-induced sweat (part -I)
STUDY_SUMMARY
Proteomic and Metabolomic analysis of exercise-induced sweat to evaluate analyte correlation with human performance parameters. Sweat was collected from participants on a treadmill at low, medium, and hard speed and incline. Sweat was analyzed by untagged HILIC LC-MS.
INSTITUTE
UES Inc, Air Force Research Laboratory
LAST_NAME
Harshman
FIRST_NAME
Sean
ADDRESS
2510 Fifth Street, Area B, Building 840, WPAFB, OH 45433
Evaluation of Seryl-leucine core 1 O-glycosylated peptide (SLC1G) in TB patient urine
STUDY_SUMMARY
Previous work detected an uncharacterized urine metabolite with a molecular mass of 874.3547 Da that showed promise as a biomarker for successful TB treatment. Using mass spectrometry combined with enzymatic digestions, the metabolite was structurally characterized as a seryl-leucine core 1 O-glycosylated peptide (SLC1G) of human origin. Examination of SLC1G in urine revealed a significant abundance increase in individuals with active TB versus their household contacts and healthy controls. Moreover, differential decreases in SLC1G levels were observed by week-one in TB patients during successful treatment versus those that failed treatment. The SLC1G levels also associated with clinical parameters used to measure bacterial burden (GeneXpert) and inflammation (PET-CT). These results demonstrate the importance of metabolite identification and provide strong evidence for applying SLC1G as a biomarker of TB treatment response.
Treatment comparison of Guinea grass planted at University of Sao Paulo Brazil 4 Ambient temp and CO2 plots, 4 elevated CO2 (600 ppm) plots, 4 elevated Temp plots (+2C),4 elevated CO2 and Temp plots. Untargeted metabolomics(GC-MS),de novo transcriptomics, and RNAseq taken at 30(A)(2014-05-22) and 50(B)(2014-07-14)days post treatment exposure. Dried leaf material sent to Uni Illinois-UC. GC-MS protocol followed as stated in Ulanov et al. (2010) J of Plant Phys.
INSTITUTE
University of Illinois at Urbana-Champaign (UIUC)
DEPARTMENT
Plant Biology
LABORATORY
Ainsworth USDA ARS Global Change and Photosynthesis Research Unit
Lipid profiling of Wnt3a-induced optic nerve regeneration
STUDY_TYPE
untargeted lipid profiling
STUDY_SUMMARY
We analyzed lipid profiles of mouse retina and optic nerve for 2 time points - 7 and 15 days post-crush, and 3 conditions - intact control, optic nerve crush + vehicle (saline) intravitreal injection, optic nerve crush + 20 ng of Wnt3a injection.
INSTITUTE
University of Miami
DEPARTMENT
Ophthalmology, Bascom Palmer Eye Institute
LABORATORY
Sanjoy K. Bhattacharya lab
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy
ADDRESS
Bascom Palmer Eye Institute (McKnight Bldg.), 1638 NW 10th Avenue, Suite 707A, University of Miami, Miami, FL, 33136
Integrated metabolome and transcriptome analyses provide novel insight into colon cancer modulation by the gut microbiota
STUDY_SUMMARY
Colon cancer onset and progression is strongly associated with the presence, absence, or relative abundances of certain microbial taxa in the gastrointestinal tract. However, specific mechanisms affecting disease susceptibility related to complex commensal bacterial mixtures are poorly understood. We used a multi-omics approach to determine how differences in the complex gut microbiome (GM) influence the metabolome and host transcriptome and ultimately affect susceptibility to adenoma development. Fecal samples collected from a preclinical rat model of colon cancer harboring distinct complex GMs were analyzed using ultra-high performance liquid chromatography mass spectrometry (UHPLC-MS). We collected samples prior to observable disease onset and identified putative metabolite profiles that predicted future disease severity, independent of GM status. Transcriptome analyses performed after disease onset from normal epithelium and tumor tissues between the high and low tumor GMs suggests that the GM is also correlated with altered host gene expression. Integrated pathway (IP) analyses of the metabolome and transcriptome based on putatively identified metabolic features indicate that bile acid biosynthesis was enriched in rats with high tumors (GM:F344) along with increased fatty acid metabolism and mucin biosynthesis. These data emphasize the utility of using untargeted metabolomics to reveal signatures of susceptibility and resistance and integrated analysis reveals common pathways that are likely to be universal targets for intervention.
Environmental exposures and inflammation in young children with cystic fibrosis
STUDY_TYPE
Untargeted high-resolution mass spectrometry profiling
STUDY_SUMMARY
Secondhand smoke exposure (SHSe) is a common environmental factor known to increase asthma severity and respiratory infections in children, as well disrupt metabolic signals and host immune responses in patients with cystic fibrosis (CF). This study defines biomarkers and metabolic profiles of SHSe (includes ENDS exposure) in the young CF population and determines how SHSe impacts regulation of infection, inflammation, and respiratory health.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Ma
FIRST_NAME
Chunyu
ADDRESS
615 Michael St. Ste 225, Atlanta, GA, 30322, USA
EMAIL
chunyu.ma@emory.edu
PHONE
(404) 727 5091
TOTAL_SUBJECTS
92
STUDY_COMMENTS
Both CHEAR and Clinical Biomarkers Laboratory pooled plasma samples were used for quality control. Study specific sample pools were not created
Dynamic labeling of intracellular metabolites in PCC 7002
STUDY_TYPE
Timecourse experiment
STUDY_SUMMARY
The experiment was conducted in a reactor at a metabolic steady state (OD 730 nm = 0.8). 13C labeled sodium bicarbonate was used as a tracer. Tracer concentration was 1 g/L. After the introduction of the tracer, the samples were collected at time point : 0, 60, 120, 180 and 240 secs. Samples were quenched with methanol and extracted using methanol-chloroform-water method. Extracts were stored at -80 degree C till LCMS analysis.
Combined NMR and MS Analysis of PC patien serum (part-II)
STUDY_SUMMARY
Serum of two groups of patients were analyzed using MS and NMR to find predictive markers of recurrence. Factor Sample Type in study design section refers to samples with Recurrence (R)/ No Recurrence (NR)/ Blank (B) / Pooled (P).
Combined NMR and MS Analysis of PC patien serum (part-III)
STUDY_SUMMARY
Serum of two groups of patients were analyzed using MS and NMR to find predictive markers of recurrence. Study Design factor Sample type represents samples with Recurrence (R) or No Recurrence (NR) of Cancer.
In this study, SETD2 null isogenic 38E/38F clones derived from 786-O cells were generated by zinc finger nucleases, and the cellular metabolic changes of 786-O (WT) and 38E/38F isogenic cell lines (n=3 per group) were analyzed by GC-MS-based targeted metabolomics.
Metabolomics and 16S rRNA sequencing of human colorectal cancers and adjacent mucosa
STUDY_SUMMARY
Colorectal cancer (CRC) is ranked the third most common cancer in human worldwide. However, the exact mechanisms of CRC are not well established. Furthermore, there may be differences between mechanisms of CRC in the Asian and in the Western populations. In the present study, we utilized a liquid chromatography-mass spectrometry (LC-MS) metabolomic approach supported by the 16S rRNA next-generation sequencing to investigate the functional and taxonomical differences between paired tumor and unaffected (normal) surgical biopsy tissues from 17 Malaysian patients. Metabolomic differences associated with steroid biosynthesis, terpenoid biosynthesis and bile metabolism could be attributed to microbiome differences between normal and tumor sites. The relative abundances of Anaerotruncus, Intestinimonas and Oscillibacter displayed significant relationships with both steroid biosynthesis and terpenoid and triterpenoid biosynthesis pathways. Metabolites involved in serotonergic synapse/ tryptophan metabolism (Serotonin and 5-Hydroxy-3-indoleacetic acid [5-HIAA]) were only detected in normal tissue samples. On the other hand, S-Adenosyl-L-homocysteine (SAH), a metabolite involves in methionine metabolism and methylation, was frequently increased in tumor relative to normal tissues. In conclusion, this study suggests that local microbiome dysbiosis may contribute to functional changes at the cancer sites. Results from the current study also contributed to the list of metabolites that are found to differ between normal and tumor sites in CRC and supported our quest for understanding the mechanisms of carcinogenesis.
INSTITUTE
University of Malaya
LAST_NAME
Loke
FIRST_NAME
Mun Fai
ADDRESS
Department of Medical Microbiology, Faculty of Medicine, Kuala Lumpur, Federal Territory, 50603, Malaysia
Physiological and metabolic response of crab megalopae and juveniles to ocean acidification
STUDY_SUMMARY
The objective of the study was to examine the physiological and metabolic response of crab megalopae and juveniles to ocean acidification treatment. Four treatments were used:high pH, high DO (dissolved oxygen); high pH, low DO; low pH, high DO; low pH, low DO
Host NLRP6 exacerbates graft-versus-host disease independent of microbial diversity
STUDY_SUMMARY
Host NLRP6 regulates innate immune responses and gastrointestinal (GI) homeostasis. It plays a protective role in pathogenic processes such as intestinal colitis and tumorigenesis in a microbiome dependent manner. Host innate immunity and changes in microbial diversity also play a role in the severity of allo-immune-mediated gastrointestinal pathogenic process, namely graft-versus-host disease (GVHD), the principal toxicity after allogeneic bone marrow transplantation (allo-BMT). Herein, we examined the role of NLRP6 in multiple murine models of allo-BMT. In contrast to its role in intestinal colitis, host NLRP6 aggravated GI GVHD. NLRP6-deficient animals showed improved intestinal barrier function, increased levels of tissue repair associated proteins and preserved Goblet and Paneth cell numbers in the GI tract after allo-BMT. The impact of host NLRP6 deficiency in mitigating GVHD was observed regardless of co-housing, antibiotic treatment, or colonizing littermate germ free wild type (WT) and NLRP6 deficient hosts with fecal microbial transplantation from SPF WT and Nlrp6-/- animals. Chimera studies were performed to assess the role of NLRP6 expression on host hematopoietic and non-hematopoietic cells. The allogeneic [B6Ly5.2āNlrp6-/-] animals demonstrated significantly improved survival compared to the allogeneic [B6Ly5.2āB6] animals, demonstrating that the absence of NLRP6 in host non-hematopoietic cells is crucial for the protection against GVHD, but did not alter the therapeutic graft-versus-tumor effects after BMT. Our results unveil a novel role for NLRP6 and demonstrate a pathogenic role in GVHD that is independent of variations in its intestinal microbiome in contrast to its well-appreciated microbiome-dependent protective role in intestinal colitis and tumorigenesis.
NMR metabolomics study of Gemcitabine resistant cancer cells
STUDY_TYPE
NMR metabolomics
STUDY_SUMMARY
1D 1H NMR spectra were collected from the samples and analyzed using MVAPACK software (http://bionmr.unl.edu/mvapack.php). Multivariate analysis was conducted to study the metabolic phenotypes of the samples.
INSTITUTE
University of Nebraska-Lincoln
DEPARTMENT
Chemistry
LABORATORY
Dr. Robert Powers and Pankaj Singh labs
LAST_NAME
Gebregiworgis
FIRST_NAME
Teklab
ADDRESS
552 Hamilton Hall, 639 N. 12th Street, Lincoln, NE, 68588, USA
Aspirin Metabolomics in Colorectal Cancer Chemoprevention (part 1 - Colon)
STUDY_TYPE
Untargeted high-resolution mass spectrometry profiling
STUDY_SUMMARY
Substantial evidence supports the effectiveness of aspirin for cancer chemoprevention in addition to its well established role in cardiovascular protection. In recent meta-analyses of randomized controlled trials in humans, daily aspirin use reduced incidence, metastasis and mortality from several common types of cancer, especially colorectal cancer. The mechanism(s) by which aspirin exerts an anticancer benefit is uncertain; numerous effects have been described involving both cyclooxygenase-dependent and -independent pathways. The goal of this research is to elucidate the key metabolic changes that are responsible for the anticancer effects of aspirin in humans using untargeted metabolomics analysis. Metabolomics, or global metabolite profiling, is an emerging discipline that has the potential to transform the study of pharmaceutical agents. Our innovative approach will use high-resolution mass spectroscopy to detect thousands of metabolites in blood plasma and normal colon mucosa biopsies that were collected from participants in the Aspirin/Folate Polyp Prevention Study, a randomized, double-blind, placebo-controlled trial of aspirin and/or folic acid supplementation for the prevention of colorectal adenomas. Participants in the trial were assigned with equal probability to three aspirin treatment arms (placebo, 81 mg, or 325 mg daily). Over the three-year treatment period, 81 mg/day of aspirin reduced the risk of adenomas, whereas the 325 mg/day dose had less effect. The aims of the current proposal are to identify metabolomic signatures, including specific metabolites and metabolic pathways, that are associated with aspirin treatment in blood and normal colon mucosal tissue of participants after three years of randomized aspirin treatment; and then to assess the associations of these metabolic signatures with adenoma risk and whether they mediate the reductions in risk due to 81 mg/day aspirin treatment. We will prioritize metabolites for study by evaluating metabolite levels in patients from the placebo and treatment arms while controlling the false discovery rate, use correlation analysis to enhance identification of relevant metabolic modules associated with these prioritized metabolites, and apply pathway mapping with post-hoc application of ion dissociation spectroscopy to representative metabolites to confirm pathway identification. Because aspirin is a multifunctional drug that is thought to modify numerous pathways with potential roles in carcinogenesis, a global discovery-based metabolomics approach is the best way to identify its key activities. The public health significance of this work is substantial because understanding the mechanism of aspirinās anticancer effects is key to optimizing its use and to the development of novel drugs targeting the metabolic pathways identified.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Uppal
FIRST_NAME
Karan
ADDRESS
615 Michael Street, Atlanta, GA, 30322, USA
EMAIL
kuppal2@emory.edu
PHONE
(404) 727 5027
NUM_GROUPS
3
TOTAL_SUBJECTS
325
NUM_MALES
214
NUM_FEMALES
111
STUDY_COMMENTS
Both pooled colon tissue samples and Clinical Biomarker Laboratory pooled plasma samples were used
Impact of genetic suppression (shRNA) of ALDH1A1 expression in human colon cancer cell line (COLO320)
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Using a multi-omics approach, we have investigated the impact of genetic suppression (shRNA) of ALDH1A1 expression on transcriptomics, proteomics and untargeted metabolomics analyses in a human colon cancer cell line (COLO320). The present study (i) generates an integrative omic profile of scramble shRNA vs. ALDH1A1 shRNA COLO320 cells, and (ii) identifies possible alterations in biological pathways caused by suppression of ALDH1A1 expression.
Early Mechanistic Events Induced by Secondhand Smoke Prevalent Low Molecular Weight Polycyclic Aromatic Hydrocarbons in Mouse Lung Epithelial Cells (part-I)
STUDY_SUMMARY
We evaluated lung epithelial cells exposed to low molecular weight polycyclic aromatic hydrocarbons and what lipid metabolites were produced following early exposure, prior to metabolism. We used 40 uM 1-methylanthracene and fluoranthene (1:1 ratio)for 30 min, 1h, and 4 h time points for the global untargeted metabolomics study (AKB study 1).
Data resource for fully 13C labelled and non-labelled plant tissues (part-I)
STUDY_SUMMARY
LC-MS/MS data files of fully 13C labelled and 12C plant tissues, which were utilized to determine the carbon element number for metabolite characterizations.
Metabolomics of Metabolic Risk in Patients Taking Atypical Antipsychotics
STUDY_TYPE
Quantitative NMR and TLC-GC fatty acid analysis
STUDY_SUMMARY
STUDY OBJECTIVE Patients with schizophrenia are known to have higher rates of metabolic disease than the general population. Contributing factors likely include lifestyle and atypical antipsychotic (AAP) use, but the underlying mechanisms are unknown. The objective of this study was to identify metabolomic variability in adult patients with schizophrenia who were taking AAPs and grouped by fasting insulin concentration, our surrogate marker for metabolic risk. DESIGN Metabolomics analysis. PARTICIPANTS Ninety-four adult patients with schizophrenia who were taking an AAP for at least 6 months, with no changes in their antipsychotic regimen for the previous 8 weeks, and who did not require treatment with insulin. Twenty age- and sex-matched nonobese (10 subjects) and obese (10 subjects) controls without cardiovascular disease or mental health diagnoses were used to match the body mass index range of the patients with schizophrenia to account for metabolite concentration differences attributable to body mass index. MEASUREMENTS AND MAIN RESULTS Existing serum samples were used to identify aqueous metabolites (to differentiate fasting insulin concentration quartiles) and fatty acids with quantitative nuclear magnetic resonance (NMR) and gas chromatography (GC) methods, respectively. To exclude metabolites from our pathway mapping analysis that were due to variability in weight, we also subjected serum samples from the nonobese and obese controls to the same analyses. Patients with schizophrenia had a median age of 47.0 (interquartile range 41.0-52.0) years. Using a false discovery rate threshold of <25%, 10 metabolites, not attributable to weight, differentiated insulin concentration quartiles in patients with schizophrenia and identified variability in one-carbon metabolism between groups. Patients with higher fasting insulin concentrations (quartiles 3 and 4) also trended toward having higher levels of saturated fatty acids compared with patients with lower fasting insulin concentrations (quartiles 1 and 2). CONCLUSION These results illustrate the utility of metabolomics to identify pathways underlying variable fasting insulin concentration in patients with schizophrenia. Importantly, no significant difference in AAP exposure was observed among groups, suggesting that current antipsychotic use may not be a primary factor that differentiates middle-aged adult patients with schizophrenia by fasting insulin concentration. This article is protected by copyright. All rights reserved. As published in Pharmacotherapy. 2018 Jun;38(6):638-650.
Metabolomics of Metabolic Risk in Patients Taking Atypical Antipsychotics (part II)
STUDY_SUMMARY
STUDY OBJECTIVE Patients with schizophrenia are known to have higher rates of metabolic disease than the general population. Contributing factors likely include lifestyle and atypical antipsychotic (AAP) use, but the underlying mechanisms are unknown. The objective of this study was to identify metabolomics variability in adult patients with schizophrenia who were taking AAPs and grouped by fasting insulin concentration, our surrogate marker for metabolic risk. DESIGN Metabolomics analysis. PARTICIPANTS Ninety-four adult patients with schizophrenia who were taking an AAP for at least 6 months, with no changes in their antipsychotic regimen for the previous 8 weeks, and who did not require treatment with insulin. Twenty age- and sex-matched nonobese (10 subjects) and obese (10 subjects) controls without cardiovascular disease or mental health diagnoses were used to match the body mass index range of the patients with schizophrenia to account for metabolite concentration differences attributable to body mass index. MEASUREMENTS AND MAIN RESULTS Existing serum samples were used to identify aqueous metabolites (to differentiate fasting insulin concentration quartiles) and fatty acids with quantitative nuclear magnetic resonance (NMR) and gas chromatography (GC) methods, respectively. To exclude metabolites from our pathway mapping analysis that were due to variability in weight, we also subjected serum samples from the nonobese and obese controls to the same analyses. Patients with schizophrenia had a median age of 47.0 (interquartile range 41.0-52.0) years. Using a false discovery rate threshold of <25%, 10 metabolites, not attributable to weight, differentiated insulin concentration quartiles in patients with schizophrenia and identified variability in one-carbon metabolism between groups. Patients with higher fasting insulin concentrations (quartiles 3 and 4) also trended toward having higher levels of saturated fatty acids compared with patients with lower fasting insulin concentrations (quartiles 1 and 2). CONCLUSION These results illustrate the utility of metabolomics to identify pathways underlying variable fasting insulin concentration in patients with schizophrenia. Importantly, no significant difference in AAP exposure was observed among groups, suggesting that current antipsychotic use may not be a primary factor that differentiates middle-aged adult patients with schizophrenia by fasting insulin concentration. This article is protected by copyright. All rights reserved. As published in Pharmacotherapy. 2018 Jun;38(6):638-650.
Untargeted LC-MS to compare blood collection tube and processing time ā HILIC & C18
STUDY_SUMMARY
Blood was collected from three healthy volunteers in 3 blood collection tubes: serum separator tube SST (serum), EDTA (plasma), and P100 (plasma) and stored at 4 degrees for 0, 0.5, 1, 2, 4, and 24 hours prior to centrifugation. Compounds were extracted using liquid-liquid extraction to obtain a hydrophilic and a hydrophobic fraction and analyzed using liquid chromatography mass spectrometry. Differences among the blood collection tubes and sample processing time were evaluated (ANOVA with Bonferroni FWER ā¤ 0.05 and ANOVA with Benjamini Hochberg FDR ā¤ 0.1, respectively).
Identification of potential sepsis biomarkers in burn injury conditions.
STUDY_SUMMARY
12 mice were subjected to one of four treatment groups: no burn injury:no injection, burn injury:no infection, no burn injury:infection, and burn injury:infection. The researchers in this study are looking for metabolites in serum that could indicate sepsis in mice with burn injuries.
Physiological and metabolic response of crab megalopae and juveniles to ocean acidification
STUDY_SUMMARY
Young crab samples were placed into 1 of 4 treatment groups to understand their metabolic response to ocean acidification and dissolved oxygen content.
Metabolomics relies on analytical methods to provide holistic information about metabolites, their distributions across samples, and their underlying dynamic properties. The latter is gaining increasing attention due to advances in modeling and new analytical methods that provide dense time-series data. We extended high-resolution-magic angle spinning (HR-MAS) NMRāan established technique to measure metabolites from tissues and live organismsāinto a flexible, untargeted, and continuous recording of in vivo metabolism. We call this technique ācontinuous in vivo metabolism by NMRā (CIVM-NMR). We used isotope-edited CIVM-NMR to reproduce a recent amino acid flux result in chronic lymphoid leukemia cells. We then collected untargeted CIVM-NMR datasets for Neurospora crassa, a classic multicellular model of biochemistry, genetics, and metabolism. CIVM-NMR requires virtually no sample preparation and allows for continuous collection of data over hours to days at ~4-min temporal resolution with little noise. CIVM-NMR provided real-time measurements that unambiguously reproduced the direction of flux of branched-chain amino acid accumulation in leukemia cells. It also revealed the dynamics of central carbon metabolism, amino acid metabolism, energy storage molecules, and lipid and cell wall precursors in N. crassa. CIVM-NMR is simple and readily adapted to different types of cells and microorganisms, making it ideally suited to experimentally complement kinetic models of metabolism for diverse biological systems.
INSTITUTE
University of Georgia
LAST_NAME
Judge
FIRST_NAME
Michael
ADDRESS
315 Riverbend Rd., Edison Lab, Athens, GA, 30605, USA
An untargeted metabolomics approach was utilized to determine urinary metabolites that could serve as small molecule biomarkers for treatment response to standard tuberculosis treatment. However, the majority of metabolites that most accurately distinguished patient samples at time of diagnosis from those at one month after the start of therapy lacked structural identification. The detection of unknown metabolite structures is a well-known limitation of untargeted metabolomics, and underscores a need for continued elucidation of novel metabolite structures. In this study, we sought to define the structure of a urine metabolite with an experimentally determined mass of 202.1326 Da, classified as molecular feature (MF) 874.3547. Using mass spectrometry combined with enzymatic digestions, the metabolite was structurally characterized as a seryl-leucine core 1 O-glycosylated peptide (SLC1G) of human origin.
The aim of this project was to characterize lipid profiles of the retinal ganglion cells (RGCs) with different regenerative capacity. RGCs were FACS-sorted (7,000 cells/sample) from mice of OPN4 Cre tdT and Thy1-CFP genotype. The treatment conditions included intact, optic nerve crush (ONC) and ONC plus CNTF to promote regeneration. Samples were then subject to lipid extraction and analyzed using LC-MS/MS, followed by identification and relative quantification in LipidSearch software.
Lipidomics of Newborn Heart Tissue Exposed to Excess Maternal Cortisol in Late Gestation (part-1)
STUDY_SUMMARY
Cardiac tissue from newborn hearts from animals exposed to excess maternal cortisol in late gestation and untreated was compared via MS lipidomic analysis
INSTITUTE
University of Florida
DEPARTMENT
Biochemsitry & Molecular Biology
LAST_NAME
Walejko
FIRST_NAME
Jacquelyn
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry and Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245
NMR Metabolomics of Newborn Heart Tissue Exposed to Excess Maternal Cortisol in Late Gestation (part -II)
STUDY_TYPE
Comparison
STUDY_SUMMARY
Cardiac tissue from newborn hearts from animals exposed to excess maternal cortisol in late gestation and untreated was compared via NMR metabolomic analysis
INSTITUTE
University of Florida
DEPARTMENT
Biochemsitry & Molecular Biology
LAST_NAME
Walejko
FIRST_NAME
Jacquelyn
ADDRESS
R3-226 Academic Research Building, Department of Biochemistry and Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245
Investigating link between metabolism and circadian rhythm in Drosophila melanogaster
STUDY_SUMMARY
We are investigating the link between metabolism, circadian rhythm, and a high-fat diet with an emphasis on the role of metabolites that affect protein post translational modifications. We've hypothesized that protein modifications, which are critical to circadian clock functions, can be affected by different metabolic profiles, such as an excess or lack of fat in the diet, that ultimately regulate changes in circadian biology.
INSTITUTE
University of California, Davis
DEPARTMENT
College of Biological Sciences
LAST_NAME
Contreras
FIRST_NAME
Adam
ADDRESS
202 Life Sciences Building, Kleiber Hall Drive, Davis, CA, 95616
Screening of Breast Cancer tissue samples for alteration in the lipidome using high resolution mass spectrometer,and mapping of these changes to clinico-pathological data to search for potential bio markers
The nutrition value of fish fillet is related to fish maturation or fish age? Integrated analysis of transcriptomics and metabolomics in blunt snout bream (Megalobrama amblycephala)
STUDY_SUMMARY
With the improvement of living standards, peopleās demand for food nutrient is getting higher and higher. Fish is one kind of protein-rich food and is increasingly favored by consumers. It has been well recognized that flesh composition of fish is closely related to its maturation and growth stages, but few researches have explored these differences. Besides, hormone residues in fish after artificial inducing reproduction also attract consumersā concern. In this study, we try to address these concerns by using a combination of transcriptomics and metabolomics analysis to identify the key pathways, genes, and metabolites regulation which may affect flesh nutrition of one typical aquaculture species in China, blunt snout bream (Megalobrama amblycephala).
INSTITUTE
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education
LAST_NAME
guan
FIRST_NAME
ningnan
ADDRESS
College of Fisheries,Huazhong Agricultural University 1 Shizishan Road Hongshan District Wuhan, Hubei
Exposure to Oral Contraceptives Alters Human Endometrial Stem Cells Culture Media Metabolomics
STUDY_SUMMARY
Although the effects of oral contraceptives (OCs) in the endometrium has been well established, its influence in the production and metabolism of endometrial mesenchymal stem cells (EnMSC) remains unclear. Therefore, we analyzed the effect of OCs in the EnMSC secretome by culture media quantitative metabolomics. The EnMSC were collected from menstrual shedding of five donors (OC group, n=3; control, non-OC group, n=2) and cultured for three passages. Cells characterization was performed by flow cytometry, and culture media was collected at the end of each passage for further quantitative metabolomics. The metabolites with higher discriminant power for sample classification were Alanine, PC aa C30:0, c4-OH-Pro, PC aa C32:2, PC ae C32:2, PC ae C40:2, glycine and PC ae C32:1 for the OC group, whereas PC aa C36:6, PC aa C34:4, SM OH C16:1, SM C26:0, PC aa C38:0, serine and PC aa C36:5 were representative of the non-OC group. This panel of metabolites showed 98% of sensitivity in sample classification according with respective groups. Altered concentrations of metabolites may be an effect of OC hormonal properties on EnMSC metabolism. Thus, this metabolomic approach could assist in the management of future stem cell therapies according to patientsā specific responses to hormone treatments.
Metabolomics profile of umbilical cord blood is associated with maternal pre-pregnant obesity in a perspective multi-ethnic cohort displaying health disparities
STUDY_SUMMARY
Maternal obesity has become a growing global health concern that impacts fetal health and subsequently predisposes the offspring to medical conditions later in life. However, the molecular link between abnormal fetal metabolomic profiles and maternal obesity has not yet been fully elucidated. In this study, we report new discoveries from the newborn cord blood metabolomes associated with a case-control maternal obesity cohort, collected from multi-ethnic populations in Hawaii, including rarely reported Native Hawaiian and other Pacific Islanders (NHPI). This cohort displays significant maternal obesity disparities by subjectsā residential area average income and health insurance. An elastic net penalized logistic regression model was constructed to associate cord blood metabolomics and demographic/physiological information to maternal obesity, with accuracy as high as 0.947. We identify 29 metabolites as early-life biomarkers manifesting intrauterine effect of maternal obesity.
Growth cone-enriched lipidome of embryonic to early postnatal mouse brain
STUDY_SUMMARY
A growth cone (GC) is a part of a neuron considered a hub for axon growth, motility and guidance functions. Unraveling the molecular composition of GCs and events through which active GCs transition to terminal synapses is of importance to developmental and regenerative neuroscience research. Here, we present a dataset on the lipid profiling of the growth cone-enriched fractions derived from E18, P0, P3, P6 and P9 C57BL/6J mouse brains. For comparison, we analyzed non-growth cone membranes.
Variability in metabolomic profiles among unique genotypes of Acropora cervicornis
STUDY_TYPE
intraspecific variability
STUDY_SUMMARY
We hypothesized that each of the three genotypes tested would have unique metabolomic profiles. These data increase our basic knowledge of the coral metabolome and represent an important step toward linking genotype, phenotype, and metabolome in reef-building corals.
INSTITUTE
University of Florida
LAST_NAME
Patterson
FIRST_NAME
Joshua
ADDRESS
Florida Aquarium Center for Conservation, 529 Estuary Shore Lane, Apollo Beach, FL 33572
Metabolomics profiles of patients with Wilson disease reveal a distinct metabolic signature
STUDY_SUMMARY
This study is comparing the plasma metabolomics profile of patients with the genetic disorder, Wilson disease, compared to healthy subjects matched by age, sex, and BMI. Wilson disease is caused by a defect in a copper transporter leading to copper accumulation in the liver and brain leading to liver and/or neuropsychiatric symptoms. Mitochondrial defects are well-documented in Wilson disease. We hypothesize the acylcarnitine and primary metabolite profile will differ between patients with Wilson disease and healthy subjects and that these differences may indicate specific metabolic abnormalities.
Quantification of microenvironmental metabolites in murine cancers reveals determinants of tumor nutrient availability
STUDY_SUMMARY
Cancer cell metabolism is heavily influenced by microenvironmental factors, including nutrient availability. Therefore, knowledge of microenvironmental nutrient levels is essential to understand tumor metabolism. To measure the extracellular nutrient levels available to tumors, we developed a quantitative metabolomics method to measure the absolute concentrations of >118 metabolites in plasma and tumor interstitial fluid, the extracellular fluid that perfuses tumors. Comparison of nutrient levels in tumor interstitial fluid and plasma revealed that the nutrients available to tumors differ from those present in circulation. Further, by comparing interstitial fluid nutrient levels between autochthonous and transplant models of murine pancreatic and lung adenocarcinoma, we found that tumor type, anatomical location and animal diet affect local nutrient availability. These data provide a comprehensive characterization of the nutrients present in the tumor microenvironment of widely used models of lung and pancreatic cancer and identify factors that influence metabolite levels in tumors.
INSTITUTE
University of Chicago
LAST_NAME
Muir
FIRST_NAME
Alexander
ADDRESS
929 E 57th St. W GCIS 306, Chicago, Illinois, 60637, USA
Reprogrammed Lipid Metabolism in Bladder Cancer with Cisplatin Resistance
STUDY_SUMMARY
This study conducted comprehensive and comparative lipidomic profiling of two isogenic human T24 bladder cell lines, which are characterized as sensitive or resistant to the cisplatin-induced cell apoptosis.
INSTITUTE
Cedars-Sinai Medical Center
DEPARTMENT
Departments of Surgery and Biomedical Sciences
LAST_NAME
Kim
FIRST_NAME
Jayoung
ADDRESS
8700 Beverly Blvd., Davis 5071 Los Angeles, CA 90048, USA
Identification of urine metabolites in patients with interstitial cystitis using untargeted metabolomics (part I)
STUDY_SUMMARY
Urine metabolites are used in many clinical and biomedical studies, but usually only for a few classic compounds. Metabolomics detects vastly more metabolic signals that may be used to precisely define the health status of individuals. However, many compounds remain unidentified, hampering biochemical conclusions. Here, we annotate all metabolites detected using an untargeted metabolomics approach with a charged surface hybrid (CSH) assay on a Q Exactive HF mass spectrometry.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
BlaženoviÄ
FIRST_NAME
Ivana
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Identification of urine metabolites in patients with interstitial cystitis using untargeted metabolomics (part II)
STUDY_SUMMARY
Urine metabolites are used in many clinical and biomedical studies, but usually only for a few classic compounds. Metabolomics detects vastly more metabolic signals that may be used to precisely define the health status of individuals. However, many compounds remain unidentified, hampering biochemical conclusions. Here, we annotate all metabolites detected using an untargeted metabolomics approach with a hydrophilic interaction liquid chromatography (HILIC) assay on a Q Exactive HF mass spectrometry.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
BlaženoviÄ
FIRST_NAME
Ivana
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Early Mechanistic Events Induced by Secondhand Smoke Prevalent Low Molecular Weight Polycyclic Aromatic Hydrocarbons in Mouse Lung Epithelial Cells
STUDY_SUMMARY
We evaluated lung epithelial cells exposed to low molecular weight polycyclic aromatic hydrocarbons and what lipid metabolites were produced following early exposure, prior to metabolism. For the targeted study (AKB study 2, we used 40uM dose of the 2PAHs (1methylanthracene and fluoranthene; 1:1 ratio)and assessed 2,4,8,and 12 hrs of treatment with the PAHs. We also examined a 24 h time point in another study at a lower dose (15 uM LMW PAH mixture; 1:1 ratio of 1-methanthrancene and fluoranthene).
WT and ĪSPT cultures of B. thetaiotaomicron and B. ovatus grown BHI liquid media (part I)
STUDY_SUMMARY
Lipid profiling was applied to WT and ĪSPT cultures of B. thetaiotaomicron and B. ovatus grown BHI liquid media revealing a greater variety of Bacteroides-derived sphingolipids than previously recognized, including ceramide phosphoinositol and deoxy-sphingolipids.
INSTITUTE
Broad Institute of MIT and Harvard;Harvard School of Public Health; University of Groningen;Novartis Institute for Biomedical Research Inc Center for the Study of Inflammatory Bowel Disease, University of Groningen and University Medical Center Groningen, Novartis Institute for Biomedical Research Inc
WT and ĪSPT cultures of B. thetaiotaomicron grown in Minimal Media (part II)
STUDY_SUMMARY
Lipid profiling was applied on WT and ĪSPT cultures of B. thetaiotaomicron grown in minimal liquid media in order to confirm the production of Bacteroides-derived sphingolipids.
INSTITUTE
Broad Institute of MIT and Harvard;Harvard School of Public Health; University of Groningen;Novartis Institute for Biomedical Research Inc Center for the Study of Inflammatory Bowel Disease, University of Groningen and University Medical Center Groningen, Novartis Institute for Biomedical Research Inc
Lipid profiling of caecal samples from GF mice colonized with B. thetaiotaomicron WT or the ĪSPT mutants (part III)
STUDY_SUMMARY
To study Bacteroidetes sphingolipids in intestinal health, we colonized germ-free mice with wild type or a sphingolipid-deficient Bacteroides thetaiotaomicron strain. A lack of Bacteroides derived sphingolipids increased intestinal inflammation, dysregulated innate immunity and altered the host ceramide pool.
WT and ĪSPT cultures of B. thetaiotaomicron grown in Minimal Media with or without d4-alanine (part IV)
STUDY_SUMMARY
Lipid profiling was applied on WT and ĪSPT cultures of B. thetaiotaomicron grown in minimal liquid media supplemented with or without deuterium (D4)-labelled alanine, in order to elucidate the production pathways of Bacteroides-derived sphingolipids.
Urea cycle and 1C/serine metabolism in the prevention of oxygen induced retinopathy
STUDY_SUMMARY
Untargeted metabolite profiling links the urea cycle and 1C/serine metabolism in the prevention of oxygen induced retinopathy by hepatic HIF stabilization. Premature infants require oxygen supplementation to survive that is simultaneously toxic to developing tissues. We have demonstrated that hypoxia inducible factor (HIF) stabilization during hyperoxia prevents oxygen induced retinopathy (OIR) and lung disease. Here, untargeted metabolite profiling coupled to XCMS systems biology analysis finds that serine/1C and urea cycles dominate pathway enrichment graphs. MS1 peak areas and MS2 library matches reveal 50% or more increased levels of plasma and retina serine, glycine, hypotaurine, methionine, and taurine. In addition, N-acetylglutamate increased 4-fold in serum, while orotate, citrulline, arginine, aspartate, glutamine were at least 50% increased after HIF stabilization. Targeted data analysis in vivo finds that retinal serine and glycine were derived from liver. HIF-1Ī±2lox/2lox; albumin-cre KO had reduced levels of serine and retinal glycine. Inhibition of 1C metabolism blocked rescue by HIF stabilization. The metabolic phenotype of mice protected from OIR by HIF stabilization is dependent on hepatic serine/1C metabolism and urea cycle.
Presentation of different serum metabolites in trauma patients versus healthy volunteers.
STUDY_SUMMARY
This study seeks to identify critical metabolic differences between patients who have undergone physical trauma and patients who have not experienced physical trauma.
Investigate the False Discovery in Biomarker Research Using LC-HRMS based Untargeted Metabolomics Profiling
STUDY_TYPE
mimicking metabolomics study
STUDY_SUMMARY
Pooled human plasma was spiked separately with two different sets of 11 metabolite standards (22 ātrue biomarkersā) to mimic plasma samples collected from diseased subjects and non-diseased subjects. Metabolite extracts of individual samples were subjected to biomarker discovery using LC-HRMS.
INSTITUTE
University of Macau, Macau, China
DEPARTMENT
Institute of Translational Medicine, Faculty of Health Sciences,
Downregulation of CENPF epigenetically remodels prostate cancer cells and alters cellular metabolism
STUDY_SUMMARY
This study aimed to determine the metabolic profile of CENPF-knockout (CENPFKO) PC cells and identify differentially expressed metabolites (DEMs) that can be used as diagnostic markers.
INSTITUTE
Cedars-Sinai Medical Center
LAST_NAME
Kim
FIRST_NAME
Jayoung
ADDRESS
8700 Beverly Blvd., Davis 5071 Los Angeles, CA 90048, USA
Different dose exposure of OPC-163493 on HepG2 cells (part-I)
STUDY_TYPE
Compound dosage test
STUDY_SUMMARY
Metabolomics analysis were on 8 samples of HepG2 cells that were treated with compound OPC-163493 (DMSO control, 1, 3, or 10ĀµM; each n=2) exposure for 30 min.
INSTITUTE
Otsuka Pharmaceuticals
LAST_NAME
Kanemoto
FIRST_NAME
Naohide
ADDRESS
463-10 Kagasuno Kawauchi-cho Tokushima 771-0192, Japan
Metabolme analysis of OPC-163493 on the Liver of ZDF rats (part-II)
STUDY_TYPE
Long-term in vivo test
STUDY_SUMMARY
Metabolome analysis were on the 24 samples of ZDF rats that were treated with OPC-163493 for 6-weeks. The 24 samples were composed of 3 different groups (Vehicles, OPC-163493 treatment, and baseline control; each n=8).
INSTITUTE
Otsuka Pharmaceuticals
LAST_NAME
Kanemoto
FIRST_NAME
Naohide
ADDRESS
463-10 Kagasuno Kawauchi-cho Tokushima 771-0192, Japan
Changes in the Canine Plasma Lipidome after Short- and Long-Term Excess Glucocorticoid Exposure
STUDY_SUMMARY
Glucocorticoids (GCs) are widely used in veterinary and human medicine. Chromic endogenous or iatrogenic GC overexposure impairs metabolic function and can result in diverse side-effects, including Cushingās syndrome. This study examines the effects of experimentally induced short-term and long-term GC excess (induced by prednisolone and tetracosactide, respectively) on the plasma lipidome of Beale dogs. Both, long- and short-term GC resulted in significant changes of the plasma lipidome.
INSTITUTE
Life Sciences Institute, National University of Singapore
Evaluation of metabolome sample preparation and extraction methodologies for oleaginous filamentous fungi Mortierella alpina
STUDY_TYPE
method optimization
STUDY_SUMMARY
In this study, based on the method of fast filtration, we evaluated the three metabolomics analysis protocols commonly used for microbial metabolomics analysis in M. alpina and systematically optimised the metabolite extraction solvent.
INSTITUTE
Jiangnan Unviersity
DEPARTMENT
School of Food Science and Technology
LABORATORY
State Key Laboratory of Food Science and Technology
928 cancer cell lines from 20 major cancer types were cultured in vitro for metabolomic profiling of 124 polar and 101 lipid species. Extracted polar and lipid metabolites were analyzed using hydrophilic interaction
INSTITUTE
Broad Institute of MIT and Harvard
LAST_NAME
Avila
FIRST_NAME
Julian
ADDRESS
415 Main Street, Rm 7175, Cambridge, MA, 02142, USA
Microbial depletion and ozone exposure - Lung tissue (part I)
STUDY_SUMMARY
Global biochemical profiles were determined in lung tissue collected from untreated control mice and mice treated for two weeks with untreated drinking water or water containing an antibiotic cocktail (ampicillin, neomycin, metronidazole, and vancomycin), followed by a three hour exposure to ambient air or ozone (2ppm). Sample collection occurred 24 hours post-ozone exposure.
Microbial depletion and ozone exposure - serum (part II)
STUDY_SUMMARY
Global biochemical profiles were determined in serum collected from untreated control mice and mice treated for two weeks with untreated drinking water or water containing an antibiotic cocktail (ampicillin, neomycin, metronidazole, and vancomycin), followed by a three hour exposure to ambient air or ozone (2ppm). Sample collection occurred 24 hours post-ozone exposure.
UPLC-MS Analysis of Lipids From Insulin Resistant Femoral Muscles of Diet-induced Obese Mice
STUDY_TYPE
Lipidomics, Basic Research
STUDY_SUMMARY
Muscle insulin resistance is a fundamental contributor in the pathogenesis of obesity-related diseases like type 2 diabetes. Increased triglyceride concentration in muscle tissue, as seen with obesity, is associated with inhibition of insulin action and decreased glucose uptake. Here we use liquid chromatography paired with mass spectrometry (LCMS) to identify patterns of lipid species in femoral muscle of mice associated with diet-induced insulin resistance. Mice were fed a standard CHOW diet for 5 weeks or HFD for 5 or 13 weeks. 806 lipids were significantly different (p ā¤ 0.05) between HFD-induced insulin resistant muscle and CHOW insulin sensitive. Of these 217 lipid species were quantified and annotated based on principle components analysis, significance (p ā¤ 0.01) and fold change of relative abundance values. CHOW insulin sensitive muscle was associated with triglycerides and phospholipids that contained higher abundance of long-chain highly unsaturated fatty acids. Serine and inositol phospholipids favored insulin sensitive femoral muscle, yet higher abundance also occurred in 13 week HFD mice compared with 5 week. Consequently, phospholipid imbalance may be indicative of cell membrane dysfunction. HFD insulin resistant femoral muscle contained triglycerides with less carbons, compared with CHOW, which were predominantly saturated. In addition, there was greater abundance of diacylglycerides and sphingomyelin, but not ceramides. Extending HFD intake to 13 weeks did not cause increased abundance of deleterious lipids with the exception of sphingomyelin. Overall, distinct lipid combinations, perhaps even ratios, should be characterized when identifying what contributes to the maintenance or dysregulation of muscle insulin sensitivity.
INSTITUTE
Colorado State University
DEPARTMENT
Food Science and Human Nutrition
LABORATORY
Adipose Tissue
LAST_NAME
Foster
FIRST_NAME
Michelle
ADDRESS
1571 Campus Delivery, Fort Collins, Colorado 80523
Here, we use global metabolomics to differentiate temporal effects (1 ā 60 d) found in nonhuman primate (NHP) urine small molecule signatures after a 4 Gy total body irradiation.
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Here, we use global metabolomics to differentiate temporal effects (1 ā 60 d) found in nonhuman primate (NHP) serum small molecule signatures after a 4 Gy total body irradiation.
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Effect of cell harvesting technique and storage on metabolic profiles in human skin fibroblasts
STUDY_TYPE
Method
STUDY_SUMMARY
Human skin fibroblasts were cultured in MEM media supplemented with 15% FBS. Cells were harvested using (i) trypsinization, (ii) scraping, (iii) methanol fixation and scraping, (iV) methanol fixation, scraping, and drying. Targeted metabolomics analysis of organic acids, amino acids, and acycarnitines was conducted at Mayo Clinic Metabolomics Core Facility.
Metabolomics of a Mouse Model for Retinitis Pigmentosa
STUDY_SUMMARY
Retinitis pigmentosa (RP) is a degenerative disease of the retina that affects approximately 1 million people worldwide. There are multiple genetic causes of this disease, for which, at present, there are no effective therapeutic strategies. We utilized broad spectrum metabolomics to identify perturbations in the metabolism of the rd10 mouse, a genetic model for RP. C57BL/6J and rd10 mice were raised in cyclic light followed by either light or dark adaptation at postnatal day (P) 18, an early stage in the degeneration process. Mice raised entirely in the dark until P18 were also evaluated. After euthanasia, retinas were removed and extracted for analysis by ultra-performance liquid chromatography-time of flight-mass spectrometry (UPLC-QTOF-MS).
4-day dietary effect of fast food vs Mediterranean diet to HDL lipidome
STUDY_TYPE
Dietary intervention study
STUDY_SUMMARY
In this randomized order cross-over study, ten healthy subjects consumed a Mediterranean (Med) and a fast food (FF) diet for 4 days, with a 4-day wash-out between treatments. Lipidomic composition was analyzed in isolated HDL fractions by an untargeted LC-MS method with 15 internal standards. HDL PE content was increased by FF diet, and 41 out of 170 lipid species were differentially affected by diet. Saturated fatty acids (FA) and odd chain FA were enriched after FF diet, while very-long chain FA and unsaturated FA were enriched after Med diet. The composition of PC, TG and CE were significantly altered to reflect the FA composition of the diet whereas the composition of SM and ceramides were generally unaffected, indicating that glycerolipids may be sensitive markers of dietary intake, whereas sphingolipids are more indicative of non-dietary factors. Results from this study indicate that the HDL lipidome is widely remodeled within 4 days of diet change and that certain lipid classes are more sensitive markers of diet whereas other lipid classes are better indicators of non-dietary factors
Metabolomic Analysis of Liver Tissues for Characterization of Hepatocellular Carcinoma
STUDY_SUMMARY
Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer causing more than half a million annual deaths world-wide. Understanding the molecular mechanisms contributing to HCC development and progression is highly desirable for improved surveillance, diagnosis and treatment. Liver tissue metabolomics has the potential to reflect the physiological changes behind HCC development. Also, it allows researchers to investigate racial disparities in HCC. The use of both gas chromatography ā mass spectrometry (GC-MS) and liquid chromatography ā mass spectrometry (LC-MS) platforms helps increase the metabolome coverage, allowing researchers to better unravel the relationships of metabolites and HCC. The objective of this study is to identify HCC-associated metabolites by analysis of liver tissues from HCC patients using both GC-MS and LC-MS platforms. Paired tumor and non-tumor tissues from 40 patients were analyzed by GC-MS and LC-MS. The patients consist of 14 African-Americans (AA), 10 Asian-Americans (AS), and 16 European-Americans (EA). The levels of the metabolites extracted from the solid liver tissue of the HCC area and adjacent non-HCC area were compared. Among the analytes detected by GC-MS and LC-MS with significant alterations, 17 were selected based on availability of putative metabolite identifications. These metabolites belong to TCA cycle, glycolysis, purines, and lipid metabolism, and have been previously reported in liver metabolomics studies where high correlation with HCC progression was implied. We demonstrated that metabolites that are related to HCC pathogenesis can be identified through metabolomics analysis of liver tissues by both GC-MS and LC-MS. In addition, this analysis has led to the identification of metabolites associated with HCC in a race-specific manner.
Background: Although the erythrocyte is the most abundant cell type in our body, acting as both a deliverer and sensor of oxygen (O2), its function and regulatory mechanism in chronic kidney disease (CKD) remain unknown. Methods: Unbiased metabolomics screening in the whole blood of mice infused with or without angiotensin II (Ang II) at 140ng/kg/min up to 14 days was conducted. Mice with specific ablation of ADORA2B in erythrocytes and patients with CKD were used to determine its function in CKD, potential mechanisms and human relevance. Results: Unbiased metabolomics revealed that 2,3-biphosphoglycerate (2,3-BPG), an erythrocyte-specific metabolite promoting O2 delivery, was significantly induced in an experimental model of CKD induced by Ang II. Mouse genetic studies revealed that erythrocyte ADORA2B signaling via AMPK-stimulated activation of BPG mutase was a key compensatory cellular response to counteract kidney hypoxia, tissue damage and disease progression in Ang II-induced CKD by promoting 2,3-BPG production and O2 delivery. Preclinical studies showed that enhancing AMPK activation offset kidney hypoxia by triggering 2,3-BPG production and O2 delivery. Human translational studies validated mouse findings that erythrocyte 2,3-BPG levels, AMPK activity and O2 delivery capacity were significantly induced in the erythrocytes of CKD patients compared to normal controls and their elevations were correlated to disease severity. Conclusion: Overall, we have provided both mouse and human evidence that ADORA2B-AMPK signaling cascade-induced 2,3-BPG production is a beneficial erythrocyte response to promote O2 delivery to counteract kidney hypoxia and progression of CKD. These findings pave a way to novel therapeutic avenues in CKD.
INSTITUTE
University of Texas Health Science Center McGovern Medical School
A comprehensive plasma metabolomics dataset for a cohort of mouse knockouts within the International Mouse Phenotyping Consortium
STUDY_SUMMARY
Untargeted and targeted metabolomics datasets were acquired for blood plasma samples of 30 mouse knockouts within the International Mouse Phenotyping Consortium (IMPC). http://www.mousephenotype.org/. West Coast Metabolomics Center at UC Davis (https://metabolomics.ucdavis.edu/) conducted the metabolomics analyses.
The Effect of Silicon on Salinity Tolerance and the Associated Metabolomics Profile Changes in Date Palm (part-I)
STUDY_SUMMARY
Silicon has a promising role in the growth and salinity tolerance in plants. While the results obtained from the current study showed that silicon enhanced growth in date palm seedlings, the mechanism behind this observation was also investigated by studying changes occurred in metabolomic profiles triggered by silicon under salinity. The global metabolomic analysis using liquid-chromatography-mass-spectrometry revealed the presence of a number significantly (p ā¤ 0.05) accumulated metabolites in leaves and roots when plants were irrigated with silicon and grown under control and salinity conditions.
INSTITUTE
Sultan Qaboos University
DEPARTMENT
Biology
LAST_NAME
Yaish
FIRST_NAME
Mahmoud
ADDRESS
Sultan Qaboos University, Department of Biology, College of Science
Comparative Metabolomic Profiling of Two Contrasting Date Palm Genotypes under Salinity Stress (part-II)
STUDY_SUMMARY
Since metabolites are the net products of the central dogma of cellular biology, this study was aimed to decipher salinity tolerance depends on the information encoded by the metabolomic profiles of the salt tolerant āUmsilaā and susceptible āZabadā cultivars when grown under salinity. Changes in the metabolomic profiles of the leaf and root tissues were determined using hydrophilic interaction liquid chromatography (HILIC) and reverse phase liquid (RPLC) mass spectrometry.
INSTITUTE
Sultan Qaboos University
LAST_NAME
Yaish
FIRST_NAME
Mahmoud
ADDRESS
Department of Biology, College of Science, Sultan Qaboos University
The gut microbiota plays a central role to modulate the plasma metabolome in response to chronic Angiotensin II infusion (part-I)
STUDY_SUMMARY
Six week old C57BL/6 conventional (mice with gut microbiota; conv, n=6) and germ-free (mice without gut microbiota; GF, n=6) mice were infused with angiotensin-II (AngII) for 4 weeks (400ng.kg-1.min-1; Alzet 1004). In parallel control groups, conv (n=6) and GF (n=6) mice received saline via minipumps. Our primary goal was to identify metabolites which were differentially regulated in conventional mice treated with AngII, but not in GF mice, indicating that these metabolites are microbial in origin. Following minipump implantation, animals were housed singly to prevent cross-contamination of microbiota. At the end of fourth week, feces and blood were collected. Both plasma and feces samples were processed and analyzed by using Liquid Chromatography-Tandem Mass Spectroscopy (LC-MS/MS) for metabolite detection (Metabolon).
The gut microbiota plays a central role to modulate the plasma metabolome in response to chronic Angiotensin II infusion (part-II)
STUDY_SUMMARY
Six week old C57BL/6 conventional (mice with gut microbiota; conv, n=6) and germ-free (mice without gut microbiota; GF, n=6) mice were infused with angiotensin-II (AngII) for 4 weeks (400ng.kg-1.min-1; Alzet 1004). In parallel control groups, conv (n=6) and GF (n=6) mice received saline via minipumps. Our primary goal was to identify metabolites which were differentially regulated in conventional mice treated with AngII, but not in GF mice, indicating that these metabolites are microbial in origin. Following minipump implantation, animals were housed singly to prevent cross-contamination of microbiota. At the end of fourth week, feces and blood were collected. Both plasma and feces samples were processed and analyzed by using Liquid Chromatography-Tandem Mass Spectroscopy (LC-MS/MS) for metabolite detection (Metabolon).
Here, we use global metabolomics to differentiate temporal effects (1 ā 60 d) found in nonhuman primate (NHP) urine small molecule signatures after a 4 Gy total body irradiation.
Here, we use global metabolomics to differentiate temporal effects (1 ā 60 d) found in nonhuman primate (NHP) serum small molecule signatures after a 4 Gy total body irradiation.
Evaluation of computational tools using serial mixtures of human plasma and vegetable juice
STUDY_SUMMARY
Mass spectrometry-based metabolomics is developed rapidly in the past few decades. There are few major vendors for LC-MS platform instruments, for example, Thermo ScientificTM LTQ Orbitrap Velos and Agilent 6510 Q-TOF mass spectrometer were used for metabolomics research. The data acquired cross different platform are rarely compared other than the comparison of the instrument itself on resolution, mass accuracy, sensitivity, dynamic range, scan speed etc., which is largely due to the foundation and principle of the instrument design. Other than this, there are many choice for data preprocessing, i.e., the data acquired from the same platform may have been processed with different feature extraction software tools. The discrepancy for the feature detections with different software will lead to the variation of the down-stream statistics analysis and metabolomics pathway interpretation. In addition, the impact of the LC-MS platform and data preprocessing software tools on the quantitative capabilities is also an interesting topic. In this research, XCMS, mzMine 2.37 and apLCMS are three tools used for the feature extraction of data acquired with Thermo ScientificTM LTQ Orbitrap Velos and Agilent 6510 Q-TOF LC-MS platform by serial dilution experiment. The quantification capability is estimated at the same time based on the linearity, accuracy, and precision
Evaluation of computational tools using serial mixtures of human plasma and vegetable juice (part - II)
STUDY_SUMMARY
Mass spectrometry-based metabolomics is developed rapidly in the past few decades. There are few major vendors for LC-MS platform instruments, for example, Thermo ScientificTM LTQ Orbitrap Velos and Agilent 6510 Q-TOF mass spectrometer were used for metabolomics research. The data acquired cross different platform are rarely compared other than the comparison of the instrument itself on resolution, mass accuracy, sensitivity, dynamic range, scan speed etc., which is largely due to the foundation and principle of the instrument design. Other than this, there are many choice for data preprocessing, i.e., the data acquired from the same platform may have been processed with different feature extraction software tools. The discrepancy for the feature detections with different software will lead to the variation of the down-stream statistics analysis and metabolomics pathway interpretation. In addition, the impact of the LC-MS platform and data preprocessing software tools on the quantitative capabilities is also an interesting topic. In this research, XCMS, mzMine 2.37 and apLCMS are three tools used for the feature extraction of data acquired with Thermo ScientificTM LTQ Orbitrap Velos and Agilent 6510 Q-TOF LC-MS platform by serial dilution experiment. The quantification capability is estimated at the same time based on the linearity, accuracy, and precision.
Variability in metabolomic profiles among unique genotypes of Acropora cervicornis (part-II)
STUDY_SUMMARY
We hypothesized that each of the three genotypes tested would have unique metabolomic profiles. These data increase our basic knowledge of the coral metabolome and represent an important step toward linking genotype, phenotype, and metabolome in reef-building corals.
Physiological and metabolic response of crab megalopae and juveniles to ocean acidification (part-III)
STUDY_SUMMARY
Young crab samples were placed into 1 of 4 treatment groups to understand their metabolic response to ocean acidification and dissolved oxygen content.
Physiological and metabolic response of pteropods to ocean acidification (part IV)
STUDY_SUMMARY
The objective of the study was to examine the physiological and metabolic response of pteropods to ocean acidification treatment. Four treatments were used:high pH, high DO (dissolved oxygen); high pH, low DO; low pH, high DO; low pH, low DO
Comprehensive Profiling by Non-targeted Stable Isotope Tracing Capillary Electrophoresis-Mass Spectrometry
STUDY_SUMMARY
We developed a stable-isotope tracing capillary electrophoresis (CE)-MS metabolomics approach to cover polar metabolites as well as isotopologues in a non-targeted way. An in-house developed software enables high throughput processing of complex multi-dimensional data. The practicability is demonstrated analysing 13C-U-glucose exposed prostate cancer and non-cancer cells.
INSTITUTE
Dalian Institute Of Chemical Physics, Chinese Academy Of Sciences
LAST_NAME
Wang
FIRST_NAME
Zhichao
ADDRESS
457, Zhongshan Road
EMAIL
wangzc05@dicp.ac.cn
PHONE
+86-15998625250
STUDY_COMMENTS
Comprehensive Profiling by Non-targeted Stable Isotope Tracing Capillary Electrophoresis-Mass Spectrometry - A Novel Tool Complementing Metabolomics Analyses of Polar Metabolites
Alterations in fecal metabolic patterns are associated with atrial fibrillation
STUDY_SUMMARY
Little evidence has been reported in characterizing the fecal alterations in metabolic patterns in atrial fibrillation (AF). We include the result of the global alterations that occur in the intestinal microbiota in a cohort of AF patients and matched controls based on a strategy of metabolomic analyses. Our findings characterize the disordered microbial metabolite profiles in AF.
Alterations in serum metabolic patterns are associated with atrial fibrillation
STUDY_SUMMARY
Little evidence has been reported in characterizing the serum alterations in metabolic patterns in atrial fibrillation (AF). We include the result of the global alterations that occur in the intestinal microbiota in a cohort of AF patients and matched controls based on a strategy of metabolomic analyses. Our findings characterize the disordered microbial metabolite profiles in AF.
Timecourse on MCF-7 cells treated with different concentration of doxorubicin
STUDY_SUMMARY
MCF-7 cells treated with 10Ī¼M doxorubicin for 4h followed by subsequent withdrawl of the drug and cultured up to 48h.Doxurubicin-treated cells and control cells were collected at 0,12,24,36,and 48h. Meanwhile,MCF-7 cells continuously exposed to a low dosage of doxorubicin at 0.1Ī¼M for 96h. Doxorubicin-treated cells and control cells were collected at 0,24,48,72 and 96h.
Metabolomics of World Trade Center Exposed New York City Firefighters
STUDY_TYPE
C18 Reversed-Phase Broad Spectrum Metabolomics
STUDY_SUMMARY
Particulate matter (PM) exposure and metabolic syndrome (MetSyn) coexist in both industrialized and developing nations. PM and MetSyn are strong risk factors for chronic obstructive pulmonary disease (COPD) and asthma. After the World Trade Center collapse in 9/11/2001, PM-exposed individuals from the Fire Department of New York City (FDNY) developed a progressively lung disease. This nested case-cohort study is composed of never smoking, WTC exposed firefighters with normal pre-9/11 lung function presenting for subspecialty pulmonary evaluation (SPE) before March 2008. Representative cohort controls with serum drawn within six months of 9/11 (n=100). FEV1 at subspecialty exam defined cases: susceptible World Trade Center Lung Injury (WTC-LI) cases (n=50) had FEV1< lower limit of normal (LLN) and resistant WTC-LI cases with FEV1 ā„107% predicted (n=50). This study will determine the metabolomics profile that differentiates firefighters with WTC-LI, firefighters resistant to WTC-LI, and similarly exposed cohort controls.
INSTITUTE
New York University
DEPARTMENT
School of Medicine
LABORATORY
Laboratory at NYU/Bellevue
LAST_NAME
Nolan
FIRST_NAME
Anna
ADDRESS
Academic office 462 1st Avenue, New Bellevue 16, S 16(Office)/ N 20 (Lab) New York, NY 10016
Deep Metabolomics of a High-Grade Serous Ovarian Cancer Triple Knockout Mouse Model.
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Metabolic alternations were investigated by applying Ultra Performance Liquid Chromatography Mass Spectrometry (UPLC-MS) to serum samples collected from triple knockout (TKO) mice at pre-malignant, early, and advanced stages of HGSC. Samples were analyzed with control mice, which have the same genetic background as TKO mice but develop no tumors. To enhance the selectivity for HGSC-specific metabolite markers, a tumor control group was also included. These were uterine tumor (UT) mice that developed uterine tumors, but no HGSC. All samples were analyzed using reverse phase (RP) and hydrophilic interaction liquid chromatography (HILIC) UPLC-MS analysis in positive and negative ion modes.
Combinatorial metabolic mixtures for encoding abstract digital data
STUDY_TYPE
MALDI MS
STUDY_SUMMARY
We present several kilobyte-scale image datasets stored in synthetic metabolomes, which are decoded with accuracy exceeding 98-99% using multi-mass logistic regression.
INSTITUTE
Brown University
DEPARTMENT
Engineering
LABORATORY
Rosenstein Lab
LAST_NAME
Kennedy
FIRST_NAME
Eamonn
ADDRESS
Barus & Holley room 353, 184 Hope St
EMAIL
eamonn_kennedy@brown.edu
PHONE
7737507192
PUBLICATIONS
E. Kennedy et al. āEncoding information in synthetic metabolomesā Plos One, accepted, 2019
Multi-omics analysis demonstrates unique mode of action of a potent new antimalarial compound, JPC-3210, against Plasmodium falciparum
STUDY_SUMMARY
The increasing incidence of antimalarial drug resistance to the first-line artemisinins, and their combination partner drugs, underpins an urgent need for new antimalarial drugs, ideally with a novel mechanism of action. The recently developed 2-aminomethylphenol, JPC-3210, (MMV 892646) is an erythrocytic schizonticide with potent in vitro antimalarial activity against multidrug-resistant Plasmodium falciparum, low cytotoxicity, potent in vivo efficacy against murine malaria, and favourable preclinical pharmacokinetics, including a lengthy plasma elimination half-life. This study demonstrates the application of a āmulti-omicsā workflow based on high resolution orbitrap mass spectrometry to investigate the impact of JPC-3210 on biochemical pathways within P. falciparum infected red blood cells. Metabolomics and peptidomics analysis revealed a perturbation in hemoglobin metabolism following JPC-3210 exposure. The metabolomics data demonstrated a depletion in short hemoglobin-derived peptides, while peptidomics analysis showed a depletion in longer hemoglobin-derived peptides. In order to further elucidate the mechanism responsible for inhibition of hemoglobin metabolism, we used in vitro Ī²-hematin polymerisation assays and showed JPC-3210 to be an intermediate inhibitor of Ī²-hematin polymerisation, about 10-fold less potent then the quinoline antimalarials. Furthermore, quantitative proteomics analysis showed that JPC-3210 treatment results in a distinct proteomic signature in comparison to other known antimalarials. Whilst JPC-3210 clustered closely with mefloquine in the metabolomics and proteomics analyses, a key differentiating signature for JPC-3210 was the significant enrichment of parasite proteins involved in regulation of translation. In conclusion, multi-omics studies using high resolution mass spectrometry revealed JPC-3210 to possess a unique mechanism of action involving inhibition of hemoglobin digestion, depletion of DNA replication and synthesis proteins, and elevation of regulators of protein translation. Importantly, this mechanism is distinct from currently-used antimalarials, suggesting that JPC-3210 warrants further investigation as a potentially useful new antimalarial agent.
Analysis of short chain phosphatidylcholine (PC) on the Golgi membrane
STUDY_TYPE
Golgi membrane lipids characterization
STUDY_SUMMARY
Studies on vesicle formation by the Coat Protein I (COPI) complex have contributed to a basic understanding of how vesicular transport is initiated. We have identified that short chain lipids promote membrane properties that are conducive for fission. Here we investigated short chain PCs on Golgi membrane. These findings will advance the understanding of how lipid geometry contributes to membrane deformation needed for vesicle fission.
Metabolomic analysis of C2C12 myoblasts induced by the transcriptional factor FOXO1
STUDY_SUMMARY
The transcriptional factor FOXO1 is considered to play roles in the regulation of energy metabolism in various tissues. To determine the metabolic changes occurring due to the activation of FOXO1, we analyzed the metabolic profile of C2C12 myoblasts expressing FOXO1-estrogen receptor fusion protein using CE-TOFMS. In the FOXO1-activated cells, the metabolite levels during glycolysis were higher. In addition, the gene expression of pyruvate dehydrogenase kinase, an enzyme that inhibits glucose utilization, increased. In the FOXO1-activated cells, the metabolite levels of numerous amino acids decreased, with increased gene expression of branched chain amino acid metabolism enzymes. Our results suggest that FOXO1 suppresses glucose utilization and promotes the use of proteins/amino acids as energy sources in muscle cells, potentially during starvation.
INSTITUTE
Kyoto Prefectural University
LAST_NAME
Kamei
FIRST_NAME
Yasutomi
ADDRESS
1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan
Metabolomic analysis of skeletal muscle in young and aged mice
STUDY_SUMMARY
Sarcopenia is the age-induced, progressive loss of skeletal muscle mass and function, which results in poor muscle performance. To better understand changes in skeletal muscles during sarcopenia, we performed a metabolomic analysis of skeletal muscle in young (8-week-old) and aged (28-month-old) mice using CE-TOFMS. Our data shows that the metabolites including glucose and polyamine metabolism were decreased in aged mice compared with young mice. In addition, neurotransmitter levels were higher in aged mice.
INSTITUTE
Kyoto Prefectural University
LAST_NAME
Kamei
FIRST_NAME
Yasutomi
ADDRESS
1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan
Metabolome Profiling of Synechococcus elogatus PCC 11802
STUDY_TYPE
Quantitative Metabolomics
STUDY_SUMMARY
Metabolomics Analysis of a novel freshwater cyanobacterium, Synechococcus elongatus PCC 11802 isolated by us from Powai Lake, Mumbai, India. PCC 11802 cells were grown under ambient and 1% CO2 conditions and metabolomics data was collected in three biological replicates and two technical replicates (n=6). The study aims to find metabolomics changes in this cyanobacterium at elevated CO2 levels.
Child Health and Development Studies womb to breast cancer F0 metabolomics
STUDY_SUMMARY
We used high resolution metabolomics to understand DDT-induced alterations of in utero environment and potential health effects. This study measured endogenous metabolites in 397 maternal perinatal serum samples collected during 1959-1967 in the Child Health and Development Studies (CHDS) and assessed associations between metabolites and envrionmental chemical concentrations in maternal serum.
INSTITUTE
Emory University
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
Correlations between LC-MS/MS-detected Glycomics and NMR-detected Metabolomics in Caenorhabditis (part -I) elegans Development.
STUDY_SUMMARY
This study examines the relationship between glycans, metabolites, and development in C. elegans. Samples of N2 animals were synchronized and grown to five different time points that ranged from L1 to a mixed population of adults, gravid adults, and offspring.
Correlations between LC-MS/MS-detected Glycomics and NMR-detected Metabolomics in Caenorhabditis elegans Development.
STUDY_SUMMARY
This study examines the relationship between glycans, metabolites, and development in C. elegans. Samples of N2 animals were synchronized and grown to five different time points that ranged from L1 to a mixed population of adults, gravid adults, and offspring.
Genetic and metabolic characterization of bioengineered human fatty liver tissue with modified SIRT1 expression
STUDY_SUMMARY
Lipidomics and metabolomics was performed three types of tissue samples to compare human normal liver tissue against human NASH liver and the bioengineered human iPS-derived fatty liver tissue-iKD-SIRT1. The purpose of this study was to show that the global lipidomics profile of iPS-derived fatty liver tissue-iKD-SIRT1 was similar to that of patients with NASH
INSTITUTE
University of Pittsburgh
DEPARTMENT
Department of Pathology
LAST_NAME
Soto-Gutierrez
FIRST_NAME
Alejandro
ADDRESS
200 Lothrop Street, 423 Biomedical Science Tower, Pittsburgh, PA 15261, USA
Untargeted metabolomics on control and compound-treated STHdhQ111 cells and control STHdhQ7 cells
STUDY_SUMMARY
Cells expressing mutant huntingtin were treated in triplicate with serum-free DMEM with vehicle (Q111SST) or serum-free DMEM with one of 14 protective compounds for 24 hours. Wild type cells were also treated with serum-free DMEM with vehicle (Q7SST) as an additional control for 24 hours. We examined the compounds' metabolomic effects on the cells using untargeted mass spectrometry, which measured lipids and polar metabolites.
Effects of selenate and cadmium exposure on the honey bee metabolome
STUDY_SUMMARY
We moved one frame of brood each from five healthy honey bee colonies with marked Italian queens and housed them in a hive body at 35Ā°C and 50% humidity under constant darkness. We then allowed the bees to emerge, mixed the newly emerged workers (NEWs) to randomize their colony of origin and placed NEWs into 13 cm x 10.5 cm x 6.5 cm wire cages equipped with feeders containing 35mL of deionized water and 35mL 50% sucrose. We also provided a pollen patty to each cage of bees consisting of 269g corn syrup, 113g sucrose and 113g of Bee Pro (Mann Lake, Hackensack, MN). To inoculate the newly emerged workers with their ācoreā microbiome, we collected 50 mL of foragers from the source hives of the NEWs, immobilized the bees at 4Ā°C, aseptically dissected out the abdomens and macerated the whole abdomens in 50% sucrose. We added 1 mL of the resulting slurry to 34 mL of 50% sucrose solution and fed it to the NEWs. We allowed the bees to feed ad libitium on the mixture for two days before replacing the feeders with 50% sucrose. We allowed the bees to feed for three more days to fully establish a microbiome. Once the bees had an established microbiome, we prepared treatment feeding solutions of 50% sucrose (as a no metal/metalloid control), 50% sucrose spiked with 0.6 mg/L sodium selenate or 50% sucrose with 0.24 mg/L cadmium chloride (Alfa Aesar, Ward Hill, MA) and pollen patties spiked with either 6.0 mg/L selenium or 0.46 mg/L cadmium as in Hladun et al 2015. We again allowed the bees to feed ad libitium. We sampled three bees from 13 cages after four days of continuous exposure to the above-mentioned treatments and immediately placed the samples on dry ice, followed by long-term storage at -80 Ā°C.
Metabolite profiling across the 1st and 2nd intraerythrocytic developmental lifecycles of the malaria parasite P. falciparum following induction of delayed death with indolmycin treatment
1H NMR spectroscopy-based metabolic profiling of Ophiocordyceps sinensis and Cordyceps militaris of water-boiled and 50% ethanol-soaked extracts
STUDY_TYPE
NMR
STUDY_SUMMARY
Introduction Ophiocordyceps sinensis, a well-known Chinese complementary herb, is a rare and valuable therapeutic resource. Cordyceps militaris (C. militaris) is a commonly used substitute for O. sinensis. A metabolomic-based approach for exploring the similarities and differences in the metabolites of O. sinensis and C. militaris in water-boiled and 50% ethanol-soaked extracts is of great significance. Objectives To determine a vital role of extraction methodologies in influencing the metabolic composition of herbs, 1HNMR-based profiling was used to characterize the metabolic fingerprints of O. sinensis and C. militaris. Methods To make a distinction between the global metabolite profiling of O. sinensis and C. militaris extracts obtained from either the water-boiled or 50% ethanol-soaked methods, we screened the herbs samples using 1HNMR-based metabolic fingerprints combined with multivariate statistical analysis. Results This study revealed that a total of 43 (82.69% of 52) metabolites were detectable in both O. sinensis and C. militaris. According to the variable importance in projection (VIP) value and p-value from the Mann-Whitney test, 7 metabolites (alanine, aspartate, glutamate, mannitol, ornithine, serine, and trehalose) differed between O. sinensis and C. militaris. Arginine, glucose, putrescine, pyroglutamate, betaine, O-phosphocholine, and xylose differed significantly between the water-boiled and 50% ethanol-soaked methods used to prepare the herb extracts. Conclusion A total of 52 primary metabolites were identified and quantified from O. sinensis and C. militaris samples. The study suggests that a water-boiled extraction is much faster method and strongly recommended over the 50% ethanol-soaked method for both O. sinensis and C. militaris.
Sepsis-related metabolic changes in ileum, jejunum, skeletal muscle, liver and lung
STUDY_SUMMARY
Rationale: Sepsis is a multi-organ disease affecting the ileum and jejunum (small intestine),liver, skeletal muscle, and lung clinically. Recently, specific alterations in circulating metabolites have been found in patients with sepsis which are thought to contribute to the pathogenesis of disease. The specific metabolic changes in the ileum, jejunum, liver, skeletal muscle, and lung have not previously been investigated. Methods: Live Pseudomonas aeruginosa isolated from a patient was given via IV catheter to pigs to induce severe sepsis. Eighteen hours later, ileum, jejunum, medial gastrocnemius skeletal muscle, liver, and lung were harvested and flash frozen. Tissues were subsequently processed for non-targeted metabolomics analysis using gas chromatography/mass spectrometry. Results: After 18 hours of sepsis, the ileum and the liver demonstrated significant changes in metabolites involved in linoleic acid metabolism, the ileum and lung had significant changes in valine/leucine/isoleucine metabolism, the jejunum, skeletal muscle, and liver had significant changes in arginine/ proline metabolism, and the skeletal muscle and lung had significant changes in aminoacyl-tRNA biosynthesis by pathway analysis. Pathway analysis also identified changes in metabolic pathways unique for different tissues, including changes in the citric acid cycle (jejunum), beta-alanine metabolism (skeletal muscle), and purine metabolism (liver). Conclusion: These findings demonstrate both overlapping metabolic pathways affected in different tissues and those that are unique to others and provide insight into the metabolic changes in sepsis leading to organ dysfunction. This may allow therapeutic interventions that focus on multiple tissues or single tissues once the relationship of the altered metabolites/metabolism to the underlying pathogenesis of sepsis is determined.
INSTITUTE
Indiana University School of Medicine
DEPARTMENT
Indiana Center for Musculoskeletal Health / Dept. of Pathology & Laboratory Medicine
A library of human gut bacterial isolates paired with longitudinal multiomics data enables mechanistic microbiome research
STUDY_TYPE
Stool metabolite profiling
STUDY_SUMMARY
Fecal microbiota transplantation (FMT) is used in the treatment of microbiome-associated diseases such as Clostridium difficile infections. In order to develop synthetic therapeutics and customized disease treatments we will need to understand the bacterial communities in the stool samples used in such treatments. For this purpose, a microbiome library was generated using human stool obtained from healthy human FMT recruited by OpenBiome, a non-profit organization that provides fecal microbiome therapeutics. In addition to characterizing the bacterial populations and obtaining bacterial isolates from FMT samples, we conducted metabolite profiling with the goal of: (1) generating a library of metabolites in FMT samples, (2) Identifying metabolites associated with defined bacterial populations, and (3) identifying microbial metabolites with immunoregulatory functions. We conducted metabolite profiling on a subset consisting of 180 stool samples from 84 donors using four nontargeted liquid chromatography mass spectrometry (LC-MS) methods. Generated data were processed, isotopes removed, and adducts and fragments clustered. The identity of known metabolites was determined based on matching retention times of neat standards run in parallel with the study.
MARBLES (Markers of Autism Risk in Babies: Learning Early Sign) Enriched Cohort Study: Internal Metabolomic Biomarker Exposome and Development Disorders (IMBEDD)
STUDY_SUMMARY
This project aims to identify internal biomarkers of drug, food and microbial exposures associated to Autism Spectrum Disorder (ASD) and neurodevelopmental outcomes in an enriched-risk cohort. Using targeted and untargeted internal exposome approaches to identify exposures in maternal blood and child cord blood, internal metabolomics biomarkers will be associated with related exposures and also associated with ASD.
Flavonoid study of Ginkgo leaves facing to different elevation and plant age
STUDY_SUMMARY
Ginkgo biloba leaves are always resources for flavonoids pharmaceutical industry. Thus, artificial planting and industrial harvesting become the vital aspect to get higher drug yields. In this research, we performed de novo transcriptome sequencing of Ginkgo leaves coupled with high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry analyses to obtain a comprehensive understanding of the influence of elevation and plant age on flavonoid synthesis. A total of 557,659,530 clean reads were assembled into 188,155 unigenes, of which 135,102 (71.80%) were successfully annotated in seven public databases. The differentially expressed genes analysis indicated DFR, LAR and ANR were significantly up-regulated with the increase of elevation in young Ginkgo trees leaves. With less strict saliency, the relative concentration of flavonoid derivatives with high parent ion signal intensity was likely to support this conclusion. Complex gene variations were observed with the plant age change. However, flavonoid derivatives analysis predicted the potential possibility that the rise of plant age is more likely to be detrimental to the biosynthesis of Ginkgo flavonoids in leaves. From the overall DEGs involved in flavonoid biosynthesis, DFRs seemed to show more considerable variability towards the variation of elevation and plant age. Furthermore, our research effectively expanded the functional genomic library of Ginkgo and provided a reference for artificial planting and industrial harvesting.
INSTITUTE
Central South University, China
LAST_NAME
Zou
FIRST_NAME
Kai
ADDRESS
Central South University, 932 Lushan South Road, Yuelu District, Changsha City, Hunan Province
Non-targeted GC-MS Analysis of Polar Soluble Fraction (part-I)
STUDY_SUMMARY
Cyanobacteria are a model photoautotroph and a chassis for the sustainable production of fuels and chemicals. Yet, knowledge of photoautotrophic metabolism in the natural environment of day/night cycles is lacking yet has implications for improved yield from plants, algae, and cyanobacteria. Here, a thorough approach to characterizing diverse metabolitesāincluding carbohydrates, lipids, amino acids, pigments, co-factors, nucleic acids and polysaccharidesāin the model cyanobacterium Synechocystis sp. PCC 6803 (S. 6803) under sinusoidal diurnal light-dark cycles was developed and applied. A custom photobioreactor and novel multi-platform mass spectrometry workflow enabled metabolite profiling every 30-120 minutes across a 24-hour diurnal sinusoidal LD (āsinLDā) cycle peaking at 1,600 mol photons m 2 s-1. We report widespread oscillations across the sinLD cycle with 90%, 94%, and 40% of the identified polar/semi-polar, non-polar, and polymeric metabolites displaying statistically significant oscillations, respectively. Microbial growth displayed distinct lag, biomass accumulation, and cell division phases of growth. During the lag phase, amino acids (AA) and nucleic acids (NA) accumulated to high levels per cell followed by decreased levels during the biomass accumulation phase, presumably due to protein and DNA synthesis. Insoluble carbohydrates displayed sharp oscillations per cell at the day-to-night transition. Potential bottlenecks in central carbon metabolism are highlighted. Together, this report provides a comprehensive view of photosynthetic metabolite behavior with high temporal resolution, offering insight into the impact of growth synchronization to light cycles via circadian rhythms. Incorporation into computational modeling and metabolic engineering efforts promises to improve industrially-relevant strain design.
Non-targeted GC-MS Analysis of Insoluble Metabolites (part-II)
STUDY_SUMMARY
Cyanobacteria are a model photoautotroph and a chassis for the sustainable production of fuels and chemicals. Yet, knowledge of photoautotrophic metabolism in the natural environment of day/night cycles is lacking yet has implications for improved yield from plants, algae, and cyanobacteria. Here, a thorough approach to characterizing diverse metabolitesāincluding carbohydrates, lipids, amino acids, pigments, co-factors, nucleic acids and polysaccharidesāin the model cyanobacterium Synechocystis sp. PCC 6803 (S. 6803) under sinusoidal diurnal light-dark cycles was developed and applied. A custom photobioreactor and novel multi-platform mass spectrometry workflow enabled metabolite profiling every 30-120 minutes across a 24-hour diurnal sinusoidal LD (āsinLDā) cycle peaking at 1,600 ?mol photons m 2 s-1. We report widespread oscillations across the sinLD cycle with 90%, 94%, and 40% of the identified polar/semi-polar, non-polar, and polymeric metabolites displaying statistically significant oscillations, respectively. Microbial growth displayed distinct lag, biomass accumulation, and cell division phases of growth. During the lag phase, amino acids (AA) and nucleic acids (NA) accumulated to high levels per cell followed by decreased levels during the biomass accumulation phase, presumably due to protein and DNA synthesis. Insoluble carbohydrates displayed sharp oscillations per cell at the day-to-night transition. Potential bottlenecks in central carbon metabolism are highlighted. Together, this report provides a comprehensive view of photosynthetic metabolite behavior with high temporal resolution, offering insight into the impact of growth synchronization to light cycles via circadian rhythms. Incorporation into computational modeling and metabolic engineering efforts promises to improve industrially-relevant strain design.
GC-MS Analysis of Insoluble/Polymeric Amino Acids (part-III)
STUDY_SUMMARY
Cyanobacteria are a model photoautotroph and a chassis for the sustainable production of fuels and chemicals. Yet, knowledge of photoautotrophic metabolism in the natural environment of day/night cycles is lacking yet has implications for improved yield from plants, algae, and cyanobacteria. Here, a thorough approach to characterizing diverse metabolitesāincluding carbohydrates, lipids, amino acids, pigments, co-factors, nucleic acids and polysaccharidesāin the model cyanobacterium Synechocystis sp. PCC 6803 (S. 6803) under sinusoidal diurnal light-dark cycles was developed and applied. A custom photobioreactor and novel multi-platform mass spectrometry workflow enabled metabolite profiling every 30-120 minutes across a 24-hour diurnal sinusoidal LD (āsinLDā) cycle peaking at 1,600 ļmol photons m 2 s-1. We report widespread oscillations across the sinLD cycle with 90%, 94%, and 40% of the identified polar/semi-polar, non-polar, and polymeric metabolites displaying statistically significant oscillations, respectively. Microbial growth displayed distinct lag, biomass accumulation, and cell division phases of growth. During the lag phase, amino acids (AA) and nucleic acids (NA) accumulated to high levels per cell followed by decreased levels during the biomass accumulation phase, presumably due to protein and DNA synthesis. Insoluble carbohydrates displayed sharp oscillations per cell at the day-to-night transition. Potential bottlenecks in central carbon metabolism are highlighted. Together, this report provides a comprehensive view of photosynthetic metabolite behavior with high temporal resolution, offering insight into the impact of growth synchronization to light cycles via circadian rhythms. Incorporation into computational modeling and metabolic engineering efforts promises to improve industrially-relevant strain design.
Targeted LC-MS/MS Analysis of Soluble Metabolites in the MeOH:H2O Phase (part-IV)
STUDY_SUMMARY
Cyanobacteria are a model photoautotroph and a chassis for the sustainable production of fuels and chemicals. Yet, knowledge of photoautotrophic metabolism in the natural environment of day/night cycles is lacking yet has implications for improved yield from plants, algae, and cyanobacteria. Here, a thorough approach to characterizing diverse metabolitesāincluding carbohydrates, lipids, amino acids, pigments, co-factors, nucleic acids and polysaccharidesāin the model cyanobacterium Synechocystis sp. PCC 6803 (S. 6803) under sinusoidal diurnal light-dark cycles was developed and applied. A custom photobioreactor and novel multi-platform mass spectrometry workflow enabled metabolite profiling every 30-120 minutes across a 24-hour diurnal sinusoidal LD (āsinLDā) cycle peaking at 1,600 ļmol photons m 2 s-1. We report widespread oscillations across the sinLD cycle with 90%, 94%, and 40% of the identified polar/semi-polar, non-polar, and polymeric metabolites displaying statistically significant oscillations, respectively. Microbial growth displayed distinct lag, biomass accumulation, and cell division phases of growth. During the lag phase, amino acids (AA) and nucleic acids (NA) accumulated to high levels per cell followed by decreased levels during the biomass accumulation phase, presumably due to protein and DNA synthesis. Insoluble carbohydrates displayed sharp oscillations per cell at the day-to-night transition. Potential bottlenecks in central carbon metabolism are highlighted. Together, this report provides a comprehensive view of photosynthetic metabolite behavior with high temporal resolution, offering insight into the impact of growth synchronization to light cycles via circadian rhythms. Incorporation into computational modeling and metabolic engineering efforts promises to improve industrially-relevant strain design.
Non-targeted LC-MS Analysis of Soluble Metabolites in the Non-Polar MTBE Phase (part-V)
STUDY_SUMMARY
Cyanobacteria are a model photoautotroph and a chassis for the sustainable production of fuels and chemicals. Yet, knowledge of photoautotrophic metabolism in the natural environment of day/night cycles is lacking yet has implications for improved yield from plants, algae, and cyanobacteria. Here, a thorough approach to characterizing diverse metabolitesāincluding carbohydrates, lipids, amino acids, pigments, co-factors, nucleic acids and polysaccharidesāin the model cyanobacterium Synechocystis sp. PCC 6803 (S. 6803) under sinusoidal diurnal light-dark cycles was developed and applied. A custom photobioreactor and novel multi-platform mass spectrometry workflow enabled metabolite profiling every 30-120 minutes across a 24-hour diurnal sinusoidal LD (āsinLDā) cycle peaking at 1,600 ļmol photons m 2 s-1. We report widespread oscillations across the sinLD cycle with 90%, 94%, and 40% of the identified polar/semi-polar, non-polar, and polymeric metabolites displaying statistically significant oscillations, respectively. Microbial growth displayed distinct lag, biomass accumulation, and cell division phases of growth. During the lag phase, amino acids (AA) and nucleic acids (NA) accumulated to high levels per cell followed by decreased levels during the biomass accumulation phase, presumably due to protein and DNA synthesis. Insoluble carbohydrates displayed sharp oscillations per cell at the day-to-night transition. Potential bottlenecks in central carbon metabolism are highlighted. Together, this report provides a comprehensive view of photosynthetic metabolite behavior with high temporal resolution, offering insight into the impact of growth synchronization to light cycles via circadian rhythms. Incorporation into computational modeling and metabolic engineering efforts promises to improve industrially-relevant strain design.
Biological Responses to Tobacco Smoke Exposure in III Children: Inflammatory Processes and the Oral Microbiome
STUDY_SUMMARY
This project evaluates the biological response to overall tobacco smoke (OTS) exposure among pediatric emergency patients enrolled in a randomized-controlled intervention trial aimed at reducing secondhand smoke exposure. The effects of OTS as measured by salivary continine on salivary metabolic profiles are measured by untargeted high resolution metabolomics, comparing higher and lower levels of OTS.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Uppal
FIRST_NAME
Karan
ADDRESS
NA
EMAIL
kuppal2@emory.edu
PHONE
(404) 727 5027
TOTAL_SUBJECTS
284
STUDY_COMMENTS
Both CHEAR pooled saliva samples and Clinical Biomarker Laboratory pooled plasma samples were used
Peroxide antimalarial treatment timecourse on trophozoite-stage P. falciparum parasites
STUDY_SUMMARY
Red blood cells (RBCs) infected with trophozoite stage P. falciparum parasites (3D7 strain) at 10% parasitaemia and 2% haematocrit were treated with OZ277 (300 nM), OZ439 (300 nM), DHA (100 nM) or vehicle (0.03% DMSO). This was a 4-timepoint study, with samples taken 0, 0.5, 1.5 and 3 h after drug or vehicle addition. Samples treated with vehicle acted as the untreated control. Samples from drug treated uninfected RBCs were also taken to ensure the observed drug effects were parasite specific.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Peroxide antimalarial treatment timecourse on ring-stage P. falciparum parasites
STUDY_SUMMARY
Red blood cells (RBCs) infected with ring stage P. falciparum parasites (3D7 strain) at 10% parasitaemia and 2% haematocrit were treated with OZ277 (1 uM), OZ439 (1 uM), DHA (300 nM) or vehicle (0.03% DMSO). This was a 5-timepoint study, with samples taken 0, 1.5, 3, 6 and 9 h after drug or vehicle addition. Samples treated with vehicle acted as the untreated control. Samples from drug treated uninfected RBCs were also taken to ensure the observed drug effects were parasite specific.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Peroxide antimalarial extended treatment timecourse on trophozoite-stage P. falciparum parasites
STUDY_SUMMARY
Red blood cells (RBCs) infected with trophozoite stage P. falciparum parasites (3D7 strain) at 10% parasitaemia and 2% haematocrit were treated with OZ277 (300 nM), OZ439 (300 nM), DHA (100 nM) or vehicle (0.03% DMSO). This was a 4-timepoint study, with samples taken 0, 3, 6 and 9 h after drug or vehicle addition. Samples treated with vehicle acted as the untreated control. Samples from drug treated uninfected RBCs were also taken to ensure the observed drug effects were parasite specific.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Peroxide antimalarial treatment of K13-mutant and -wildtype P. falciparum parasites
STUDY_SUMMARY
Red blood cells (RBCs) infected with trophozoite stage P. falciparum parasites (Cam3.IIR539T or Cam3.IIrev lines) at 4% parasitaemia and 2% haematocrit were treated with 100 nM of DHA, OZ277 or OZ439 for a duration of 1, 3 and 5 h, respectively. The K13-mutant artemisinin resistant parasite line used was Cam3.IIR539T. The K13-wildtype artemisinin sensitive parasite line used was Cam3.IIrev. The Samples treated with vehicle (DMSO) acted as the untreated control.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Lipidomics in the serum of cold exposed mice treated with 12-LOX inhibitor LOXBlock-1
STUDY_SUMMARY
We aimed to investigate whether the cold-induced release of 12-LOX products into the circulation were dependent on 12-LOX activation. We pre-treated C57BL6/J mice with the pharmacological inhibitor LOXBlock-1 or its vehicle (DMSO), and after 15 minutes we placed them under cold temperature (5C) for 4 hours. A control group was injected with DMSO and kept at room temperature for the same 4 hours. After this period of time we, collected the blood, and obtained the serum fraction that was immediately frozen and submitted for untargeted lipidomics.
Characterization of feces in Atrial Fibrillation (AF) patients
STUDY_SUMMARY
Atrial Fibrillation (AF), an abnormal heart rhythm characterized by the rapid and irregular beating of the atria, is the most common arrhythmia with heavy global burdens. The present project aimed to characterized the feature of metabolites in feces of AF patients by using LC-MS.
MARBLES (Markers of Autism Risk in Babies: Learning Early Sign) Enriched Cohort Study:Internal Metabolomic Biomarker Exposome and Development Disorders (IMBEDD)
STUDY_SUMMARY
This project aims to identify internal biomarkers of drug, food and microbial exposures associated to Autism Spectrum Disorder (ASD) and neurodevelopmental outcomes in an enriched-risk cohort. Using targeted and untargeted internal exposome approaches to identify exposures in maternal blood and child cord blood, internal metabolomics biomarkers will be associated with related exposures and also associated with ASD.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Uppal
FIRST_NAME
Karan
ADDRESS
NA
EMAIL
kuppal2@emory.edu
PHONE
(404) 727 5027
TOTAL_SUBJECTS
269
STUDY_COMMENTS
Both CHEAR pooled plasma samples and Clinical Biomarker Laboratory pooled plasma samples were used
Metabolomic Markers of Methotrexate Response, In Vitro
STUDY_SUMMARY
Erythroblastoid cells (K562) maintained in logarithmic growth phase were treated with 1000 nM methotrexate or vehicle alone (i.e. D-PBS) under standard culture conditions for 24 hours. Cellular response to methotrexate was measured based on anti-proliferative activity by cell counting. Cells were washed, flash frozen in liquid nitrogen and submitted for metabolomics analysis to the NIH West Coast Metabolomics Center.
Fish-oil supplementation in pregnancy, child metabolomics and asthma risk
STUDY_SUMMARY
We investigated potential metabolic mechanisms using untargeted liquid chromatography-mass spectrometry-based metabolomics on 577 plasma samples collected at age 6 months in the offspring of mothers participating in the n-3 LCPUFA randomized controlled trial. First, associations between the n-3 LCPUFA supplementation groups and child metabolite levels were investigated using univariate regression models and data-driven partial least square discriminant analyses (PLS-DA). Second, we analyzed the association between the n-3 LCPUFA metabolomic profile and asthma development using Cox-regression. Third, we conducted mediation analyses to investigate whether the protective effect of n-3 LCPUFA on asthma was mediated via the metabolome
INSTITUTE
Copenhagen Prospective Studies on Asthma in Childhood, Herlev and Gentofte Hospital, University of Copenhagen
DEPARTMENT
Copenhagen Prospective Studies on Asthma in Childhood, Herlev and Gentofte Hospital
Serum lipidomic profile of cold-exposed Ucp1cre/12-LOX KO mice
STUDY_SUMMARY
We aimed to evaluate whether specific deletion of 12-Lipoxygenase (12-LOX) in brown fat can affect the serum concentrations of 12-LOX products under cold exposure.
We aimed to determine the levels of the cold-induced 12-LOX products in patients with different degrees of body mass index (BMI). This analysis allowed us to infer about the role of these oxylipins in the pathophysiology of obesity.
Vitamin D regulates the microbiota to induce RORgt/FoxP3+ regulatory T cells
STUDY_SUMMARY
The active form of vitamin D (1,25(OH)2D) suppresses experimental models of inflammatory bowel disease in part by regulating the microbiota. In this study, the role of vitamin D in the regulation of microbe induced RORgt/FoxP3+ T regulatory (reg) cells in the colon was determined. Vitamin D sufficient (D+) mice had significantly higher frequencies of FoxP3+ and RORgt/FoxP3+ T reg cells in the colon compared to vitamin D deficient (D-) mice. The higher frequency of RORgt/FoxP3+ T reg cells in D+ colon correlated with higher numbers of bacteria from the Clostridium XIVa and Bacteroides in D+ compared to D- cecum. D- mice with fewer RORgt/FoxP3+ T reg cells were significantly more susceptible to colitis than D+ mice. Transfer of the cecal bacteria from D+ or D- mice to germfree recipients phenocopied the higher numbers of RORgt/FoxP3+ cells and reduced susceptibility to colitis in D+ versus D- recipient mice. 1,25(OH)2D treatment of the D- mice beginning at 3 weeks of age did not completely recover RORgt/FoxP3+ T reg cells or the Bacteriodes, Bacteriodes thetaiotaomicron, and Clostridium XIVa numbers to D+ values. Early vitamin D status shapes the microbiota to optimize the population of colonic RORgt/FoxP3+ T reg cells important for resistance to colitis.
INSTITUTE
The Pennsylvania State University (Penn State)
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
917 Old Boalsburg Road, State College, Pennsylvania, 16801, USA
Effects of selenate and cadmium exposure on the honey bee metabolome (part-II)
STUDY_SUMMARY
We moved one frame of brood each from five healthy honey bee colonies with marked Italian queens and housed them in a hive body at 35Ā°C and 50% humidity under constant darkness. We then allowed the bees to emerge, mixed the newly emerged workers (NEWs) to randomize their colony of origin and placed NEWs into 13 cm x 10.5 cm x 6.5 cm wire cages equipped with feeders containing 35mL of deionized water and 35mL 50% sucrose. We also provided a pollen patty to each cage of bees consisting of 269g corn syrup, 113g sucrose and 113g of Bee Pro (Mann Lake, Hackensack, MN). To inoculate the newly emerged workers with their ācoreā microbiome, we collected 50 mL of foragers from the source hives of the NEWs, immobilized the bees at 4Ā°C, aseptically dissected out the abdomens and macerated the whole abdomens in 50% sucrose. We added 1 mL of the resulting slurry to 34 mL of 50% sucrose solution and fed it to the NEWs. We allowed the bees to feed ad libitium on the mixture for two days before replacing the feeders with 50% sucrose. We allowed the bees to feed for three more days to fully establish a microbiome. Once the bees had an established microbiome, we prepared treatment feeding solutions of 50% sucrose (as a no metal/metalloid control), 50% sucrose spiked with 0.6 mg/L sodium selenate or 50% sucrose with 0.24 mg/L cadmium chloride (Alfa Aesar, Ward Hill, MA) and pollen patties spiked with either 6.0 mg/L selenium or 0.46 mg/L cadmium as in Hladun et al 2015. We again allowed the bees to feed ad libitium. We sampled three bees from 13 cages after four days of continuous exposure to the above-mentioned treatments and immediately placed the samples on dry ice, followed by long-term storage at -80 Ā°C.
Lyme disease is a tick-borne bacterial illness that occurs in areas of North America, Europe, and Asia. Early infection typically presents as generalized symptoms with an erythema migrans (EM) skin lesion. Bacterial dissemination can result in multiple EM skin lesions or in extracutaneous manifestations such as Lyme neuroborreliosis. Metabolic biosignatures of patients with early Lyme disease can potentially provide diagnostic targets, as well as highlight metabolic pathways that contribute to pathogenesis. Sera from well-characterized patients diagnosed with either early localized Lyme disease (ELL) or early disseminated Lyme disease (EDL), plus healthy individuals (HC), from the United States were analyzed by liquid chromatography-mass spectrometry (LC-MS). Comparative analyses were performed between ELL, or EDL, or ELL combined with EDL, and the HC to develop biosignatures present in early Lyme disease. A direct comparison between ELL and EDL was also performed to develop a biosignature for stages of early Lyme disease. Metabolic pathway analysis and chemical identification of metabolites with LC-tandem mass spectrometry (LC-MS/MS) demonstrated alterations of eicosanoid, bile acid, sphingolipid, glycerophospholipid, and acylcarnitine metabolic pathways during early Lyme disease . These metabolic alterations were confirmed using a separate set of serum samples for validation. The findings demonstrated the metabolic pathways altered in the host during early Lyme disease and provide evidence that the diversity in the type of early Lyme disease manifestations may be associated with particular metabolic alterations.
INSTITUTE
Colorado State University
DEPARTMENT
MIP
LABORATORY
Belisle
LAST_NAME
Belisle
FIRST_NAME
John
ADDRESS
200 West Lake, Campus Delivery 0922, Colorado State University, Fort Collins, CO, 80523
Vaginal swab lipidome profiles at 48 h reflect the fat composition of neonatal diet during first two days postnatal
STUDY_TYPE
MRM-profiling
STUDY_SUMMARY
In this study, we further investigated the efficacy of using MRM-profiling of vaginal lipids to differentiate PND 2 vaginal swabs between gilts suckled by sow or fed milk replacer. Secondly, we tested the effect of a lard based supplement on vaginal lipid profiles of gilts.
INSTITUTE
Purdue University
DEPARTMENT
Animal Sciences
LABORATORY
Metabolite Profiling Facility - Purdue University
LAST_NAME
Ferreira
FIRST_NAME
Christina
ADDRESS
1203 W. State St, West Lafayette, IN, 47906, USA
EMAIL
cferrei@purdue.edu
PHONE
7654095924
NUM_GROUPS
colostrum suckled (S, n=8); S plus fat supplement (SF, n=5); bottle-fed milk replacer (B, n=8); or B plus fat supplement (BF, n=7
We propose to evaluate microbial and metabolic profiles in baseline and endpoint colonic mucosal, fecal, and serum samples from human patients at risk for CRC and enrolled in a 90-day phase I clinical trial. Patients will receive daily supplementation with calcium alone, a calcium-rich multimineral (Aquamin?), or placebo (maltodextrin) (n=10 per group). We hypothesize that dietary supplementation will correlate with CRC-protective metabolic profiles and that multimineral supplementation will generate more favorable profiles than calcium supplementation alone.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
NUM_GROUPS
24
TOTAL_SUBJECTS
18
STUDY_COMMENTS
Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancer related death when both genders are combined. Epidemiologically, calcium intake has been protective against colonic adenomas and even colon cancer. Calcium supplementation has reduced the risk of colon adenoma formation in subjects with a history of previous colon polyps. The utility of calcium supplementation for colon cancer prevention is somewhat modulated by the modest or inconsistent level of protection afforded. Our preliminary data in mice and human enterocyte models shows that dietary supplementation with a multimineral supplement (Aquamin?) containing calcium in combination with 72 measureable trace minerals is more protective against tumors and epithelial growth dysregulation than calcium alone. One potential mechanism, supported by our rodent data, is that multimineral supplementation alters gut microbial populations to generate bile acid and short chain fatty acid (SCFA) profiles that are CRC-protective.
We propose to evaluate microbial and metabolic profiles in baseline and endpoint colonic mucosal, fecal, and serum samples from human patients at risk for CRC and enrolled in a 90-day phase I clinical trial. Patients will receive daily supplementation with calcium alone, a calcium-rich multimineral (Aquamin?), or placebo (maltodextrin) (n=10 per group). We hypothesize that dietary supplementation will correlate with CRC-protective metabolic profiles and that multimineral supplementation will generate more favorable profiles than calcium supplementation alone.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
NUM_GROUPS
24
TOTAL_SUBJECTS
18
STUDY_COMMENTS
Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancer related death when both genders are combined. Epidemiologically, calcium intake has been protective against colonic adenomas and even colon cancer. Calcium supplementation has reduced the risk of colon adenoma formation in subjects with a history of previous colon polyps. The utility of calcium supplementation for colon cancer prevention is somewhat modulated by the modest or inconsistent level of protection afforded. Our preliminary data in mice and human enterocyte models shows that dietary supplementation with a multimineral supplement (Aquamin?) containing calcium in combination with 72 measureable trace minerals is more protective against tumors and epithelial growth dysregulation than calcium alone. One potential mechanism, supported by our rodent data, is that multimineral supplementation alters gut microbial populations to generate bile acid and short chain fatty acid (SCFA) profiles that are CRC-protective.
We propose to evaluate microbial and metabolic profiles in baseline and endpoint colonic mucosal, fecal, and serum samples from human patients at risk for CRC and enrolled in a 90-day phase I clinical trial. Patients will receive daily supplementation with calcium alone, a calcium-rich multimineral (Aquamin?), or placebo (maltodextrin) (n=10 per group). We hypothesize that dietary supplementation will correlate with CRC-protective metabolic profiles and that multimineral supplementation will generate more favorable profiles than calcium supplementation alone.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
NUM_GROUPS
24
TOTAL_SUBJECTS
18
STUDY_COMMENTS
Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancer related death when both genders are combined. Epidemiologically, calcium intake has been protective against colonic adenomas and even colon cancer. Calcium supplementation has reduced the risk of colon adenoma formation in subjects with a history of previous colon polyps. The utility of calcium supplementation for colon cancer prevention is somewhat modulated by the modest or inconsistent level of protection afforded. Our preliminary data in mice and human enterocyte models shows that dietary supplementation with a multimineral supplement (Aquamin?) containing calcium in combination with 72 measureable trace minerals is more protective against tumors and epithelial growth dysregulation than calcium alone. One potential mechanism, supported by our rodent data, is that multimineral supplementation alters gut microbial populations to generate bile acid and short chain fatty acid (SCFA) profiles that are CRC-protective.
We propose to evaluate microbial and metabolic profiles in baseline and endpoint colonic mucosal, fecal, and serum samples from human patients at risk for CRC and enrolled in a 90-day phase I clinical trial. Patients will receive daily supplementation with calcium alone, a calcium-rich multimineral (Aquamin?), or placebo (maltodextrin) (n=10 per group). We hypothesize that dietary supplementation will correlate with CRC-protective metabolic profiles and that multimineral supplementation will generate more favorable profiles than calcium supplementation alone.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
NUM_GROUPS
24
TOTAL_SUBJECTS
18
STUDY_COMMENTS
Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancer related death when both genders are combined. Epidemiologically, calcium intake has been protective against colonic adenomas and even colon cancer. Calcium supplementation has reduced the risk of colon adenoma formation in subjects with a history of previous colon polyps. The utility of calcium supplementation for colon cancer prevention is somewhat modulated by the modest or inconsistent level of protection afforded. Our preliminary data in mice and human enterocyte models shows that dietary supplementation with a multimineral supplement (Aquamin?) containing calcium in combination with 72 measureable trace minerals is more protective against tumors and epithelial growth dysregulation than calcium alone. One potential mechanism, supported by our rodent data, is that multimineral supplementation alters gut microbial populations to generate bile acid and short chain fatty acid (SCFA) profiles that are CRC-protective.
We propose to evaluate microbial and metabolic profiles in baseline and endpoint colonic mucosal, fecal, and serum samples from human patients at risk for CRC and enrolled in a 90-day phase I clinical trial. Patients will receive daily supplementation with calcium alone, a calcium-rich multimineral (Aquamin?), or placebo (maltodextrin) (n=10 per group). We hypothesize that dietary supplementation will correlate with CRC-protective metabolic profiles and that multimineral supplementation will generate more favorable profiles than calcium supplementation alone.
We propose to evaluate microbial and metabolic profiles in baseline and endpoint colonic mucosal, fecal, and serum samples from human patients at risk for CRC and enrolled in a 90-day phase I clinical trial. Patients will receive daily supplementation with calcium alone, a calcium-rich multimineral (Aquamin?), or placebo (maltodextrin) (n=10 per group). We hypothesize that dietary supplementation will correlate with CRC-protective metabolic profiles and that multimineral supplementation will generate more favorable profiles than calcium supplementation alone.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
NUM_GROUPS
24
TOTAL_SUBJECTS
18
STUDY_COMMENTS
Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancer related death when both genders are combined. Epidemiologically, calcium intake has been protective against colonic adenomas and even colon cancer. Calcium supplementation has reduced the risk of colon adenoma formation in subjects with a history of previous colon polyps. The utility of calcium supplementation for colon cancer prevention is somewhat modulated by the modest or inconsistent level of protection afforded. Our preliminary data in mice and human enterocyte models shows that dietary supplementation with a multimineral supplement (Aquamin?) containing calcium in combination with 72 measureable trace minerals is more protective against tumors and epithelial growth dysregulation than calcium alone. One potential mechanism, supported by our rodent data, is that multimineral supplementation alters gut microbial populations to generate bile acid and short chain fatty acid (SCFA) profiles that are CRC-protective.
Plasma untargeted metabolomics study of pulmonary tuberculosis
STUDY_SUMMARY
In this study, differentially abundant plasma metabolites were screened by using the ultra-high performance liquid chromatography coupled with Q Exactive mass spectrometry in pulmonary tuberculosis(TB) patients and normal controls(NC) or patients with other pulmonary diseases such as, community-acquired pneumonia(CAP) and lung cancer(LC).
INSTITUTE
Zhengjiang University
DEPARTMENT
School of Medicine
LABORATORY
Institute of Cell Biology
LAST_NAME
Li
FIRST_NAME
Ji-Cheng
ADDRESS
866 Yuhangtang Road, West Lake District, Hangzhou, Zhejiang Province, 310058, PR China
The effect of weaning stress, sex and temperament on fecal microbiota and metabolites in Brahman calves.
STUDY_SUMMARY
The objective of this study was to conduct a cross-sectional study to 1) investigate the effect of weaning on the fecal microbiota as well as serum metabolites in Bos indicus (Brahman) calves, and 2) compare the fecal microbiota as well as serum metabolites between males (bulls) and females (heifers) as well as between calm and temperamental animals at weaning (d0) and 4 days post weaning (d4). Equal numbers of animals were present in each category (5 calm female, 5 temperamental female, 5 calm male, and 5 temperamental male animals).
Metabolic responses to PD1 immune-checkpoint blockade and association with therapeutic benefits - Part I
STUDY_SUMMARY
Inhibition of immune-checkpoint targets including PD1 is clinically effective in a variety of cancers. However, only a subset of patients respond and complete response remains uncommon. Given the known role of metabolites in modulating immunity, we sought to understand how individual patientsā metabolic activities adapt to PD1 immune checkpoint blockade and how they associate with therapeutic benefits. To this end, we profiled metabolites in pre- and multiple on-treatment patient serum samples from three independent immunotherapy trials using hydrophilic interaction liquid chromatography coupled with either triple quadrupole MS multiple reaction monitoring or high resolution full scan MS detection. The study consisted of two Phase I trials (CA209-038, NCT01621490; CA209-009, NCT01358721) which included 78 patients with advanced melanoma and 91 patients with metastatic renal cell carcinoma (RCC) treated with nivolumab. To investigate the generalizability of our results, we also analyzed a large randomized Phase III trial (CheckMate 025, NCT01668784) with 743 RCC patients, among which 394 received nivolumab and 349 received everolimus. V600E is the most common BRAF mutation in melanoma and "BRAF_V600E" indicates the mutation status.
Metabolic responses to PD1 immune-checkpoint blockade and association with therapeutic benefits - Part II
STUDY_SUMMARY
Inhibition of immune-checkpoint targets including PD1 is clinically effective in a variety of cancers. However, only a subset of patients respond and complete response remains uncommon. Given the known role of metabolites in modulating immunity, we sought to understand how individual patientsā metabolic activities adapt to PD1 immune checkpoint blockade and how they associate with therapeutic benefits. To this end, we profiled metabolites in pre- and multiple on-treatment patient serum samples from three independent immunotherapy trials using hydrophilic interaction liquid chromatography coupled with either triple quadrupole MS multiple reaction monitoring or high resolution full scan MS detection. The study consisted of two Phase I trials (CA209-038, NCT01621490; CA209-009, NCT01358721) which included 78 patients with advanced melanoma and 91 patients with metastatic renal cell carcinoma (RCC) treated with nivolumab. To investigate the generalizability of our results, we also analyzed a large randomized Phase III trial (CheckMate 025, NCT01668784) with 743 RCC patients, among which 394 received nivolumab and 349 received everolimus. V600E is the most common BRAF mutation in melanoma and "BRAF_V600E" indicates the mutation status.
Metabolic responses to PD1 immune-checkpoint blockade and association with therapeutic benefits - Part III
STUDY_SUMMARY
Inhibition of immune-checkpoint targets including PD1 is clinically effective in a variety of cancers. However, only a subset of patients respond and complete response remains uncommon. Given the known role of metabolites in modulating immunity, we sought to understand how individual patientsā metabolic activities adapt to PD1 immune checkpoint blockade and how they associate with therapeutic benefits. To this end, we profiled metabolites in pre- and multiple on-treatment patient serum samples from three independent immunotherapy trials using hydrophilic interaction liquid chromatography coupled with either triple quadrupole MS multiple reaction monitoring or high resolution full scan MS detection. The study consisted of two Phase I trials (CA209-038, NCT01621490; CA209-009, NCT01358721) which included 78 patients with advanced melanoma and 91 patients with metastatic renal cell carcinoma (RCC) treated with nivolumab. To investigate the generalizability of our results, we also analyzed a large randomized Phase III trial (CheckMate 025, NCT01668784) with 743 RCC patients, among which 394 received nivolumab and 349 received everolimus.
P falciparum asexual metabolomics following drug treatment (part-I)
STUDY_SUMMARY
P falciparum infected human red blood cells were treated with 10X IC50 drug for 2.5 hours, followed by extraction and analysis of polar metabolites using HPLC-MS or HPLC-MS/MS
INSTITUTE
Penn State
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA
Uropathogenic versus Urocolonizing Escherichia coli
STUDY_SUMMARY
Urinary tract infections (UTIs) represent a major burden across the population, although key facets of their pathogenesis challenge physicians and investigators alike. Escherichia coli epitomizes these obstacles: this Gram-negative bacterial species is the most prevalent agent of UTIs worldwide and can also colonize the urogenital tract in a phenomenon known as asymptomatic bacteriuria (ASB). Unfortunately, at the level of the organism, the relationship between symptomatic UTI and ASB is poorly defined, confounding our understanding of microbial pathogenesis and strategies for clinical management. Unlike diarrheagenic pathotypes of E. coli, the definition of uropathogenic E. coli (UPEC) remains phenomenologic, without conserved phenotypes and (known) genetic determinants that rigorously distinguish UTI- and ASB-associated strains. This manuscript provides a cross-disciplinary review of the current issues ā from interrelated mechanistic and diagnostic perspectives ā and describes new opportunities by which clinical resources can be leveraged to overcome molecular challenges. Specifically, we present our work harnessing a large collection of patient-derived isolates to identify features that do (and do not) distinguish UTI- from ASB-associated E. coli strains. Analyses of biofilm formation, previously reported to be higher in ASB strains, revealed extensive phenotypic heterogeneity that did not correlate with symptomatology. However, metabolomic experiments revealed distinct signatures between ASB and cystitis isolates, including species in the purine pathway (previously shown to be critical for intracellular survival during acute infection). Together, these studies demonstrate how large-scale, wild-type approaches can help dissect the physiology of colonization-versus-infection, suggesting that the molecular definition of UPEC may rest at the level of global bacterial metabolism.
INSTITUTE
Vanderbilt University
LAST_NAME
Rutledge
FIRST_NAME
Alexandra
ADDRESS
7330 Stevenson Center Lane, NASHVILLE, TENNESSEE, 37235, USA
Luteal lipids regulate progesterone production and may modulate immune cell function during the estrous cycle and pregnancy
STUDY_SUMMARY
Despite data indicating an important functional role for bioactive lipids in luteal function, little is known about the patterns of abundance of these lipids in corpus luteum (CL) during luteal development, maintenance, and rescue, in any species. Therefore, the abundance of lipid mediators, including endocannabinoids and oxylipins from cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P450 (CYP)-dependent metabolism were profiled in the CL on days 4, 11, and 18 of the estrous cycle and on day 18 of pregnancy. The objectives of this study were to identify lipid mediators that regulate luteal function during these transitions, to integrate the lipid profile with a previously published mRNA profile of CL during maternal recognition of pregnancy, and to determine the effect of a subset of lipids on in vitro progesterone production.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome and Biomedical Sciences Facility
LABORATORY
WCMC Metabolomics Core
LAST_NAME
Fiehn
FIRST_NAME
Oliver
ADDRESS
1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
TFPa/HADHA is required for fatty acid beta-oxidation and cardiolipin re-modeling in human cardiomyocytes (part-I)
STUDY_SUMMARY
Mitochondrial trifunctional protein deficiency, due to mutations in hydratase subunit A (HADHA), results in sudden infant death syndrome (SIDS) with no cure. To reveal the disease etiology, we generated stem cell-derived cardiomyocytes from HADHA-deficient hiPSCs and accelerated their maturation via a novel, engineered MicroRNA Maturation Cocktail (MiMaC) that upregulated the epigenetic regulator, HOPX. Fatty acid challenged MiMaC treated HADHA mutant cardiomyocytes manifested the disease phenotype: defective calcium dynamics and repolarization kinetics which resulted in a pro-arrhythmic state. Single cell RNA-seq revealed a novel cardiomyocyte developmental intermediate, based on metabolic gene expression. This intermediate gave rise to mature-like cardiomyocytes in control cells but, mutant cells transitioned to a pathological state with reduced fatty acid beta-oxidation (FAO), reduced mitochondrial proton gradient, disrupted cristae structure and defective cardiolipin remodeling. This study reveals that TFPa/HADHA, a MLCL-AT-like enzyme, is required for FAO and cardiolipin remodeling, essential for functional mitochondria in human cardiomyocytes.
INSTITUTE
UC Davis
LAST_NAME
Showalter
FIRST_NAME
Megan
ADDRESS
UC Davis Genome Center, room 1313, 451 Health Sci Drive
Longitudinal Characterization of the Fecal Metabolome in Dogs with Idiopathic Inflammatory Bowel Disease
STUDY_SUMMARY
Thirteen dogs diagnosed with idiopathic IBD, that previously failed to respond to treatment with elimination diets and metronidazole, were enrolled. Stool samples were collected from all dogs before initiating therapy with prednisone, after 3 and 8 weeks, and more than one year after beginning treatment. Thirteen healthy dogs were enrolled in the study as a control group.
The cecal and fecal metabolomes of horses before and after metronidazole administration
STUDY_SUMMARY
Metronidazole (15mg/kg BID PO) was given to horses (n=5) with in-dwelling cecal cannulas. The study was suspended after the fifth dose (day 3) due to adverse gastrointestinal effects. Cecal and fecal samples were obtained before and after (Days days -52, -28, -14, 0, 7, 14, 28 and 52) metronidazole administration. The metabolome was characterized by mass spectrometry-based methods. Fecal, but not cecal metabolites were affected by metronidazole. The fecal metabolites affected represented diverse metabolic pathways such as nucleic acid metabolism, secondary bile metabolism, fatty acid synthesis/degradation/elongation or metabolism and sugar metabolism.
Stool samples are acquired from mice models after biliary diversion surgery. Metabolite extractions are then performed on the stool samples and run through an optimized RPLC-IM-MS method.
The effects of a training program encompassing cold exposure, breathing exercises, and meditation on plasma metabomics during experimental human endotoxemia
STUDY_TYPE
Experimental human endotoxemia study
STUDY_SUMMARY
Study to investigate the effects of a training program encompassing cold exposure, breathing exercises, and meditation on plasma metabolomic during experimental human endotoxemia.
INSTITUTE
Radboud University Medical Centre Nijmegen
DEPARTMENT
Intensive Care Medicine
LABORATORY
Metabolomic Discoveries (acquired by Metabolon in September 2017)
LAST_NAME
Kox
FIRST_NAME
Matthijs
ADDRESS
Intensive Care Medicine (710), Geert Grooteplein 10
The sphingolipid, ceramide-1-phosphate (C1P), directly binds and activates Group IVA cytosolic phospholipase A2 (cPLA2) to generate eicosanoids. Due to the role of eicosanoids in wound healing, we choose to use our novel genetic mouse model expressing cPLA2 with an ablated C1P interaction site (KI) to examine the cPLA2/C1P interaction in wound healing. Wound closure rate was not affected, but wound maturation was enhanced by loss of the C1P/cPLA2Ī± interaction based on the following findings. Wounds in KI mice displayed: i) increased infiltration of dermal fibroblasts into the wound environment; ii) increased wound tensile strength; and iii) higher Type I/Type III collagen ratios. These findings were recapitulated in vitro as primary dermal fibroblasts (pDFs) from KI mice showed significantly increased collagen deposition and migration velocity compared to WT and KO pDFs. Additionally, the KI showed an altered eicosanoid profile of reduced pro-inflammatory prostaglandins (PGE2) and increased levels of specific HETE species (5-HETE). Elevated 5-HETE levels promoted increased dermal fibroblast migration and collagen deposition. This āgain of functionā role for the mutant cPLA2 was also linked to differential cellular localization of cPLA2Ī± and 5-HETE biosynthetic factors. These studies demonstrate regulation of key biological mechanisms by a defined protein:lipid interaction in vivo and provide new insights into cPLA2 function.
Phenotyping Mouse blood metabolites in day and night in type 2 diabetes
STUDY_TYPE
time course in a 24hr period
STUDY_SUMMARY
This experiment compares the metabolites in control db/m and diabetic db/db in day and night.
INSTITUTE
Indiana University School of Medicine
DEPARTMENT
Ophthalmology
LABORATORY
Maria Grant Laboratory
LAST_NAME
Beli
FIRST_NAME
Eleni
ADDRESS
97 Lisburn Road
EMAIL
e.beli@qub.ac.uk
PHONE
5176144409
NUM_GROUPS
4
TOTAL_SUBJECTS
15
NUM_MALES
15
PUBLICATIONS
Nutrients.2019. Beli E. Loss of diurnal oscillatory rhythms in gut microbiota correlates with changes in circulating metabolites in type 2 diabetic, db/db mice
Eicosanoid profiles of dermal fibroblasts (part-II)
STUDY_SUMMARY
The sphingolipid, ceramide-1-phosphate (C1P), directly binds and activates Group IVA cytosolic phospholipase A2 (cPLA2) to generate eicosanoids. Due to the role of eicosanoids in wound healing, we choose to use our novel genetic mouse model expressing cPLA2 with an ablated C1P interaction site (KI) to examine the cPLA2/C1P interaction in wound healing. Wound closure rate was not affected, but wound maturation was enhanced by loss of the C1P/cPLA2Ī± interaction based on the following findings. Wounds in KI mice displayed: i) increased infiltration of dermal fibroblasts into the wound environment; ii) increased wound tensile strength; and iii) higher Type I/Type III collagen ratios. These findings were recapitulated in vitro as primary dermal fibroblasts (pDFs) from KI mice showed significantly increased collagen deposition and migration velocity compared to WT and KO pDFs. Additionally, the KI showed an altered eicosanoid profile of reduced pro-inflammatory prostaglandins (PGE2) and increased levels of specific HETE species (5-HETE). Elevated 5-HETE levels promoted increased dermal fibroblast migration and collagen deposition. This āgain of functionā role for the mutant cPLA2 was also linked to differential cellular localization of cPLA2Ī± and 5-HETE biosynthetic factors. These studies demonstrate regulation of key biological mechanisms by a defined protein:lipid interaction in vivo and provide new insights into cPLA2 function.
Immunomodulatory activity of hyaluronidase is associated with metabolic adaptations during acute inflammation
STUDY_SUMMARY
Objective and design: Investigate survival outcomes, and immunological and metabolomic effects of hyaluronidase (Hz) treatment during mouse models of acute inflammation and sepsis. Methods: Survival of C57Bl/6 mice was monitored after lethal challenge with lipopolysaccharide (LPS) or cecal and ligation puncture (CLP)-induced sepsis and treated with Hz or saline. Mice were also challenged with LPS and treated with Hz for leukocyte counting, cytokine quantification and determination of metabolomic profiles in the peritoneal fluid. Results: Hz treatment improved survival outcomes after lethal challenge with LPS or CLPinduced sepsis. LPS challenge promoted acute neutrophil accumulation and production of interleukin-1Ī² (IL-1Ī²) and IL-6 in the peritoneum, whereas Hz treatment suppressed neutrophil infiltration and cytokine production. We further characterized the metabolomic alterations caused by LPS challenge, which predicted activity of metabolic pathways related to fatty acids and eicosanoids. Hz treatment had a profound effect over the metabolic response, reflected by reductions of the relative levels of fatty acids. Conclusion: Collectively, these data demonstrate that Hz treatment is associated with metabolic reprogramming of pathways that sustain the inflammatory response.
INSTITUTE
Sao Paulo University
DEPARTMENT
School of Pharmaceutical Sciences of Ribeirao Preto
Metabolic landscape remodeling in dystrophic muscle through glucocorticoid steroid regimens
STUDY_SUMMARY
Duchenne muscular dystrophy is caused by genetic defects in the gene encoding dystrophin and leads to progressive muscle degeneration. Glucocorticoid steroids are current mainstay pharmacological regimen to decrease muscle inflammation and prolong the ambulatory period in these patients, but daily intake of glucocorticoids like prednisone and deflazacort causes adverse side effects like osteoporosis, adrenal suppression, insulin resistance and obesity. Intermittent steroid dosing has been proposed as alternative to maintain benefits and limit side effects, but a detailed understanding of the mechanisms underpinning the regimen-specific effects in muscle is still missing. Here we explore how once-daily versus once-weekly prednisone (4 week-long treatment) affect the metabolomic landscape in mdx mouse muscle (genetic model of Duchenne muscular dystrophy; DBA/2J background) through metabolomics profiling.
INSTITUTE
Northwestern University
LAST_NAME
Quattrocelli
FIRST_NAME
Mattia
ADDRESS
303 East Superior St, SQBRC 5-500, Chicago, IL, 60611, USA
Although health benefits of the Dietary Approaches to Stop Hypertension (DASH) diet are established, it is not understood which food compounds result in these benefits. We used a step-wise approach to identify unique compounds from individual foods of a DASH-style diet, determined if these Food-Specific Compounds (FSC) are detectable in urine, and then examined relationships between urinary FSC and blood pressure (BP). Nineteen subjects were randomized into 6-week controlled DASH-style diet interventions. Untargeted, LC/MS-based metabolomics was performed on 24-hour urine samples collected before and after each intervention and on 12 representative DASH-style foods.
Modeling the metabolic interplay between a parasitic worm and its bacterial endosymbiont allows the identification of novel drug targets
STUDY_SUMMARY
The filarial nematode Brugia malayi represents a leading cause of disability in the developing world, causing lymphatic filariasis in nearly 40 million people. Currently available drugs are not well-suited to mass drug administration efforts, so new treatments are urgently required. One potential vulnerability is the endosymbiotic bacteria Wolbachiaāpresent in many filariaeāwhich is vital to the worm. Genome scale metabolic networks have been used to study prokaryotes and protists and have proven valuable in identifying therapeutic targets, but only recently have been applied to eukaryotic organisms. Here, we present iDC625, the first compartmentalized metabolic model of a parasitic worm. We used this model to show how metabolic pathway usage allows the worm to adapt to different environments, and predict a set of 99 reactions essential to the survival of B. malayi. We validated three of those reactions with drug tests and demonstrated novel antifilarial properties for all three compounds.
INSTITUTE
Hospital for Sick Children, University of Toronto, NYU Langone Health
Targeted Metabolomic Analysis in Patients with Wilson Disease Reveals Dysregulated Choline, Methionine and Aromatic Amino Acid Metabolism: Implications for Hepatic and Neurological Phenotypes
STUDY_TYPE
Cross-sectional
STUDY_SUMMARY
This study is comparing the plasma metabolomics profile of patients with the genetic disorder, Wilson disease, compared to healthy subjects matched by age, sex, and BMI. Wilson disease (WD) is a genetic copper overload condition characterized by hepatic and neuropsychiatric symptoms with a pathogenesis not well-understood. Choline is essential for lipid metabolism and the methionine cycle; a dysregulated methionine cycle is reported in animal models of WD, though not verified in humans. Defects in neurotransmitters, acetylcholine, and biogenic amines are reported in WD patients with neurological presentations. Precursors of these neuromodulators include choline, phenylalanine, tyrosine, and histidine. Less is known about the circulating levels of these precursors in WD. We aimed to study choline, methionine, aromatic amino acids, and phospholipids in serum profiles of WD subjects compared to healthy subjects (HC).
Metabolic changes of Fusobacterium nucleatum when co-cultured with other oral microbes (part-I)
STUDY_SUMMARY
We used membrane-separated co-culture systems to globally assess metabolomic changes of Fusobacterium nucleatum when co-cultured with Streptococcus gordonii and/or Veillonella parvula.
Metabolic changes of culture supernatants of Fusobacterium nucleatum co-cultured with other oral microbes (part-II)
STUDY_SUMMARY
We used membrane-separated co-culture systems to globally assess extracellular metabolomic changes of Fusobacterium nucleatum co-cultured with Streptococcus gordonii and/or Veillonella parvula.
The impact of tobacco smoke exposure and environmental exposures on the pulmonary microbiome of critically ill children
STUDY_SUMMARY
This project evaluates the effects of tobacco smoke exposure (TSE) on the pediatric lung microbiome in critically ill children. The impact of TSE on the airway microbiome of critically ill, mechanically ventilated pediatric patients will be determined by through clinical outcomes and analysis of urinary and plasma metabolomes to identify other environmental exposures contributing to the alteration of the pediatric microbiome.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Uppal
FIRST_NAME
Karan
ADDRESS
NA
EMAIL
kuppal2@emory.edu
PHONE
(404) 727 5027
TOTAL_SUBJECTS
367
STUDY_COMMENTS
Both CHEAR pooled urine samples and Clinical Biomarker Laboratory pooled plasma samples were used
Metabolomic Profiling of Oxalate-Degrading Probiotic Lactobacillus acidophilus and Lactobacillus gasseri
STUDY_TYPE
Metabolomic/Lipidomic Profiling
STUDY_SUMMARY
Metabolomic and lipidomic profiling of Lactobacillus acidophilus and Lactobacillus gasseri to identify unique differences in their biochemistry that could potentially influence their ability to serve as probiotic agents for oxalate diseases.
INSTITUTE
University of Florida
DEPARTMENT
Pathology, Immunology and Laboratory Medicine
LABORATORY
Timothy J. Garrett, PhD
LAST_NAME
Chamberlain
FIRST_NAME
Casey
ADDRESS
UF Dept of Pathology PO Box 100275 Gainesville, FL 32610
Plasma samples from Wistar rats fed a control or High-sodium and low-potassium HNaLK diet with or without antibiotic treatment (n = 7 each, a total of 28) were subjected to lipidomics analysis.The HNaLK diet interacts with gut bacteria to alter plasma lipid profiles, which may be related to its health effects.
Comparative metabolomics of MCF-7 breast cancer cells using different extraction solvents assessed by mass spectroscopy
STUDY_TYPE
Analysing metabolomics using GC Mass Spectroscopy
STUDY_SUMMARY
Metabolic profiling of cancer cells can play a vital role in revealing the molecular bases of cancer development and progression. In this study, gas chromatography coupled with mass spectrometry (GC-MS) was employed for the determination of signatures found in ER+/ PR+ breast cancer cells derived from MCF-7 using different extraction solvents including: A, formic acid in water; B, ammonium hydroxide in water; C, ethyl acetate; D, methanol: water (1:1, v/v); and E, acetonitrile: water (1:1, v/v). The greatest extraction rate and diversity of metabolites occurs with extraction solvents A and E. Extraction solvent D showed moderate extraction efficiency, whereas extraction solvent B and C showed inferior metabolite diversity. Metabolite set enrichment analysis results showed energy production pathways to be key in MCF-7 cell lines. This study showed that mass spectrometry could identify key metabolites associated with cancers. The highest enriched pathways were related to energy production as well as Warburg effect pathways, which may shed light on how energy metabolism has been hijacked to encourage tumour progression and eventually metastasis in breast cancer.
Metabolomic Profiles of Pancreatic Ī²-Cells and Islets Exposed to Arsenic, part I Ī²-Cells
STUDY_SUMMARY
Type-2 diabetes (T2D) is a complex metabolic disorder that affects hundreds of millions of people world-wide and is a growing public health concern. Despite recent advances in T2D research, the etiology of this disease and the mechanisms underlying the metabolic defects remain poorly understood. While obesity is thought to be the main cause for the rising prevalence of T2D, obesity alone cannot explain differences in the trends of T2D among different geographical regions and populations. Growing evidence suggests that environmental exposures to toxic and diabetogenic substances must play important roles. Inorganic arsenic (iAs) is a naturally occurring toxic metalloid. Hundreds of millions of people worldwide are exposed to unsafe levels of iAs in drinking water and food. iAs is a potent carcinogen, but iAs exposure has also been linked to increase risk of T2D. While the link between iAs exposure and T2D is well-established, the mechanisms underlying the diabetogenic effects of iAs exposure remain unclear. Results of our previously published and ongoing studies suggest that pancreatic Ī²-cells are a primary target for iAs and its metabolites and that impaired insulin secretion by Ī²-cells is the mechanism by which iAs exposure leads to diabetes. The proposed project will use metabolomics to identify metabolic pathways in Ī²-cells that are targeted by iAs and its metabolites, monomethyl-As (MAs) and dimethyl-As (DMAs). The metabolomics data combined with results of our ongoing mechanistic studies will provide a comprehensive picture of the metabolic dysfunction leading to the development of diabetes in individuals exposed to iAs and of the molecular mechanisms that underlie this dysfunction. Identifying the affected pathways and mechanisms will ultimately help to improve strategies for prevention and/or treatment of T2D associated with chronic exposure to iAs.
Mass spectrometry-based lipidomics of oral squamous cell carcinoma tissue reveals aberrant cholesterol and glycerophospholipid metabolism
STUDY_TYPE
Case study
STUDY_SUMMARY
Comparison between the lipid profile in oral squamous cell carcinoma of the tongue and healthy epithelial tissue from the contralateral side of the tongue of the same patient.
INSTITUTE
University of Helsinki
DEPARTMENT
Department of Otorhinolaryngology-Head and Neck Cancer
Department of Otorhinolaryngology - Head and Neck Surgery, University of Helsinki and Helsinki University Hospital, PO Box 263, FI-00029 HUS, Helsinki, Finland
Plasma metabolomics from HIV subjects and controls was incorporated with microbiome data to develop Annotation of Metabolite Origins via Networks (AMON).
INSTITUTE
University of Colorado Denver|Anschutz Medical Campus
Exosomal lipids for classifying early and late stage non-small cell lung cancer
STUDY_TYPE
Biomarker Discovery
STUDY_SUMMARY
Lung cancer is the leading cause of cancer deaths in the United States. Patients with early stage lung cancer have the best prognosis with surgical removal of the tumor, but the disease is often asymptomatic until advanced disease develops, and there are no effective blood-based screening methods for early detection of lung cancer in at-risk populations. We have explored the lipid profiles of blood plasma exosomes using ultra high-resolution Fourier transform mass spectrometry (UHR-FTMS) for early detection of the prevalent non-small cell lung cancers (NSCLC). Exosomes are nanovehicles released by various cells and tumor tissues to elicit important biofunctions such as immune modulation and tumor development. Plasma exosomal lipid profiles were acquired from 39 normal and 91 NSCLC subjects (44 early stage and 47 late stage). We have applied two multivariate statistical methods, Random Forest (RF) and Least Absolute Shrinkage and Selection Operator (LASSO) to classify the data. For the RF method, the Gini importance of the assigned lipids was calculated to select 16 lipids with top importance. Using the LASSO method, 7 features were selected based on a grouped LASSO penalty. The Area Under the Receiver Operating Characteristic curve for early and late stage cancer versus normal subjects using the selected lipid features was 0.85 and 0.88 for RF and 0.79 and 0.77 for LASSO, respectively. These results show the value of RF and LASSO for metabolomics data-based biomarker development, which provide robust an independent classifiers with sparse data sets. Application of LASSO and Random Forests identifies lipid features that successfully distinguish early stage lung cancer patient from healthy individuals.
INSTITUTE
University of Kentucky
DEPARTMENT
Center for Environmental and Systems Biochemistry
LAST_NAME
Thompson
FIRST_NAME
Patrick
ADDRESS
789 South Limestone, Lexington, Kentucky, 40536, USA
Necrotizing soft-tissue infections (NSTIs) have multiple causes, risk factors, anatomical locations, and pathogenic mechanisms. In patients with NSTI, circulating metabolites may serve as substrate having impact on bacterial adaptation at the site of infection. Metabolic signatures associated with NSTI may reveal potential be useful as diagnostic and prognostic markers, as well as novel targets for therapy. This study used untargeted metabolomics analyses of plasma from NSTI patients (n=34) and healthy (non-infected) controls (n=24) to identify the metabolic signatures and connectivity patterns among metabolites associated with NSTI.
Colorectal cancer before and after surgery metabolomics data integration
STUDY_TYPE
Pilot study
STUDY_SUMMARY
In this study 40 urine samples were analyzed in 5 batches. There are 3 labels: control group volunteers (8 pcs., CG), colorectal cancer patients before surgical operation (20 pcs., pre) and after surgical operation (12 pcs., post).
Growth cone memebrane and growth cone particulate lipidomics
STUDY_TYPE
untargeted LC-MS/MS lipidomics
STUDY_SUMMARY
We performed high performance liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis of the proteome and lipidome of GC from C57BL/6 mice across five age groups (E18, P0, P3, P6, and P9) from two growth cone fractions: growth cone membrane (GCM) and growth cone particulate (GCP)
Lipidomics Dataset of Sonication-Induced Traumatic Optic Neuropathy in Mice
STUDY_SUMMARY
Traumatic optic neuropathy (TON) is the loss of vision secondary to trauma. Approximately two weeks after traumatic damage, diffuse retinal ganglion cell loss and axon degeneration of the optic nerve are exhibited. Here we present the changes that occur in the optic nerve lipidome of two-month-old C57BL/6J mice following sonication-induced TON (SI-TON), which closely models the indirect clinical mechanism in TON. Optic nerves were harvested at three time points following injury: 1-day, 7-days, and 14-days for comparison with the control group (uninjured optic nerves from 2-month-old mice). Lipidomic changes were observed at each of the various time points, with a pattern of progression present in multiple lipid classes. This data demonstrates the distinct lipidomic changes at each time point following indirect trauma to the optic nerve.
INSTITUTE
Bascom Palmer Eye Institute, Miller School of Medicine at University of Miami, Miami, FL 33136, USA
DEPARTMENT
Ophthalmology, Bascom Palmer Eye Institute
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy K
ADDRESS
1638 NW 10th Avenue, Suite 707A University of Miami Miami, FL, 33136
Metabolomics-based profiling of the mode of action of Pathogen Box compounds in Trypanosoma brucei (part-I)
STUDY_SUMMARY
The mode of action of anti-Trypanosomal compounds from the Pathogen Box was investigated using an unbiased metabolomics approach. Trypanosoma brucei parasites were incubated for five hours with the test compounds at 0.5 ĀµM concentration. Analysis of cell pellets allowed reproducible detection of diverse metabolites from a range of metabolic pathways.
INSTITUTE
Monash University
LAST_NAME
Creek
FIRST_NAME
Darren
ADDRESS
Monash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Metabolomics-based profiling of the mode of action of Pathogen Box compounds in Trypanosoma brucei (part-II)
STUDY_SUMMARY
The mode of action of anti-Trypanosomal compounds from the Pathogen Box was investigated using an unbiased metabolomics approach. Trypanosoma brucei parasites were incubated for five hours with the test compounds at 0.5 ĀµM concentration. Analysis of cell pellets allowed reproducible detection of diverse metabolites from a range of metabolic pathways.
INSTITUTE
Monash University
LAST_NAME
Creek
FIRST_NAME
Darren
ADDRESS
Monash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Development and Characterisation of a Novel Class of Aroyl Guanidine Containing Anti-Trypanosomal Compounds
STUDY_SUMMARY
The mode of action of a novel class of aroyl guanidine containing anti-Trypanosomal compounds was investigated using an unbiased metabolomics approach. Trypanosoma brucei parasites were incubated for five hours with the test compounds at 1 ĀµM concentration. Analysis of cell pellets allowed reproducible detection of diverse metabolites from a range of metabolic pathways.
INSTITUTE
Monash University
LAST_NAME
Creek
FIRST_NAME
Darren
ADDRESS
Monash Institute of Pharmaceutical Sciences, 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Pseudoexfoliation Aqueous Humor Metabolites from Veterans Affairs Patients
STUDY_SUMMARY
We performed high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) Isotopic Ratio Outlier Analysis of aqueous humor samples from patients from patients with pseudoefoliation syndrome (PEX).
POAG and Control Aqueous Humor IROA Metabolites from Veterans Affairs Patients
STUDY_TYPE
untargeted LC-MS/MS metabolomics IROA
STUDY_SUMMARY
We performed high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of the metabolome of POAG and Control patients. Samples were collected asynchronously.
K13 mutations driving artemisinin resistance rewrite Plasmodium falciparumās programmed intra-erythrocytic development and transform mitochondrial physiology
STUDY_SUMMARY
The emergence of artemisinin resistance in Southeast Asia, dictated by mutations in the Plasmodium falciparum k13 gene, has compromised antimalarial efficacy and created a core vulnerability in the global malaria elimination campaign. Applying quantitative transcriptomics, proteomics, and metabolomics to a panel of isogenic K13 mutant or wild-type P. falciparum lines, we observe that K13 mutations reprogram multiple aspects of intra-erythrocytic parasite biology. These changes impact its cell cycle periodicity, the unfolded protein response and protein degradation, vesicular trafficking and endocytosis, and mitochondrial functions including the TCA cycle, the electron transport chain, and redox regulation. Ring-stage artemisinin resistance mediated by the K13 R539T mutation was neutralized using atovaquone, an electron transport chain inhibitor. Our data suggest that modification of mitochondrial physiology, accompanied by other processes to reduce artemisininās proteotoxic effects, help protect parasites against this pro-oxidant drug, allowing resumption of growth once the rapidly-cleared artemisinins have reached sub-therapeutic levels.
INSTITUTE
Pennsylvania State University
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA
Macrophage-Mediated Clofazimine Sequestration is Accompanied by a Shift in Host Energy Metabolism
STUDY_TYPE
4 timepoints for urine collection, whole blood collected at sacrifice (8 weeks)
STUDY_SUMMARY
Examined the effects of long-term dosing with CFZ of mice, metabolic data on CFZ and sham mice was collected at 0, 2, 4, and 8 weeks, at 8 weeks mice were sacrificed and WB was collected from 6 CFZ treated mice and 7 sham mice
INSTITUTE
University of Michigan
DEPARTMENT
Clinical Pharmacy
LABORATORY
NMR Metabolomics Laboratory, University of Michigan
LAST_NAME
Stringer
FIRST_NAME
Kathleen
ADDRESS
College of Pharmacy, 428 Church St, Ann Arbor, MI, 48109
Transgenic Parkinson's Mice Following Immunotherapy
STUDY_SUMMARY
An UHPLC-HRMS Metabolomics and Lipidomics Study of Stool from Transgenic Parkinson's disease Mice Following Immunotherapy. Parkinsonās disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta of the brain as well as degeneration of motor and non-motor circuitry. The cause of neuronal death is currently unknown, although chronic neuroinflammation, aggregated Ī±-synuclein, mitochondrial dysfunction and oxidative stress have all been implicated. Gliosis has been shown to exacerbate neuroinflammation via secretion of pro-inflammatory cytokines, and there is a subsequent infiltration of T lymphocytes (T-cells), into the brain of PD patients. Using liquid chromatography-high resolution mass spectrometry (LC-HRMS), we have observed metabolomic changes in stool samples, thought to be associated with the potential disease-modifying effect of an immunotherapy administered to transgenic Parkinsonian (A53T) mice. Significant elevations (p<0.05) in metabolites associated with immune response (taurine, histamine and its methylated product, 3-methylhistamine) are identified as being higher in the mice undergoing immunotherapy. Furthermore, a reduction in triacylglycerols (TG) and diacylglyceols (DG) expression in stool following immunotherapy suggests a regulation of lipid breakdown or biosynthesis with the vaccine. These āomicsā markers (among others reported in this article) along with weight gain and increased life expectancy suggest that the immunotherapy is positively modifying the disease state.
Lipid composition of isolated lipid droplets from the functional bovine corpus luteum
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Establishment and maintenance of pregnancy is dependent on progesterone synthesized by the corpus luteum (CL). The CL is known for the prominent presence of intracellular lipid droplets (LDs). However relatively little is known about the composition and function of these luteal LDs. Our objective was to identify the lipid composition of LDs from fully functional bovine CLs. Luteal LDs were isolated by flotation through a discontinuous sucrose gradient, lipids were then extracted using a standard Bligh and Dyer protocol, dried, and sent to Avanti Polar Lipids for lipidomics analysis. The samples were provided for lipidomic profiling of free sterols, cholesteryl esters, triglycerides, diacylglycerols, phospholipids, and sphingolipids. Molecular species were resolved by reversed-phase liquid chromatography in the presence of class and sub-class specific internal standard compounds added to each sample. The compounds were detected by tandem mass spectrometry (MS/MS) with scheduled multiple reaction monitoring (MRM) for mass-specific fragment ions according to the lipid class and molecular weight of the compound. Quantification of cholesterol, cholesteryl esters, triglycerides, and diglycerides were directly calculated with standards and internal standards from calibration response curves. The remaining lipid species were semi-quantization using the integrated area of each analyteās MRM peak, divided by the appropriate internal standard peak area, and multiplied by the standardās known concentration. Lipid concentrations were normalized to the corresponding protein concentration of each sample and as a mol % relative to total lipids or within each lipid class. Isolated luteal LDs were composed primarily of triglyceride (88%, mol% of lipid class to total lipids). Other neutral lipids included diacylglycerol, 2.9%; and cholesteryl esters, 1.5%. Polar lipids were primarily composed of phosphatidylcholine (3.1%), sphingomyelin (1.5%), phosphatidylinositol (0.9%), phosphatidylethanolamine (0.8%) and phosphatidylserine (0.4%). A number of other minor lipids representing less than 0.32% of the total lipid pool were also detected including phosphatidylglycerol, lysophospholipids, ceramides, and glycosylated ceramides. Lipid composition of bovine luteal LDs are distinct from LDs isolated from other tissues and in other species.
INSTITUTE
University of Nebraska Medical Center
DEPARTMENT
Obstetrics and Gynecology
LABORATORY
John S. Davis
LAST_NAME
Davis
FIRST_NAME
John
ADDRESS
983255 Nebraska Medical Center Omaha, NE 68198-3255
Luminal succinate in UC-HMA (human microbiota-associated) mice.
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Patients with ulcerative colitis (UC) are known to be at higher risk for Clostridium difficile (C. difficile) infection (CDI), and CDI in UC patients is recognized as a major clinical problem because it worsens UC outcome. In order to assess the role of gut dysbiosis, seen in UC patients, we have established a human microbiota-associated mouse model in which germ-free mice are colonized with gut microbiota from UC patients. Utilizing this model, we found that UC microbiota colonized HMA mice (UC-HMA mice) were susceptible to CDI. To address the mechanism by which UC-HMA mice are unable to acquire the colonization resistance against C. difficile, we analyzed the luminal metabolites in UC-HMA mice, especially in succinate which is a crucial metabolite doe the growth of C. difficile.
Subcellular organelle lipidomics in TLR-4-activated macrophages
STUDY_SUMMARY
Lipids orchestrate biological processes by acting remotely as signaling molecules or locally as membrane components that modulate protein function. Detailed insight into lipid function requires knowledge of the subcellular localization of individual lipids. We report an analysis of the subcellular lipidome of the mammalian macrophage, a cell type that plays key roles in inflammation, immune responses, and phagocytosis. Nuclei, mitochondria, endoplasmic reticulum (ER), plasmalemma, and cytoplasm were isolated from RAW 264.7 macrophages in basal and activated states. Subsequent lipidomic analyses of major membrane lipid categories identified 229 individual/isobaric species, including 163 glycerophospholipids, 48 sphingolipids, 13 sterols, and 5 prenols. Major subcellular compartments exhibited substantially divergent glycerophospholipid profiles. Activation of macrophages by the Toll-like receptor 4-specific lipopolysaccharide Kdo2-lipid A caused significant remodeling of the subcellular lipidome. Some changes in lipid composition occurred in all compartments (e.g. increases in the levels of ceramides and the cholesterol precursors desmosterol and lanosterol). Other changes were manifest in specific organelles. For example, oxidized sterols increased and unsaturated cardiolipins decreased in mitochondria, whereas unsaturated ether-linked phosphatidylethanolamines decreased in the ER. We speculate that these changes may reflect mitochondrial oxidative stress and the release of arachidonic acid from the ER in response to cell activation.
INSTITUTE
LIPID MAPS
DEPARTMENT
Multiple
LABORATORY
Multiple
LAST_NAME
Fahy
FIRST_NAME
Eoin
ADDRESS
9500 Gilman, La Jolla, CA, 92093, USA
EMAIL
efahy@ucsd.edu
PHONE
858-534-4076
PUBLICATIONS
Andreyev AY, Fahy E, Guan Z, Kelly S, Li X, McDonald JG, Milne S, Myers D, Park H, Ryan A, Thompson BM, Wang E, Zhao Y, Brown HA, Merrill AH, Raetz CR, Russell DW, Subramaniam S, Dennis EA. Subcellular organelle lipidomics in TLR-4-activated macrophages. J Lipid Res. 2010 Sep;51(9):2785-97. doi: 10.1194/jlr.M008748. Epub 2010 Jun 23. PMID: 20574076; PMCID: PMC2918461.
Regulated accumulation of desmosterol integrates macrophage lipid metabolism and inflammatory responses
STUDY_SUMMARY
To investigate the relationship between hypercholesterolemia, foam cell formation and inflammation, we performed lipidomic and transcriptomic analyses of elicited peritoneal macrophages in wild type (WT) or LDL receptor knockout (LDLR KO) mice fed either a normal cholesterol, normal fat (NCNF) diet or a high cholesterol, high fat (HCHF) 'Western' style diet. The combination of the LDLR KO genotype and the HCHF diet results in the formation of macrophage foam cells in the elicited peritoneal macrophage population. Analysis of macrophages from the above four experimental groups revealed massive reprogramming of the lipidome in response to both diet and genotype. These studies confirmed and extended prior knowledge regarding the roles of SREBP and LXR signaling in cholesterol and fatty acid homeostasis. Unexpectedly, peritoneal macrophage foam cells exhibited a strongly 'deactivated' phenotype, with marked suppression of pro-inflammatory mediators that are normally characteristic of the inflammatory responses associated with atherosclerotic lesions. Many of these changes in gene expression and lipid metabolism appear to be related to the paradoxical accumulation of high levels of desmosterol, the last intermediate in the Bloch pathway of cholesterol biosynthesis. WT or LDLR KO mice were fed either a NCNF diet or a HCHF diet for twelve weeks to establish four experimental groups (WT-NCNF diet, WT-HCHF diet, KO-NCNF diet, and KO-HCHF diet). As expected, the combination of the HCHF diet and LDLR KO genotype resulted in a synergistic effect on serum lipid levels. Elicited peritoneal macrophages (92-96% F4/80-positive) were immediately prepared for analysis, thereby preserving in vivo gene expression and lipid profiles. Macrophages derived from LDLR KO mice fed the HCHF diet contained nearly four-fold more total cholesterol than cells from WT mice fed the same diet. Quantitative analysis of 245 lipid species revealed significant changes in nearly all major lipid classes. Using a two-way ANOVA model, we found that 176 (72%) of the lipids analyzed were significantly affected by the HCHF diet, 133 (54%) by the LDLR KO genotype, and 114 (46%) by interactions between the HCHF diet and LDLR KO genotype. Many of the observed interactions (60%) were synergistic.
INSTITUTE
LIPID MAPS
DEPARTMENT
Multiple
LABORATORY
Multiple
LAST_NAME
Fahy
FIRST_NAME
Eoin
ADDRESS
9500 Gilman, La Jolla, CA, 92093, USA
EMAIL
efahy@ucsd.edu
PHONE
858-534-4076
PUBLICATIONS
Spann NJ, Garmire LX, McDonald JG, Myers DS, Milne SB, Shibata N, Reichart D, Fox JN, Shaked I, Heudobler D, Raetz CR, Wang EW, Kelly SL, Sullards MC, Murphy RC, Merrill AH Jr, Brown HA, Dennis EA, Li AC, Ley K, Tsimikas S, Fahy E, Subramaniam S, Quehenberger O, Russell DW, Glass CK. Regulated accumulation of desmosterol integrates macrophage lipid metabolism and inflammatory responses. Cell. 2012 Sep 28;151(1):138-52. doi: 10.1016/j.cell.2012.06.054. PMID: 23021221; PMCID: PMC3464914.
Lipidome Signatures of Metastasis in a Transgenic Mouse Model of Sonic Hedgehog Medulloblastoma.
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Metabolic alternations were investigated by applying Ultra Performance Liquid Chromatography Mass Spectrometry (UPLC-MS) to mice brain tissue samples collected from SmoA1-Math-GFP mice with (n=18) and without (n=7) metastasis. All samples were analyzed using reverse phase (RP) UPLC-MS analysis in positive and negative ion modes.
Metabolomic study of disease progression in scrapie prion infected (RML) mice
STUDY_TYPE
Timecourse experiment
STUDY_SUMMARY
We used NMR spectroscopy to investigate metabolic perturbations in the brains of RML infected mice along the progression of prion disease at 30, 60, 90, 120, and 125 days post-inoculation.
Metabolomics of blood plasma and kidney tissue from control (db/m) and diabetic (db/db) mice.
STUDY_TYPE
MS/MSMS analysis
STUDY_SUMMARY
Plasma and kidney from control (db/m) and diabetic (db/db) mice on regular or eNOS -/- background. Some mice received treatment (20 mg/kg/d lisinopril + 30 mg/kg/d losartan) for 12 wk (starting at 12 wk age). Samples paired (plasma/tissue from individual mice). Samples snap frozen and stored at -80 until use.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
1000 wall St., Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
STUDY_COMMENTS
The wrong mouse number was recorded for P_29 and P_59 so they were excluded from analysis.
Determining secondary metabolite profile of Ilyonectria spp. pathogenic to ginseng root
STUDY_SUMMARY
This experiment aims to ascertain a profile of secondary metabolites produced by Ilyonectria species capable of causing disappearing root rot in ginseng. Ilyonectria isolates were grown on potato dextrose agar for 20 days, then plugs were taken from the cultures and extracted with ethyl acetate. Extracts were analyzed by LC-HRMS and tandem HRMS. Data were analyzed by Principal component analysis and molecular networking with GNPS.
Estimating Platelet Mitochondrial Function in Patients with Sepsis - Platelet NMRs (part-I)
STUDY_TYPE
single timepoint
STUDY_SUMMARY
Relationships between platelet mitochondrial oxygen consumption rates (mOCR) and metabolites in platelets as measured by quantitative 1H-NMR metabolomics. Samples collected in ED at a single timepoint. WB and platelets isolated from the same blood samples. Comparison of mitochondrial function and metabolomics in patients with sepsis and non-sepsis ED patients
INSTITUTE
University of Michigan, University of Mississippi, University of Minnesota
DEPARTMENT
Clinical Pharmacy (UMich); Emergency Medicine (UMiss)
Estimating Platelet Mitochondrial Function in Patients with Sepsis - WB NMRs (part-II)
STUDY_TYPE
single timepoint
STUDY_SUMMARY
Relationships between platelet mitochondrial oxygen consumption rates (mOCR) and metabolites in platelets as measured by quantitative 1H-NMR metabolomics in WB. Comparison of mitochondrial function and metabolomics in patients with sepsis and non-sepsis ED patients
INSTITUTE
University of Michigan, University of Mississippi, University of Minnesota
DEPARTMENT
Clinical Pharmacy (UMich); Emergency Medicine (UMiss)
Metabolomics and Hormonomics to Crack the Code of Filbert Growth
STUDY_SUMMARY
Introduction: Plants respond to changes in their environments through hormonal activation of a physiological cascade that redirects metabolic resources and growth. In filberts (Corylus sp.), chelated iron promotes the growth of new shoots but the mechanism(s) are not understood. Objectives: To use untargeted metabolomics and hormonomics approaches to generate novel hypotheses for the morphoregulatory role of ferric ethylenediamine-N,N'-di-(ortho-hydroxyphenyl) acetic acid (Fe-EDDHA) in filbert shoot organogenesis in vitro. Methods: Data were generated using previously optimized standardized untargeted metabolomics protocols with time of flight mass spectrometry. Multivariate statistical tools (principal component and partial least squares discriminant analysis) did not detect significant differences. Discovery tools Significance Analysis of Microarrays (SAM), multiple linear regression analysis, Bayesian analysis, logical algorithms, machine learning, synthetic biotransformations, targeted hormonomics, and online resources including MetaboAnalyst were used. Results: Starch/sucrose metabolism and shikimate pathway metabolites were increased. Dose dependent decreases were found in polyphenol metabolism, specifically ellagic acid and its methylated derivative 3,4,3'-tri-O-methylellagic acid. Hormonomics analysis revealed significant differences in phytohormones and their conjugates. FeEDDHA treatment reduced indole-3-acetic acid, abscisic acid, salicylic acid, jasmonic acid conjugates (JA-Trp, JA-Ile, OH-JA) and dihydrozeatinglucoside in regenerating explants. Serotonin (5HT) was decreased in FeEDDHA-treated regenerating tissues while the related metabolite melatonin was increased. Eight phenolic conjugates of 5HT and eight catabolites were affected by FeEDDHA indicating that metabolism to sequester, deactivate and metabolize 5HT was induced by Fe(III). Tryptophan was metabolized through kynurenine but not anthranilate. Conclusion: Seven novel hypotheses were generated to guide future studies to understand the regulatory control(s) of shoot organogenesis.
Metabolome Profiling of Synechococcus elongatus PCC 11801 strains engineered for Succinate Production
STUDY_TYPE
Measurement of relative metabolite pools of wild type and engineered Synechococcus elongatus PCC 11801 strains using Isotopic Ratio Method
STUDY_SUMMARY
Experiments to measure relative metabolite pools of wild type Synechococcus elongatus PCC 11801 and its recombinants producing succinate. The wild type and the engineered strains producing succinate were cultivated at 1% CO2 and their metabolome data was collected in three biological and three technical replicates (n=9). The study aims to find metabolomics changes between the wild type and the engineered to identify potential rate-limiting steps that be used as targets for improved production.
INSTITUTE
Indian Institute of Technology Bombay (IIT Bombay)
DEPARTMENT
Department of Chemical Engineering
LABORATORY
Bio systems Engineering Lab
LAST_NAME
Wangikar
FIRST_NAME
Pramod P
ADDRESS
Department of Chemical Engineering, IIT Bombay, Powai, Mumbai, Maharashtra, India - 400076
Metatranscriptomic Analysis of the Mouse Gut Microbiome Response to the Persistent Organic Pollutant 2,3,7,8-Tetrachlorodibenzofuran
STUDY_SUMMARY
Persistent organic pollutants (POPs) are important environmental chemicals and continued study of their mechanism of action remains a high priority. POPs, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), and polychlorinated biphenyls (PCBs), are widespread environmental contaminants that are agonists for the aryl hydrocarbon receptor (AHR). Activation of the AHR modulates the gut microbiome community structure and function, host immunity, and the host metabolome. In the current study, male C57BL6/J mice were exposed, via the diet, to 5 ug/kg body weight (BW) TCDF or 24 ug/kg BW of TCDF every day for 5 days. The functional and structural changes imparted by TCDF exposure to the gut microbiome and host metabolome were explored via 16S rRNA gene amplicon sequencing, metabolomics, and bacterial metatranscriptomics. Significant changes included increases in lipopolysaccharide (LPS) biosynthesis gene expression after exposure to 24 ug/kg BW of TCDF. Increases in LPS biosynthesis were confirmed with metabolomics and LPS assays using serum obtained from TCDF-treated mice. Significant increases in gene expression within aspartate and glutamate metabolism were noted after exposure to 24 ug/kg BW of TCDF. Together, these results suggest that after exposure to 24 ug/kg BW of TCDF, the gut microbiome increases the production of LPS and glutamate to promote localized gut inflammation, potentially using glutamate as a stress response.
INSTITUTE
The Pennsylvania State University (Penn State)
LAST_NAME
Nichols
FIRST_NAME
Robert
ADDRESS
917 Old Boalsburg Road, State College, Pennsylvania, 16801, USA
Metabolome Profiling of a Fast-growing Cyanobacterium Synechococcus elongatus PCC 11801 under Diurnal Cycle
STUDY_TYPE
Measurement of relative metabolite pools under diurnal cycle using isotopic ratio method
STUDY_SUMMARY
Experiments were carried out by growing Synechococcus elongatus PCC 11801 cells in multicultivators designed to provide sinusoidal light. The period was 14:10 (light-dark). The samples for metabolomics analysis were collected in the second diurnal cycle at an interval of 6 hours starting from 25th hour (25, 31, 37 and 43 hours). The light intensity amplitude was 600 Āµmole photons.m-2. s-1. Additionally samples were also collected using cells grown under continuous light illumination of 600 Āµmole photons.m-2. s-1 to compare the difference in metabolite levels compared to that in the diurnal cycle.
INSTITUTE
Indian Institute of Technology Bombay
DEPARTMENT
Department of Chemical Engineering
LABORATORY
Biosystems Engineering Lab
LAST_NAME
Wangikar
FIRST_NAME
Pramod P
ADDRESS
Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai, Maharashtra, India - 400076
Transforming growth factor Ī² (TGFĪ²) family comprises the main player in the development of fibrosis including three isoforms: TGFĪ²1, TGFĪ²2 and TGFĪ²3. TGFĪ²3 may play an antifibrotic role at the renal level, counteracting the role of TGFĪ²1, using a mouse model heterozygous for the TGFĪ²3 gene (TGFĪ²3+/-). Partial deletion of TGFĪ²3 causes in the mice albuminuria, loss of glomerular filtration rate, accelerated fibrosis, epithelial-to-mesenchymal transition and increment of glomerular basement membrane thickening.
Multi-omics analysis delineates the distinct functions of sub-cellular acetyl-CoA pools in Toxoplasma gondii
STUDY_SUMMARY
Acetyl-CoA is a key metabolite in all organisms, implicated in transcriptional regulation, post-translational modification as well as fuelling the TCA-cycle and the synthesis and elongation of fatty acids (FAs). The obligate intracellular parasite Toxoplasma gondii possesses two enzymes which produce acetyl-CoA in the cytosol and nucleus: acetyl-CoA synthetase (ACS) and ATP-citrate lyase (ACL), while the branched-chain Ī±-keto acid dehydrogenase-complex (BCKDH) generates acetyl-CoA in the mitochondrion. To obtain a global and integrative picture of the role of distinct sub-cellular acetyl-CoA pools, we measured the acetylome, transcriptome, proteome and metabolome of parasites lacking ACL/ACS or BCKDH. Loss of ACL/ACS results in the hypo-acetylation of nucleo-cytosolic and secretory proteins, alters gene expression broadly and is required for the synthesis of parasite-specific FAs. In contrast, loss of BCKDH causes few specific changes in the acetylome, transcriptome and proteome which allow these parasites to rewire their metabolism to adapt to the obstruction of the TCA-cycle.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Biomolecular analyses of hypospadias according to severity
STUDY_SUMMARY
Hypospadias, characterized by the displacement of the opening of the urethra at any point in the medial-ventral side of the penis, is classified upon severity as mild (Type I) and severe (Type II and Type III) hypospadias. Hypospadiasā etiology is idiopathic in the majority of cases, and underlying causes seem of multifactorial origin. Studies regarding genetic variants support this notion. It is unknown whether downstream gene products fit this profile. This study evaluated the metabolome of hypospadias by using the emerging technology of metabolomics in the search for distinct cellular processes associated with hypospadiasā etiology according to the severity of this congenital urogenital condition. Foreskin samples were collected during urethroplasty from boys with Type I, II, and III hypospadias or undergoing elective circumcision (N=28) between 5 to 28 months of age. Samples were processed and submitted to gas chromatography-mass spectrometry (GC/MS). MetaboloAnalyst (http://www.metaboanalyst.ca/) online platform was used for bioinformatic analyses. The metabolome of Type II and Type III hypospadias patients differs from the metabolome of Type I hypospadias and control patients. Thirty-five metabolites were identified by GC/MS. Of those, 14 metabolites, amino acids, were found in significantly low concentrations in Type II and Type III hypospadias in comparison to Type I hypospadias and controls. Amino acids comprised asparagine, aspartate, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, and tyrosine. The difference observed in the metabolome between severe and mild hypospadias supports previous research work of plausible severity-dependent etiologies for hypospadias. The observed downregulation of specific amino acids in severe hypospadias provides alternative routes for future research aiming to identify disrupted networks and pathways while considering the severity of hypospadias.
INSTITUTE
University of Puerto Rico, Medical Sciences Campus
DEPARTMENT
Anatomy & Neurobiology
LAST_NAME
PiƱeyro-Ruiz
FIRST_NAME
Coriness
ADDRESS
University of Puerto Rico, Medical Sciences Campus, Department of Anatomy & Neurobiology, Main Building, 5th Floor, Room A-521 PO BOX 365067 San Juan, PR 00936-5067
1H NMR metabolomics corroborates serine hydroxymethyltransferase as the primary target of 2-aminoacrylate in a ridA mutant of Salmonella enterica
STUDY_TYPE
NMR metabolomics on Salmonella enterica
STUDY_SUMMARY
The reactive intermediate deaminase RidA (EC: 3.5.99.10) is conserved across all domains of life and deaminates reactive enamine species. When S. enterica ridA mutants are grown in minimal medium, 2-aminoacrylate (2AA) accumulates, damages several pyridoxal 5ā-phosphate (PLP)- dependent enzymes, and elicits an observable growth defect. Genetic studies suggested that damage to serine hydroxymethyltransferase (GlyA), and the resultant depletion of 5,10-methelenetetrahydrofolate (5,10-mTHF), was responsible for the observed growth defect. However, the downstream metabolic consequence from GlyA damage by 2AA remains relatively unexplored. This study sought to use untargeted 1H NMR metabolomics to determine whether the metabolic state of a S. enterica ridA mutant was accurately reflected by characterizing growth phenotypes. The data supported the conclusion that metabolic changes in a ridA mutant were due to the IlvA-dependent generation of 2AA, and that the majority of these changes were a consequence of damage to GlyA. While many of the shifts in the metabolome of a ridA mutant could be explained, changes in some metabolites were not easily modeled, suggesting that additional levels of metabolic complexity remain to be unraveled.
INSTITUTE
University of Georgia
DEPARTMENT
Microbiology, Biochemistry, Complex Carbohydrate Research Center
LABORATORY
Edison Lab and Downs lab
LAST_NAME
Gouveia
FIRST_NAME
Goncalo
ADDRESS
315 riverbend road, Complex Carbohydrate Research Centre, ATHENS, GA, 30605, USA
Metabolite expression in liver after early life exposure to an endocrine disruptor at 240 days postnatal (part-I)
STUDY_TYPE
Metabolite expression after chemical exposure versus control.
STUDY_SUMMARY
Our early-life environment has a profound influence on developing organs that impact metabolic function and determines disease susceptibility across the life-course. Using a rat model for exposure to an endocrine disrupting chemical (EDC), we show that early-life exposure causes metabolic dysfunction in adulthood and reprograms histone marks in the developing liver to accelerate acquisition of an adult epigenomic signature. This epigenomic reprogramming persists long after the initial exposure, but many reprogrammed genes remain transcriptionally silent with their impact on metabolism not revealed until a later life exposure to a Western-style diet. Diet-dependent metabolic disruption was largely driven by reprogramming of the Early Growth Response 1 (EGR1) transcriptome and production of metabolites in pathways linked to cholesterol, lipid and one-carbon metabolism. These findings demonstrate the importance of epigenome: environment interactions, which early in life accelerate epigenomic aging, and later in adulthood unlock metabolically restricted epigenetic reprogramming to drive metabolic dysfunction.
Placental trophoblast cells are potentially at risk from circulating endocrine-disrupting chemicals, such as bisphenol A (BPA). To understand how BPA and the reputedly more inert bisphenol S (BPS) affect the placenta, C57BL6J mouse dams were fed 200 Ī¼g/kg body weight BPA or BPS daily for 2 wk and then bred. They continued to receive these chemicals until embryonic day 12.5, whereupon placental samples were collected and compared with unexposed controls. BPA and BPS altered the expression of an identical set of 13 genes. Both exposures led to a decrease in the area occupied by spongiotrophoblast relative to multinucleated giant cells (GCs) within the junctional zone, markedly reduced placental serotonin (5-HT) concentrations, and lowered 5-HT GC immunoreactivity. Concentrations of dopamine and 5-hydroxyindoleacetic acid, the main metabolite of serotonin, were increased. GC dopamine immunoreactivity was increased in BPA- and BPS-exposed placentas. A strong positive correlation between 5-HT+ GCs and reductions in spongiotrophoblast to GC area suggests that this neurotransmitter is essential for maintaining cells within the junctional zone. In contrast, an inverse correlation existed between dopamine+ GCs and reductions spongiotrophoblast to GC area. These outcomes lead to the following conclusions. First, BPS exposure causes almost identical placental effects as BPA. Second, a major target of BPA/BPS is either spongiotrophoblast or GC within the junctional zone. Third, imbalances in neurotransmitter-positive GC and an observed decrease in docosahexaenoic acid and estradiol, also occurring in response to BPA/BPS exposure, likely affect the placentalābrain axis of the developing mouse fetus.
INSTITUTE
University of Missouri
DEPARTMENT
Life Sciences Center
LABORATORY
Univ. of Missouri Metabolomics Center
LAST_NAME
Sumner
FIRST_NAME
Lloyd
ADDRESS
1201 Rollins Street Columbia, Missouri 65211-7310
EMAIL
sumnerlw@missouri.edu
PHONE
573-882-5486
NUM_GROUPS
3 treatment X 2 sex = 6
TOTAL_SUBJECTS
40
PUBLICATIONS
Mao et al, Proceedings National Academy of Science, USA, 2020
Lipid expression in liver after early lifer exposure to an endocrine disruptor at 70 days postnatal in the liver (part-II)
STUDY_TYPE
Lipid expression after chemical exposure versus control
STUDY_SUMMARY
Our early-life environment has a profound influence on developing organs that impact metabolic function and determines disease susceptibility across the life-course. Using a rat model for exposure to an endocrine disrupting chemical (EDC), we show that early-life exposure causes metabolic dysfunction in adulthood and reprograms histone marks in the developing liver to accelerate acquisition of an adult epigenomic signature. This epigenomic reprogramming persists long after the initial exposure, but many reprogrammed genes remain transcriptionally silent with their impact on metabolism not revealed until a later life exposure to a Western-style diet. Diet-dependent metabolic disruption was largely driven by reprogramming of the Early Growth Response 1 (EGR1) transcriptome and production of metabolites in pathways linked to cholesterol, lipid and one-carbon metabolism. These findings demonstrate the importance of epigenome:environment interactions, which early in life accelerate epigenomic aging, and later in adulthood unlock metabolically restricted epigenetic reprogramming to drive metabolic dysfunction.
Lipid expression in serum after early lifer exposure to an endocrine disruptor at 70 days postnatal (part-III)
STUDY_TYPE
Lipid expression after chemical exposure versus control.
STUDY_SUMMARY
Our early-life environment has a profound influence on developing organs that impact metabolic function and determines disease susceptibility across the life-course. Using a rat model for exposure to an endocrine disrupting chemical (EDC), we show that early-life exposure causes metabolic dysfunction in adulthood and reprograms histone marks in the developing liver to accelerate acquisition of an adult epigenomic signature. This epigenomic reprogramming persists long after the initial exposure, but many reprogrammed genes remain transcriptionally silent with their impact on metabolism not revealed until a later life exposure to a Western-style diet. Diet-dependent metabolic disruption was largely driven by reprogramming of the Early Growth Response 1 (EGR1) transcriptome and production of metabolites in pathways linked to cholesterol, lipid and one-carbon metabolism. These findings demonstrate the importance of epigenome:environment interactions, which early in life accelerate epigenomic aging, and later in adulthood unlock metabolically restricted epigenetic reprogramming to drive metabolic dysfunction.
Lipid expression in serum after early life exposure to an endocrine disruptor and a Western Diet at 240 days postnatal (part-IV)
STUDY_TYPE
Lipid expression after chemical exposure versus control
STUDY_SUMMARY
Our early-life environment has a profound influence on developing organs that impact metabolic function and determines disease susceptibility across the life-course. Using a rat model for exposure to an endocrine disrupting chemical (EDC), we show that early-life exposure causes metabolic dysfunction in adulthood and reprograms histone marks in the developing liver to accelerate acquisition of an adult epigenomic signature. This epigenomic reprogramming persists long after the initial exposure, but many reprogrammed genes remain transcriptionally silent with their impact on metabolism not revealed until a later life exposure to a Western-style diet. Diet-dependent metabolic disruption was largely driven by reprogramming of the Early Growth Response 1 (EGR1) transcriptome and production of metabolites in pathways linked to cholesterol, lipid and one-carbon metabolism. These findings demonstrate the importance of epigenome:environment interactions, which early in life accelerate epigenomic aging, and later in adulthood unlock metabolically restricted epigenetic reprogramming to drive metabolic dysfunction.
Metabolomic Profiles of Pancreatic Ī²-Cells and Islets Exposed to Arsenic, Islets (part-II)
STUDY_SUMMARY
Type-2 diabetes (T2D) is a complex metabolic disorder that affects hundreds of millions of people world-wide and is a growing public health concern. Despite recent advances in T2D research, the etiology of this disease and the mechanisms underlying the metabolic defects remain poorly understood. While obesity is thought to be the main cause for the rising prevalence of T2D, obesity alone cannot explain differences in the trends of T2D among different geographical regions and populations. Growing evidence suggests that environmental exposures to toxic and diabetogenic substances must play important roles. Inorganic arsenic (iAs) is a naturally occurring toxic metalloid. Hundreds of millions of people worldwide are exposed to unsafe levels of iAs in drinking water and food. iAs is a potent carcinogen, but iAs exposure has also been linked to increase risk of T2D. While the link between iAs exposure and T2D is well-established, the mechanisms underlying the diabetogenic effects of iAs exposure remain unclear. Results of our previously published and ongoing studies suggest that pancreatic islets are a primary target for iAs and its metabolites and that impaired insulin secretion by islets is the mechanism by which iAs exposure leads to diabetes. The proposed project will use metabolomics to identify metabolic pathways in Ī²-cells that are targeted by iAs and its metabolites, monomethyl-As (MAs) and dimethyl-As (DMAs). The metabolomics data combined with results of our ongoing mechanistic studies will provide a comprehensive picture of the metabolic dysfunction leading to the development of diabetes in individuals exposed to iAs and of the molecular mechanisms that underlie this dysfunction. Identifying the affected pathways and mechanisms will ultimately help to improve strategies for prevention and/or treatment of T2D associated with chronic exposure to iAs.
Retargeting azithromycin-like compounds as antimalarials with dual modality
STUDY_SUMMARY
Resistance to front-line antimalarials (artemisinin combination therapies) is spreading, and development of new drug treatment strategies to rapidly kill Plasmodium parasites that cause malaria are urgently needed. Here, we show that azithromycināa clinically used macrolide antibiotic that targets the bacterium-like ribosome of the malaria parasites apicoplast organelle and causes a slow-killing ādelayed deathā phenotypeācan also rapidly kill parasites throughout the asexual blood-stages of the lifecycle via a āquick-killingā mechanism of action. Investigation of 84 azithromycin analogues revealed nanomolar quick-killing potency that is directed against the very earliest stage of parasite development within red blood cells. Indeed, the best analogue exhibited 1600-fold higher potency than azithromycin for in vitro treatment windows less than 48 hours. Analogues were also effective against the zoonotic malaria parasite P. knowlesi, and against both multi-drug and artemisinin resistant P. falciparum lines. Metabolomic profiles of azithromycin analogue treated parasites were similar to those of chloroquine treated parasites, suggesting that the quick-killing mechanism of action may in part be localised to the parasite food vacuole. However, metabolomic signatures associated with mitochondrial disruption were also present. In addition, unlike chloroquine, azithromycin and analogues were active across blood stage development, including merozoite invasion, suggesting that these macrolides have a multi-factorial mechanism of quick-killing activity. The positioning of functional groups added to azithromycin and its quick-killing analogues altered their activity against bacterial-like ribosomes but had minimal change on quick-killing activity, which suggests that apicoplast-targeting, delayed-death activity can either be preserved or removed independently of quick-killing. Apicoplast minus parasites remained susceptible to both azithromycin and its analogues, further demonstrating that quick-killing is independent of apicoplast-targeting, delayed-death activity. Therefore, development of azithromycin and analogues as antimalarials offers the possibility of targeting parasites through both a quick-killing and delayed death mechanism of action in a single, multifactorial chemotype.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Time-course experiment of Microchloropsis gaditana cells supplemented with CO2
STUDY_TYPE
Time-course experiment
STUDY_SUMMARY
Experiments were conducted with Microchloropsis gaditana supplemented with very-low CO2 and high CO2. Sampling was done on the following time points: Day 3, 6 and 9.
INSTITUTE
International Centre for Genetic Engineering and Biotechnology
DEPARTMENT
Integrative Biology
LABORATORY
Omics of Algae
LAST_NAME
Jutur
FIRST_NAME
Pannaga Pavan
ADDRESS
2nd Floor, New Building, ICGEB, Aruna Asaf Ali Marg, New Delhi - 110067
EMAIL
jppavan@icgeb.res.in
PHONE
+91 11 26741358
STUDY_COMMENTS
Former name of species: Nannochloropsis gaditana LubiƔn
Dynamics of Exposure, Phthalates, and Asthma in a Randomized Trial (DEPART)
STUDY_TYPE
CHEAR Study
STUDY_SUMMARY
project will investigate relationships between phthalate exposure, pediatric asthma health, and underlying biological pathways of toxicity among a rural, underserved Latino population located in Yakima Valley, WA. DEPART will benefit from the original studyās (HAPIās) robust longitudinal repeat-measure design and community-engaged framework. DEPART will add new measurements including concentrations of urinary phthalate monoester metabolites and biomarkers of oxidative stress to better characterize exposure-response associations. This projectās primary goal is to deepen the understanding of pathophysiological phenomena underlying exposure-response relationships between phthalates and asthma health. Our specific aims are: (1) Characterize associations between urinary phthalate metabolite concentrations and short-term asthma morbidity, and (2) Determine individual relationships between urinary phthalate metabolite concentrations, short-term asthma morbidity, and biomarkers for oxidative stress to assess the potential for a mediating effect by oxidative stress. Covariates of interest will include atopic status, randomized intervention grouping, and the caregiver psychosocial stress assessment.
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Department of Environmental Medicine and Public Health
Maternal adiposity alters the human milk metabolome: a link between non-glucose monosaccharides and infant adiposity
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
An untargeted metabolomics analysis of human milk was performed to test the hypothesis that a unique human milk metabolome would emerge based on maternal adiposity (maternal fat mass and body mass index). This study also aimed to identify differentially expressed milk metabolites that are associated with fat mass in the infant. To our knowledge this study reports on the largest cohort to date examining the metabolomic differences in human milk composition between normal weight and obese women. Data generated from this study indicate the need for further research in the area of human milk metabolomics and the potential role for human milk small molecules in contributing to offspring growth and development.
INSTITUTE
University of California, Davis
DEPARTMENT
West Coast Metabolomics Center
LAST_NAME
Paglia
FIRST_NAME
Kelly
ADDRESS
451 Health Sciences Drive, Room 1313, Davis, CA, 95616, USA
We analyzed mouse serum samples from a mouse dietary intervention experiment. Briefly, C57BL/6 mice (n=44) were divided into 4 groups (n=11 per group) and fed High-fat diet (HFD), 1% deoxycholic acid (DCA) in drinking water, both, or left as control for 9 months. For quality control, 12 TQC samples and 2 blanks were also included in the analysis (total 58 samples and 6 groups). The two treatments were selected to demonstrate the ability of lipidomics to detect gross changes induced by HFD in the serum lipidome, as well as specific/minor changes induced by the secondary bile acid (DCA) through regulation of liver lipid metabolism.
Metabolomics Adaptation of Juvenile Pacific Abalone Haliotis discus hannai to Heat Stress
STUDY_SUMMARY
We compared two groups of abalones (from the same population) with different temperature acclimation history, through their survival at acute heat treatment and metabolites changes. The results indicated significantly higher survival for the high temperature acclimation group. Both groups experienced mitochondrial homeostasis break down during heat treatment, while the higher temperature acclimation group accumulated more metabolites beneficial to metabolic homeostasis, including various dipeptides, antioxidants, and neuroprotective substances.
INSTITUTE
Institute of Oceanology, Chinese Academy of Sciences
Obesity and Poor Diet as Susceptibility Factors for Secondhand Smoke in Childhood Asthma
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Aim 1: To investigate the effect of SHS exposure on asthma morbidity, systemic inflammation and oxidative stress among inner-city children with asthma. Hypotheses 1.Increasing SHS exposure will be associated with increases in asthma morbidity, systemic inflammation and oxidative stress. Aim 2: To determine if being overweight/obese modifies the effect of SHS exposure on respiratory symptoms, inflammation and oxidative stress responses among inner-city children with asthma.Hypothesis 2. SHS exposure will be associated with a worsened asthma and increases in systemic inflammation and oxidative stress among overweight/obese children compared to normal weight children. Aim 3: To determine if diet quality modifies the effect of SHS exposure on respiratory symptoms, inflammation and oxidative stress responses among inner-city children with asthma. Hypothesis 3. SHS smoke exposure will associated with worsened respiratory symptoms and increases in inflammation and oxidative stress among children with poor quality diet compared those with better quality diet inner-city children with asthma. (Diet will be assessed by dietary inflammatory index, healthy eating index, and additional serum markers proposed in this application).
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Kachman
FIRST_NAME
Maureen
ADDRESS
Ann Arbor, MI
EMAIL
mkachman@med.umich.edu
PHONE
734-232-0842
TOTAL_SUBJECTS
185
STUDY_COMMENTS
Low-income minority children in urban areas often inhabit an environment that has excessive pollution and these children are at risk for obesity and poor diet. Secondhand smoke (SHS) is a common exposure in homes of children in Baltimore City and approximately 50-80% of households have a smoker. Secondhand smoke has been associated with worse asthma outcomes. We hypothesize that secondhand smoke exposure will be associated with increases in asthma morbidity and increases in systemic markers of inflammation and oxidative stress and that these responses will be exaggerated among overweight and obese children compared with normal weight and among those with poor diets compared to better diets among children with asthma.
Untargeted lipidome changes in Chlamydomonas reinhardtii treated with small molecules containing adamantane structures
STUDY_SUMMARY
A study to investigate the effect of small molecule lipid inducing compounds that leads to hyper accumulation of lipids in N replete cells of Chlamydomonas reinhardtii. These compounds were identified through a high throughput screening designed for that purpose. During that screening, we screened 43,783 compounds and identified 367 primary hits. These 367 hits were further retested using a 8-point dilution series (from 0.25 to 30 uM) and verified the activity of 250 compounds that induce the hyper lipid accumulating phenotype in algae. Once the hit compounds were identified and confirmed, we then performed extensive chemoinformatics analysis to look for common scaffolds and identified several common substructures. We then selected 15 top performing compounds from 5 diverse structural groups and tested biochemical parameters such as growth, lipid accumulating capacity, effect on photosynthetic rates, respiration rates, oxygen consumption rates, analysis of different lipid species to quantify and identify fatty acid species using GC-MS. To understand the global changes in the lipidome, 2 structurally similar compounds were selected and compared with cells grown without compounds as control for untargeted lipidome analysis.
Mitochondrial Lipid Profiles in Traumatic Optic Neuropathy
STUDY_SUMMARY
C57Bl/6J Mice were exposed to sonication to induce a traumatic optic neuropathy model. Optic nerves were harvested and mitochondrial isolation was performed. Lipids were then extracted from isolated mitochondria. Mass spectrometry was used to analyze mitochondrial lipids.
INSTITUTE
University of Miami
DEPARTMENT
Ophthalmology, Bascom Palmer Eye Institute
LAST_NAME
Bhattacharya
FIRST_NAME
Sanjoy K
ADDRESS
1638 NW 10th Avenue, Suite 707A University of Miami Miami, FL, 33136
Multiplatform, non-targeted analysis of lung extracts of uninfected and Mtb-infected C57BL/6 mice at 4 and 9 weeks p.i.
STUDY_SUMMARY
The goal of this multiplatform, non-targeted metabolomics study was to explore the metabolic alterations occurring during the natural progression of pulmonary tuberculosis in a murine model of disease (C57BL/6 genotype). For this purpose, we used gas chromatography, capillary electrophoresis, and reversed-phase liquid chromatography coupled to high-resolution mass analyzers (GC-EI-QTOF/MS, CE-ESI(+)-QTOF/MS, LC-ESI(+)-QTOF/MS and LC-ESI(-)-QTOF/MS to analyze lung extracts of age and sex-matched uninfected mice (UW, n=4), Mycobacterium tuberculosis-infected mice at 4 weeks post-infection (4W, n=4) and Mycobacterium tuberculosis-infected mice at 9 weeks post-infection. All data were acquired in MS1 mode, following a canonical non-targeted workflow.
INSTITUTE
Universidad San Pablo-CEU, CEU Universities
DEPARTMENT
Departamento de Quimica y Bioquimica
LABORATORY
Centro de Metabolomica y Bioanalisis (CEMBIO)
LAST_NAME
Fernandez Garcia
FIRST_NAME
Miguel
ADDRESS
Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, UrbanizaciĆ³n MonteprĆncipe, 28660 Boadilla del Monte, Spain
SS-31 and NMN: Two Paths to Improve Metabolism and Function in Aged Hearts
STUDY_SUMMARY
The effects of two different mitochondrial-targeted drugs, SS-31 and NMN, were tested on Old mouse hearts. After treatment with the drugs, individually or Combined, heart function was examined by echocardiography. SS-31 partially reversed an age-related decline in diastolic function while NMN fully reversed an age-related deficiency in systolic function at a higher workload. Metabolomic analysis revealed that both NMN and the Combined treatment increased nicotinamide and 1-methylnicotinamide levels, indicating greater NAD+ turnover, but only the Combined treatment resulted in significantly greater steady state NAD(H) levels. A novel magnetic resonance approach was used to assess how metabolite levels responded to changing workload. PCr/ATP decreased in response to increased workload in Old Control, but not Young, hearts, indicating an age-related decline in energetic capacity. Both drugs were able to normalize the PCr/ATP dynamics. SS-31 and NMN treatment also increased mitochondrial NAD(P)H production under the higher workload while only NMN increased NAD+ in response to the work jump. These measures did not shift in hearts given the Combined treatment, which may be owed to the enhanced NAD(H) levels in the resting state after this treatment. Overall, these results indicate that both drugs are effective at restoring different aspects of mitochondrial and heart health and that combining them results in a synergistic effect that rejuvenates Old hearts and best recapitulates the Young state.
Multi-omics of OsGF14b-mediated innate immunity against panicle blast in rice
STUDY_SUMMARY
In the present study, we used a multi-omics approach to decipher the molecular mechanisms of OsGF14b in governing panicle resistance to Magnaporthe oryzae.Results revealed OsGF14b mediated panicle blast resistance was involved in the activation of auxin and JA signaling pathways, resulting in reprogramming of the phenylpropanoid and diterpenoid pathway.
INSTITUTE
Agro-biological Gene Research Center , Guangdong Academy of Agricultural Sciences
Multi-omics of OsGF14b-mediated innate immunity against panicle blast in rice (part-II)
STUDY_SUMMARY
In the present study, we used a multi-omics approach to decipher the molecular mechanisms of OsGF14b in governing panicle resistance to Magnaporthe oryzae.Results revealed OsGF14b mediated panicle blast resistance was involved in the activation of auxin and JA signaling pathways, resulting in reprogramming of the phenylpropanoid and diterpenoid pathway.
INSTITUTE
Agro-biological Gene Research Center , Guangdong Academy of Agricultural Sciences
Huntingtonās Disease Genotype Suppresses Global Manganese-Responsive Processes
STUDY_TYPE
Untargeted metabolomics analysis
STUDY_SUMMARY
Global untargeted metabolomics studies performed in the striatum tissue, the brain region most sensitive to neurodegeneration in Huntingtonās Disease, to investigate global Mn-dependent and Mn-responsive biology following various Mn exposures in a mouse model of HD.
Deadly Duality of PEBP1: Shutting off Necroptosis, Turning on Ferroptosis
STUDY_TYPE
Observation study
STUDY_SUMMARY
Necroptosis and ferroptosis are two pathways of regulated cell death executed in several major cardiovascular and neurological acute and degenerative diseases. While the necroptosis program relies on activation of RIP1, RIP3 kinases and MLKL, ferroptotic death is triggered by 15- Lipoxygenase (15LO) catalyzed oxidation of arachidonoyl- (AA) or adrenoyl- (AdA) phosphatidylethanolamines (PE) controlled by the phosphatidylethanolamine-binding protein 1 (PEBP1). PEBP1 displays āregulatoryā promiscuity towards multiple protein partners, including RAF1 kinase. Given a distinct structural homology between RAF1 kinase and RIP3 kinase, we hypothesized that PEBP1 may interact with RIP3 and act as a switch from necroptosis to ferroptosis. Using computational, genetic and redox lipidomics approaches, we show that PEBP1 liberated from RAF1 kinase binds and sterically inhibits RIP3 thus turning-off necroptosis. Highly expressed 15LO may outcompete and bind PEBP1 to promote AA-PE/AdA-PE oxidation and ferroptosis. Using cell- based and animal models, we identified the conditions disrupting PEBP1ās interactions with RAF1 kinase to alternatively bind/inhibit RIP3 kinase or bind/activate 15LO. We further established that PEBP1 knockdown sensitizes cells to RIP3-mediated necroptosis. These newly established regulatory functions of PEBP1 serve multiple and diverse roles across various human disease states.
Air Pollution, Placenta Function, and Birth Outcomes in Los Angeles in the Placental Assessment in Response to ENvironmenTal pollution study (PARENTs) cohort
STUDY_SUMMARY
This project aims to evaluate the internal environmental exposome of mother/fetus pairs within the PARENTs cohort, assessing a wide range of biomarkers/internal exposure measures for environmental toxins including air pollutants. Focusing on advanced imaging data on placental development and robust, clinically-confirmed birth outcomes, this a hypothesis-generating, untargeted pilot metabolomics study aims to identify potential predictors of placental insufficiencies and adverse birth outcomes. Measurements of exposures which include residential, occupational, and behavioral exposures, along with personalized air pollution measures, will help us in identifying related metabolomics patterns.
Effect of high-fat diet and bile acid treatment on serum and tissue lipidomes in mice
STUDY_SUMMARY
We analyzed mouse serum and esophageal tissue samples from a mouse dietary intervention experiment. Briefly, C57BL/6 mice (n=44) were divided into 4 groups (n=11 per group) and fed High-fat diet (HFD), 0.2% deoxycholic acid (DCA) in drinking water, both, or left as control for 9 months. For quality control,TQC samples and blanks were also included in the analysis. The two treatments were selected to demonstrate the ability of lipidomics to detect gross changes induced by HFD in the serum and tissue lipidomes, as well as specific/minor changes induced by the secondary bile acid (DCA) through regulation of liver lipid metabolism. The serum and tissue samples were analyzed using targeted and untargeted lipidomics methods. The targeted serum lipidomics data has previously been uploaded as part of study ST001323.
The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that responds to a variety of structurally diverse exogenous and endogenous small molecules. Gut microbiota utilizing tryptophan and indole metabolism as a reservoir, has been demonstrated to provide an abundant source of AHR ligands. So untargeted global profiling was performed to find the potential candidates of AHR activator in human feces.
INSTITUTE
The Pennsylvania State University
LAST_NAME
DONG
FIRST_NAME
FANGCONG
ADDRESS
314 Life Sciences Building, University Park, PA, 16802
The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that responds to a variety of structurally diverse exogenous and endogenous small molecules. Gut microbiota utilizing tryptophan and indole metabolism as a reservoir, has been demonstrated to provide an abundant source of AHR ligands. So differential analysis was performed to find the potential candidates of AHR activator in cecal contents between conventional and germ-free mice with the help of untargeted global profiling.
INSTITUTE
Pennsylvania State University
LAST_NAME
DONG
FIRST_NAME
FANGCONG
ADDRESS
314 Life Sciences Building, University Park, PA 16802
Disruption of Redox Balance Enhances the Effects of BRAF-inhibitors in Melanoma
STUDY_SUMMARY
Specifically, we report that drug-insensitive melanoma cells can maintain higher levels of antioxidant metabolites to withstand the lethal effects of drugs. By extending our analysis to other melanoma subtypes in the TCGA, we show that elevated redox capacity could indeed be a general feature of melanoma. Our results suggest that redox vulnerabilities could be exploited for therapeutic benefits and identify unsuspected combination targets to enhance the effects of BRAFi in pan-melanoma.
C56BL6 BMDM stimulated with different TLRs W/O acetylated LDL (part-II)
STUDY_TYPE
Acetylated LDL
STUDY_SUMMARY
C57BL6 bone marrow derived macrophages were preloaded with 50 ug/mL of acetylated LDL and stimulated with TLR2, TLR3, TLR4, TLR7, and TLR9 ligands for 48 h.
Metabolomic changes in mouse liver on a protein restricted diet
STUDY_SUMMARY
The data provided here are in support of the publication āRestriction of essential amino acids dictates the systemic metabolic response to dietary protein dilutionā Yann W. Yap et al. Nature Communications 2020 (accepted for publication). Here we provide untargeted metabolomics LC-MS data from liver from C57Bl/6NCrl mice fed a diet in which dietary protein was restricted and corresponding unrestricted controls. Specifically, liver from animals on a low-protein diet following a week of diet adaptation and correspond controls with n = 5 for each group.
Metabolomic changes in mouse plasma on a protein restricted diet
STUDY_SUMMARY
The data provided here are in support of the publication āRestriction of essential amino acids dictates the systemic metabolic response to dietary protein dilutionā Yann W. Yap et al. Nature Communications 2020 (accepted for publication). Here we provide untargeted metabolomics LC-MS data from plasma from C57Bl/6NCrl mice fed a diet in which dietary protein was restricted and corresponding unrestricted controls. Specifically, plasma from animals on a low-protein diet following a week of diet adaptation and correspond controls with n = 5 for each group.
Metabolomic changes in mouse liver on a threonine restricted diet (part-III)
STUDY_SUMMARY
The data provided here are in support of the publication āRestriction of essential amino acids dictates the systemic metabolic response to dietary protein dilutionā Yann W. Yap et al. Nature Communications 2020 (accepted for publication). The data in this study corresponds to liver from C57Bl/6JMarp mice subject to a 3wk treatment with diets either containing 18% digestible energy from amino acids with either a normal distribution of amino acids or with restricted amounts of threonine. This followed prior treatment with adeno-associated viruses to transduce the liver to express yeast threonine biosynthetic enzymes (AAV-yTHR1+THR4) or a negative control (AAV-GFP). n= 6 individual mice per group.
NMR Metabolomic Analysis of Bacterial Resistance Pathways Using Multivalent Quaternary Ammonium Functionalized Macromolecules
STUDY_TYPE
NMR Hydrophilic Metabolomics
STUDY_SUMMARY
Multivalent antimicrobial dendrimers are an exciting new system that is being developed to address the growing problem of drug resistant bacteria. Nuclear Magnetic Resonance (NMR) metabolomics is a quantitative and reproducible method for the determination of bacterial response to environmental stressors and for visualization of perturbations to biochemical pathways. NMR metabolomics is used to elucidate metabolite differences between wild type and antimicrobially mutated Escherichia coli (E. coli) samples. Proton (1H) NMR hydrophilic metabolite analysis was conducted on samples of E. coli after 33 growth cycles of a minimum inhibitory challenge to E. coli by poly(amidoamine) dendrimers functionalized with mannose and with C16-DABCO quaternary ammonium endgroups and compared to the metabolic profile of wild type E. coli. The wild type and mutated E. coli samples were separated into distinct sample sets by hierarchical clustering, principal component analysis (PCA) and sparse partial least squares discriminate analysis (sPLS-DA). Metabolite components of membrane fortification and energy related pathways had a significant p-value and fold change between the wild type and mutated E. coli. Amino acids commonly associated with membrane fortification from cationic antimicrobials, such as lysine, were found to have a higher concentration in the mutated E. coli than the wild type E. coli. N-acetylglucosamine, a major component of peptidoglycan synthesis, was found to have a 25 fold higher concentration in the mid log phase of the mutated E. coli than the mid log phase of the wild type.The metabolic profile suggests that E. coli change their peptidoglycan composition in order to garner protection from the highly positively charged and multivalent C16-DABCO and mannose functionalized dendrimer.
INSTITUTE
Montana State University
DEPARTMENT
Chemistry and Biochemistry
LABORATORY
Cloninger
LAST_NAME
Aries
FIRST_NAME
Michelle
ADDRESS
Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717 USA
Untargeted metabolomics in skeletal muscle of mice with chronic kidney disease
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
This study performed untargeted metabolomics analysis of skeletal muscle obtained form mice with and without chronic kidney disease.
INSTITUTE
University of Florida
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
EMAIL
ryant@ufl.edu
PHONE
352-294-1700
NUM_GROUPS
4
TOTAL_SUBJECTS
18
NUM_MALES
8
NUM_FEMALES
10
STUDY_COMMENTS
two control male samples processed mistakenly were from soles muscles, while all other samples were gastrocnemius muscles. Due to differences in fiber type proportions, soleus muscles were not used in final analysis
Exposure to paraben was associated with allergic outcomes, partially through the metabolomics changes. Urinary metabolomic analysis can be useful to elucidate the mechanisms underlying the associations between exposure to paraben and allergic outcomes.
INSTITUTE
Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital
Diel Metabolites in the North Pacific Subtropical Gyre (KM1513)
STUDY_TYPE
Diel metabolomics
STUDY_SUMMARY
Diverse organisms within the marine microbial communities show 24-hour cycles of gene expression, likely driven by the need to harness energy from sunlight and to cope with dramatic fluctuations in solar radiation over the course of the day. Metabolites are the direct product of metabolic activity; they are therefore expected to both reflect and influence the daily cycle of the microbial community. Here we measure the intracellular metabolome of the microbial community of the North Pacific Subtropical Gyre, sampled at 4-hour intervals for 8 days. Concentrations of some metabolites common to many organisms exhibit diel periodicity, revealing synchrony of community-level metabolism. Comparing these data to gene expression data reveals temporal offsets between gene transcription and cellular activity, and ties some metabolites to the activities of specific organisms. For example, the dramatic fluctuations of the disaccharide trehalose likely reflect the daily cycles of {Crocosphaera}, a photosynthesizing cyanobacteria that needs to store energy during the day to fuel nighttime nitrogen-fixation. This study illustrates how pairing multiple types of 'omics and environmental data can provide insight into how the activities of individual organisms lead to community functions such as net primary productivity and nitrogen fixation.
Longitudinal wastewater sampling and untargeted metabolomics of three buildings
STUDY_SUMMARY
Direct sampling of building wastewater has the potential to enable "precision public health" observations and interventions. Temporal sampling offers additional dynamic information that can be used to increase the informational content of individual metabolic āfeaturesā, but few studies have focused on high-resolution sampling. Here, we sampled three spatially close buildings, revealing individual metabolomics features, retention time (rt) and mass-to-charge ratio (mz) pairs, that often possess similar stationary statistical properties, as expected from aggregate sampling. However, the temporal profiles of featuresāproviding orthogonal information to physicochemical propertiesāillustrate that many possess different feature temporal dynamics (fTDs) across buildings, with large and unpredictable single day deviations from the mean. Internal to a building, numerous and seemingly unrelated features, with mz and rt differences up to hundreds of Daltons and seconds, display highly correlated fTDs, suggesting non-obvious feature relationships. Data-driven building classification achieves high sensitivity and specificity, and extracts building-identifying features found to possess unique dynamics. Analysis of fTDs from many short-duration samples allows for tailored community monitoring with applicability in public health studies.
Monophasic lipidomics extraction in cancer cell line
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
We performed a comprehensive characterization of a monophasic extraction method in cancer cell lines. We used pharmacological perturbation on HepG2 cells to identify changes in different lipid families. We optimized the MS parameters, the chromatography and the data analysis to perform rapid and robust lipidomics analysis from cell lines.
Serum was processed using a targeted metabolomics platform for quantifying tryptophan metabolites as a number of these metabolites are well establish uremic toxins.
INSTITUTE
University of Florida
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Effect of BPA and genistein exposure on the fecal metabolome
STUDY_SUMMARY
Xenoestrogens are found in plant products, such as genistein (GEN), or industrial chemicals, such as bisphenol A (BPA), present in consumer products that are also pervasive in the environment. Early exposure to such endocrine disrupting chemicals (EDC) may affect neural development by inducing direct neural effects and/or through the microbiome-gut-brain axis. To test this hypothesis, California mice (Peromyscus californicus) offspring were exposed through the maternal diet to GEN (250 mg/kg feed weight) or BPA (5 mg/kg feed weight, low dose- LD and 50 mg/kg, upper dose-UD), and dams were placed on these diets two weeks prior to breeding, throughout gestation, and lactation. Various behaviors, gut microbiome, and fecal metabolome were assessed starting at 90 days of age. The LD but not UD of BPA resulted in individuals spending more time engaging in repetitive behaviors. GEN exposed individuals were more likely to exhibit such behaviors and showed socio-communicative disturbances. BPA and GEN exposed females had increased number of metabolites involved in carbohydrate metabolism and synthesis.. Males exposed to BPA or GEN showed alterations in lysine degradation and phenylalanine and tyrosine metabolism. Current findingsindicate cause for concern that developmental exposure to BPA or GEN might affect the microbiome-gut-brain axis.
INSTITUTE
University of Missouri
DEPARTMENT
MU Metabolomics Center
LAST_NAME
Sarma
FIRST_NAME
Saurav
ADDRESS
1201 Rollins street, 243 Bond Life Science Center, University of Missouri, Columbia, MO 65211, USA
Monophasic lipidomics extraction in cancer cell lines
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
We performed a comprehensive characterization of a monophasic extraction method in cancer cell lines. We used pharmacological perturbation on HepG2 cells to identify changes in different lipid families. We optimized the MS parameters, the chromatography and the data analysis to perform rapid and robust lipidomics analysis from cell lines.
INSTITUTE
IGMM
LAST_NAME
Rodriguez Blanco
FIRST_NAME
Giovanny
ADDRESS
Crewe Road South, Edinburgh, Midlothian, EH42XU, United Kingdom
Core Functional Nodes and Sex-Specific Pathways in Human Ischemic and Dilated Cardiomyopathy
STUDY_SUMMARY
Restricted access to human left ventricular myocardium is a significant limitation in the study of heart failure (HF). Here, we utilise a large human heart biobank of carefully procured, cryopreserved left ventricular myocardium to obtain direct molecular insights into ischaemic (ICM) and dilated cardiomyopathy (DCM), the most common causes of HF worldwide1. We performed unbiased, deep proteomic and metabolomic analyses of 51 left ventricular (LV) samples from 44 cryopreserved human ICM and DCM hearts, including age-matched, histopathologically normal, donor controls of both genders for comparison. For the first time, we report perturbed thyroid hormone signalling pathways in the myocardium of both types of HF, and unveil the interaction of gender with HF, including increased nitric oxide-related arginine metabolism in male hearts, and many gender-specific mitochondrial and X chromosome-linked protein and metabolite changes. We provide all raw data, in addition to an interactive online application, as a publicly-available resource.
RP-UPLC-FTMS (+/- ion detection) were conducted on hippocampi from healthy (CON) and malnourished (MAL) mice, and MAL-BG (malnutrition plus E.coli/Bacteroidales exposure) mice.
INSTITUTE
University of British Columbia
DEPARTMENT
Microbiology and Immunology
LABORATORY
B. Brett Finlay
LAST_NAME
Bauer
FIRST_NAME
Kylynda
ADDRESS
Michael Smith Laboratories, #301 ā 2185 East Mall, University of British Columbia Vancouver, British Columbia Canada, V6T 1Z4
Malnutrition and Liver Metabolomics Pre-Intervention (part-I)
STUDY_SUMMARY
RP-UPLC-FTMS (+/- ion detection) and HILIC-FTMS (+/- ion detection) were conducted on murine livers from healthy (CON) and malnourished (MAL) mice. To examine the impact of gut microbes and malnutrition, data was also collected from a third group (MBG).
INSTITUTE
University of British Columbia
DEPARTMENT
Microbiology and Immunology
LABORATORY
B. Brett Finlay
LAST_NAME
Bauer
FIRST_NAME
Kylynda
ADDRESS
Michael Smith Laboratories, #301 ā 2185 East Mall, University of British Columbia Vancouver, British Columbia Canada, V6T 1Z4
Malnutrition and Liver Metabolomics Intervention (part-II)
STUDY_SUMMARY
RP-UPLC-FTMS (+/- ion detection) and HILIC-FTMS (+/- ion detection) were conducted on murine livers from healthy (CON) and malnourished model (MBG) mice. To examine the impact of dietary intervention the same untargeted metabolomics was conducted following an intervention treatment (CON, MBG, C-MBG, MBG-R groups). C-MBG = healthy to malnourished, MBG-R = malnourished model reversed on a healthy diet.
INSTITUTE
University of British Columbia
DEPARTMENT
Microbiology and Immunology
LABORATORY
B. Brett Finlay
LAST_NAME
Bauer
FIRST_NAME
Kylynda
ADDRESS
Michael Smith Laboratories, #301 ā 2185 East Mall, University of British Columbia Vancouver, British Columbia Canada, V6T 1Z4
Grass pollen sublingual immunotherapy treatment induces transcriptomic and metabolic changes due to AIT treatment
STUDY_SUMMARY
47 patients were enrolled in a double-blind, placebo-controlled, multicenter trial using with GRAZAXĀ® (Phleum pretense) during 2 years of therapy (T2). Immunological assays such as sIgE, sIgG4 and ISAC were carried out to the 31 patients who finished the trial. Additionally, serum and PBMCs samples from these samples were analyzed by metabolomics and transcriptomics, respectively. Based on their sensitization level, 22 patients were grouped in Mono and Poli groups, excluding epithelial allergic patients. Individuals were studied based on their treatment in Active and Placebo and their sensitization level. For metabolomics, samples were analyzed by Liquid and Gas Chromatography coupled to Mass Spectrometry (LC-MS and GC-MS, respectively).
INSTITUTE
Institute of Applied Molecular Medicine, CEU; The Centre of Metabolomics and Bioanalysis, CEU
LAST_NAME
Obeso Montero
FIRST_NAME
David
ADDRESS
Avda. Monteprincipe s/n 28668 Boadilla del Monte, Madrid, EspaƱa
EMAIL
david.obesomontero@beca.ceu.es
PHONE
Tlf: 91 372 47 00 ext. 4662
NUM_GROUPS
2 main groups: Active and Placebo, and 2 subgroups: Monosensitized and Polisensitized patients.
Metabolomic profiling of Canadian species of Alternaria
STUDY_SUMMARY
This study aims to determine the predominant Alternaria species present in Canadian crops, their subsequent substrate distribution and which secondary metabolites are produced. 131 isolates obtained from the Canadian Collection of Fungal Cultures (CCFC) were grown as three-point inoculations on potato dextrose agar (PDA) and grown in the dark for seven days at 25Ā°C. Each strain was extracted with ethyl acetate containing 1% formic acid, and analyzed by high resolution mass spectrometry (HRMS) in full MS mode in both positive and negative ionization modes at 140K resolution. Data were analyzed using principal component analysis (PCA), and groups were assigned based on k-means clustering analysis. All metabolites detected in the peak lists were investigated for significance (P<0.001) between groups using the Kruskal-Wallace test using Benjamini Hochberg false discovery rate (FDR) correction.
UPLC-MSE analysis of samples from Quercus ilex acorns flour. The objective of the study is to obtain a metabolomic profile of several acorns from different trees. This phytochemical analysis and characterization will be a base for the identification of bioactive, antinutritional, or toxic compounds and traceability analysis.
INSTITUTE
Universidad de CĆ³rdoba
DEPARTMENT
Department Biochemistry and Molecular Biology
LABORATORY
Agroforestry and Plant Biochemistry, Proteomics and Systems Biology
Patterns in metabolite pools show that phytoplankton leave a taxon-specific signature on particulate carbon: North Pacific Subtropical Gyre depth profile
STUDY_TYPE
Marine metabolomics depth profile
STUDY_SUMMARY
In the surface ocean, carbon is fixed by phytoplankton and respired by the entire marine community at an astonishingly high rate. At any point in time, the difference between these two processes yields a carbon pool in surface particles that is a combination of both freshly fixed and partially degraded material. On a molecular level, we have a limited knowledge of the small molecules, or metabolites, within this pool. Specific metabolites have been shown to be responsible for fueling respiration, maintaining organismal interactions, and transferring energy throughout the microbial community. Metabolomics, or the direct observation and quantification of the small molecules that are the result of cellular activity, provides an important lens through which we can begin to assess the standing stocks of small compounds that likely fuel a great deal of heterotrophic activity in the surface ocean. Here we describe community metabolomes of particulate material into the North Pacific Ocean and compare the metabolomes to a variety of phytoplankton grown in the lab. Using both targeted and untargeted metabolomics, we identify metabolites in the particulate carbon pool and explore their latitudinal and phylogenetic distributions. This analysis reveals several compounds that have not been previously recognized as abundant components of the marine organic carbon pool. We found that the community metabolome showed distinct differences between the regimes that likely reflects the phytoplankton community present. The community metabolome in surface waters of the subtropical domain was remarkably consistent even when sampled weeks apart, while the northern regions showed a patichier and less reproducible community metabolome. Some individual compounds showed distinct patterns between oceanographic regimes, including homarine, an abundant molecule that can contribute up to 4% of the total particulate carbon pool in marine surface waters. Glutamic acid and glutamine showed opposite patterns in the oceanographic regimes, suggesting differences in community-level nitrogen assimilation in these different regimes. Overall, this study offers a new perspective into particulate carbon composition in oceanographic research, reveals important carbon pools that may fuel the microbial loop, and suggests an altered community-level nitrogen assimilation capacity over the North Pacific transition zone.
INSTITUTE
University of Washington
DEPARTMENT
School of Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Heal
FIRST_NAME
Katherine
ADDRESS
1501 NE Boat Street, Marine Science Building, Room G
Targeting Sirt2 reprograms T cell metabolism for effective immune response
STUDY_TYPE
Targeted Metabolomics
STUDY_SUMMARY
There is a growing evidence that metabolism is a key driver of T cell functions. A switch from oxidative phosphorylation to aerobic glycolysis is a hallmark of T cell activation and is required to meet metabolic demands of proliferation and effector functions. However the mechanisms underlying the metabolic switch in T cells remain unclear. Here we identify Sirt2 as a crucial immune checkpoint coordinating metabolic and functional fitness of T cells. Sirt2 is induced upon T cells activation and increases in late maturation stages. Sirt2 negatively regulates glycolysis by targeting key glycolytic enzymes. Remarkably, Sirt2 knockout T cells exhibit profound upregulation of aerobic glycolysis with enhanced proliferation and effector function and thus effectively reject tumor challenge in vivo. Furthermore pharmacologic inhibition of Sirt2 in human tumor infiltrating lymphocytes demonstrated similar phenotype. Taken together our results demonstrate Sirt2 as an actionable target to reprogram T cell metabolism to augment immunotherapy.
Use of Information Dependent Acquisition mass spectra and Sequential Window Acquisition of all Theoretical fragment-ion mass spectra for fruit juices metabolomics and authentication
STUDY_SUMMARY
Introduction LC-MS based untargeted metabolomics are the main untargeted methods used for juice metabolomics to solve the authentication problem faced in fruit juice industry. Objectives To evaluate the performances of different untargeted metabolomics methods on fruit juices metabolomics and authentication, orange and apple fruit juices were selected for this study. Methods IDA-MS and SWATH-MS based on UHPLC-QTOF were used for the metabolomics and authenticity determination of apple and orange juices, including the lab-made samples of oranges (Citrus sinensis Osb.) from Jiangxi Province, apples (Malus domestica Borkh) from Shandong Province, and different brands of commercial orange and apple juice samples from markets. Results IDA-MS and SWATH-MS could both acquire numerous MS1 features and MS2 information of juice components, while SWATH-MS excels at the acquisition rate of MS2. Distinctive separation between authentic orange juice and not authentic orange juice could be seen from principal component analysis and hierarchical clustering analysis based on both IDA-MS and SWATH-MS. After analysis of variance, fold change analysis and orthogonal projection to latent structures discriminant mode, 53 and 46 potential markers were defined by IDA-MS and SWATH-MS (with 77.4% and 100% MS2 acquisition rate) separately. Subsequently, these potential markers were putatively annotated using general chemical databases with 6 more annotated by SWATH-MS. Furthermore, 7 of the potential markers, l-asparagine, umbelliferone, glucosamine, phlorin, epicatechin, phytosphingosine and chlorogenic acid, were identified with standards. For the consideration of model simplicity, two determined makers (umbelliferone and chlorogenic acid) were selected to construct the DD-SIMCA model in commercial samples because of their good correlation with apple adulteration proportion, and the sensitivity and specificity of the model were 100% and 95%. Conclusion SWATH-MS excels at the MS2 acquisition of juice components and potential markers. This study provides an overall performance comparison between IDA-MS and SWATH-MS, and guidance for the method selection on fruit juice metabolomics and juice authenticity determination. Two of the potential markers determined, umbelliferone and chlorogenic acid, could be used as apple juice indicators in orange juice.
INSTITUTE
China agricultural university
LAST_NAME
Xu
FIRST_NAME
Lei
ADDRESS
No. 17 Qinghua East Road, Haidian District, Beijing, Beijing, 100083, China
Fructosamine-3-kinase (FN3K) KO in HepG2 liver cancer cells
STUDY_SUMMARY
Fructosamine-3-kinases (FN3Ks) are a family of metabolic kinases which are evolutionarily related to eukaryotic protein kinases. Aberrant regulation of these kinases by altered redox homeostasis is a major contributing factor in aging and disease. However, the mechanisms of regulation and cellular functions of these kinases are not known. Bioinformatic analyses of cancer cell lines identified significant overexpression of FN3K in liver and eye cancer cells. To assess the functional significance of this increased expression, a CRISPR knockout of FN3K (FN3K-KO) was generated in the HepG2 liver cancer cell line. The metabolome was compared between FN3K-KO and WT HepG2 cells using untargeted 1H NMR metabolomics. This revealed significant differences in several metabolites that suggest a role for FN3K in regulating redox and energy balance in HepG2 cells.
INSTITUTE
University of Georgia
DEPARTMENT
Complex Carbohydrate Research Center
LABORATORY
Edison
LAST_NAME
Colonna
FIRST_NAME
Maxwell
ADDRESS
315 Riverbend Rd, Athens, GA 30602
EMAIL
maxwellbaca@uga.edu
PHONE
7065420257
NUM_GROUPS
2
PUBLICATIONS
A redox-active switch in Fructosamine-3-kinases expands the regulatory repertoire of the protein kinase super-family
2,3,7,8 -Tetrachlorodibenzo-p-dioxin Mediated Effects on Hepatic Coenzyme A (CoA) and Acetyl-CoA Levels (part-I)
STUDY_TYPE
Dioxin
STUDY_SUMMARY
Experiment to study TCDD-elicited effects on hepatic Coenzyme A (CoA) and Acetyl-CoA levels in liver of male mice treated with sesame oil vehicle or 0.01-30 ug/kg TCDD every 4 days for 28 days.
Stirred suspension bioreactors maintain naĆÆve pluripotency of human pluripotent stem cells (hPSCs)
STUDY_SUMMARY
Although cell therapies require large numbers of quality-controlled hPSCs, existing technologies are limited in their ability to efficiently grow and scale stem cells. We report here that cell-state conversion from primed-to-naĆÆve pluripotency enhances the biomanufacturing of hPSCs. NaĆÆve hPSCs exhibit superior growth kinetics and aggregate formation characteristics in stirred suspension bioreactors compared to their primed counterparts. Moreover, we demonstrate the role of the bioreactor mechanical environment in the maintenance of naĆÆve pluripotency, through transcriptomic enrichment of mechano-sensing signaling for cells in the bioreactor along with a decrease in expression of lineage-specific and primed pluripotency hallmarks. Bioreactor-cultured, naĆÆve hPSCs express epigenetic regulatory transcripts associated with naĆÆve pluripotency, and display hallmarks of X-chromosome reactivation. They exhibit robust production of naĆÆve pluripotency metabolites and display reduced expression of primed pluripotency cell surface markers. We also show that these cells retain the ability to undergo targeted differentiation into beating cardiomyocytes, hepatocytes, and neural rosettes. They additionally display faster kinetics of teratoma formation compared to their primed counterparts. NaĆÆve bioreactor hPSCs also retain structurally stable chromosomes. Our research corroborates that converting hPSCs to the naĆÆve state enhances hPSC manufacturing and indicates a potentially important role for the bioreactorās mechanical environment in maintaining naĆÆve pluripotency.
To discover distinctive endogenous metabotype of patients with COPD associated with TB from those originated from Tabaco smoking. Cross-sectional metabolomic analyses of serum samples were performed for subjects including TB-associated COPD (T-COPD), smoking-associated COPD (S-COPD) and healthy subjects. To retain a broad spectrum of metabolites, technically distinct analyses (global metabolomic profiling using liquid chromatography quadrupole time-of-flight mass spectrometry) were employed. Frozen samples were diluted with either an acetonitrile:methanol:water (3:3:4) mixture. Each sample (5 Ī¼L) was loaded onto an C18 column and was analyzed using an Agilent 6530 QTOF mass spectrometer (Agilent Technologies). Detailed protocol is obtained in this project
2,3,7,8 -Tetrachlorodibenzo-p-dioxin Mediated Effects on Hepatic Coenzyme A (CoA) and Acetyl-CoA Levels (part-II)
STUDY_TYPE
Dioxin
STUDY_SUMMARY
Experiment to study TCDD-elicited effects on hepatic Coenzyme A (CoA) and Acetyl-CoA levels in liver of male mice treated with sesame oil vehicle or 0.01-30 ug/kg TCDD every 4 days for 28 days.
Fast and sensitive flow-injection mass spectrometry metabolomics by analyzing sample specific ion distributions
STUDY_SUMMARY
Mass spectrometry based metabolomics is a widely used approach in biotechnology and biomedical research. However, current methods coupling mass spectrometry with chromatography are time-consuming and not suitable for high-throughput analysis of thousands of samples. An alternative approach is flow-injection mass spectrometry (FI-MS) in which samples are directly injected to the ionization source. Here, we show that the sensitivity of Orbitrap FI-MS metabolomics methods is limited by ion competition effect in the detection system. We describe an approach for overcoming this effect by analyzing the distribution of ion m/z values and computationally determining a series of optimal scan ranges. This enables reproducible detection of ~9,000 and ~10,000 m/z features in metabolomics and lipidomics analysis of serum samples, respectively, with a sample scan time of ~15 seconds and duty time of ~30 seconds; a ~50% increase versus current spectral-stitching FI-MS. This approach facilitates high-throughput metabolomics for a variety of applications, including biomarker discovery and functional genomics screens.
Lipid profile Dataset of optogenetics induced optic nerve regeneration
STUDY_SUMMARY
Using the transgenic Chr2 mouse (Thy1-ChR2-EYFP) as a model of regeneration, we present the profile the lipid changes that occur after optic nerve crush, light stimulation and RGC growth. Thy1-ChR2-EYFP mice and controls (C57BL/6) were divided in four groups each, no crush and no stimulation, no crush and stimulation, crush and no stimulation, crush and stimulation.
Distinct metabolic states of a cell guide alternate fates of mutational buffering through altered proteostasis
STUDY_SUMMARY
Changes in metabolism can alter the cellular milieu; can this also change intracellular proteostasis? Since proteostasis can modulate mutational buffering, if change in metabolism has the ability to change proteostasis, arguably, it should also alter mutational buffering. Building on this, we find that altered cellular metabolic states in E. coli buffer distinct mutations. Buffered-mutants had folding problems in vivo and were differently chaperoned in different metabolic states. Notably, this assistance was dependent upon the metabolites and not on the increase in canonical chaperone machineries. Additionally, we were able to reconstitute the folding assistance afforded by metabolites in vitro and propose that changes in metabolite concentrations have the potential to alter proteostasis. Collectively, we unravel that the metabolite pools are bona fide members of proteostasis and aid in mutational buffering. Given the plasticity in cellular metabolism, we posit that metabolic alterations may play an important role in the positive or negative regulation of proteostasis.
INSTITUTE
CSIR National Chemical Laboratory
LAST_NAME
Shanmugam
FIRST_NAME
Dhanasekaran
ADDRESS
Dr. Homi Bhabha Road, Pune, maharashtra, 411008, India
Investigating exposures and health impacts of endocrine disrupting chemicals among inner-city children.
STUDY_TYPE
Observational study
STUDY_SUMMARY
Our CHEAR project will conduct untargeted metabolomics analyses among children with and without asthma to gain insight into potential mechanisms related to asthma development. Metabolomics analysis will be conducted in a subset of participants from DISCOVER, a panel study enriched with low-income African American children in Baltimore, MD enrolled from 2009 to 2013. We will conduct untargeted metabolomics in serum collected from 15 children with non-atopic asthma and 22 non-atopic children without asthma. For each child, we will conduct analyses in two serum samples collected 3-9 months apart (total = 74 samples). We will use these pilot data to explore differences in metabolomic profiles among non-atopic children with asthma compared to non-asthmatic controls, and leverage repeated sample measures to assess differences in metabolomic profiles among asthmatic children by symptom frequency or severity. Finally, these data will be used to explore differences in metabolomic profiles by exposure status after urinary organophosphate flame retardant biomarkers have been quantified by CHEAR. Together, these analyses will provide pilot data for future work in our Center to assess potential mechanisms related to asthma development and severity as well as effects of organophoshate flame retardant exposures.
Plasmodium falciparum increased time in circulation underlies persistent asymptomatic infection in the dry season
STUDY_SUMMARY
The dry season is a major challenge for Plasmodium falciparum parasites in many malaria endemic regions, where water availability limits mosquitoes to only part of the year. How P. falciparum bridges two transmission seasons months apart, without being cleared by the host or compromising host survival is poorly understood. Here we show that low levels of P. falciparum parasites persist in the blood of asymptomatic Malian individuals during the 5- to 6-month dry season, rarely causing symptoms and minimally affecting the host immune response. Parasites isolated during the dry season are transcriptionally distinct from those of subjects with febrile malaria in the transmission season, reflecting longer circulation within each replicative cycle, of parasitized erythrocytes without adhering to the vascular endothelium. Low parasite levels during the dry season are not due to impaired replication, but rather increased splenic clearance of longer-circulating infected erythrocytes. We propose that P. falciparum virulence in areas of seasonal malaria transmission is regulated so that the parasite decreases its endothelial binding capacity, allowing increased splenic clearance and enabling several months of subclinical parasite persistence.
INSTITUTE
Penn State
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA
To perform an untargeted metabolomics analysis of urine samples, matrix blanks and quality control samples. The metabolomics approach will be performed using both reverse phase (RP) and HILIC chromatography (ZHP) separations coupled to high-resolution mass spectrometry. Samples were received and stored at -80Ā°C until processing. In total, 315 samples had sufficient sample volume for metabolomics analysis.
INSTITUTE
Icahn School of Medicine at Mount Sinai
LAST_NAME
Petrick
FIRST_NAME
Lauren
ADDRESS
Department of Environmental Medicine and Public Health, Atran Building 3rd floor, 101st St. between Madison and 5th Ave, New York, New York, 10029, USA
Primary metabolites were quantified in human plasma from the 1:3 matched TEDDY case-control subjects. Information on the nested case-control study design can found in: Biomarker discovery study design for type 1 diabetes in The Environmental Determinants of Diabetes in the Young (TEDDY) study. Lee HS, Burkhardt B, McLeod W, Smith S, Eberhard C, Lynch K, Hadley D, Rewers M, Simell O, She JX, Hagopian W, Lernmark A, Akolkar B, Ziegler AG, Krischer J, and the TEDDY Study Group. Diabetes/Metabolism Research and Reviews. Epub 2013 December 15. doi: 10.1002/dmrr.2510 (PubMed ID: 24339168). Primary metabolites were extracted from 30 Āµl plasma aliquots by adding 1 ml of a carefully degassed -20 Ā°C cold isopropanol/acetonitrile/water mixture (3:3:2, v/v/v) for 5 min at 4 Ā°C which simultaneously precipitates proteins. After centrifugation, half of the extract was dried and cleaned up from triglycerides by a 50% acetonitrile mixture. After drying, internal standards were added as C08-C30 fatty acid methyl esters in chloroform as retention index markers (Kind et. al, 2009). Primary metabolites were derivatized by methoximation and trimethylsilylation. Primary metabolites were analyzed by cold injection/automatic liner exchange gas chromatography time-of- flight mass spectrometry (CIS/ALEX GC-TOF MS) (Fiehn et. al, 2008). In order to limit buildup of involatile material in the GC system and to prevent any carry over, an automatic liner exchange with multi-baffled liners and cold injection procedures was used instead of classic hot injections into standard s/sl liners. Multi-baffled inert glass liners were used because classic glass wool liners might hamper derivatization of amino groups for amino acid analysis. Robotic derivatization was further used to control reaction times (Ji et. al, 2011); and GC-columns were employed with integrated 10 meter guard columns, which could cut multiple times in 10 cm increments whenever quality control samples determined out-of-control situations. A temperature of 280 Ā°C was determined to be the optimum transfer line temperature at which even higher-boiling compounds did not show tailing effects and at which the electron ionization filaments could still be operated at their optimal temperature of 250 Ā°C and -70 eV. The mass spectrometer was operated using daily mass calibration auto-tuning using FC43 (perfluorotributyl-amine) and acquired 17 spectra per second and 1850-1950 V detector voltage. This high spectral acquisition rate was necessary to obtain enough data for mass spectral deconvolution of co-eluting compounds. Under these conditions, the system was around 10-times more sensitive than classic quadrupole GC-MS instruments and also clearly outperformed GC-triple quadrupole mass spectrometers. For select compounds, even lower limits of detection were achieved than for optimized MRM conditions in UPLC-QTRAP MS analysis. Around 144 unique metabolites were detectable in blood plasma (Fiehn & Kind, 2007); in addition to 221 unidentified compounds that were captured in the BinBase database system and hence, were comparable across studies. References: 1) Kind T, Wohlgemuth G, Lee DY, Lu Y, Palazoglu M, Shahbaz S, Fiehn O: FiehnLib: mass spectral and retention index libraries for metabolomics based on quadrupole and time-of-flight gas chromatography/mass spectrometry. Anal Chem 2009, 81(24):10038-10048. 2) Fiehn O, Wohlgemuth G, Scholz M, Kind T, Lee DY, Lu Y, Moon S, Nikolau B: Quality control for plant metabolomics: reporting MSI-compliant studies. Plant J 2008, 53(4):691-704. 3) Ji Y, Hebbring S, Zhu H, Jenkins GD, Biernacka J, Snyder K, Drews M, Fiehn O, Zeng Z, Schaid D et al: Glycine and a glycine dehydrogenase (GLDC) SNP as citalopram/escitalopram response biomarkers in depression: pharmacometabolomics-informed pharmacogenomics. Clin Pharmacol Ther 2011, 89(1):97-104. 4) Fiehn O, Kind T: Metabolite profiling in blood plasma. Methods Mol Biol 2007, 358:3-17. An explanation of the study design variables are explained in detail in a data dictionary provided in the raw data download section.
In this study, we investigated changes in hepatic lipid profiles of little brown bats (Myotis lucifugus) and big brown bats (Eptesicus fuscus) at an early stage (70 d) of infection with the etiological agent, Pseudogymnoascus destructans (Pd).
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Natural history of the systemic responses to a renal inoculation of uropathogenic E. coli in swine
STUDY_SUMMARY
Background: The pathogenesis of systemic infection and its progression to sepsis remains poorly understood. Progress in the field has been stifled by the shortcomings of experimental models which include poor replication of the human condition. To address these challenges, we developed a novel large animal model of systemic infection that is capable of generating high-dimensional clinically relevant data. Methods: Male swine (n=5) were anesthetized, mechanically ventilated, and surgically instrumented for continuous hemodynamic monitoring and serial blood sampling. Animals were inoculated with uropathogenic E. coli by direct injection into the renal parenchyma and were maintained under anesthesia for up to 24 hours. The natural history of the infection was studied, animals were not resuscitated. Multi-dimensional data were collected hourly to every 6 hours; all animals were euthanized when at predetermined physiologic endpoints. Results: Core body temperature progressively increased from mean (SD) 37.9(0.8) ĢC at baseline to 43.0(1.2) ĢC at experiment termination (p=0.006). While mean arterial pressure did not begin to decline until 6h post inoculation, dropping from 86(9) mmHg at baseline to 28(5) mmHg (p=0.005) at termination. Blood glucose progressively declined but lactate levels did not elevate until the last hours of the experiment. There were also temporal changes in whole blood concentrations of a number of metabolites including increases in the catecholamine precursors, tyrosine (p=0.005) and phenylalanine (p=0.005). Lung, liver, and kidney function parameters worsened as infection progressed and at study termination there was histopathological evidence of injury in these end-organs. Conclusion: We demonstrate a versatile, multi-dimensional, longitudinal, swine model of systemic infection that could be used to further our understanding of the mechanisms that underlie infection-induced multi-organ dysfunction and failure, optimize resuscitation protocols and test therapeutic interventions. Such a model could improve translation of findings from the bench to the bedside, circumventing a significant obstacle in sepsis research.
Metabolic Response in Patients with Post-Treatment Lyme Disease Symptoms/Syndrome
STUDY_SUMMARY
Post-treatment Lyme Disease Symptoms/Syndrome (PTLDS) occurs in approximately 10% of Lyme disease patients following antibiotic treatment. Objective biomarkers or specific clinical symptoms to identify PTLDS patients do not currently exist and the PTLDS classification is based on the report of persistent subjective symptoms for ā„ 6 months following antibiotic treatment for Lyme disease. Untargeted liquid chromatography-mass spectrometry metabolomics was used to define metabolic changes that occurred longitudinally in PTLDS and clinically cured non-PTLDS Lyme patients from two separate cohorts. An elastic net regularization model was applied to define the metabolites that classified PTLDS and non-PTLDS patients at different time points, and the PTLDS defining metabolites were evaluated in two sample cohorts using linear discriminant analysis. This study determined that observable metabolic alterations occur between PTLDS and non-PTLDS patients at multiple time points. These metabolic alterations discriminated between PTLDS and non-PTLDS patients and consisted of metabolites of glycerophospholipid, bile acid and acylcarnitine metabolism. Longitudinal analyses showed distinct patterns in metabolite abundance changes that indicated a greater variability in PTLDS vs non-PTLDS patients. These data provide evidence that an objective metabolite-based measurement can distinguish patients with PTLDS and help understand the underlying biochemistry of PTLDS.
INSTITUTE
Colorado State University
DEPARTMENT
MIP
LABORATORY
Belisle
LAST_NAME
Belisle
FIRST_NAME
John
ADDRESS
200 West Lake, Campus Delivery 0922, Colorado State University, Fort Collins, CO, 80523
Compatible solutes were quantified in sea-ice diatoms
STUDY_SUMMARY
Sea-ice algae provide an important source of primary production in polar regions, yet we have limited understanding of their responses to the seasonal cycling of temperature and salinity. Using a targeted liquid chromatography-mass spectrometry-based metabolomics approach, we found that axenic cultures of the Antarctic sea-ice diatom, Nitzschia lecointei, displayed large differences in their metabolomes when grown in a matrix of conditions that included temperatures of ā1 and 4Ā°C, and salinities of 32 and 41, despite relatively small changes in growth rate. Temperature exerted a greater effect than salinity on cellular metabolite pool sizes, though the N- or S-containing compatible solutes, 2,3-dihydroxypropane-1-sulfonate (DHPS), glycine betaine (GBT), dimethylsulfoniopropionate (DMSP), and proline responded strongly to both temperature and salinity, suggesting complexity in their control. We saw the largest (> 4 fold) response to salinity for proline. DHPS, a rarely studied but potential compatible solute, reached the highest intracellular compatible solute concentrations of ~ 85 mM. When comparing the culture findings to natural Arctic sea-ice diatom communities, we found extensive overlap in metabolite profiles, highlighting the relevance of culture-based studies to probe environmental questions. Large changes in sea-ice diatom metabolomes and compatible solutes over a seasonal cycle could be significant components of biogeochemical cycling within sea ice.
INSTITUTE
University of Washington
DEPARTMENT
School of Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Dawson
FIRST_NAME
Hannah
ADDRESS
1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA 98195
Metabolomic study of Escherichia coli K-12 MG1655 and mutants
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Metabolomic analysis of Wildtype, crp mutant and its five adaptively evolved populations evolved in glucose minimal media with 40 mM MOPS during its exponential phase of growth. Three biological and two technical replicate samples (n=6) were harvested for each of the strains while growing in a bioreactor aerobically at 37 degree Celsius and 700 rpm. This study aims to characterize and compare the metabolic profile of all these strains.
INSTITUTE
IIT Bombay
DEPARTMENT
Department of Chemical Engineering
LABORATORY
Systems Biology and Metabolic Engineering Laboratory
LAST_NAME
Pal
FIRST_NAME
Ankita
ADDRESS
IIT Bombay, Powai, Mumbai - 400076, Maharashtra, India
Time-course experiment of Microchloropsis gaditana cells supplemented with CO2 (part-II)
STUDY_TYPE
Time-course experiment
STUDY_SUMMARY
Experiments were conducted with Microchloropsis gaditana supplemented with very-low CO2 and high CO2. Sampling was done on the following time points: Day 3, 6 and 9.
INSTITUTE
International Centre for Genetic Engineering and Biotechnology
DEPARTMENT
Integrative Biology
LABORATORY
Omics of Algae
LAST_NAME
Jutur
FIRST_NAME
Pannaga Pavan
ADDRESS
2nd Floor, New Building, ICGEB, Aruna Asaf Ali Marg, New Delhi - 110067
EMAIL
jppavan@icgeb.res.in
PHONE
+91 11 26741358
STUDY_COMMENTS
Former name of species: Nannochloropsis gaditana LubiƔn
Quantitative Lipids study on total murine liver tissue from mice at different age
STUDY_TYPE
Liver tissue/Primary tissue
STUDY_SUMMARY
Following birth, the neonatal intestine is exposed to maternal and environmental bacteria that successively form a dense and highly dynamic intestinal microbiota. Whereas the effect of exogenous factors has been extensively investigated, endogenous, host-mediated mechanisms have remained largely unexplored. Concomitantly with microbial colonization, the liver undergoes functional transition from a hematopoietic organ to a central organ of metabolic regulation and immune surveillance. The aim of the present study was to analyze the influence of the developing hepatic function and liver metabolism on the early intestinal microbiota. Using metabolomic and microbial profiling in combination with multivariate analysis we characterized the colonization dynamics and liver metabolism in the murine gastrointestinal tract (n=6-10 per age group). We observed major age-dependent microbial and metabolic changes and identified bile acids as potent drivers of the early intestinal microbiota maturation. Consistently, oral administration of tauro-cholic acid or Ī²-tauro-murocholic acid to newborn mice (n= 7-14 per group) accelerated postnatal microbiota maturation. Lipids in total liver tissue from healthy C57BL/6 mice at 1, 7, 14, 21, 28 and 56 day after birth was analyzed.
Quantitative Hexose study on total murine liver tissue from mice at different age
STUDY_TYPE
Liver tissue/Primary tissue
STUDY_SUMMARY
Following birth, the neonatal intestine is exposed to maternal and environmental bacteria that successively form a dense and highly dynamic intestinal microbiota. Whereas the effect of exogenous factors has been extensively investigated, endogenous, host-mediated mechanisms have remained largely unexplored. Concomitantly with microbial colonization, the liver undergoes functional transition from a hematopoietic organ to a central organ of metabolic regulation and immune surveillance. The aim of the present study was to analyze the influence of the developing hepatic function and liver metabolism on the early intestinal microbiota. Using metabolomic and microbial profiling in combination with multivariate analysis we characterized the colonization dynamics and liver metabolism in the murine gastrointestinal tract (n=6-10 per age group). We observed major age-dependent microbial and metabolic changes and identified bile acids as potent drivers of the early intestinal microbiota maturation. Consistently, oral administration of tauro-cholic acid or Ī²-tauro-murocholic acid to newborn mice (n= 7-14 per group) accelerated postnatal microbiota maturation.Summed hexoses in total liver tissue from healthy C57BL/6 mice at 1, 7, 14, 21, 28 and 56 day after birth was analyzed.
Mechanism of Trichloroethylene (TCE) toxicity in the placenta
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Cells in culture will be exposed to S-(1,2-dichlorovinyl)-L-cysteine (DCVC), a relevant metabolite of TCE. The first cell line used will be HTR-8/SVneo, originally derived from first trimester female human cytotrophoblast cells and immortalized with simian virus 40 large T antigen, this cell line models first-trimester placental extravillous trophoblasts in vitro. HTR-8/SVneo cells will be cultured for 24 hours followed by exposure to cell culture media (control) or 20 ĀµM DCVC for 6 or 12 hours. Cells (cells frozen in cell culture dish) will then be collected (n=5) In the TCE aim of this study, we will utilizeone cell model that phenotypically represents an important placental cell population.
Activation of ectopic olfactory receptor 544 induces GLP-1 secretion, alters gut microbiome, and improves intestinal permeability.
STUDY_TYPE
Fecal metabolome
STUDY_SUMMARY
Metabolome data set from mouse fecal samples Group - WT_AZA: fecal samples from wild type mouse fed with high fat diet and azelaic acid (0.05%, w/w) Group - WT_DW: fecal samples from wild type mouse fed with high fat diet Group - KO_AZA: fecal samples from Olfr544 receptor knock out mouse fed with high fat diet and azelaic acid (0.05%, w/w) Group - KO_DW: fecal samples from Olfr544 receptor knock out mouse fed with high fat diet
INSTITUTE
Korea University
LAST_NAME
Kim
FIRST_NAME
Jungyeon
ADDRESS
145, Anam-ro, Seongbuk-gu, Seoul, Seoul, 02841, Korea, South
Identification of distinct metabolic perturbations and associated immunomodulatory events during intra-erythrocytic development stage of pediatric Plasmodium falciparum malaria
STUDY_SUMMARY
The goal of this study was to interrogate biochemical profiles manifested in human serum samples originating from a cohort of West African children, collected before and during P. falciparum malarial infection, with the aim of characterizing metabolic migration associated with severity of malarial infection.
Steady-state metabolomics time course of Saccharomyces cerevisiae mitochondrial fatty acid synthesis (mtFAS) mutants
STUDY_TYPE
Steady-state targeted and untargeted metabolomics time course
STUDY_SUMMARY
The goal of this work was to analyze metabolic changes in yeast at various time points with either the oar1 KO or the mct1 knock-out conditions when compared to a time-matched wild-type samples using gas chromatography-mass spectrometry (GC-MS).
INSTITUTE
University of Utah
DEPARTMENT
Biochemistry
LABORATORY
Rutter
LAST_NAME
Berg
FIRST_NAME
Jordan
ADDRESS
15 N Medical Drive East RM 5520, Salt Lake City, UT 84112-5650 USA
Ontogeny related changes in the pediatric liver metabolome
STUDY_SUMMARY
A major challenge in implementing personalized medicine in pediatrics is identifying appropriate drug dosages for children. The majority of drug dosing studies have been based on adult populations, often with modification of the dosing for children based on size and weight. However, the growth and development experienced by children between birth and adulthood represents a dynamically changing biological system, with implications for effective drug dosing, efficacy as well as potential drug toxicity. The purpose of this study was to apply a metabolomics approach to gain preliminary insights into the ontogeny of liver function from newborn to adolescent.
Ontogeny related changes in the pediatric liver metabolome (part-II)
STUDY_SUMMARY
A major challenge in implementing personalized medicine in pediatrics is identifying appropriate drug dosages for children. The majority of drug dosing studies have been based on adult populations, often with modification of the dosing for children based on size and weight. However, the growth and development experienced by children between birth and adulthood represents a dynamically changing biological system, with implications for effective drug dosing, efficacy as well as potential drug toxicity. The purpose of this study was to apply a metabolomics approach to gain preliminary insights into the ontogeny of liver function from newborn to adolescent.
Ontogeny related changes in the pediatric liver metabolome (part-III)
STUDY_SUMMARY
A major challenge in implementing personalized medicine in pediatrics is identifying appropriate drug dosages for children. The majority of drug dosing studies have been based on adult populations, often with modification of the dosing for children based on size and weight. However, the growth and development experienced by children between birth and adulthood represents a dynamically changing biological system, with implications for effective drug dosing, efficacy as well as potential drug toxicity. The purpose of this study was to apply a metabolomics approach to gain preliminary insights into the ontogeny of liver function from newborn to adolescent.
MDM2-Dependent Rewiring of Metabolomic and Lipidomic Profiles in Dedifferentiated Liposarcoma Models
STUDY_SUMMARY
Dedifferentiated liposarcoma (DDLPS) is an aggressive mesenchymal cancer marked by amplification of MDM2, an inhibitor of the tumor suppressor TP53. DDLPS patients with higher MDM2 amplification have lower chemotherapy sensitivity and worse outcome than patients with lower MDM2 amplification. We hypothesized that MDM2 amplification levels may be associated with changes in DDLPS metabolism. Six patient-derived DDLPS cell line models were subject to comprehensive metabolomic (Metabolon) and lipidomic (SCIEX 5600 TripleTOF-MS) profiling to assess associations with MDM2 amplification and their responses to metabolic perturbations. Comparing metabolomic profiles between MDM2 higher and lower amplification cells yielded a total of 23 differentially abundant metabolites across both panels (FDR < 0.05, log2 FC < 0.75), including ceramides, glycosylated ceramides, and sphingomyelins. Disruption of lipid metabolism through statin administration resulted in a chemo-sensitive phenotype in MDM2 lower cell lines only, suggesting that lipid metabolism may be a large contributor to the more aggressive nature of MDM2 higher DDLPS tumors. This study is the first to provide comprehensive metabolomic and lipidomic characterization of DDLPS cell lines and provides evidence for MDM2-dependent differential molecular mechanisms that are critical factors in chemoresistance and could thus affect patient outcome.
Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach (part-II)
Background: Advances in untargeted metabolomic technologies have great potential for insight into adverse metabolic effects underlying exposure to environmental chemicals. However, important challenges need to be addressed, including how biological response corresponds to the environmental chemical burden in different target tissues. Aim: We performed a pilot study using state-of-the-art ultra-high-resolution mass spectrometry (UHRMS) to characterize the burden of lipophilic persistent organic pollutants (POPs) in metabolic tissues and associated alterations in the plasma metabolome. Methods: We studied 11 adolescents with severe obesity at the time of bariatric surgery. We measured 18 POPs that can act as endocrine and metabolic disruptors (i.e. 2 dioxins, 11 organochlorine compounds [OCs] and 5 polybrominated diphenyl ethers [PBDEs]) in visceral and subcutaneous abdominal adipose tissue (vAT and sAT), and liver samples using gas chromatography with UHRMS. Biological pathways were evaluated by measuring the plasma metabolome using high-resolution metabolomics. Network and pathway enrichment analysis assessed correlations between the tissue-specific burden of three frequently detected POPs (i.e. p,pā-dichlorodiphenyldichloroethene [DDE], hexachlorobenzene [HCB] and PBDE-47) and plasma metabolic pathways. Results: Concentrations of 4 OCs and 3 PBDEs were quantifiable in at least one metabolic tissue for >80% of participants. All POPs had the highest median concentrations in adipose tissue, especially sAT, except for PBDE-154, which had comparable average concentrations across all tissues. Pathway analysis showed high correlations between tissue-specific POPs and metabolic alterations in pathways of amino acid metabolism, lipid and fatty acid metabolism, and carbohydrate metabolism. Conclusions: Most of the measured POPs appear to accumulate preferentially in adipose tissue compared to liver. Findings of plasma metabolic pathways potentially associated with tissue-specific POPs concentrations merit further investigation in larger populations.
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Environmental Medicine and Public Health
LABORATORY
High Resolution Exposomics Research Group
LAST_NAME
Walker
FIRST_NAME
Doug
ADDRESS
One Gustave L. Levy Place, Box 1057, New York, NY 10029
EMAIL
douglas.walker@mssm.edu
PHONE
212-241-9891
NUM_GROUPS
4
TOTAL_SUBJECTS
11
NUM_MALES
1
NUM_FEMALES
10
STUDY_COMMENTS
Upload #1: Visceral and subcutaneous abdominal adipose tissue, liver tissue. Plasma metabolomics are in upload #2
PUBLICATIONS
Valvi D, Walker DI, Inge T, Bartell SM, Jenkins T, Helmrath M, Ziegler TR, La Merrill MA, Eckel SP, Conti D, Liang Y, Jones DP, McConnell R, Chatzi L. (2020). Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach. Environment International. In Press.
Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach
Background: Advances in untargeted metabolomic technologies have great potential for insight into adverse metabolic effects underlying exposure to environmental chemicals. However, important challenges need to be addressed, including how biological response corresponds to the environmental chemical burden in different target tissues. Aim: We performed a pilot study using state-of-the-art ultra-high-resolution mass spectrometry (UHRMS) to characterize the burden of lipophilic persistent organic pollutants (POPs) in metabolic tissues and associated alterations in the plasma metabolome. Methods: We studied 11 adolescents with severe obesity at the time of bariatric surgery. We measured 18 POPs that can act as endocrine and metabolic disruptors (i.e. 2 dioxins, 11 organochlorine compounds [OCs] and 5 polybrominated diphenyl ethers [PBDEs]) in visceral and subcutaneous abdominal adipose tissue (vAT and sAT), and liver samples using gas chromatography with UHRMS. Biological pathways were evaluated by measuring the plasma metabolome using high-resolution metabolomics. Network and pathway enrichment analysis assessed correlations between the tissue-specific burden of three frequently detected POPs (i.e. p,pā-dichlorodiphenyldichloroethene [DDE], hexachlorobenzene [HCB] and PBDE-47) and plasma metabolic pathways. Results: Concentrations of 4 OCs and 3 PBDEs were quantifiable in at least one metabolic tissue for >80% of participants. All POPs had the highest median concentrations in adipose tissue, especially sAT, except for PBDE-154, which had comparable average concentrations across all tissues. Pathway analysis showed high correlations between tissue-specific POPs and metabolic alterations in pathways of amino acid metabolism, lipid and fatty acid metabolism, and carbohydrate metabolism. Conclusions: Most of the measured POPs appear to accumulate preferentially in adipose tissue compared to liver. Findings of plasma metabolic pathways potentially associated with tissue-specific POPs concentrations merit further investigation in larger populations.
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Environmental Medicine and Public Health
LABORATORY
High Resolution Exposomics Research Group
LAST_NAME
Walker
FIRST_NAME
Doug
ADDRESS
One Gustave L. Levy Place, Box 1057, New York, NY 10029
EMAIL
douglas.walker@mssm.edu
PHONE
212-241-9891
NUM_GROUPS
1
TOTAL_SUBJECTS
11
NUM_MALES
1
NUM_FEMALES
10
STUDY_COMMENTS
Upload #1: Visceral and subcutaneous abdominal adipose tissue, liver tissue. Plasma metabolomics are in upload #2
PUBLICATIONS
Valvi D, Walker DI, Inge T, Bartell SM, Jenkins T, Helmrath M, Ziegler TR, La Merrill MA, Eckel SP, Conti D, Liang Y, Jones DP, McConnell R, Chatzi L. (2020). Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach. Environment International. In Press.
Metabolomic profiling of baseline plasmas from a longitudinal prospective cohort of 491 active surveillance (AS) participants
STUDY_TYPE
Biomarker study
STUDY_SUMMARY
Untargeted metabolomics analyses were performed on clinically matched baseline plasma samples (n = 16 per group) prospectively collected from patients with clinically low-risk early stage prostate cancer undergoing AS who exhibited early disease progression (DP) (defined as upgrading of Gleason score (GS) and/or increased tumor volume on surveillance biopsy within 18 months after start of AS) or indolent disease (no progression for five or more years after start of AS) as well as 459 baseline plasma samples prospectively collected from patients with early-stage prostate cancer undergoing AS.
Patterns in metabolite pools show that phytoplankton leave a taxon-specific signature on particulate carbon: North Pacific Transition Zone depth profile
STUDY_TYPE
Marine metabolomics depth profile
STUDY_SUMMARY
In the surface ocean, carbon is fixed by phytoplankton and respired by the entire marine community at an astonishingly high rate. At any point in time, the difference between these two processes yields a carbon pool in surface particles that is a combination of both freshly fixed and partially degraded material. On a molecular level, we have a limited knowledge of the small molecules, or metabolites, within this pool. Specific metabolites have been shown to be responsible for fueling respiration, maintaining organismal interactions, and transferring energy throughout the microbial community. Metabolomics, or the direct observation and quantification of the small molecules that are the result of cellular activity, provides an important lens through which we can begin to assess the standing stocks of small compounds that likely fuel a great deal of heterotrophic activity in the surface ocean. Here we describe community metabolomes of particulate material into the North Pacific Ocean and compare the metabolomes to a variety of phytoplankton grown in the lab. Using both targeted and untargeted metabolomics, we identify metabolites in the particulate carbon pool and explore their latitudinal and phylogenetic distributions. This analysis reveals several compounds that have not been previously recognized as abundant components of the marine organic carbon pool. We found that the community metabolome showed distinct differences between the regimes that likely reflects the phytoplankton community present. The community metabolome in surface waters of the subtropical domain was remarkably consistent even when sampled weeks apart, while the northern regions showed a patchier and less reproducible community metabolome. Some individual compounds showed distinct patterns between oceanographic regimes, including homarine, an abundant molecule that can contribute up to 4% of the total particulate carbon pool in marine surface waters. Glutamic acid and glutamine showed opposite patterns in the oceanographic regimes, suggesting differences in community-level nitrogen assimilation in these different regimes. Overall, this study offers a new perspective into particulate carbon composition in oceanographic research, reveals important carbon pools that may fuel the microbial loop, and suggests an altered community-level nitrogen assimilation capacity over the North Pacific transition zone.
Patterns in metabolite pools show that phytoplankton leave a taxon-specific signature on particulate carbon: Surface samples from the North Pacific Subtropical Gyre to North Pacific Transition Zone
STUDY_TYPE
Marine metabolomics surface samples from the North Pacific Subtropical Gyre to North Pacific Transition Zone
STUDY_SUMMARY
In the surface ocean, carbon is fixed by phytoplankton and respired by the entire marine community at an astonishingly high rate. At any point in time, the difference between these two processes yields a carbon pool in surface particles that is a combination of both freshly fixed and partially degraded material. On a molecular level, we have a limited knowledge of the small molecules, or metabolites, within this pool. Specific metabolites have been shown to be responsible for fueling respiration, maintaining organismal interactions, and transferring energy throughout the microbial community. Metabolomics, or the direct observation and quantification of the small molecules that are the result of cellular activity, provides an important lens through which we can begin to assess the standing stocks of small compounds that likely fuel a great deal of heterotrophic activity in the surface ocean. Here we describe community metabolomes of particulate material into the North Pacific Ocean and compare the metabolomes to a variety of phytoplankton grown in the lab. Using both targeted and untargeted metabolomics, we identify metabolites in the particulate carbon pool and explore their latitudinal and phylogenetic distributions. This analysis reveals several compounds that have not been previously recognized as abundant components of the marine organic carbon pool. We found that the community metabolome showed distinct differences between the regimes that likely reflects the phytoplankton community present. The community metabolome in surface waters of the subtropical domain was remarkably consistent even when sampled weeks apart, while the northern regions showed a patchier and less reproducible community metabolome. Some individual compounds showed distinct patterns between oceanographic regimes, including homarine, an abundant molecule that can contribute up to 4% of the total particulate carbon pool in marine surface waters. Glutamic acid and glutamine showed opposite patterns in the oceanographic regimes, suggesting differences in community-level nitrogen assimilation in these different regimes. Overall, this study offers a new perspective into particulate carbon composition in oceanographic research, reveals important carbon pools that may fuel the microbial loop, and suggests an altered community-level nitrogen assimilation capacity over the North Pacific transition zone.
INSTITUTE
University of Washington
DEPARTMENT
School of Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Heal
FIRST_NAME
Katherine
ADDRESS
1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA, 98195, USA
Plasma metabolites of lipid metabolism associate with diabetic polyneuropathy in a cohort with screen-tested type 2 diabetes: ADDITION-Denmark
STUDY_SUMMARY
The global rise in type 2 diabetes (T2D) is associated with a concomitant increase in diabetic complications. Diabetic polyneuropathy (DPN), the most frequent T2D complication, is characterized by sensory peripheral nerve damage. Although managing glucose effectively slows DPN progression in type 1 diabetes patients, it has limited efficacy in neuropathic T2D patients. The metabolic syndrome (MetS) recently emerged as a major risk factor for DPN; however, the metabolites associated with MetS that correlate with DPN are unknown. We conducted a global plasma metabolomics analysis from a cohort of patients enrolled in the Anglo-Danish-Dutch study of Intensive Treatment of Diabetes in Primary Care (ADDITION), including healthy control subjects, T2D patients, and T2D DPN patients. We identified 15 total plasma metabolites that were altered in T2D DPN patients, including lipids, amino acids, and energy-related metabolites. We evaluated the correlation between these metabolites and all lipid species to identify major changes in both plasma free fatty acids and complex lipids in T2D DPN patients, and found significant alterations in the abundance of long-chain saturated fatty acids, acylcarnitines, and sphingolipids. Our study suggests that DPN in T2D is associated with novel alterations in plasma metabolites related to lipid metabolism.
INSTITUTE
University of Michigan
LAST_NAME
Feldman
FIRST_NAME
Eva
ADDRESS
5017 AATBSRB, 109 Zina Pitcher Place, Ann Arbor, MI 48109-2200
Metabolomics study in Plasma of Obese Patients with Neuropathy Identifies Potential Metabolomics Signatures
STUDY_SUMMARY
The goal of this study was to characterize biochemical profiles observed in human plasma samples originating from an obese cohort stratified by a diagnosis of neuropathy as well as a cohort of lean control subjects without a clinical manifestation of neuropathy.
INSTITUTE
University of Michigan
LAST_NAME
Feldman
FIRST_NAME
Eva
ADDRESS
5017 AATBSRB, 109 Zina Pitcher Place Ann Arbor, MI 48109-2200
Metabolomic and Transcriptomic Signatures of Prenatal Excessive Methionine in Mice
STUDY_TYPE
MS Analysis
STUDY_SUMMARY
Micronutrients are key regulators of prenatal one-carbon (C1) metabolism. We show here that prenatal excessive methionine (MET), a principal micronutrient and methyl-donor, produces in early life stages significant changes in the components of brain C1 pathways and other metabolic pathways including glutamate transmission, mitochondrial function, and lipid metabolism.
INSTITUTE
University of California, Irvine
DEPARTMENT
Department of Pharmaceutical Sciences, School of Medicine. Department of Computer Science, School of Information and Computer Sciences
Untargeted metabolomics analysis of in vitro headspace volatiles from 81 Pseudomonas aeruginosa bacterial isolates from individuals with cystic fibrosis. Headspace volatiles were collected using solid-phase microextraction (SPME) (in triplicate) and comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (GCxGC-TOFMS). 15 replicates of un-inoculated media were prepared and analyzed in parallel, for a total of 258 samples.
Multi-omic profiling of primary mouse neutrophils reveals a pattern of sex and age-related functional regulation
STUDY_SUMMARY
Neutrophils are the most abundant white blood cells in humans and constitute one of the first lines of defense in the innate immune response. Neutrophils are extremely short-lived cells, which survive less than a day after reaching terminal differentiation. Thus, little is known about how organismal aging, rather than the daily cellular aging process, may impact neutrophil biology. In addition, accumulating evidence suggests that both immunity and organismal aging are extremely sex-dimorphic. Here, we describe a multi-omic resource of mouse primary bone marrow neutrophils from young and old female and male animals, at the transcriptomic, metabolomic and lipidomic levels. Importantly, we identify widespread age-related and sex-dimorphic regulation of āomicsā in neutrophils, specifically regulation of chromatin metabolism. We leverage machine-learning and identify candidate molecular drivers of age-related and sex-dimorphic transcriptional regulation of neutrophils. We leverage our resource to predict increased levels/release of neutrophil elastase in male mice. To date, this dataset represents the largest multi-omic resource for the study of neutrophils across biological sex and ages. This resource identifies molecular states linked to neutrophil characteristics linked to organismal age or sex, which could be leveraged to improve immune responses across individuals.
Metabolomic analysis of patients with recurrent angina
STUDY_TYPE
Case and control studies
STUDY_SUMMARY
This is a prospective study. After an overnight fast, venous blood was collected from patients with PCI. The patients were discharged after the blood draw. They were followed up every 30 days for angina recurrence up to 270 days (9 months). The samples were grouped into two groups: recurrent angina and angina-free based on the follow-up results at 9 months.
Metabolomic study of Escherichia coli K-12 MG1655 WT and its transcriptional regulator mutants under anaerobic fermentation conditions
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Metabolomic analysis of Wildtype, fnr, arcA and ihf mutants in glucose minimal media under anaerobic fermentation conditions during its exponential phase of growth. Three biological and two technical replicate samples (n=6) were harvested for each of the strains while growing in a bioreactor anaerobically at 37 degrees Celsius and 150 rpm. This study aims to characterize and compare the metabolic profiles of all these strains.
INSTITUTE
IIT Bombay
DEPARTMENT
Department of Chemical Engineering
LABORATORY
Systems Biology and Metabolic Engineering Laboratory (SBMEL)
LAST_NAME
Pal
FIRST_NAME
Ankita
ADDRESS
Department of Chemical Engineering, IIT Bombay, Mumbai, Maharashtra, 400076, India
Aspirin Metabolomics in Colorectal Cancer Chemoprevention - blood (part-II)
STUDY_SUMMARY
Substantial evidence supports the effectiveness of aspirin for cancer chemoprevention in addition to its well established role in cardiovascular protection. In recent meta-analyses of randomized controlled trials in human, daily aspirin use reduced incidence, metastasis and mortality from several common types of cancer, especially colorectal cancer. The mechanism(s) by which aspirin exerts an anticancer benefit is uncertain; numerous effects have been described involving both cyclooxygenase-dependent and -independent pathways. The goal of this research is to elucidate the key metabolic changes that are responsible for the anticancer effects of aspirin in humans using untargeted metabolomics analysis. Metabolomics, or global metabolite profiling, is an emerging discipline that has the potential to transform the study of pharmaceutical agents. Our innovative approach will use high-resolution mass spectroscopy to detect thousands of metabolites in blood plasma that were collected from participants in the Aspirin/Folate Polyp Prevention Study, a randomized, double-blind, placebo-controlled trial of aspirin for the prevention of colorectal adenomas. Participants in the trial were assigned with equal probability to three aspirin treatment arms (placebo, 81mg, or 325mg daily). Over the three-year period, 81mg/day of aspirin reduced the risk of adenomas, whereas the 325 mg/day dose had less effect. The aims of the current proposal are to identify metabolomic signatures, including specific metabolites and metabolic pathways, that are associated with aspirin treatment in blood of participants after three years of randomized aspirin treatment; and then to assess the associations of these metabolic signatures with adenoma risk and whether they mediate the reductions in risk due to 81 mg/day aspirin treatment. We will prioritize metabolites for study by evaluating metabolite levels in patients from the placebo and treatment arms while controlling the false discovery rate, use correlation analysis to enhance identification of relevant metabolic modules associated with these prioritized metabolites, and apply pathway mapping with post-hoc application of ion dissociation spectroscopy to representative metabolites to confirm pathway identification. Because aspirin is a multifunctional drug that is thought to modify numerous pathways with potential roles in carcinogenesis, a global discovery-based metabolomics approach is the best way to identify its key activities. The public health significance of this work is substantial because understanding the mechanism of aspirin's anticancer effects is key to optimizing its use and to the development of novel drugs targeting the metabolic pathways identified.
Aspirin Metabolomics in Colorectal Cancer Chemoprevention - blood (part-II)
STUDY_SUMMARY
Substantial evidence supports the effectiveness of aspirin for cancer chemoprevention in addition to its well established role in cardiovascular protection. In recent meta-analyses of randomized controlled trials in human, daily aspirin use reduced incidence, metastasis and mortality from several common types of cancer, especially colorectal cancer. The mechanism(s) by which aspirin exerts an anticancer benefit is uncertain; numerous effects have been described involving both cyclooxygenase-dependent and -independent pathways. The goal of this research is to elucidate the key metabolic changes that are responsible for the anticancer effects of aspirin in humans using untargeted metabolomics analysis. Metabolomics, or global metabolite profiling, is an emerging discipline that has the potential to transform the study of pharmaceutical agents. Our innovative approach will use high-resolution mass spectroscopy to detect thousands of metabolites in blood plasma that were collected from participants in the Aspirin/Folate Polyp Prevention Study, a randomized, double-blind, placebo-controlled trial of aspirin for the prevention of colorectal adenomas. Participants in the trial were assigned with equal probability to three aspirin treatment arms (placebo, 81mg, or 325mg daily). Over the three-year period, 81mg/day of aspirin reduced the risk of adenomas, whereas the 325 mg/day dose had less effect. The aims of the current proposal are to identify metabolomic signatures, including specific metabolites and metabolic pathways, that are associated with aspirin treatment in blood after three years of randomized aspirin treatment; and then to assess the associations of these metabolic signatures with adenoma risk and whether they mediate the reductions in risk due to 81 mg/day aspirin treatment. We will prioritize metabolites for study by evaluating metabolite levels in patients from the placebo and treatment arms while controlling the false discovery rate, use correlation analysis to enhance identification of relevant metabolic modules associated with these prioritized metabolites, and apply pathway mapping with post-hoc application of ion dissociation spectroscopy to representative metabolites to confirm pathway identification. Because aspirin is a multifunctional drug that is thought to modify numerous pathways with potential roles in carcinogenesis, a global discovery-based metabolomics approach is the best way to identify its key activities. The public health significance of this work is substantial because understanding the mechanism of aspirin's anticancer effects is key to optimizing its use and to the development of novel drugs targeting the metabolic pathways identified.
Dependence of the Staphylococcal Volatilome Composition on Microbial Nutrition
STUDY_TYPE
Untargeted MS
STUDY_SUMMARY
Introduction: In vitro cultivation of staphylococci is fundamental to both clinical and research microbiology, and selection of growth medium will substantially influence staph growth rates, genetic integrity, pathogenicity, and metabolic capacity. Few studies, to-date, have investigated how the differences in rich media can influence the volatilome of cultivated bacteria. Objectives: The objective of this study was to determine the influence of rich media composition on the chemical characteristics of the volatilomes of Staphylococcus aureus and S. epidermidis. Methods: S. aureus (ATCC 12600) and S. epidermidis (ATCC 12228) were cultured in triplicate in four rich complex media (BHI, LB, MHB, and TSB), and the volatile metabolites produced by each culture were analyzed using headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography ā time-of-flight mass spectrometry (HS-SPME-GCĆGC-TOFMS). Results: When comparing the chemical compositions of the staph volatilomes produced in each medium, we observed few differences when the presence versus absence of volatiles were compared. However, when the relative abundances of volatiles were included in the analyses, we observed that culturing staph in media containing free glucose (BHI and TSB) resulted in volatilomes dominated by acids and esters (67%). The low-glucose media (LB and MHB) produced ketones in greatest relative abundances, but the volatilome compositions in these two media were highly dissimilar. Conclusion: The staphylococcal volatilome is strongly influenced by the nutritional composition of the growth medium, especially the availability of free glucose, which is much more evident when the relative abundances of the volatiles are analyzed, compared to the presence versus absence.
INSTITUTE
Arizona State University
DEPARTMENT
School of Life Sciences
LABORATORY
Heather D. Bean Lab
LAST_NAME
Bean, Ph.D.
FIRST_NAME
Heather
ADDRESS
PO Box 874501 Tempe, AZ 85287
EMAIL
heather.d.bean@asu.edu
PHONE
4807273395
STUDY_COMMENTS
Staphylococcus aureus (ATCC 12600) and Stpahylococcus epidermidis (ATCC 12228) grown in four complex media
PUBLICATIONS
Jenkins, C. L., H. D. Bean (2020). Influence of media on the differentiation of Staphylococcus spp. by volatile compounds. Journal of breath research 14, 016007 doi:10.1088/1752-7163/ab3e9d
HPLC-(Q)-TOF-MS based study of plasma metabolic profiles differences associated with age in paediatric population using animal model
STUDY_SUMMARY
A deep knowledge about the biological development of children is essential for an appropriate drug administration and dosage in paediatrics. Even though the advances made in developmental biology the information available about organ maturation in the early stages of life is limited. This fact, together with the scarcity of clinical trials in children, sometimes leads to the use of off-label drugs. The best approximation to study organ maturation is analysing tissue samples but their collection requires a very invasive method. For this reason, a surrogate matrix such as plasma, which represents a snapshot of global organ/tissue metabolism, may be a suitable alternative. To test this hypothesis, plasma metabolic profiles from piglets of different ages (newborns, infants, and children) obtained by HPLC-(Q)-TOF-MS at positive and negative ionization modes were here studied. The multiblock principal component analysis used in this work proved to be a useful tool to improve the clustering within groups compared to classical principal component analysis. Furthermore, the separation observed among groups was better resolved by using partial least squares-discriminant analysis, which was validated by bootstrapping and permutation testing. Finally, 27 relevant features in positive and 74 features in negative ionization mode were selected by univariate analysis. Among the significant metabolies, an acylcarnitine and eight glycerophospholipids were annotated. The findings indicate that changes with age in the lipid metabolism, where lysophosphatidylcholine and lysophoshatidylethanolamine are included, might be related with the organ maturation state.
Role of environmental toxicants in modulating disease severity in children with NAFLD
STUDY_SUMMARY
This project aims to further understand about how the environment impacts nonalcoholic fatty liver disease NAFLD and nonalcoholic steatohepatitis (NASH) in children. At present the NASH CRN has the largest, well characterized cohort of children with NAFLD, including clinical data, labs, cytokines, genetic polymorphisms, but no proteomics, metabolomics, lipidomics or toxicant assessment. Exposure and untargeted metabolomics analyses will examine the role of environmental toxicants in modulating disease severity and the endogenous response to exposures in children with NAFLD.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Tran
FIRST_NAME
ViLinh
ADDRESS
615 Michael St, suite 225, Atlanta GA 30322
EMAIL
vtran6@emory.edu
PHONE
9122281788
TOTAL_SUBJECTS
427
STUDY_COMMENTS
Both CHEAR pooled plasma samples and Clinical Biomarker Laboratory pooled plasma samples were used
MYC regulates ribosome biogenesis and mitochondrial gene expression programs through its interaction with Host Cell Factor-1
STUDY_SUMMARY
MYC is an oncoprotein transcription factor that is overexpressed in the majority cancers. Although MYC itself is considered undruggable, it may be possible to inhibit MYC by targeting the co-factors it uses to drive oncogenic gene expression patterns. Here, we use loss- and gain- of function approaches to interrogate how one MYC co-factorāHost Cell Factor (HCF)-1ācontributes to MYC activity in a Burkitt lymphoma setting. We identify high-confidence direct targets of the MYCāHCF-1 interaction that are regulated through a recruitment-independent mechanism, including genes that control mitochondrial function and rate-limiting steps for ribosome biogenesis and translation. We describe how these gene expression events impact cell growth and metabolism, and demonstrate that the MYCāHCF-1 interaction is essential for tumor maintenance in vivo. This work highlights the MYCāHCF-1 interaction as a focal point for development of novel anti-cancer therapies.
Metabolic dynamics and prediction og gestational ange and time to delivery in pregant women
STUDY_SUMMARY
Metabolism during pregnancy is a constantly changing yet precisely programmed process, the failure of which may have devastating consequences for the fetus. To capture in high resolution the sequence of metabolic events underlying the normal human pregnancy, we carried out an untargeted metabolome investigation on 784 weekly blood samples collected from 30 Danish pregnant women. The study revealed extensive metabolome alterations over the course of normal pregnancy: of 9,651 detected metabolic features, 4,995 were significantly changed (FDR < 0.05). Many metabolic changes were timed precisely according to pregnancy progression so that the overall metabolic profile demonstrated a highly choreographed pattern. Using machine-learning methods, we were able to build a linear models with five metabolites (four steroids and one phospholipid) that predicts gestational age with high accuracy (Pearson correlation coefficient, R = 0.95).
INSTITUTE
Stanford University
LAST_NAME
Liang
FIRST_NAME
Liang
ADDRESS
Alway M339, 300 Pasteur Drive, Palo Alto, California, 94305, USA
A Compromised Developmental Trajectory of the Infant Gut Microbiome and Metabolome in Atopic Eczema -(targeted SCFAs profiling)
STUDY_SUMMARY
Evidence is accumulating that the establishment of the gut microbiome in early life influences the development of atopic eczema. In this longitudinal study, we used integrated multi-omics analyses to infer functional mechanisms by which the microbiome modulates atopic eczema risk.
INSTITUTE
National University of Singapore
LAST_NAME
Ta
FIRST_NAME
Le Duc Huy
ADDRESS
MD1 - Tahir Foundation Building (MD1), Level 15, Department of Paediatrics, Allergy & Immunology Division, National University of Singapore (NUS), 12 Science Drive 2. Singapore 117549
A Compromised Developmental Trajectory of the Infant Gut Microbiome and Metabolome in Atopic Eczema - (untargeted global metabolomics profiling) )
STUDY_SUMMARY
Evidence is accumulating that the establishment of the gut microbiome in early life influences the development of atopic eczema. In this longitudinal study, we used integrated multi-omics analyses to infer functional mechanisms by which the microbiome modulates atopic eczema risk.
INSTITUTE
National University of Singapore (NUS)
LAST_NAME
Ta
FIRST_NAME
Le Duc Huy
ADDRESS
MD1 - Tahir Foundation Building (MD1), Level 15, Department of Paediatrics, Allergy & Immunology Division, National University of Singapore (NUS), 12 Science Drive 2. Singapore 117549
Evidence for proline utilisation by oral bacterial biofilms grown in saliva
STUDY_TYPE
Research study
STUDY_SUMMARY
Within the mouth bacteria are starved of saccharides as their main nutrient source between meals and it is unclear what drives their metabolism. Previously oral in vitro biofilms grown in saliva have shown proteolytic degradation of salivary proteins and increased extracellular proline. Although arginine and glucose have been shown before to have an effect on oral biofilm growth and activity, there is limited evidence for proline. Nuclear magnetic resonance (NMR) spectroscopy was used to identify extracellular metabolites produced by bacteria in oral biofilms grown on hydroxyapatite discs. Biofilms were inoculated with whole mouth saliva and then grown for 7 days using sterilised whole mouth saliva supplemented with proline, arginine and glucose as a growth-medium. Overall proline had a beneficial effect on biofilm growth ā with significantly fewer dead bacteria present by biomass and surface area of the biofilms (p <0.05). Where arginine and glucose significantly increased and decreased pH, respectively, the pH of proline supplemented biofilms remained neutral at pH 7.3-7.5. SDS-polyacrylamide gel electrophoresis of the spent saliva from proline and arginine supplemented biofilms showed inhibition of salivary protein degradation of immature biofilms. NMR analysis of the spent saliva revealed that proline supplemented biofilms were metabolically similar to unsupplemented biofilms, but these biofilms actively metabolised proline to 5-aminopentanoate, butyrate and propionate, and actively utilised glycine. This study shows that in a nutrient limited environment, proline has a beneficial effect on in vitro oral biofilms grown from a saliva inoculum.
INSTITUTE
King's College London
DEPARTMENT
Centre for Host Microbiome Interactions
LAST_NAME
Cleaver
FIRST_NAME
Leanne
ADDRESS
Floor 17, Tower Wing, Guy's Hospital, Great Maze Pond
Untargeted lipidomics of liver to assess the potential protective role in atherosclerosis progression of A12 antibodies infusion into LDLR-/-mice
STUDY_TYPE
LC-MS Untargeted Lipidomics
STUDY_SUMMARY
In order to assess the therapeutic potential of A12 antibodies in atherosclerosis, untargeted lipidomics of liver samples was performed. LDLR-/-mice were treated with a fully murine version of the A12 antibody (mA12-IgG2b), with the isotype control antibody mB1.8-IgG2b (n=16) or with PBS as controls.
INSTITUTE
Centro Nacional de Investigaciones Cardiovasculares Carlos III
LAST_NAME
Ferrarini
FIRST_NAME
Alessia
ADDRESS
Calle de Melchor FernƔndez Almagro, 3, 28029 Madrid
Sap samples from sugar maple trees across the Canadian province of Ontario were collected in 2019. These samples were minimally prepared and analyzed in both positive ESI and negative ESI by C18 and HILIC chromatography. This was done to uncover the chemical changes that occurred in the sap over the season. This will serve as the base for future analysis of maple syrup where compounds that may be responsible for specific organoleptic properties can be linked back to precursors found here in the sap.
Transkingdom interactions between Lactobacilli and hepatic mitochondria attenuate western diet induced diabetes
STUDY_TYPE
Supplementation of mice with probiotic bacteria
STUDY_SUMMARY
For WD + Microbes 1 and WD + Microbes 3 experiments, C57BL/6 mice were fed western diet or western diet supplemented with Lactobacillus gasseri or Lactobacillus johnsonii for 8 weeks and serum was collected from each mice. For Pooled_1 (control group, western diet only), and Pooled_2 (western diet + Lactobacillus gasseri) groups, serum from 5 mice each was pooled. For Pooled_3 (western diet + Lactobacillus johnsonii) group, serum from 4 mice was pooled. For Pooled_7 (control group, western diet only), Pooled_8 (western diet + Lactobacillus gasseri) and Pooled_9 (western diet + Lactobacillus johnsonii) groups, serum from 6 mice each was pooled. For WD + Microbes 6 experiment, C57BL/6 mice were fed western diet for 8 weeks. Then, one group (n = 5) of mice was supplemented with Lactobacillus gasseri for 12 weeks along with continuation of western diet and serum was collected. For Pooled_10 (control group, western diet only) and Pooled_11 (western diet + Lactobacillus gasseri) groups, serum from 5 mice each was pooled. For GF + WD + LG experiment, C57BL/6 germ free mice were fed western diet or western diet supplemented with Lactobacillus gasseri for 2 weeks. For Pooled_12 (control, western diet only) and Pooled_13 (western diet + Lactobacillus gasseri) groups, serum from 2 mice each was pooled.
Sub-nanoliter metabolomics via mass spectrometry to characterize volume-limited samples
STUDY_SUMMARY
The human metabolome provides a window into the mechanisms and biomarkers of various diseases. However, because of limited availability, many sample types are still difficult to study by metabolomic analyses. Here, we present a new mass spectrometry (MS)-based metabolomics strategy that only consumes sub-nanoliter sample volumes. The approach consists of combining a customized metabolomics workflow with a pulsed MS ion generation method, known as triboelectric nanogenerator inductive nanoelectrospray ionization (TENGi nanoESI) MS. The first set of samples tested for this approach included exhaled breath condensates (EBC) collected from cystic fibrosis (CF) patients with impaired glucose tolerance to study the metabolome changes before and after the oral glucose tolerance test.
The human metabolome provides a window into the mechanisms and biomarkers of various diseases. However, because of limited availability, many sample types are still difficult to study by metabolomic analyses. Here, we present a new mass spectrometry (MS)-based metabolomics strategy that only consumes sub-nanoliter sample volumes. The approach consists of combining a customized metabolomics workflow with a pulsed MS ion generation method, known as triboelectric nanogenerator inductive nanoelectrospray ionization (TENGi nanoESI) MS. A second example to illustrate TENGi MS capabilities involves rare cell metabolomics of cultured mesenchymal stromal cells (MSCs), a cell type that has shown potential for treating a variety of chronic diseases. Examination of metabolic changes of MSCs cultured under conditions that may impact in vitro therapeutic activity, such as aggregate culture, or preconditioning with interferon gamma (IFN- Ī³)13, is critical for identifying attributes of cell quality. Reducing cell numbers required to perform MSC metabolomic analysis is essential for improving the manufacturing of highly therapeutic MSCs without significantly impeding production.
Metabolites in contents of small intestine in wild type and DAOG181R/G181R mice
STUDY_SUMMARY
To investigate if DAO regulates metabolites in intestinal lumen, we compared metabolites in contents of small intestine in wild type and DAOG181R/G181R mice. All mice have C57BL/6 background and 8 weeks of age.
Developing preliminary blood metabolomics-based biomarkers of insufficient sleep in humans
STUDY_SUMMARY
Study Objective: Identify small molecule biomarkers of insufficient sleep using untargeted plasma metabolomics in humans undergoing experimental insufficient sleep. Methods: We conducted a cross-over laboratory study where 16 normal weight participants (8 men; age 22 Ā± 5 years; body mass index < 25 kg/m2) completed three baseline days (BL; 9h sleep opportunity per night) followed by five day insufficient (5H; 5h sleep opportunity per night) and adequate (9H; 9h sleep opportunity per night) sleep conditions. Energy balanced diets were provided during baseline, with ad libitum energy intake provided during the insufficient and adequate sleep conditions. Untargeted plasma metabolomics analyses were performed using blood samples collected every 4h across the final 24h of each condition. Biomarker models were developed using logistic regression and linear support vector machine algorithms. Results: The top performing biomarker model was developed by linear support vector machine modeling, consisted of 65 compounds, and discriminated insufficient versus adequate sleep with 74% overall accuracy and a Matthewās Correlation Coefficient of 0.39. The compounds in the top performing biomarker model were associated with ATP Binding Cassette Transporters in Lipid Homeostasis, Phospholipid Metabolic Process, Plasma Lipoprotein Remodeling, and sphingolipid metabolism. Conclusion: We identified potential metabolomics-based biomarkers of insufficient sleep in humans. Further development and validation of omics-based biomarkers of insufficient sleep will advance our understanding of the negative consequences of insufficient sleep, improve diagnosis of poor sleep health, and identify targets for countermeasures designed to mitigate the negative health consequences of insufficient sleep.
7 control fibroblasts samples and 7 patient-derived fibroblasts samples were collected at day 0 and day 5. Intracellular metabolites were extracted from cells cultured in 6 well plate while acyl-CoA and 5-methyltetrahydrofolate were extracted from cells cultured in 60 mm dish.
The NIH sponsored multicenter Genetic Epidemiology of COPD (COPDGene (ClinicalTrials.gov Identifier: NCT01969344) study was approved and reviewed by the institutional review board at all participating centers (1). All study participants provided written informed consent. This study enrolled 10,198 non-Hispanic white (NHW) and African American (AA) individuals from January 2008 until April 2011 (Phase 1) who were aged 45-80 with ā„10 pack-year smoking history and no exacerbations for >30 days. In addition, 465 age and gender matched healthy individuals with no history of smoking were enrolled as controls (mostly at Phase 2). From July 2013 to July 2017, 5,697 subjects returned for an in-person 5-year visit. Each in-person visit included spirometry before and after albuterol, quantitative CT imaging of the chest, and blood sampling. From two clinical centers (National Jewish Health and University of Iowa) 162 subjects at Phase 1 (all NHW) and 1,136 subjects (1,040 NHW, 96 AA) participated in an ancillary study in which they provided fresh frozen plasma collected using an 8.5 ml p100 tube (Becton Dickinson) at Phase 2.
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Colon NMR 1D
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Colon NMR HSQC
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Colon ICMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Colon DI-FTMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Five Easy Metrics of Data Quality for LC-MS based Global Metabolomics
STUDY_SUMMARY
Data quality in global metabolomics is of great importance for biomarker discovery and systems biology studies. However, comprehensive metrics and methods to evaluate and compare the data quality of global metabolomics data sets are lacking. In this work, we combine newly developed metrics, along with well-known measures, to comprehensively and quantitatively characterize the data quality across two similar LC-MS platforms, with the goal of providing an efficient and improved ability to evaluate the data quality in global metabolite profiling experiments. A pooled human serum sample was run 50 times on two high-resolution LC-QTOF-MS platforms to provide profile and centroid MS data. These data were processed using Progenesis Qi software and then analyzed using five important data quality measures, including retention time drift, compound coverage, missing values and MS reproducibility (2 measures). The coverage was fit to a Gamma distribution versus compound abundance, which was normalized to allow comparison of different platforms. To evaluate missing values, characteristic curves were obtained by plotting the compound detection percentage versus extraction frequency. To characterize reproducibility, the accumulative coefficient of variation (CV) versus percentage of total compounds detected and CV vs intra-class correlation coefficient (ICC) were investigated. Key findings include significantly better performance using profile mode data compared to centroid mode as well quantitatively better performance from the newer, higher resolution instrument. A summary of the results given in tabulated form gives a snapshot of the experimental results and provides a template to evaluate the global metabolite profiling workflow. In total, these measures give a good overall view of data quality in global profiling and allow comparisons of data acquisition strategies and platforms as well as optimization of parameters.
INSTITUTE
University of Washington
DEPARTMENT
Anesthesiology and Pain
LABORATORY
Daniel Raftery
LAST_NAME
Zhang
FIRST_NAME
Xinyu
ADDRESS
850 Republican Street, Seattle, Washington 98109, United States
Quantification of lipid species from cultured human MDA-MB-231 and BT549 cells with or without siRNA knockdown of ACSL1 and treated or not with eleostearic acid
Targeted metabolomic analysis on hexosamine biosynthetic pathway in flies on time restricted feeding
STUDY_TYPE
metabolomic identification
STUDY_SUMMARY
The integration of circadian and metabolic signals is essential for maintaining robust circadian rhythms and ensuring efficient metabolism and energy use. Using Drosophila as an animal model, we showed observed strong correlation between daily daily rhythms of protein O-linked N-acetylglucosaminylation (O-GlcNAcylation) and clock-controlled feeding-fasting cycles, suggesting that O-GlcNAcylation rhythms are primarily driven by nutrient input. Interestingly, daily O-GlcNAcylation rhythms were severely dampened when we subjected flies to time-restricted feeding (TRF) at unnatural feeding time. This suggests the presence of a clock-regulated buffering mechanism that prevents excessive O-GlcNAcylation at non-optimal times of the day-night cycle, which could disrupt circadian health. We performed targeted metabolomic analysis on hexosamine biosynthetic pathway (HBP), which produces UDP-GlcNAc (the substrate for O-GlcNAcylation), to evaluate the daily activity of HBP enzymes under TRF conditions. We found glutamine--fructose-6-phosphate amidotransferase (GFAT) mediates this buffering mechanism.
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Liver ICMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Liver NMR 1D
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Liver NMR HSQC
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Lung NMR 1D
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Lung NMR HSQC
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Plasma NMR 1D
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Plasma NMR HSQC
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Small Intenstines NMR 1D
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Spleen NMR 1D
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation Spleen - NMR HSQC
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolites changes related to glucose-mediated energy production in chemotheraphy-induced cachexia
STUDY_SUMMARY
Targeted metabolomics platforms included amino acids and metabolites related to glucose-mediated energy production. The targeted metabolome changed with chemotheraphy-induced cachexia, and the changes were reversed with potential treatment of the cachexia. .rdb files were included as raw data files where detailed information regarding MRM transitions and internal standards can be found. Several amino acids (Gly, Pro, Gln, Taurine) were analyzed after dilution because their peak intensities were too high. Thus their analysis was performed separately from other amino acids, and their rdb files were saved in separate rdb files.
INSTITUTE
Asan medical Center, University of Ulsan College of Medicine
LAST_NAME
Yoo
FIRST_NAME
Hyun Ju
ADDRESS
88, Olympic-ro, 43-gil, Songpa-gu, Seoul 05505, South Korea
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Small Intenstines NMR HSQC
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Lung DI-FTMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Lung ICMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Small Intestines DI-FTMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Small Intestines ICMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Spleen DI-FTMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Spleen ICMS
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Metabolomics of lung injury after allogeneic hematopoietic cell transplantation - Liver DI-FTMS
STUDY_TYPE
preliminary data
STUDY_SUMMARY
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment option for a variety of hematological malignancies. Interactions between the donor immune system and the patient tissue result in a disease, called GVHD. The pathophysiology of acute GVHD can be hypothesized in three sequential phases: cytokine storm and activation of the antigen-presenting cells (APC), donor T cell activation and effector cell phase. Idiopathic pneumonia syndrome (IPS) is one of the most deleterious complications after allogeneic HCT and is considered not only to be related to conditioning regimen toxicity but also represents an end organ damage caused by allo-reactive T cells, therefore making the lung susceptible to a two-pronged attack, one of which overlaps with GVHD causing other target organ injury. IPS results in mortality of up to 90% of patients. We will use a murine model of IPS and GVHD which is well established in our group, and in which disease evolves either across disparities in major histocompatibility complex (MCH) class I and II, minor histocompatibility antigens (miHags) or both. Metabolomics changes following syngeneic and allogeneic HCT at post-transplantation Days +7 (cytokine storm phase) and Days +42 (cellular effector phase) are compared to baseline wild-type (naive) controls. Prior to analysis, naĆÆve - and experimental mice (N=3 from each group) were fed with semi-liquid diet supplemented with tracers (13C6-glucose ) over 24 hours. At the end of 7 days or 42 days, respectively, feces and aGVHD target organs (colon, liver and lung) were collected from all groups and further processed and / or analyzed. We expect to reveal metabolic pathways affected after allo-HCT which contribute to immune cell mediated lung injury (IPS) and will potentially identify different metabolic pathways in other GVHD target organs.
INSTITUTE
University of Kentucky
DEPARTMENT
MCC
LAST_NAME
Hildebrandt
FIRST_NAME
Gerhard
ADDRESS
CTW-453, 900 South Limestone street. UKY. Lexington, Kentucky-40536
Design of Experiments for Maximizing T cell endpoints
STUDY_SUMMARY
Purified T cells from a single healthy donor were expanded in vitro with either magnetic beads or degradable micro-scaffold (DMS) particles. Magnetic bead cultures were expanded according to manufacturerās instructions, while DMS cultures were expanded with varying DMS particle concentration, IL2 concentration, and antigen surface density on the particles, according to a design of experiments. Media of each culture of was sampled repeatedly over the course of a 14 day period. Quantities and characteristics of activated T cells were assessed at the end of the culture period, and media was analyzed by 1H-NMR. Analysis of spectra resulted in a set of 20 features that was used in predictive modeling of T cell endpoints, along with culture parameters described above and cytokine data. A second validation experiment was performed with different values for culture parameters, using the same culture, sampling, and NMR analysis methods.
Lipidomics dataset of PTEN deletion-induced nerve regeneration mouse model
STUDY_SUMMARY
We present the lipidome of adult PTENloxP/loxP mice subjected to intravitreal injection of adeno-associated viruses expressing Cre (AAV-Cre) as a model of regeneration. At 4-week-old, PTENloxP/loxP mice were intravitreally-injected with 2-3 Ī¼l of either AAV-Cre (KO) or AAV-PLAP (control), and two weeks later optic nerve crush was performed. At indicated time-points after crush (0 days, 7 days, 14 days), mice were euthanized and optic nerves were immediately dissected out, and then flash frozen on dry ice. The Bligh and Dyer method was used for lipid extraction, followed by mass spectrometry lipid profiling with a Q-Exactive Orbitrap Liquid Chromatography-Mass Spectrometer (LC MS-MS). The raw scans were analysed with LipidSearch 4.2 and the statistical analysis was conducted through Metaboanalyst 4.0
NMR spectroscopy analysis reveals altered metabolic homeostasis in Arabidopsis seedlings treated with a cytokinesis inhibitor
STUDY_SUMMARY
In plant cytokinesis, de novo formation of a cell plate that evolves into the new cell wall, partitions the cytoplasm of the dividing cell. Cell plate formation involves highly orchestrated vesicle accumulation, fusion, and membrane network maturation supported by the temporary integration of elastic and pliable callose. The small molecule, Endosidin 7 (ES7), arrests late cytokinesis in Arabidopsis by inhibiting callose deposition at the cell plate. Its effect is specific, as it does not broadly affect endomembrane trafficking or cytoskeletal organization. It has emerged as a very valuable tool for dissecting this essential plant process. In order to gain deeper insights regarding its mode of action and the effects of cytokinesis inhibition on overall plant growth, we investigated the effect of ES7 through a nuclear magnetic resonance (NMR) spectroscopy metabolomics approach. In this case study, profiles of Arabidopsis leaf and root tissues were analyzed at different growth stages and ES7 exposure levels. The results show tissue-specific changes in the plant metabolic profile across a developmental gradient, and the effect that ES7 treatment has on the corresponding metabolome. The ES7 induced profile suggests metabolic compensations in central metabolism pathways in response to cytokinesis inhibition. Further, this study shows that long-term treatment of ES7 disrupts the homeostasis of primary metabolism in Arabidopsis seedlings, likely via alteration of hormonal regulation.
Large diversity in nitrogen- and sulfur-containing compatible solute profiles in polar and temperate diatoms
STUDY_TYPE
Intracellular metabolites were quantified in diatom species
STUDY_SUMMARY
Intense bottom-ice algal blooms, often dominated by diatoms, are an important source of food for grazers, organic matter for export during sea ice melt, and dissolved organic carbon. Sea-ice diatoms have a number of adaptations, including accumulation of compatible solutes, that allow them to inhabit this highly variable environment characterized by extremes in temperature, salinity, and light. In addition to protecting them from extreme conditions, these compounds present a labile, nutrient-rich source of organic matter and include precursors to climate active compounds (e.g. DMS), which are likely regulated with environmental change. Here, intracellular concentrations of 45 metabolites were quantified in three sea-ice diatom species and were compared to two temperate diatom species, with a focus on compatible solutes and free amino acid pools. There was a large diversity of metabolite concentrations between diatoms with no clear pattern identifiable for sea-ice species. Concentrations of some compatible solutes (isethionic acid, homarine) approached 1 M in the sea-ice diatoms, Fragilariopsis cylindrus and Navicula cf. perminuta, but not in the larger sea-ice diatom, Nitzschia lecointei or in the temperate diatom species. The differential use of compatible solutes in sea-ice diatoms suggest different adaptive strategies and highlights which small organic compounds may be important in polar biogeochemical cycles.
INSTITUTE
University of Washington
DEPARTMENT
Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Dawson
FIRST_NAME
Hannah
ADDRESS
1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA, 98195, USA
EMAIL
hmdawson@uw.edu
PHONE
206-543-0744
PUBLICATIONS
Dawson et al, 2020, Integrative and Comparative Biology
Metabolomics by UHPLC-HRMS reveals the impact of heat stress on pathogen-elicited immunity in maize
STUDY_SUMMARY
Studies investigating crop resistance to biotic and abiotic stress have largely focused on plant responses to singular forms of stress and individual biochemical pathways that only partially represent stress responses. Thus, combined biotic and abiotic stress treatments and the global assessment of their elicited metabolic expression remains largely unexplored. In this study, we employed targeted and untargeted metabolomics to investigate the metabolic responses of maize (Zea mays) to both individual and combinatorial stress treatments using heat (abiotic) and Cochliobolus heterostrophus infection (biotic) experiments. Ultra-high-performance liquid chromatography-high-resolution mass spectrometry revealed significant metabolic responses to C. heterostrophus infection and heat stress, and comparative analyses between these individual forms of stress demonstrated differential elicitation between the two global metabolomes. In combinatorial experiments, treatment with heat stress prior to fungal inoculation negatively impacted maize disease resistance against C. heterostrophus, and distinct metabolome separation between combinatorial stressed plants and the non-heat stressed infected controls was observed. Targeted analysis revealed inducible primary and secondary metabolite responses to biotic/abiotic stress, and combinatorial experiments indicated that deficiency in the hydroxycinnamic acid, p-coumaric acid, may lead to the heat-induced susceptibility of maize to C. heterostrophus. Collectively, these findings demonstrate that abiotic stress can predispose crops to more severe disease symptoms, underlining the increasing need to investigate defense chemistry in plants under combinatorial stress.
INSTITUTE
Agricultural Research Service, United States Department of Agriculture
DEPARTMENT
Center of Medical, Agricultural, and Veterinary Entomology
Plasma lipidomic profiles after a low and high glycemic load dietary pattern in a randomized controlled cross over feeding study
STUDY_SUMMARY
Background: Dietary patterns low in glycemic load are associated with reduced risk of cardiometabolic diseases. Improvements in serum lipid concentrations may play a role in these observed associations. Objective: We investigated how dietary patterns differing in glycemic load affect a clinical lipid panel and plasma lipidomics profiles. Methods: In a crossover, controlled feeding study, 80 healthy participants (n=40 men, n=40 women), 18-45 y were randomized to receive low-glycemic load (LGL) or high glycemic load (HGL) diets for 28 days each with at least a 28-day washout period between controlled diets. Fasting plasma samples were collected at baseline and end of each diet period. A clinical lipid panel including total-, VLDL-, LDL-, and HDL-cholesterol and triglycerides were measured using an auto-analyzer. Lipidomics analysis using mass-spectrometry provided the concentrations of 863 species. Linear mixed models were used to test for a diet effect. Results: Lipids from the clinical panel were not significantly different between diets. Lipidomics analysis showed that 67 lipid species, predominantly in the triacylglycerol class, differed between diets (FDR<0.05). A majority of these were higher after the LGL diet compared to the HGL. Conclusion: While the clinical lipid measures did not differ between diets, some lipid species were higher after the LGL diet in the lipidomics analysis. The two diets were eucaloric and had similar percentage of energy from carbohydrate, protein and fat. Thus, the difference in macronutrient, particularly carbohydrate, quality of the LGL diet is likely affecting the composition of lipid species.
Global Urine Metabolic Profiling to Predict Gestational Age in Term and Preterm Pregnancies
STUDY_SUMMARY
Assessment of gestational age (GA) is key to provide optimal care during pregnancy. However, its accurate determination remains challenging in low- and middle-resource countries, where access to obstetric ultrasound is limited. Hence, there is an urgent need to develop clinical approaches that allow accurate and inexpensive estimation of GA. We investigated the ability of urinary metabolites to predict GA at time of collection in a diverse multi-site cohort (n = 99) using a broad-spectrum liquid chromatography coupled with mass spectrometry (LC-MS) platform. Our approach detected a myriad of steroid hormones and their derivatives including estrogens, progesterones, corticosteroids and androgens that associated with pregnancy progression. We developed a prediction model that predicted GA with high accuracy using the levels of three metabolites (rho = 0.87, .RMSE = 1.58 weeks). These predictions were robust irrespective of whether the pregnancy went to term or ended prematurely. Overall, we demonstrate the feasibility of implementing urine collection for metabolomics analysis in large-scale multi-site studies and we report a predictive model of GA with a potential clinical value.
Global metabolomics of IFNy cued neurogenic NSCs seeded on hydrogel
STUDY_SUMMARY
Neural stem cells (NSCs) provide a strategy to replace damaged neurons following traumatic central nervous system injuries. A major hurdle to translation of this therapy is that direct application of NSCs to CNS injury does not support sufficient neurogenesis due to lack of proper cues. To provide prolonged spatial cues to NSCs IFN-Ī³ was immobilized to biomimetic hydrogel substrate to supply physical and biochemical signals to instruct the encapsulated NSCs to be neurogenic. However, the immobilization of factors, including IFN-Ī³, versus soluble delivery of the same factor, has been incompletely characterized especially with respect to activation of signaling and metabolism in cells over longer time points. In this study, protein and metabolite changes in NSCs induced by immobilized versus soluble IFN-Ī³ at 7 days were evaluated. Soluble IFN-Ī³, refreshed daily over 7 days, elicited stronger responses in NSCs compared to immobilized IFN-Ī³ indicating that immobilization may not sustain signaling or has altered ligand/receptor interaction and integrity. However, both IFN-Ī³ delivery types supported increased Ī²III tubulin expression in parallel with canonical and non-canonical receptor-signaling compared to no IFN-Ī³. Global metabolomics and pathway analysis revealed that soluble and immobilized IFN-Ī³ altered metabolic pathway activities including energy, lipid and amino acid synthesis, with soluble IFN-Ī³ having the greatest metabolic impact overall.
INSTITUTE
University of Akron
DEPARTMENT
Chemistry
LABORATORY
Shriver lab
LAST_NAME
Baumann
FIRST_NAME
Hannah
ADDRESS
190 E. Buchtel Common
EMAIL
hjb17@zips.uakron.edu
PHONE
4198864033
NUM_GROUPS
3
PUBLICATIONS
Metabolomic and Signaling Programs Induced by Immobilized versus Soluble IFN Ī³ in Neural Stem Cells
Dynamic binning peak detection and assessment of various lipidomics liquid chromatography-mass spectrometry pre-processing platforms
STUDY_SUMMARY
Liquid chromatography-mass spectrometry (LC-MS) based lipidomics generate a large dataset, which requires high-performance data pre-processing tools for their interpretation such as XCMS, mzMine and Progenesis. These pre-processing tools rely heavily on accurate peak detection, which depends on setting the peak detection mass tolerance (PDMT) properly. The PDMT is usually set with a fixed value in either ppm or Da units. However, this fixed value may result in duplicates or missed peak detection. Therefore, we developed the dynamic binning method for accurate peak detection, which takes into account the peak broadening described by well-known physics laws of ion separation and set dynamically the value of PDMT as a function of m/z. Namely, in our method, the PDMT is proportional to for FTICR, to for Orbitrap, to m/z for Q-TOF and is a constant for Quadrupole mass analyzer, respectively. The dynamic binning method was implemented in XCMS. Our further goal was to compare the performance of different lipidomics pre-processing tools to find differential compounds. We have generated set samples with 43 lipids internal standards differentially spiked to aliquots of one human plasma lipid sample using Orbitrap LC-MS/MS. The performance of the various pipelines using aligned parameter sets was quantified by a quality score system which reflects the ability of a pre-processing pipeline to detect differential peaks spiked at various concentration levels. The quality score indicates that the dynamic binning method improves the performance of XCMS (maximum p-value 9.8Ā·10-3 of two-sample Wilcoxon test). The modified XCMS software was further compared with mzMine and Progenesis. The results showed that modified XCMS and Progenesis had a similarly good performance in the aspect of finding differential compounds. In addition, Progenesis shows lower variability as indicated by lower CVs, followed by XCMS and mzMine. The lower variability of Progenesis improve the quantification, however, provide an incorrect quantification abundance order of spiked-in internal standards.
Metabolomics of murine embryos cultured in a microfluidic device and comparison with traditional microdrops culture
STUDY_SUMMARY
Global untargeted metabolomics study to analyse culture media extracted from an innovative microfluidic device or traditional microdrops in presence or absence of murine embryos to investigate PDMS-release of biomolecules and embryo metabolic activity.
To apply this method in a pilot study, we spiked either taurine or sulfoquinovose into an in vitro simplified human microbiota model with and without Bilophila wadsworthia, a known sulfonate utilizer.
Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Sample Type: Heart, Ion Mode: Pos/Neg (part- Heart_ Pos/Neg)
STUDY_SUMMARY
Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis. Mice are randomly assigned to 4 groups in study design. Control: saline + saline Model: saline + LPS; Treatment: STV-Na + LPS; Positive: dexamethasone (Dex) + LPS. Drugs were administered i.p. Six hours after LPS injection, mice were sacrificed. And blood and tissues (heart, lung, liver, spleen and kidney) were subjected to UHPLC-TIMS TOF MS/MS-based metabolomics analyses.
INSTITUTE
Guangdong University of Technology
LAST_NAME
Wang
FIRST_NAME
Shanping
ADDRESS
No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,
Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Liver
STUDY_SUMMARY
Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.
INSTITUTE
Guangdong University of Technology
LAST_NAME
Wang
FIRST_NAME
Shanping
ADDRESS
No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,
Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Lung
STUDY_SUMMARY
Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.Mice are randomly assigned to 4 groups in study design. Control: saline + saline Model: saline + LPS; Treatment: STV-Na + LPS; Positive: dexamethasone (Dex) + LPS. Drugs were administered i.p. Six hours after LPS injection, mice were sacrificed. And blood and tissues (heart, lung, liver, spleen and kidney) were subjected to UHPLC-TIMS TOF MS/MS-based metabolomics analyses.
INSTITUTE
Guangdong University of Technology
LAST_NAME
Wang
FIRST_NAME
Shanping
ADDRESS
No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,
Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Kidney
STUDY_SUMMARY
Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.
INSTITUTE
Guangdong University of Technology
LAST_NAME
Wang
FIRST_NAME
Shanping
ADDRESS
No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,
Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Spleen
STUDY_SUMMARY
Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.
INSTITUTE
Guangdong University of Technology
LAST_NAME
Wang
FIRST_NAME
Shanping
ADDRESS
No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,
Metabolomics reveals the protective effect of isosteviol sodium against multiple organ injury in septic mice - Plasma
STUDY_SUMMARY
Sepsis is a severe inflammatory disorder that can lead to multiple organ injury. Isosteviol sodium (STV-Na) is a terpenoid derived from stevioside that exerts anti-inflammatory, antioxidant and anticancer activities. However, the influence of STV-Na on sepsis remains unknown. Here, we assessed the potential effects of STV-Na on sepsis and multiple organ injury induced by lipopolysaccharide (LPS). We found that STV-Na increased the survival rate of mice treat with LPS, significantly improved the functions of the heart, lung, liver, and kidney, and reduced the production of inflammatory cytokines. Moreover, Multiorgan metabolomics analysis demonstrated that glutathione metabolism, purine metabolism, glycerophospholipid metabolism and pantothenate and CoA biosynthesis, were significantly altered by STV-Na. This study provides novel insights into the metabolite changes of multiple organ injury in septic mice, which may help characterize the underlying mechanism and provide an improved understanding of the therapeutic effects of STV-Na on sepsis.
INSTITUTE
Guangdong University of Technology
LAST_NAME
Wang
FIRST_NAME
Shanping
ADDRESS
No. 100, Waihuan Xilu, Guangzhou Higher Education Mega Center, Panyu District,
Hepatic [U-13C]Lactate tracing and metabolomics in young and old WT and SIRT6 overexpressing mice
STUDY_SUMMARY
Our previous data had suggested that gluconeogenesis capacity from the precursors lactate and glycerol declines in old C57BL/6 mice. This decline was rescued by whole body SIRT6 overexpression. Since the liver is the main gluconeogenic organ, we performed here liver metabolomics in young (6 months) versus old (20-24 months) WT and SIRT6-transgenic mice. We used liver tissues of 6h morning-fasted mice, either in naĆÆve state or 15 minutes after intraperitoneal injection of 1mg/kg [U-13C]Lactate.
INSTITUTE
Bar Ilan University
DEPARTMENT
The Mina & Everard Goodman Faculty of Life Sciences
Liver metabolomics in old liver-specific SIRT6 overexpressing mice
STUDY_SUMMARY
Our previous data had suggested that gluconeogenesis capacity from the precursors lactate and glycerol declines in old C57BL/6 mice. This decline was rescued by whole body SIRT6 overexpression, but not in liver-specific SIRT6 overexpression. Here we preformed liver metabolomics in old liver-specific SIRT6 overexpression and control mice.
INSTITUTE
Bar Ilan University
DEPARTMENT
The Mina & Everard Goodman Faculty of Life Sciences
Metabolomic profiles of six bacteria and K. oxytoca were compared to identify candidate metabolites that may prevent B. bassiana infection of D. antiqua.
INSTITUTE
Qilu University of Technology (Shandong Academy of Sciences)
LAST_NAME
Zhou
FIRST_NAME
Fangyuan
ADDRESS
No. 28789 Jingshidong Road, Jinan, Shandong, 250103, China
Diel investments in phytoplankton metabolite production influenced by associated heterotrophic bacteria
STUDY_TYPE
Incubation experiment
STUDY_SUMMARY
Organic substrate transfer between photoautotrophic and heterotrophic microbes in the surface ocean is a central but poorly understood process in the global carbon cycle. This study developed a co-culture system of marine diatom Thalassiosira pseudonana and heterotrophic bacterium Ruegeria pomeroyi, and addressed diel changes in phytoplankton endometabolite production using nuclear magnetic resonance (NMR) and bacterial metabolite consumption using gene expression. Here we deposit data for NMR analysis from the study. Samples were collected every 6 hours over two days under a diel light cycle. During the course of the study, we observed an increase in some phytoplankton endometabolites presumably due to the effects of the associated bacteria. We introduced an additional experiment and tested this possibility by comparing phytoplankton endometabolite accumulation between axenic treatments and bacteria coculture treatments.
INSTITUTE
University of Georgia
DEPARTMENT
Department of Marine Sciences; Complex Carbohydrate Research Center
Community metabolomes reflect taxon-specific fingerprints of phytoplankton in the ocean
STUDY_TYPE
Metabolomic survey of 21 phytoplankton species
STUDY_SUMMARY
Phytoplankton transform inorganic carbon into thousands of biomolecules, including polar metabolites that represent an important pool of labile fixed carbon, nitrogen, and sulfur. Metabolite production is not identical among phytoplankton, and the flux of these molecules through the microbial loop depends on compound-specific bioavailability to a wider microbial community. Yet relatively little is known about the diversity or concentration of polar metabolites within marine plankton. Here we evaluate 313 metabolites in 21 phytoplankton species and in natural marine particles across environmental gradients to show that bulk community metabolomes reflect the phytoplankton community on a chemical level.
INSTITUTE
University of Washington
DEPARTMENT
Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Heal
FIRST_NAME
Katherine
ADDRESS
1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA, 98195, USA
A Metabolomic Signature of Glucagon Action in Healthy Individuals with Overweight/Obesity Humans
STUDY_SUMMARY
Objective: We sought to identify the circulating metabolites that would serve as markers of glucagon action. Design: In this study, we performed a continuous 72-hour glucagon infusion in healthy individuals with overweight/obesity. Participants were randomized to either glucagon (12.5 ng/kg/min) (GCG 12.5) or glucagon (25 ng/kg/min) GCG 25 or a placebo control were included. A comprehensive metabolomics analysis was then performed from plasma isolated at several time points during the infusion to identify markers of glucagon activity.
INSTITUTE
Translational Research Institute- AdventHealth Orlando
Identification of distinct metabolic perturbations and associated immunomodulatory events during intra-erythrocytic development stage of pediatric Plasmodium falciparum malaria (part-II)
STUDY_SUMMARY
The goal of this study was to interrogate biochemical profiles manifested in human serum samples originating from a cohort of West African children, collected before and during P. falciparum malarial infection, with the aim of characterizing metabolic migration associated with severity of malarial infection.
Identification of distinct metabolic perturbations and associated immunomodulatory events during intra-erythrocytic development stage of pediatric Plasmodium falciparum malaria (part-III)
STUDY_SUMMARY
The goal of this study was to interrogate biochemical profiles manifested in human serum samples originating from a cohort of West African children, collected before and during P. falciparum malarial infection, with the aim of characterizing metabolic migration associated with severity of malarial infection.
Metabolome analysis in the diagnosis of childhood cerebellar ataxia
STUDY_SUMMARY
Metabolome studies to aid in the diagnosis and molecular elucidation of a child presenting chronic progressive cerebellar ataxia and an undiagnosed condition.
INSTITUTE
CEMBIO
LAST_NAME
SƔiz
FIRST_NAME
Jorge
ADDRESS
km 0, Universidad CEU-San Pablo UrbanizaciĆ³n MonteprĆncipe. M-501, 28925 AlcorcĆ³n
Stool metabolites of known identity profiled using hybrid nontargeted methods (part-I)
STUDY_SUMMARY
We determined the effect of diet on the composition and metabolic function of human gut microbiome using a controlled feeding experiment with three divergent diets (vegan, omnivore, and an exclusive enteral nutrition diet (EEN) devoid of dietary fiber) in the Food And Resulting Microbial Metabolites (FARMM) study. The study included an antibiotic and polyethylene glycol (Abx/PEG) intervention to dynamically assess the effect of diet on the reconstitution of the gut microbiota and its associated fecal and plasma metabolome. Samples from thirty healthy volunteers between the ages of 18 and 60, 10 mper diet group were analyzed. Self-reported vegans were required to have followed a vegan diet for a minimum of 6 months prior to enrollment. Key exclusion criteria included inflammatory bowel disease, celiac disease, or other chronic intestinal disorders; prior bowel resection surgery other than appendectomy; baseline bowel frequency less than every 2 days or greater than 3 times daily; creatinine concentration greater than the upper limit of normal; diabetes mellitus; currently smoking; body mass index (BMI) <18.5 or >35; and use of antibiotics and probiotics in the prior 6 months. The 10 vegans continued to follow their usual diet as outpatients. All participants completed the Diet History Questionnaire II (DHQ II), a food frequency questionnaire developed by the Risk Factor Monitoring and Methods Branch of the National Cancer Institute. The vegan participants also completed three 24 hour diet recalls with a dietitian in the week prior to starting antibiotics. We randomly assigned the 20 omnivores to receive an omnivore diet or EEN (ModulenĀ® IBD) while residing in an inpatient research unit. The macronutrient composition of ModulenĀ® is protein 36g, fat 47g, and carbohydrate 110g per 1000 Kcal. The two omnivore diets were engineered to have a similar composition to EEN. All the subjects consumed the menu A on day 11 and menu B on days 4 and 14. Diets for omnivores were constructed to provide the expected total calories required per day for the participant to maintain their current weight and were adjusted if there was weight gain or loss of more than 2.5 pounds. On days 6, 7, and 8, inpatients participants received vancomycin 500mg orally every 6 hours and neomycin 1000mg orally every 6 hours. On day 7, participants consumed 4L of polyethylene glycol (PEG) based bowel purgative (GoLytelyĀ®). Participants in the omnivore and EEN arms left the inpatient research unit only under the direct supervision of a research staff member. The vegan outpatients reported to the hospital twice on days 6,7 and 8 to receive antibiotics and to consume PEG on day 7. The first stool sample of each day of the inpatient groups was collected, aliquoted and frozen immediately at -80oC. Blood was collected on days 1, 5, 9, 12 and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. The outpatient participants following the vegan diet also had blood collected on days 1, 5, 9, 12, and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. Among these participants, the first stool of the day was collected at home daily and kept on ice packs until it was brought to the research unit where it could be aliquoted and frozen. Samples from vegans were received for aliquoting within 24 hours and on average within 4 hours (Wu et al., 2010). Day 0 stool was not collected from vegans since their diet did not change. The aliquoted amounts for all samples ranged from 500 mg to 1 g. The aliquots were taken from different areas of the sample. Remaining sample was then collected into one residual 50 ml conical tube with a tongue depressor. Any remaining stool was discarded. The University of Pennsylvania Institutional Review Board (IRB) approved the research protocol.
INSTITUTE
Broad Institute of MIT and Harvard
LAST_NAME
Clish
FIRST_NAME
Clary
ADDRESS
415 Main Street, Cambridge, MA, 02142, USA
EMAIL
clary@broadinstitute.org
PHONE
617-714-7654
NUM_GROUPS
3
TOTAL_SUBJECTS
30
NUM_MALES
20
NUM_FEMALES
10
STUDY_COMMENTS
stool samples collected at baseline and 3-4 timepoints
Stool unknowns profiled using hybrid nontargeted methods (part-II)
STUDY_SUMMARY
We determined the effect of diet on the composition and metabolic function of human gut microbiome using a controlled feeding experiment with three divergent diets (vegan, omnivore, and an exclusive enteral nutrition diet (EEN) devoid of dietary fiber) in the Food And Resulting Microbial Metabolites (FARMM) study. The study included an antibiotic and polyethylene glycol (Abx/PEG) intervention to dynamically assess the effect of diet on the reconstitution of the gut microbiota and its associated fecal and plasma metabolome. Samples from thirty healthy volunteers between the ages of 18 and 60, 10 mper diet group were analyzed. Self-reported vegans were required to have followed a vegan diet for a minimum of 6 months prior to enrollment. Key exclusion criteria included inflammatory bowel disease, celiac disease, or other chronic intestinal disorders; prior bowel resection surgery other than appendectomy; baseline bowel frequency less than every 2 days or greater than 3 times daily; creatinine concentration greater than the upper limit of normal; diabetes mellitus; currently smoking; body mass index (BMI) <18.5 or >35; and use of antibiotics and probiotics in the prior 6 months. The 10 vegans continued to follow their usual diet as outpatients. All participants completed the Diet History Questionnaire II (DHQ II), a food frequency questionnaire developed by the Risk Factor Monitoring and Methods Branch of the National Cancer Institute. The vegan participants also completed three 24 hour diet recalls with a dietitian in the week prior to starting antibiotics. We randomly assigned the 20 omnivores to receive an omnivore diet or EEN (ModulenĀ® IBD) while residing in an inpatient research unit. The macronutrient composition of ModulenĀ® is protein 36g, fat 47g, and carbohydrate 110g per 1000 Kcal. The two omnivore diets were engineered to have a similar composition to EEN. All the subjects consumed the menu A on day 11 and menu B on days 4 and 14. Diets for omnivores were constructed to provide the expected total calories required per day for the participant to maintain their current weight and were adjusted if there was weight gain or loss of more than 2.5 pounds. On days 6, 7, and 8, inpatients participants received vancomycin 500mg orally every 6 hours and neomycin 1000mg orally every 6 hours. On day 7, participants consumed 4L of polyethylene glycol (PEG) based bowel purgative (GoLytelyĀ®). Participants in the omnivore and EEN arms left the inpatient research unit only under the direct supervision of a research staff member. The vegan outpatients reported to the hospital twice on days 6,7 and 8 to receive antibiotics and to consume PEG on day 7. The first stool sample of each day of the inpatient groups was collected, aliquoted and frozen immediately at -80oC. Blood was collected on days 1, 5, 9, 12 and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. The outpatient participants following the vegan diet also had blood collected on days 1, 5, 9, 12, and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. Among these participants, the first stool of the day was collected at home daily and kept on ice packs until it was brought to the research unit where it could be aliquoted and frozen. Samples from vegans were received for aliquoting within 24 hours and on average within 4 hours (Wu et al., 2010). Day 0 stool was not collected from vegans since their diet did not change. The aliquoted amounts for all samples ranged from 500 mg to 1 g. The aliquots were taken from different areas of the sample. Remaining sample was then collected into one residual 50 ml conical tube with a tongue depressor. Any remaining stool was discarded. The University of Pennsylvania Institutional Review Board (IRB) approved the research protocol.
Plasma metabolites of known identity profiled using hybrid nontargeted methods (part-III)
STUDY_SUMMARY
We determined the effect of diet on the composition and metabolic function of human gut microbiome using a controlled feeding experiment with three divergent diets (vegan, omnivore, and an exclusive enteral nutrition diet (EEN) devoid of dietary fiber) in the Food And Resulting Microbial Metabolites (FARMM) study. The study included an antibiotic and polyethylene glycol (Abx/PEG) intervention to dynamically assess the effect of diet on the reconstitution of the gut microbiota and its associated fecal and plasma metabolome. Samples from thirty healthy volunteers between the ages of 18 and 60, 10 mper diet group were analyzed. Self-reported vegans were required to have followed a vegan diet for a minimum of 6 months prior to enrollment. Key exclusion criteria included inflammatory bowel disease, celiac disease, or other chronic intestinal disorders; prior bowel resection surgery other than appendectomy; baseline bowel frequency less than every 2 days or greater than 3 times daily; creatinine concentration greater than the upper limit of normal; diabetes mellitus; currently smoking; body mass index (BMI) <18.5 or >35; and use of antibiotics and probiotics in the prior 6 months. The 10 vegans continued to follow their usual diet as outpatients. All participants completed the Diet History Questionnaire II (DHQ II), a food frequency questionnaire developed by the Risk Factor Monitoring and Methods Branch of the National Cancer Institute. The vegan participants also completed three 24 hour diet recalls with a dietitian in the week prior to starting antibiotics. We randomly assigned the 20 omnivores to receive an omnivore diet or EEN (ModulenĀ® IBD) while residing in an inpatient research unit. The macronutrient composition of ModulenĀ® is protein 36g, fat 47g, and carbohydrate 110g per 1000 Kcal. The two omnivore diets were engineered to have a similar composition to EEN. All the subjects consumed the menu A on day 11 and menu B on days 4 and 14. Diets for omnivores were constructed to provide the expected total calories required per day for the participant to maintain their current weight and were adjusted if there was weight gain or loss of more than 2.5 pounds. On days 6, 7, and 8, inpatients participants received vancomycin 500mg orally every 6 hours and neomycin 1000mg orally every 6 hours. On day 7, participants consumed 4L of polyethylene glycol (PEG) based bowel purgative (GoLytelyĀ®). Participants in the omnivore and EEN arms left the inpatient research unit only under the direct supervision of a research staff member. The vegan outpatients reported to the hospital twice on days 6,7 and 8 to receive antibiotics and to consume PEG on day 7. The first stool sample of each day of the inpatient groups was collected, aliquoted and frozen immediately at -80oC. Blood was collected on days 1, 5, 9, 12 and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. The outpatient participants following the vegan diet also had blood collected on days 1, 5, 9, 12, and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. Among these participants, the first stool of the day was collected at home daily and kept on ice packs until it was brought to the research unit where it could be aliquoted and frozen. Samples from vegans were received for aliquoting within 24 hours and on average within 4 hours (Wu et al., 2010). Day 0 stool was not collected from vegans since their diet did not change. The aliquoted amounts for all samples ranged from 500 mg to 1 g. The aliquots were taken from different areas of the sample. Remaining sample was then collected into one residual 50 ml conical tube with a tongue depressor. Any remaining stool was discarded. The University of Pennsylvania Institutional Review Board (IRB) approved the research protocol.
INSTITUTE
Broad Institute of MIT and Harvard
LAST_NAME
Clish
FIRST_NAME
Clary
ADDRESS
415 Main Street, Cambridge, MA, 02142, USA
EMAIL
clary@broadinstitute.org
PHONE
617-714-7654
NUM_GROUPS
3
TOTAL_SUBJECTS
30
NUM_MALES
20
NUM_FEMALES
10
STUDY_COMMENTS
plasma samples collected at baseline and 3-4 timepoints
Plasma unknowns profiled using hybrid nontargeted methods (part-IV)
STUDY_SUMMARY
We determined the effect of diet on the composition and metabolic function of human gut microbiome using a controlled feeding experiment with three divergent diets (vegan, omnivore, and an exclusive enteral nutrition diet (EEN) devoid of dietary fiber) in the Food And Resulting Microbial Metabolites (FARMM) study. The study included an antibiotic and polyethylene glycol (Abx/PEG) intervention to dynamically assess the effect of diet on the reconstitution of the gut microbiota and its associated fecal and plasma metabolome. Samples from thirty healthy volunteers between the ages of 18 and 60, 10 mper diet group were analyzed. Self-reported vegans were required to have followed a vegan diet for a minimum of 6 months prior to enrollment. Key exclusion criteria included inflammatory bowel disease, celiac disease, or other chronic intestinal disorders; prior bowel resection surgery other than appendectomy; baseline bowel frequency less than every 2 days or greater than 3 times daily; creatinine concentration greater than the upper limit of normal; diabetes mellitus; currently smoking; body mass index (BMI) <18.5 or >35; and use of antibiotics and probiotics in the prior 6 months. The 10 vegans continued to follow their usual diet as outpatients. All participants completed the Diet History Questionnaire II (DHQ II), a food frequency questionnaire developed by the Risk Factor Monitoring and Methods Branch of the National Cancer Institute. The vegan participants also completed three 24 hour diet recalls with a dietitian in the week prior to starting antibiotics. We randomly assigned the 20 omnivores to receive an omnivore diet or EEN (ModulenĀ® IBD) while residing in an inpatient research unit. The macronutrient composition of ModulenĀ® is protein 36g, fat 47g, and carbohydrate 110g per 1000 Kcal. The two omnivore diets were engineered to have a similar composition to EEN. All the subjects consumed the menu A on day 11 and menu B on days 4 and 14. Diets for omnivores were constructed to provide the expected total calories required per day for the participant to maintain their current weight and were adjusted if there was weight gain or loss of more than 2.5 pounds. On days 6, 7, and 8, inpatients participants received vancomycin 500mg orally every 6 hours and neomycin 1000mg orally every 6 hours. On day 7, participants consumed 4L of polyethylene glycol (PEG) based bowel purgative (GoLytelyĀ®). Participants in the omnivore and EEN arms left the inpatient research unit only under the direct supervision of a research staff member. The vegan outpatients reported to the hospital twice on days 6,7 and 8 to receive antibiotics and to consume PEG on day 7. The first stool sample of each day of the inpatient groups was collected, aliquoted and frozen immediately at -80oC. Blood was collected on days 1, 5, 9, 12 and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. The outpatient participants following the vegan diet also had blood collected on days 1, 5, 9, 12, and 15 from which plasma aliquots were immediately isolated and frozen at -80oC. Among these participants, the first stool of the day was collected at home daily and kept on ice packs until it was brought to the research unit where it could be aliquoted and frozen. Samples from vegans were received for aliquoting within 24 hours and on average within 4 hours (Wu et al., 2010). Day 0 stool was not collected from vegans since their diet did not change. The aliquoted amounts for all samples ranged from 500 mg to 1 g. The aliquots were taken from different areas of the sample. Remaining sample was then collected into one residual 50 ml conical tube with a tongue depressor. Any remaining stool was discarded. The University of Pennsylvania Institutional Review Board (IRB) approved the research protocol.
INSTITUTE
Broad Institute of MIT and Harvard
LAST_NAME
Clish
FIRST_NAME
Clary
ADDRESS
415 Main Street, Cambridge, MA, 02142, USA
EMAIL
clary@broadinstitute.org
PHONE
617-714-7654
NUM_GROUPS
3
TOTAL_SUBJECTS
30
NUM_MALES
20
NUM_FEMALES
10
STUDY_COMMENTS
plasma samples collected at baseline and 3-4 timepoints
The effect of fasting and sirtuin overexpression on serum metabolome.
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
The levels of the sirtuins SIRT1 and SIRT6 are regulated by nutrient availability in several mammalian tissues. In addition, overexpressing SIRT1 in mice improves healthspan, while SIRT6 overexpression increases both healthspan and lifespan. However, little is known about the impact of these sirtuins at the in vivo metabolomics level. Thus, we performed metabolomics on serum taken from 15 months old male mice overexpressing SIRT1, SIRT6 and SIRT1+SIRT6 as well as WT controls. Sera were obtained from mice at three nutritional time points - at fed state, after 4h morning fast and after 16h fast.
INSTITUTE
Bar Ilan University
DEPARTMENT
The Mina & Everard Goodman Faculty of Life Sciences
Prochlorococcus extracellular vesicles: Molecular composition and adsorption to diverse microbial cells
STUDY_TYPE
Characterizing the metabolome of Prochlorococcus cells and vesicles
STUDY_SUMMARY
Extracellular vesicles are small (~50ā200 nm diameter) membrane-bound structures released by cells from all domains of life. While extremely abundant in the oceans, our understanding of their functions, both for cells and the emergent ecosystem, is in its infancy. To advance this understanding, we analyzed the lipid, metabolite, and protein content of vesicles produced by two strains of the most abundant phytoplankton cell in the ocean, the cyanobacterium Prochlorococcus. We show that Prochlorococcus exports an enormous array of cellular compounds into their surroundings via extracellular vesicles. The vesicles produced by the two different strains contained some materials in common, but also displayed numerous strain-specific differences, reflecting functional complexity within natural vesicle populations. Prochlorococcus vesicles contain active enzymes, indicating that they can mediate biogeochemically relevant extracellular reactions in the wild. Interaction assays demonstrate that vesicles from Prochlorococcus and multiple genera of heterotrophic bacteria can associate with other marine microbes, including Pelagibacter, the most abundant heterotrophic group in the oceans. Our observations suggest that vesicles may play diverse functional roles in the oceans, including but not limited to mediating energy and nutrient transfers, catalyzing extracellular biochemical reactions, and mitigating toxicity of reactive oxygen species. These findings further indicate that a portion of the ādissolvedā compounds in the oceans are not truly dissolved, but are instead packaged within locally structured, colloidal vesicles.
INSTITUTE
University of Washington
DEPARTMENT
Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Carlson
FIRST_NAME
Laura
ADDRESS
1501 NE Boat Street, Marine Science Building, Room G, Seattle, WA 98195
Perfluorooctanesulfonic acid (PFOS) and perfluorohexanesulfonic acid (PFHxS) alter the blood lipidome and the hepatic proteome in a murine model of diet-induced obesity
STUDY_SUMMARY
Perfluorooctanesulfonic acid (PFOS) and perfluorohexanesulfonic acid (PFHxS) alter the blood lipidome and the hepatic proteome in a murine model of diet-induced obesity
Mitochondrial health is enhanced in rats with higher vs. lower intrinsic exercise capacity and extended lifespan
STUDY_SUMMARY
The intrinsic aerobic capacity of an organism is thought to play a role in aging and longevity. Maximal respiratory rate capacity, a metabolic performance measure, is one of the best predictors of cardiovascular- and all-cause mortality. Rats selectively bred for high-(HCR) vs. low-(LCR) intrinsic running-endurance capacity have up to 31% longer lifespan. We found that positive changes in indices of mitochondrial health in cardiomyocytes (respiratory reserve, maximal respiratory capacity, resistance to mitochondrial permeability transition, autophagy/mitophagy, higher lipids-over-glucose utilization) are uniformly associated with the extended longevity in HCR vs. LCR female rats. Cross-sectional heart metabolomics revealed pathways from lipid metabolism in the heart which were significantly enriched by a select group of strain dependent metabolites, consistent with enhanced lipids utilization by HCR cardiomyocytes. Heart-liver-serum metabolomics further revealed shunting of lipidic substrates between liver and heart via serum during aging. Thus, mitochondrial health in cardiomyocytes is associated with extended longevity in rats with higher intrinsic exercise capacity, and likely these findings can be translated to other populations as predictors of outcomes of health and survival.
INSTITUTE
National Institute on Aging
DEPARTMENT
Cardioprotection Section
LABORATORY
Laboratory of Cardiovascular Science
LAST_NAME
Sollott
FIRST_NAME
Steven
ADDRESS
Laboratory of Cardiovascular Science, National Institute on Aging, NIH, Baltimore, MD 21224, USA
This study explored models predictive of staging and chemotherapy response based on metabolomic analysis of fresh, patient-derived non-small cell lung cancer (NSCLC) core biopsies. Prospectively collected tissue samples before initial treatment were evaluated with high-resolution 2DLC-MS/MS and 13C-glucose enrichment, and the data were comprehensively analyzed with machine learning techniques. Patients were categorized as Disease-Control (DC) [encompassing complete-response (CR), partial-response (PR), and stable-disease (SD)] and Progressive-Disease (PD). Four major types of learning methods (partial least squares discriminant analysis (PLS-DA), support vector machines (SVM), artificial neural networks, and random forests) were applied to differentiate between positive (DC and CR/PR) and poor (PD and SD/PD) responses, and between stage I/II/III and stage IV disease. Models were trained with forward feature selection based on variable importance and tested on validation subsets.
Ī²-Adrenergic regulation of metabolism in macrophages
STUDY_SUMMARY
Macrophages have important roles in the immune system including clearing pathogens and wound healing. Metabolic phenotypes have been associated with functional phenotypes, where pro-inflammatory macrophages have an increased rate of glycolysis and anti-inflammatory macrophages primarily use oxidative phosphorylation. Ī²-adrenoceptor (Ī²AR) signalling in macrophages has been implicated in disease states such as cancer, atherosclerosis and rheumatoid arthritis. The impact of Ī²-adrenoceptor signalling on macrophage metabolism has not been defined. Here we expand on defining the phenotype of macrophages treated with isoprenaline and describe the impact that Ī²AR signalling has on the metabolome and proteome. We found that Ī²AR signalling alters proteins involved in cytoskeletal rearrangement and redox control of the cell. We showed that Ī²AR signalling in macrophages shifts glucose metabolism from glycolysis towards the tricarboxylic acid cycle and pentose phosphate pathways. We also show that Ī²AR signalling perturbs purine metabolism by accumulating adenylate pools. Taken together these results indicate that Ī²AR signalling shifts metabolism to support redox perturbations and upregulate proteins involved in cytoskeletal changes that may impact migration and phagocytosis processes.
INSTITUTE
Monash University
LAST_NAME
Peterson
FIRST_NAME
Amanda
ADDRESS
Drug delivery, disposition and dynamics, Pharmacy and Pharmaceutical Sciences, 381 Royal Parade, Parkville, Victoria, 3052, Australia
Ī²-Adrenergic regulation of metabolism in macrophages (part-II)
STUDY_SUMMARY
Macrophages have important roles in the immune system including clearing pathogens and wound healing. Metabolic phenotypes have been associated with functional phenotypes, where pro-inflammatory macrophages have an increased rate of glycolysis and anti-inflammatory macrophages primarily use oxidative phosphorylation. Ī²-adrenoceptor (Ī²AR) signalling in macrophages has been implicated in disease states such as cancer, atherosclerosis and rheumatoid arthritis. The impact of Ī²-adrenoceptor signalling on macrophage metabolism has not been defined. Here we expand on defining the phenotype of macrophages treated with isoprenaline and describe the impact that Ī²AR signalling has on the metabolome and proteome. We found that Ī²AR signalling alters proteins involved in cytoskeletal rearrangement and redox control of the cell. We showed that Ī²AR signalling in macrophages shifts glucose metabolism from glycolysis towards the tricarboxylic acid cycle and pentose phosphate pathways. We also show that Ī²AR signalling perturbs purine metabolism by accumulating adenylate pools. Taken together these results indicate that Ī²AR signalling shifts metabolism to support redox perturbations and upregulate proteins involved in cytoskeletal changes that may impact migration and phagocytosis processes.
INSTITUTE
Monash University
LAST_NAME
Peterson
FIRST_NAME
Amanda
ADDRESS
Drug delivery, disposition and dynamics, Pharmacy and Pharmaceutical Sciences, 381 Royal Parade, Parkville, Victoria, 3052, Australia
Ī²-Adrenergic regulation of metabolism in macrophages (part-III)
STUDY_SUMMARY
Macrophages have important roles in the immune system including clearing pathogens and wound healing. Metabolic phenotypes have been associated with functional phenotypes, where pro-inflammatory macrophages have an increased rate of glycolysis and anti-inflammatory macrophages primarily use oxidative phosphorylation. Ī²-adrenoceptor (Ī²AR) signalling in macrophages has been implicated in disease states such as cancer, atherosclerosis and rheumatoid arthritis. The impact of Ī²-adrenoceptor signalling on macrophage metabolism has not been defined. Here we expand on defining the phenotype of macrophages treated with isoprenaline and describe the impact that Ī²AR signalling has on the metabolome and proteome. We found that Ī²AR signalling alters proteins involved in cytoskeletal rearrangement and redox control of the cell. We showed that Ī²AR signalling in macrophages shifts glucose metabolism from glycolysis towards the tricarboxylic acid cycle and pentose phosphate pathways. We also show that Ī²AR signalling perturbs purine metabolism by accumulating adenylate pools. Taken together these results indicate that Ī²AR signalling shifts metabolism to support redox perturbations and upregulate proteins involved in cytoskeletal changes that may impact migration and phagocytosis processes.
INSTITUTE
Monash University
LAST_NAME
Peterson
FIRST_NAME
Amanda
ADDRESS
Drug delivery, disposition and dynamics, Pharmacy and Pharmaceutical Sciences, 381 Royal Parade, Parkville, Victoria, 3052, Australia
Aromatic amino acid metabolism by the anaerobic gut bacterium Clostridium sporogenes
STUDY_SUMMARY
Germ-free mice were mono-colonized with wild-type Clostridium sporogenes or its isogenic phenyllactate dehydratase (fldC) mutant. Feces, cecal contents, serum, and urine were collected for untargeted GC-TOF analysis.
Genetic background shapes phenotypic response to diet for adiposity in the Collaborative Cross
STUDY_TYPE
Diet challenge
STUDY_SUMMARY
Defined as chronic excessive accumulation of adiposity, obesity results from long-term imbalance between energy intake and expenditure. The mechanisms behind how caloric imbalance occurs are complex and influenced by numerous biological and environmental factors, especially genetics and diet. Population-based diet recommendations have had limited success partly due to the wide variation in physiological responses across individuals when they consume the same diet. Thus, it is necessary to broaden our understanding of how individual genetics and diet interact relative to the development of obesity for improving weight loss treatment. To determine how consumption of diets with different macronutrient composition alter adiposity and other obesity-related traits in a genetically diverse population, we analyzed body composition, metabolic rate, clinical blood chemistries, and circulating metabolites in 22 strains of mice from the Collaborative Cross (CC), a highly diverse recombinant inbred mouse population, before and after 8 weeks of feeding either a high protein or high fat high sucrose diet. At both baseline and post-diet, adiposity and other obesity-related traits exhibited a broad range of phenotypic variation based on CC strain; diet-induced changes in adiposity and other traits also depended largely on CC strain. In addition to estimating heritability at baseline, we also quantified the effect size of diet for each trait, which varied by trait and experimental diet. Our findings identified CC strains prone to developing obesity, demonstrate the genotypic and phenotypic diversity of the CC for studying complex traits, and highlight the importance of accounting for genetic differences when making dietary recommendations.
Comparing gas chromatography with time-of-flight, quadrupole time-of-flight and quadrupole mass spectrometry for stable isotope tracing (part-I)
STUDY_SUMMARY
Stable isotope tracers are applied in vivo and in vitro studies to reveal the activity of enzymes and intracellular metabolic pathways. Most often, such tracers are used with gas chromatography coupled to mass spectrometry (GC-MS) due to its ease of operation and reproducible mass spectral databases. Differences in isotope tracer performance of classic GC-quadrupole MS instrument and newer time-of-flight instruments are not well-studied. Here, we used three commercially available instruments for the analysis of identical samples from a stable isotope labeling study that used [U-13C6] d-glucose to investigate the metabolism of Rothia mucilaginosa with respect to 29 amino acids and hydroxyl acids involved in primary metabolism. Overall, all three GC-MS instruments (low-resolution GC-SQ-MS, low-resolution GC-TOF-MS, and high-resolution GC-Q-TOF-MS) can be used to perform stable isotope tracing studies for glycolytic intermediates, TCA metabolites and amino acids, yielding similar biological results, with high-resolution GC-Q-TOF-MS offering additional capabilities to identify chemical structures of unknown compounds that might show significant isotope enrichments in biological studies.
INSTITUTE
University of California, Davis
LABORATORY
Oliver Fiehn
LAST_NAME
Zhang
FIRST_NAME
Ying
ADDRESS
West Coast Metabolomics Center, University of California, Davis, 95616, CA, USA
Comparative metabolomics analysis of two Saccharomyces cerevisiae strains: the wild type and mtl1Ī, which carries a deletion of the mechanosensor Mtl1p.
STUDY_SUMMARY
To gather more in-depth knowledge of the Mtl1p mechanosensor's role in Saccharomyces cerevisiae metabolism, we conducted a comparative metabolomic analysis of two Saccharomyces cerevisiae strains: the wild type and mtl1Ī, which carries a deletion of the mechanosensor Mtl1p. Both strains were grown under normal conditions at 27Ā°C. The most significant metabolic changes between these strains were related to amino acid metabolism, purine metabolism, and carboxylic acid metabolism.
INSTITUTE
University of Puerto Rico, Medical Sciences Campus
LAST_NAME
Chorna
FIRST_NAME
Nataliya
ADDRESS
University of Puerto Rico, Medical Sciences Campus, Department of Biochemistry, Main Building, 6th Floor, Room A-632, San Juan, PR 00935
Control (DMSO 0.1%; v/v) and 10 ĀµM DRB18 treated A549 lung cancer cells in vitro for 48 hours
STUDY_TYPE
Anticancer compound treatment experiment
STUDY_SUMMARY
Control (DMSO 0.1%; v/v) and 10 ĀµM DRB18 were used to treated 5 million A549 lung cancer cells in vitro for 48 hours. The untargeted metabolomics analysis was performed on the cell lysates. The main objective of the study was to determine changes in metabolite abundances in lung cancer after treatment with DRB18, an inhibitor of glucose transporter proteins.
INSTITUTE
Ohio University
DEPARTMENT
Biological Sciences
LABORATORY
Dr. Xiaozhuo Chen, Edison biotechnology Institute
LAST_NAME
Shriwas
FIRST_NAME
Pratik
ADDRESS
172 Water Tower, Building 25, The Ridges, Konnekar Research Centerm Athens Ohio - 45701, USA
Comparative metabolomics analysis of two Saccharomyces cerevisiae strains: the wild type and mtl1Ī, which carries a deletion of the mechanosensor Mtl1p (part-II)
STUDY_SUMMARY
To gather more in-depth knowledge of the Mtl1p mechanosensor's role in Saccharomyces cerevisiae metabolism, we conducted a comparative metabolomic analysis of two Saccharomyces cerevisiae strains: the wild type and mtl1Ī, which carries a deletion of the mechanosensor Mtl1p. Both strains were grown under normal conditions at 27Ā°C. The most significant metabolic changes between these strains were related to amino acid metabolism, purine metabolism, and carboxylic acid metabolism.
INSTITUTE
University of Puerto Rico, Medical Sciences Campus
DEPARTMENT
Biochemistry
LAST_NAME
Chorna
FIRST_NAME
Nataliya
ADDRESS
University of Puerto Rico, Medical Sciences Campus, Department of Biochemistry, Main Building, 6th Floor, Room A-632, San Juan, PR 00935
Comparing gas chromatography with time-of-flight, quadrupole time-of-light and quadrupole mass spectrometry for stable isotope tracing
STUDY_SUMMARY
Stable isotope tracers are applied in vivo and in vitro studies to reveal the activity of enzymes and intracellular metabolic pathways. Most often, such tracers are used with gas chromatography coupled to mass spectrometry (GC-MS) due to its ease of operation and reproducible mass spectral databases. Differences in isotope tracer performance of classic GC-quadrupole MS instrument and newer time-of-flight instruments are not well-studied. Here, we used three commercially available instruments for the analysis of identical samples from a stable isotope labeling study that used [U-13C6] d-glucose to investigate the metabolism of Rothia mucilaginosa with respect to 29 amino acids and hydroxyl acids involved in primary metabolism. Overall, all three GC-MS instruments (low-resolution GC-SQ-MS, low-resolution GC-TOF-MS, and high-resolution GC-Q-TOF-MS) can be used to perform stable isotope tracing studies for glycolytic intermediates, TCA metabolites and amino acids, yielding similar biological results, with high-resolution GC-Q-TOF-MS offering additional capabilities to identify chemical structures of unknown compounds that might show significant isotope enrichments in biological studies.
INSTITUTE
University of California, Davis
LAST_NAME
Zhang
FIRST_NAME
Ying
ADDRESS
451 East Health Science Drive, Davis, CA, 95616, USA
NMR metabolomics analysis of ricin-induced and fasting hypoglycemia (part-I)
STUDY_TYPE
Targeted NMR
STUDY_SUMMARY
Mice were subject to ricin exposure or fasting conditions for 2 hours, 8 hours, or an overnight time period. Following treatment, livers were removed and metabolites were extracted and analyzed by NMR.
Unique metabolomic profile of skeletal muscle in chronic limb threatening ischemia (organic phase samples)
STUDY_TYPE
NMR
STUDY_SUMMARY
This project is focused on a cross-sectional analysis of non-PAD controls and CLTI patients undergoing either a vascular intervention or undergoing limb amputation was performed and involved a detailed assessment of the limb muscle metabolome using HR-MAS and solution state NMR spectroscopy. It was hypothesized that patients undergoing limb amputation would present with altered muscle metaolite features compared with non-PAD controls.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
University of Florida, Applied Physiology and Kinesiology, 1864 stadium RD, Gainesville, FL 32611
EMAIL
ryant@ufl.edu
PHONE
352-294-1700
NUM_GROUPS
3
TOTAL_SUBJECTS
30
NUM_MALES
25
NUM_FEMALES
5
STUDY_COMMENTS
PAD metabolomic study via NMR. A detailed assessment of the limb muscle metabolome using semi-solid and solution state NMR spectroscopy
Unique metabolomic profile of skeletal muscle in chronic limb threatening ischemia (aqueous phase samples )
STUDY_TYPE
NMR
STUDY_SUMMARY
This project is focused on a cross-sectional analysis of non-PAD controls and CLTI patients undergoing either a vascular intervention or undergoing limb amputation was performed and involved a detailed assessment of the limb muscle metabolome using HR-MAS and solution state NMR spectroscopy. It was hypothesized that patients undergoing limb amputation would present with altered muscle metaolite features compared with non-PAD controls.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
University of Florida, Applied Physiology and Kinesiology, 1864 stadium RD, Gainesville, FL 32611
EMAIL
ryant@ufl.edu
PHONE
352-294-1700
NUM_GROUPS
3
TOTAL_SUBJECTS
30
NUM_MALES
25
NUM_FEMALES
5
STUDY_COMMENTS
PAD metabolomic study via NMR. A detailed assessment of the limb muscle metabolome using semi-solid and solution state NMR spectroscopy
Unique metabolomic profile of skeletal muscle in chronic limb threatening ischemia (HR-MAS study)
STUDY_SUMMARY
This project is focused on a cross-sectional analysis of non-PAD controls and CLTI patients undergoing either a vascular intervention or undergoing limb amputation was performed and involved a detailed assessment of the limb muscle metabolome using HR-MAS and solution state NMR spectroscopy. It was hypothesized that patients undergoing limb amputation would present with altered muscle metaolite features compared with non-PAD controls.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
EMAIL
ryant@ufl.edu
PHONE
352-294-1700
NUM_GROUPS
3
TOTAL_SUBJECTS
30
NUM_MALES
25
NUM_FEMALES
5
STUDY_COMMENTS
PAD metabolomic study via NMR. A detailed assessment of the limb muscle metabolome using semi-solid and solution state NMR spectroscopy.
Drug addiction is a major threat to the public health in the US and many other countries. Opioid abuse is associated with increased risks for cancer, psychological complications, heart and lung disease, and infections of the liver and blood. Because metabolites are intrinsically involved in multiple metabolic pathways in vivo, the relative quantification of metabolites in body fluids (for example urine) can provide a profile of the metabolic state of an organism. Metabolomics is a powerful technique for revealing the impact of exposure on the overall biochemistry of an individual or system. Opioids can modify the output of urinary metabolites through many integrated neural and hormonal mechanisms within the periphery, central nervous system, and kidneys. Opioids modulate the expression of genes involved in neuroplasticity through epigenetic and possibly RNA modifications, ultimately change the intracellular signaling cascades and dysfunction, and cause long-lasting changes in metabolome. The objective of this study is to identify how opium impacts metabolic pathways to provide markers of abuse, long-term opium addiction, the addiction molecular pathway, and unknown metabolites that are important to differentiation of the study phenotypes. To reach these goals in the present study, the urine specimens of opium abusers and non-users as controls will be profiled using an untargeted liquid chromatography mass spectrometric (LC-MS/MS) at University of North Carolina at Chapel Hill. The Golestan Cohort Study is conducted in Northeast of Iran to primarily study the risk factors for upper gastrointestinal cancers in this high-risk region, in which about 50,000 volunteers were analyzed for opium users and their mortality. More than 8,000 of participants (17%) age 40-75 reported opium use with a mean duration of 12.7 years. Opium was either smoked or orally consumed. The participants were selected from the cohort stratified by opium use patterns and tobacco use.
Drug addiction is a major threat to the public health in the US and many other countries. Opioid abuse is associated with increased risks for cancer, psychological complications, heart and lung disease, and infections of the liver and blood. Because metabolites are intrinsically involved in multiple metabolic pathways in vivo, the relative quantification of metabolites in body fluids (for example urine) can provide a profile of the metabolic state of an organism. Metabolomics is a powerful technique for revealing the impact of exposure on the overall biochemistry of an individual or system. Opioids can modify the output of urinary metabolites through many integrated neural and hormonal mechanisms within the periphery, central nervous system, and kidneys. Opioids modulate the expression of genes involved in neuroplasticity through epigenetic and possibly RNA modifications, ultimately change the intracellular signaling cascades and dysfunction, and cause long-lasting changes in metabolome. The objective of this study is to identify how opium impacts metabolic pathways to provide markers of abuse, long-term opium addiction, the addiction molecular pathway, and unknown metabolites that are important to differentiation of the study phenotypes. To reach these goals in the present study, the urine specimens of opium abusers and non-users as controls was profiled using an untargeted nuclear magnetic resonance spectroscopy (NMR) metabolomics platform at University of North Carolina at Chapel Hill. The Golestan Cohort Study is conducted in Northeast of Iran to primarily study the risk factors for upper gastrointestinal cancers in this high-risk region, in which about 50,000 volunteers were analyzed for opium users and their mortality. More than 8,000 of participants (17%) age 40-75 reported opium use with a mean duration of 12.7 years. Opium was either smoked or orally consumed. The participants were selected from the cohort stratified by opium use patterns and tobacco use.
Dietary composition analysis of chow diet and purified diet using untargeted metabonomics
STUDY_SUMMARY
Dietary patterns and psychosocial factors, ubiquitous part of modern lifestyle, critically shape the gut microbiota and human health. However, it remains obscure how dietary and psychosocial inputs coordinately modulate the gut microbiota and host impact.Ā Here, we show that dietary raffinose metabolism to fructose couples stress-induced gut microbial remodeling to intestinal stem cells (ISC) renewal and epithelial homeostasis. Chow diet (CD) and purified diet (PD) confer distinct vulnerability to gut epithelial injury, microbial alternation and ISC dysfunction in chronically restrained mice. CD preferably enriches Lactobacillus reuteri, and its colonization is sufficient to rescue stress-triggered epithelial injury.
Untargeted metabolomics of intestinal organoids treated with fructose
STUDY_TYPE
Metabolite profile and pathway analysis
STUDY_SUMMARY
We performed untargeted metabolomics of intestinal organoids treated with fructose (10 mM) for 24 hr.At the 5th day of secondary organoid culture, the organoids were treated with fructose (10 mM D-fructose)-containing IntestiCult organoid growth medium for 24 hr and washed with pre-cooled PBS before harvesting.
Isotope Tracing Analysis in Intestinal Organoids with fructose
STUDY_SUMMARY
We isolated and cultured Intestinal Organoids from Mouse. To trace the metabolism of fructose, intestinal organoids were cultured in unlabeled fructose-containing DMEM/F12 medium (10 mM D-fructose, 0 mM D-glucose, 2.5 mM L-glutamine) for 12 hr, and then switched into 13C labeled fructose-containing medium (10 mM U-[13C] D-fructose, Sigma-aldrich) for 6 hr.
Targeted metabolomics analysis with raffinose-rich diet in mouse ileum
STUDY_TYPE
Analysis of fructose level in ileum of stressed mouse fed with raffinose-rich diet
STUDY_SUMMARY
Purified diet (AIN-93G) supplemented with raffinose was prepared. The mice were maintained on the separate diet for at least 1 week before the initiation of experiment.Chronic restraint stress (RS) in mice was performed 14 days.After sacrifice, the ileum of mice were used to analysis.
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - aqueous phase Kidney (part-I)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - organic phase Kidney (part-II)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - aqueous phase Heart (part-III)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - organic phase Heart (part-IV)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - aqueous phase Liver (part-V)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - organic phase Liver (part-VI)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - aqueous phase Quadricep (part-VII)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - organic phase Quadricep (part-VIII)
STUDY_TYPE
Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR
STUDY_SUMMARY
This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Aromatic amino acid metabolism by the anaerobic gut bacterium Clostridium sporogenes (part-II)
STUDY_SUMMARY
Germ-free MCAD-/- mice or MCAD+/+ controls were mono-colonized with wild-type Clostridium sporogenes. Urine was collected for untargeted LC-QTOF analysis.
A combinatorial action of GmMYB176 and bZIP controls isoflavonoid biosynthesis in soybean.
STUDY_SUMMARY
This study identified how the GmMYB176 protein complex affects the metabolome of soybean hairy roots using non-targeted high resolution mass spectrometry.
Higher Dietary Carbohydrates Detrimentally Impact Obesity-Associated Breast Cancer Chemoresistance
STUDY_TYPE
Untargeted UPLC-MS Metabolomics Analysis
STUDY_SUMMARY
This study is a part of an ongoing project entitled, āHigher Dietary Carbohydrates Detrimentally Impact Obesity-Associated Breast Cancer Chemoresistance.ā (See project). Untargeted LCMS metabolomics analysis was performed to identify metabolic markers of nutritionally dependent and obesity-associated chemoresistance in female C57LB/6 mice fed either a high carbohydrate plus high fat (HCHF) diet or a high protein diet (HP). Mice were fed 15 weeks on either diet, then injected with a MMTV-Wnt-1 mouse mammary basal-like breast cancer cell line, for tumor formation up to 6 weeks. Three mice on either diet were treated for 24 hr with Doxorubicin or Saline as a vehicle control. All samples were analyzed in the positive mode for untargeted metabolomics analysis. Liver samples were selected for analysis because of an observation of fatty liver development in mice only on the HCHF diet after 15 weeks. Samples were extracted and analyzed by UPLS-MS analysis using Q-Exactive HFX mass spectrometer.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Nutrition
LABORATORY
Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel Hill
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
500 Laureate Way, Nutrition Research Institute, UNC Chapel Hill
Lipids were quantified in human plasma from the 1:3 matched TEDDY case-control subjects (Lee et al., 2013). Blood plasma was extracted following the methyl-tert-butyl ether (MTBE) extraction protocols. The choice of internal standards and chromatographic conditions was optimized by using toluene in the reconstitution solvent mixture to ensure that very lipophilic components like cholesteryl esters (CEs) and triacylglycerols (TAGs) are efficiently transferred to the ultrahigh-pressure liquid chromatography (UHPLC) column in the injection process. Lipids were analyzed by charged surface hybrid column electrospray ionization quadrupole time of flight tandem mass spectrometer (CSH-ESI QTOF MS/MS). The analytical ultra-high-pressure liquid chromatography (UHPLC) column is protected by a short guard column which was replaced after 400 injections while the UHPLC column was replaced after 1,200 serum (or plasma) extract injections. The sequence of column replacements were evaluated to ensure no detrimental effects were detected with respect to peak shapes, absolute or relative lipid retention times or reproducibility of quantifications. Automatic valve switching was used after each injection to reduce sample carryover for highly lipophilic compounds. This valve switching employed a dual solvent wash, first with a water/acetonitrile mixture (1:1, v/v) and subsequently with a 100% isopropanol wash. LC-BinBase was used as an untargeted approach for annotating chromatographic peaks and spectra against a dynamically built library of compounds. The quantified raw dataset was normalized by the SERRF bioinformatics pipeline [1]. In the LC-QTOF data, samples were removed before normalization if they failed the laboratoryās QC standards or were missing data for more than half of the compounds. The SERRF normalized data have been made available for identified compounds. Results data for unidentified compounds are available un-normalized. An explanation of the study design variables are explained in detail in a data dictionary provided in the raw data download section. References: 1) Lee HS, Burkhardt B, McLeod W, Smith S, Eberhard C, Lynch K, Hadley D, Rewers M, Simell O, She JX, Hagopian W, Lernmark A, Akolkar B, Ziegler AG, Krischer J, and the TEDDY Study Group: Biomarker discovery study design for type 1 diabetes in The Environmental Determinants of Diabetes in the Young (TEDDY) study. Diabetes/Metabolism Research and Reviews. Epub 2013 December 15. doi: 10.1002/dmrr.2510 (PubMed ID: 24339168). 2) Fan S, Kind T, Cajka T, Hazen SL, Tang WHW, Kaddurah-Daouk R, Irvin MR, Arnett DK, Barupal DK, Fiehn O: Systematic Error Removal Using Random Forest for Normalizing Large-Scale Untargeted Lipidomics Data. Anal Chem 2019, 91(5):3590-3596.
The mammalian brain relies on neurochemistry to fulfill its functions. Yet, the complexity of the brain metabolome and its changes during diseases or aging remains poorly understood. To start bridging this gap, we generated a metabolome atlas of the aging mouse brain from 10 anatomical regions spanning from adolescence to late adulthood. We combined data from three chromatography-based mass spectrometry assays and structurally annotated 1,709 metabolites to reveal the underlying architecture of aging-induced changes in the brain metabolome. Overall differences between sexes were minimal. We found 94% of all metabolites to significantly differ between brain sections in at least one age group. We also discovered that 90% of the metabolome showed significant changes with respect to age groups. For example, we identified a shift in sphingolipid patterns during aging that is related to myelin remodeling in the transition from adolescent to adult brains. This shift was accompanied by large changes in overall signature in a range of other metabolic pathways. We found clear metabolic similarities in brain sections that were functionally related such as brain stem, cerebrum and cerebellum. In cerebrum, metabolic correlation patterns got markedly weaker in the transition from adolescent to ear adults, whereas correlation patterns between cerebrum and brainstem regions decreased from early to late adulthood. We were also able to map metabolic changes to gene and protein brain atlases to link molecular changes to metabolic brain phenotypes. Metabolic profiles can be investigated via https://atlas.metabolomics.us/. This new resource enables brain researchers to link new metabolomic studies to a foundation data set.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome Center
LABORATORY
West Coast Metabolomics Center
LAST_NAME
Ding
FIRST_NAME
Jun
ADDRESS
451 East Health Science Drive, Davis, CA, 95616, USA
Metabolomics analysis of Vehicle (DMSO/PBS; (1:1) v/v) and DRB18 (10 mg/kg) treated A549 xenograft tumors
STUDY_TYPE
Anticancer compound treatment experiment in vivo
STUDY_SUMMARY
A549 xenograft tumors were treated with (DMSO/PBS; (1:1) v/v) and DRB18 (10 mg/kg) for 5 weeks. The mice were then sacrificed and tumors were then excised. Tumors were then subjected to untargeted metabolomics analysis.
INSTITUTE
Ohio University
DEPARTMENT
Biological Sciences
LABORATORY
Dr. Xiaozhuo Chen, Edison biotechnology Institute
LAST_NAME
Shriwas
FIRST_NAME
Pratik
ADDRESS
172 Water Tower, Building 25, The Ridges, Konnekar Research Centerm Athens Ohio - 45701, USA
Plasma Metabolomic signatures of COPD in a SPIROMICS cohort
STUDY_SUMMARY
The Subpopulations and Intermediate Outcome Measures in COPD Study (SPIROMICS) (ClinicalTrials.gov Identifier: NCT01969344) includes 2,771 subjects, aged 40-80 years with at least 20 pack-years of smoking. An additional 202 subjects were never smokers. Fasting blood drawn at the enrollment visit using a p100 tube. The first 649 subjects who returned for a 5-7 year visit (Visit 5) were selected for this study. The blood profiled were from the year 1 visit.
Remodeling Lipids in the Transition from Chronic Liver Disease to Hepatocellular Carcinoma (Blood) - part I
STUDY_SUMMARY
Comparing blood lipidomics of healthy volunteers to patients with chronic liver disease (CLD), and to patients with HCC caused by viral infections. We contrasted our findings in blood to lipid alterations in liver tumor and nontumor tissue samples from HCC patients.
INSTITUTE
University of California, Davis
LAST_NAME
Ismail
FIRST_NAME
Israa
ADDRESS
451 Health Sciences Drive, Room 1313, Davis, CA, 95616, USA
Remodeling Lipids in the Transition from Chronic Liver Disease to Hepatocellular Carcinoma (Liver) - part II
STUDY_SUMMARY
Comparing blood lipidomics of healthy volunteers to patients with chronic liver disease (CLD), and to patients with HCC caused by viral infections. We contrasted our findings in blood to lipid alterations in liver tumor and nontumor tissue samples from HCC patients.
INSTITUTE
University of California, Davis
DEPARTMENT
West coast metabolomics center
LABORATORY
Fiehn lab
LAST_NAME
Ismail
FIRST_NAME
Israa
ADDRESS
451 health science drive, 95616 ,Davis, California, USA.
Lipidomics in high-risk subjects for the identification of integrated biomarker signatures of type 1 diabetes
STUDY_SUMMARY
We present the lipidome of plasma collected from high-risk type 1 diabetes subjects. The methyl tert-butyl ether (MTBE) method was used for lipid extraction, followed by high performance liquid chromatography (HPLC) tandem mass spectrometry (LC-MS/MS) using a Q Exactive Orbitrap mass spectrometer and an Accela 600 HPLC. Lipid species were identified and quantified by analyzing the raw files in LipidSearch 4.2. Further analysis was conducted using Graphpad Prism and Ingenuity Pathway Analysis (IPA).
Deletion of the diabetes candidate gene Slc16a13 in mice
STUDY_SUMMARY
The metabolome of plasma and liver lysates of Slc16a13 knockout mice was analyzed. Genome-wide association studies identified SLC16A13 as novel target gene in type 2 diabetes. The SLC16A13 gene encodes for SLC16A13/MCT13, a member of the solute carrier 16 family of monocarboxylate transporters. So far, biology and physiological function of SLC16A13 is unknown. Deletion of Slc16a13 is hypothezised to affect intrahepatocellular monocarboxylate availability, that drives increased oxidative phosphorylation, while reducing hepatic lipid content, thereby attenuating hepatic insulin resistance.
In Vitro Characterization and Metabolomic Analysis of Cold-Stored Platelets
STUDY_SUMMARY
Platelet concentrates are currently stored at room temperature (RPs) under constant agitation for up to 5-7 days depending on national regulations. However, platelet quality deteriorates during storage and room temperature storage also increases the risk of bacterial growth. Previous studies have shown that cold-stored platelets (CPs) have higher hemostatic function and can be stored for up to three weeks. While these studies have compared the metabolic phenotypes of CPs and RPs, they have not compared the impact of storage temperature and cold agitation (CPAs) on platelet function, nor have they identified metabolic correlates to such parameters. In vitro analysis showed CPAs and CPs had reduced count, faster CD62P expression and increased lactadherin binding. Furthermore, CPAs and CPs had higher maximal aggregation and a reduced aggregation lag phase compared to RPs. Metabolomic analysis revealed CPAs and CPs exhibited lower oxidative stress shown by preserved glutathione and pentose phosphate pools. CPAs and CPs also had reduced markers of beta-oxidation and amino acid catabolism demonstrating reduced needs for energy. Agitation did not significantly impact in vitro function or metabolomic parameters of cold-stored platelets. Correlation of in vitro and metabolomic results highlighted important metabolites that may contribute to stored platelet functions.
Variability in metabolomic profiles among unique genotypes of Acropora cervicornis (part -II)
STUDY_TYPE
intraspecific variability
STUDY_SUMMARY
This project aims to identify differences in metabolomic profiles among seven known, unique genotypes of the threatened staghorn coral Acropora cervicornis.
INSTITUTE
University of Florida
DEPARTMENT
SECIM
LAST_NAME
Patterson
FIRST_NAME
Joshua
ADDRESS
Florida Aquarium Center for Conservation, 529 Estuary Shore Lane, Apollo Beach, FL 33572
Comprehensive dissection of primary metabolites in response to diverse abiotic stress in barley at seedling stage
STUDY_TYPE
Artical
STUDY_SUMMARY
Plants will meet various abiotic stresses during their growth and development. One of the important strategies for plants to deal with the stress is involved in metabolic regulation, causing the dramatic changes of metabolite profiles. Metabolomic studies have been intensively conducted to reveal the responses of plants to abiotic stress, but most of them were limited to one or at most two abiotic stresses in a single experiment. In this study, we compared the metabolite profiles of barley seedlings exposed to seven abiotic stresses simultaneously, including drought, salt stress, aluminum (Al), cadmium (Cd), deficiency of nitrogen (N), phosphorus (P) and potassium (K). The results showed that metabolite profiles of barley under these stresses could be classified into three types: osmotic stresses (drought and salt); metal stresses (Al and Cd) and nutrient deficiencies (N, P and K deficiencies). Compared with the control, some metabolites (including polyamines, raffinose and piperonic acid) in plants exposed to all abiotic stresses changed significantly, while some other metabolites showed the specific change only under a certain abiotic stress, such as proline being largely increased by osmotic stress (drought and salinity), the P-containing metabolites being largely decreased under P deficiency, some amino acids (lysine, tyrosine, threonine, ornithineļ¼ glutamine and so on) showing the dramatic reduction in the plants exposed to N deficiencies, respectively. The current meta-analysis obtained a comprehensive view on the metabolic responses to various abiotic stress, and improved the understanding of the mechanisms for tolerance of barley to abiotic stress.
Urinary microbiota and metabolome in pediatric vesicoureteral reflux and scarring
STUDY_TYPE
Observational/cross-sectional
STUDY_SUMMARY
We enrolled girls and boys aged zero to nine years presenting to a pediatric urologist for recurrent urinary tract infection (UTI) or renal scarring (decreased uptake on a nuclear renal scan) or grade 3-5 vesico ureteral reflux (VUR). Exclusion criteria included other urogenital abnormalities, medical renal disease, immunodeficiency, syndromes associated with VUR, acute UTI, persistent UTI (ongoing positive urine culture 1-3 weeks after completing a treatment course), or global renal atrophy on imaging. At one patient visit, a urine specimen was collected for 16S and metabolomic analysis.
Atypical Molecular Basis for Drug Resistance to Mitochondrial AQ: A Function Inhibitors in Plasmodium falciparum
STUDY_SUMMARY
In this study, we present a clear genotype for the P. falciparum SB1-A6 acridone-resistant clonal parasite strain and, through a combination of targeted and whole-cell methods, establish that the mechanism of resistance to both cytochrome bc1 and DHODH inhibitors results from the contribution of multiple genetic polymorphisms. We find that P. falciparum SB1-A6 accumulates both a copy number variation and a specific mutation in PfDHODH, and both of these genetic polymorphisms contribute to the panresistant phenotype. This study uncovers a mechanism of cross-resistance between PfDHODH and mtETC inhibitors and serves as a cautionary note to future antimalarial combination therapy formulations containing such drugs.
INSTITUTE
U.S. Food & Drug Administration
LAST_NAME
Painter
FIRST_NAME
Heather
ADDRESS
10903 New Hampshire Ave., WO 52/72-5324, Silver Spring, MD 20993
Mucosal-Associated-Invariant-T (MAIT) cells stimulated with IL-12/IL-12, anti-CD3/CD28 or both for 16 hours
STUDY_SUMMARY
The untargeted metabolomics analysis was performed after metabolite extraction from vital cells. The main object of the study was to define the activation of MAIT cells on the molecular level.
Metabolic profiling of plasma collected from Peromyscus leucopus and Mus musculus following LPS treatment
STUDY_TYPE
Untargeted Metabolomic profiling
STUDY_SUMMARY
Small molecule metabolites were extracted from plasma from Peromyscus leucopus and Mus musculus BALB/c and analyzed by liquid chromatography couple with mass spectrometry (LC-MS). XCMS software was used for peak picking, grouping and retention time correction from LC-MS data and generated a molecular feature spreadsheet. Pathway way enrichment analysis was carried out using Mummichog pathway analysis software using data from feature table.
Characterization of anaphylaxis reveals different metabolic changes depending on severity and triggers.
STUDY_SUMMARY
Background: Despite its increasing incidence, the underlying molecular processes of anaphylaxis remain unclear and there are not known biomarkers for appropriate diagnosis. The mechanism associated to the reactions still needs to be clarified in humans. The rapid onset and potentially fatal outcome in the absence of managed treatment, prevent its study and prompt obvious technical and ethical implications. Methods: Twenty episodes of anaphylaxis were analyzed. Sera was collected at different times: during the acute phase (T1), the recovery phase (T2) and around 2-3 months after the anaphylactic reaction (T0). The analysis included untargeted metabolomics combining liquid chromatography coupled to mass spectrometry (LC-MS) and proton-nuclear magnetic resonance (1H-NMR). Reactions were classified according to the trigger (food and/or drug) and severity (moderate and severe). Results: āFood T1 vs T2ā and āmoderate T1 vs T2ā anaphylaxis comparisons showed clear metabolic patterns during the onset of an anaphylactic reaction, which differed from those induced by drugs, food+drug or severe anaphylaxis āT1 vs T2ā. Moreover, the model of food anaphylaxis was able to distinguish the well-characterized IgE (beta-lactam) from non-IgE- mediated anaphylaxis (NSAIDs), suggesting a differential metabolic pathway associated with the mechanism of action. Moreover, metabolic differences between āmoderate vs severeā at T1 and T0 were studied. Among the metabolites, glucose, lipids, cortisol, betaine and oleamide were observed altered. Conclusions: The results of the study provide the first evidence that different anaphylactic triggers, induce differential metabolic changes. Besides, the basal status might identify high risk patients, thus opening new ways to understand, diagnose and treat anaphylaxis.
Characterization of anaphylaxis reveals different metabolic changes depending on severity and triggers - MS (part-II)
STUDY_SUMMARY
Background: Despite its increasing incidence, the underlying molecular processes of anaphylaxis remain unclear and there are not known biomarkers for appropriate diagnosis. The mechanism associated to the reactions still needs to be clarified in humans. The rapid onset and potentially fatal outcome in the absence of managed treatment, prevent its study and prompt obvious technical and ethical implications. Methods: Twenty episodes of anaphylaxis were analyzed. Sera was collected at different times: during the acute phase (T1), the recovery phase (T2) and around 2-3 months after the anaphylactic reaction (T0). The analysis included untargeted metabolomics combining liquid chromatography coupled to mass spectrometry (LC-MS) and proton-nuclear magnetic resonance (1H-NMR). Reactions were classified according to the trigger (food and/or drug) and severity (moderate and severe). Results: āFood T1 vs T2ā and āmoderate T1 vs T2ā anaphylaxis comparisons showed clear metabolic patterns during the onset of an anaphylactic reaction, which differed from those induced by drugs, food+drug or severe anaphylaxis āT1 vs T2ā. Moreover, the model of food anaphylaxis was able to distinguish the well-characterized IgE (beta-lactam) from non-IgE- mediated anaphylaxis (NSAIDs), suggesting a differential metabolic pathway associated with the mechanism of action. Moreover, metabolic differences between āmoderate vs severeā at T1 and T0 were studied. Among the metabolites, glucose, lipids, cortisol, betaine and oleamide were observed altered. Conclusions: The results of the study provide the first evidence that different anaphylactic triggers, induce differential metabolic changes. Besides, the basal status might identify high risk patients, thus opening new ways to understand, diagnose and treat anaphylaxis.
E.coli K-12 treated by IPL_analysis of organic phase
STUDY_SUMMARY
In this study, E.coli K-12 was treated by intense pulsed light (IPL) for 0-20 seconds. Then the organic/lipid phase of the cellular metabolome was extracted and submitted to untargeted LC-MS based metabolomic study.
Control of Topoisomerase II Activity and Chemotherapeutic Inhibition by TCA Cycle Metabolites
STUDY_SUMMARY
Topoisomerase II (topo II) is essential for disentangling newly-replicated chromosomes. DNA unlinking involves the physical passage of one DNA duplex through another and depends on the transient formation of double-strand DNA breaks, a step exploited by frontline chemotherapies to kill cancer cells. Although anti-topo II drugs are efficacious, they also elicit cytotoxic side effects in normal cells; insights into how topo II is regulated in different cellular contexts is essential to improve their targeted use. Using chemical fractionation and mass spectrometry, we have discovered that topo II is subject to metabolic control through the TCA cycle. We show that TCA metabolites stimulate topo II activity in vitro and that levels of TCA flux modulate cellular sensitivity to anti-topo II drugs in vivo. Our works reveals an unanticipated connection between the control of DNA topology and cellular metabolism, a finding with important ramifications for the clinical use of anti-topo II therapies.
Identify putative volatile biomarkers of Coccidioides spp. grown in vitro
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Valley fever (coccidioidomycosis) is an endemic fungal pneumonia of the North and South American deserts. The causative agents of Valley fever are the dimorphic fungi Coccidioides immitis and C. posadasii, which grow as mycelia in the environment and spherules within the lungs of vulnerable hosts. The current diagnostics for Valley fever are severely lacking due to poor sensitivity and invasiveness, contributing to a 23-day median time-to-diagnosis, and therefore new diagnostic tools are needed. We are working toward the development of a breath-based diagnostic for coccidioidomycosis, and in this initial study we characterized the volatile metabolomes (or volatilomes) of in vitro cultures of Coccidioides. Using solid-phase microextraction and comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GCĆGCāTOFMS), we characterized the VOCs produced by six strains of each species during mycelial or spherule growth. We detected a total of 353 VOCs that were at least two-fold more abundant in a Coccidioides culture versus medium controls and found the volatile metabolome of Coccidioides is more dependent on growth phase (spherule versus mycelia) than on the species. The volatile profiles of C. immitis and C. posadasii have strong similarities, indicating that a single suite of Valley fever breath biomarkers can be developed to detect both species.
INSTITUTE
Arizona State University
DEPARTMENT
School of Life Sciences
LABORATORY
Bean Laboratory
LAST_NAME
Bean
FIRST_NAME
Heather
ADDRESS
PO Box 874501 Tempe, AZ 85287
EMAIL
Heather.D.Bean@asu.edu
PHONE
4807273395
PUBLICATIONS
Lifecycle dominates the volatilome character of the dimorphic fungus Coccidioides spp Emily A. Higgins Keppler, Heather L. Mead, Bridget M. Barker, Heather D. Bean bioRxiv 2021.01.15.426916; doi: https://doi.org/10.1101/2021.01.15.426916
Plasmodium falciparum metabolomics as a result of treatment with putative acetyl-CoA synthetase inhibitors
STUDY_SUMMARY
Plasmodium falciparum cells in culture were treated with respective compounds for 2.5 hours at 10xIC50 values. Metabolites were isolated using 90% methanol, dried, reconstituted in HPLC-grade water, and analyzed by HPLC/MS. Resulting data were analyzed and compiled to generate study data.
INSTITUTE
Pennsylvania State University
DEPARTMENT
Chemistry
LABORATORY
LlinƔs Laboratory
LAST_NAME
LlinƔs
FIRST_NAME
Manuel
ADDRESS
W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA
Extension of Diagnostic Fragmentation Filtering for Automated Discovery in DNA Adductomics
STUDY_SUMMARY
Development of high resolution/accurate mass liquid chromatography-coupled tandem mass spectrometry (LC-MS/MS) methodology enables the characterization of covalently modified DNA induced by interaction with genotoxic agents in complex biological samples. Constant neutral loss monitoring of 2Ā“-deoxyribose or the nucleobases using data-dependent acquisition represents a powerful approach for the unbiased detection of DNA modifications (adducts). The lack of available bioinformatics tools necessitates manual processing of acquired spectral data and hampers high throughput application of these techniques. To address this limitation, we present an automated workflow for the detection and curation of putative DNA adducts by using diagnostic fragmentation filtering of LC-MS/MS experiments within the open-source software MZmine. The workflow utilizes a new feature detection algorithm, DFBuilder, which employs diagnostic fragmentation filtering using a user-defined list of fragmentation patterns to reproducibly generate feature lists for precursor ions of interest. The DFBuilder feature detection approach readily fits into a complete small molecule discovery workflow and drastically reduces the processing time associated with analyzing DNA adductomics results. We validate our workflow using a mixture of authentic DNA adduct standards and demonstrate the effectiveness of our approach by reproducing and expanding the results of a previously published study of colibactin-induced DNA adducts. The reported workflow serves as a technique to assess the diagnostic potential of novel fragmentation pattern combinations for the unbiased detection of chemical classes of interest.
INSTITUTE
University of Minnesota
DEPARTMENT
School of Public Health, Division of Environmental Health Sciences
LABORATORY
Balbo Research Group
LAST_NAME
Murray
FIRST_NAME
Kevin
ADDRESS
2-210 CCRB, 2231 6th St SE, Minneapolis, MN 55455
EMAIL
murra668@umn.edu
PHONE
612-626-2182
NUM_GROUPS
1
TOTAL_SUBJECTS
3
STUDY_COMMENTS
Synthetic samples of authentic standards for workflow testing and validation.
PUBLICATIONS
Murray K.J.; Carlson E.S.; Stornetta A.; Balskus E.P.; Villalta P.W.; Balbo S. Extension of Diagnostic Fragmentation Filtering for Automated Discovery in DNA Adductomics. Anal. Chem. 2021. (In Revision).
LC-MS Metabolomics of Urine Reveals Distinct Profiles for Low- and High-Grade Bladder Cancer
STUDY_SUMMARY
Bladder cancer (BC) is among the most frequent malignancies worldwide. Novel non-invasive markers are needed to diagnose and stage BC with more accuracy than invasive procedures such as cystoscopy. Our aim was to discover novel urine metabolomic profiles to diagnose and stage non-muscle invasive (NMIBC) and muscle-invasive (MIBC) patients using ultra-performance liquid chromatography analysis (UPLC)-based metabolomics. We prospectively recruited 64 BC patients (19 TaG1, 11 TaG3, 20 T1G3, 12 T2G3, 1 T2G2, 1 T3G3) and 20 age- and sex-matched healthy volunteers without evidence of renal or bladder condition confirmed by ultrasound, from whom we collected a first morning urine sample (before surgery in patients). We conducted a UPLC-quadrupole-time-of-flight mass spectrometry (UPLC-Q-ToF MS) untargeted metabolomic analysis in all urine samples. We selected the discriminant variables between groups with a supervised orthogonal-least-squares discriminant analysis (OPLS-DA) analysis and we identified them by querying their exact mass against those presented in online databases through a mediator platform. Subsequently, we confirmed the dysregulated metabolites when chemical standards were commercially available. We compared all clinical groups of patients with controls and we identified dysregulated metabolites in every comparison. Of these, we confirmed p-cresol glucuronide as potential diagnostic biomarker, and potential staging tool for NMIBC patients. Among NMIBC patients, we identified p-coumaric acid as a potential staging biomarker for milder NMIBC stages (TaG1). Additionally, we confirmed spermine and adenosine as potential staging biomarkers for MIBC. This is the first study conducted in urine samples of most stages of NMIBC and MIBC and healthy controls to identify non-invasive biomarkers. Once confirmed, these may improve BC management thus reducing the use of current harmful diagnostic techniques.
INSTITUTE
Health Research Institute Hospital La Fe
LABORATORY
Analytical Unit
LAST_NAME
Roca MarugƔn
FIRST_NAME
Marta
ADDRESS
Avenida Fernando Abril Martorell 106, Torre A, Valencia, Valencia, 46026, Spain
E.coli K-12 treated by IPL - analysis of polar phase (part-I)
STUDY_SUMMARY
E.coli K-12 cells were treated by IPL, extracted and separated into organic/lipid phase and polar phase. The polar phase was analyzed by HILIC-MS, in both positive and negative ionization mode.
E.coli K-12 treated by IPL - analysis of polar phase (part-II)
STUDY_SUMMARY
E.coli K-12 cells were treated by IPL, extracted and separated into organic/lipid phase and polar phase. Chemical derivatization with dansyl chloride was applied for analysis of amino acids in the polar phase extraction.
Branched-chain alpha-ketoacids are preferentially reaminated and activate protein synthesis in the rat heart
STUDY_SUMMARY
Branched-chain amino acids (BCAA) and their cognate Ī±-ketoacids (BCKA) are elevated in an array of cardiometabolic diseases. Here we demonstrate that the major metabolic fate of uniformly-13C-labeled Ī±-ketoisovalerate ([U-13C]KIV) in the heart is reamination to valine. Activation of cardiac branched-chain Ī±-ketoacid dehydrogenase (BCKDH) by treatment with the BCKDH kinase inhibitor, BT2, does not impede the strong flux of [U-13C]KIV to valine.
Branched-chain alpha-ketoacids are preferentially reaminated and activate protein synthesis in the mouse heart
STUDY_SUMMARY
Branched-chain amino acids (BCAA) and their cognate Ī±-ketoacids (BCKA) are elevated in an array of cardiometabolic diseases. Here we demonstrate that sequestration of BCAA and BCKA away from mitochondrial oxidation is likely due to low levels of expression of the mitochondrial BCAA transporter SLC25A44 in the heart, as its overexpression significantly lowers accumulation of [13C]-labeled valine from [U-13C]KIV.
LC-MS metabolomics analysis of ricin-induced and fasting hypoglycemia (part-II)
STUDY_SUMMARY
Mice were subjected to ricin exposure or fasting conditions for 2 hours, 8 hours, or an overnight period. Following treatment, livers were removed and metabolites were extracted and analyzed by LC-MS.
INSTITUTE
Montana State University
LAST_NAME
Kempa
FIRST_NAME
Jake
ADDRESS
103 Chemistry and Biochemistry Building, Bozeman, Montana, 59717, USA
Serum metabolome of Guangzhou Nutrition and Health Study (GNHS)
STUDY_SUMMARY
Our study was based on the Guangzhou Nutrition and Health Study (GNHS). This study aims to investigate the relationships among human nutrition, enviromental factors, gut microbiome and human diseases.
INSTITUTE
Westlake University
LAST_NAME
Ju-Sheng
FIRST_NAME
Zheng
ADDRESS
Westlake University,18 Shilongshan Rd, Cloud Town, Hangzhou, China
Four different treatment groups were used for metabolite characterization: 5 dpf larvae with/without beta-cell ablation and with/without folinic acid treatment.
A gut microbe-focused metabolomics pipeline enables mechanistic interrogation of microbiome metabolism
STUDY_SUMMARY
Gut microbes modulate host phenotypes and are associated with numerous health effects in humans, ranging from cancer immunotherapy response to metabolic disease and obesity. However, difficulty in accurate and high-throughput functional analysis of human gut microbes has hindered defining mechanistic connections between individual microbial strains and host phenotypes. One key way the gut microbiome influences host physiology is through the production of small molecules1ā3, yet progress in elucidating this chemical interplay has been hindered by limited tools calibrated to detect products of anaerobic biochemistry in the gut. Here we construct a microbiome-focused, integrated mass-spectrometry pipeline to accelerate the identification of microbiota-dependent metabolites (MDMs) in diverse sample types. We report the metabolic profiles of 178 gut microbe strains using our library of 833 metabolites. Leveraging this metabolomics resource we establish deviations in the relationships between phylogeny and metabolism, use machine learning to discover novel metabolism in Bacteroides, and employ comparative genomics-based discovery of candidate biochemical pathways. MDMs can be detected in diverse body fluids in gnotobiotic and conventional mice and traced back to corresponding metabolomic profiles of cultured bacteria. Collectively, our microbiome-focused metabolomics pipeline and interactive metabolomics profile explorer are a powerful tool for characterizing microbe and microbe-host interactions.
INSTITUTE
Stanford University
DEPARTMENT
Microbiology and Immunology
LABORATORY
Justin Sonnenburg
LAST_NAME
Van Treuren
FIRST_NAME
Will
ADDRESS
Sherman Fairchild Building, 299 Campus Drive, Stanford CA, 94305
Targeted Sphingolipid analysis of human Fibroblasts silenced for or overexpressing GOLPH3 (part-I)
STUDY_SUMMARY
A group of sequentially-acting enzymes operating at the branchpoint among sphingolipid synthetic pathways binds the Golgi-localised oncoprotein GOLPH3. GOLPH3 sorts these enzymes into vesicles for intra-Golgi retro-transport, acting as a component of the cisternae inter-conversion mechanisms. Through these effects, GOLPH3 controls the sub-Golgi localisation, and the lysosomal degradation rate of specific enzymes. Here we evaluated the impact of overexpressing or silencing GOLPH3 on the sphingolipid composition of dermal human fibroblasts by targeted lipid analysis.
Targeted Sphingolipid analysis of HeLa silenced for or overexpressing GOLPH3 or LCS
STUDY_SUMMARY
A group of sequentially-acting enzymes operating at the branchpoint among sphingolipid synthetic pathways binds the Golgi-localised oncoprotein GOLPH3. GOLPH3 sorts these enzymes into vesicles for intra-Golgi retro-transport, acting as a component of the cisternae inter-conversion mechanisms. Through these effects, GOLPH3 controls the sub-Golgi localisation, and the lysosomal degradation rate of specific enzymes. Here we evaluated the impact of overexpressing or silencing GOLPH3 or its client enzyme lactosylceramide synthase (LCS) on the sphingolipid composition of HeLa cells by targeted lipid analysis.
Untargeted phospholipid analysis of HeLa silenced for or overexpressing GOLPH3
STUDY_SUMMARY
A group of sequentially-acting enzymes operating at the branchpoint among sphingolipid synthetic pathways binds the Golgi-localised oncoprotein GOLPH3. GOLPH3 sorts these enzymes into vesicles for intra-Golgi retro-transport, acting as a component of the cisternae inter-conversion mechanisms. Through these effects, GOLPH3 controls the sub-Golgi localisation, and the lysosomal degradation rate of specific enzymes. Here we evaluated the impact of overexpressing or silencing GOLPH3 on the glycerophospholipid composition of HeLa cells by untargeted lipid analysis.
Quantitative measurements of ceramides and glycosphingolipids in Th17 and iTreg cells (part-II)
STUDY_TYPE
MS: Targeted analysis
STUDY_SUMMARY
Part 2/5: It includes quantitative targeted measurements of sphingolipids (ceramides and glycosphingolipids) in Th17, iTreg, and their paired controls (Th0 cells).
Quantitative targeted measurements of sphingolipids in Th17 cells before and after the triple knockdown of SPTLC1,2,3 genes (SPT, de novo pathway: sphingolipid metabolism) (part-III)
STUDY_TYPE
MS: Targeted analysis
STUDY_SUMMARY
Part 3/5: It includes quantitative targeted measurements of sphingolipids (ceramides and glycosphingolipids)in Th17 cells before(scrambled / control)and after the triple knockdown of SPTLC1,2,3 genes (SPT de novo pathway: sphingolipid metabolism).
INSTITUTE
University of Turku
DEPARTMENT
Systems Medicine, Turku Bioscience
LABORATORY
Metabolomics
LAST_NAME
Sen
FIRST_NAME
Partho
ADDRESS
Tykistƶkatu 6B, BioCity, 5th Floor, Turku, Southwest, 20521, Finland
Quantitative measurements of sphingolipids in Th17 cells before and after the knockdown of UGCG gene (GCS pathway: sphingolipid metabolism) (part-IV)
STUDY_TYPE
MS: Targeted analysis
STUDY_SUMMARY
Part 4/5: It includes quantitative targeted measurements of sphingolipids (ceramides, glycosphingolipids) in Th17 cells before (scrambled / control) and after the knockdown of UGCG gene (GCS pathway: sphingolipid metabolism).
INSTITUTE
University of Turku
DEPARTMENT
Systems Medicine, Turku Bioscience
LABORATORY
Metabolomics
LAST_NAME
Sen
FIRST_NAME
Partho
ADDRESS
Tykistƶkatu 6B, BioCity, 5th Floor, Turku, Southwest, 20521, Finland
Quantitative measurements of sphingomyelins in Th17 cells before and after the knockdown of UGCG gene (GCS pathway: sphingolipid metabolism) (part-V)
STUDY_TYPE
MS: Untargeted and targeted analysis
STUDY_SUMMARY
Part 5/5: It includes measurements of sphingolipids (sphingomyelins) in Th17 cells before (scrambled / control) and after the knockdown of UGCG gene (GCS pathway: sphingolipid metabolism).
INSTITUTE
University of Turku
DEPARTMENT
Systems Medicine, Turku Bioscience
LABORATORY
Metabolomics
LAST_NAME
Sen
FIRST_NAME
Partho
ADDRESS
Tykistƶkatu 6B, BioCity, 5th Floor, Turku, Southwest, 20521, Finland
We investigated the effects of a RAR beta 2 agonist, AC261066, on the hepatic metabolites changed in high fat fed NAFLD mouse model. We suggest that AC261066 has potential therapeutic relevance for the prevention/treatment of NAFLD and NASH.
Integrated trajectories of the maternal metabolome, proteome, and immunome predict labor onset
STUDY_SUMMARY
Estimating the time of delivery is of high clinical importance as pre- and post-term deviations are associated with complications for the mother and her offspring. However, current estimation approaches are inaccurate. As pregnancy progresses towards labor, major transitions occur in fetomaternal immune, metabolic, and endocrine systems that culminate in the delivery of the fetus. The comprehensive characterization of metabolic, proteomic and immune cell events that precede the spontaneous onset of labor is a key step to understanding these physiological transitions and identifying predictive biomarkers of parturition. Here, over 7,000 circulating plasma analytes and peripheral immune cell responses collected during the last 100 days of pregnancy were integrated into a multi-omic model that accurately predicted the time to spontaneous onset of labor (R = 0.85, p-value = 1.2e-40, training set; R = 0.81, p-value = 3.9e-7, independent test set). Coordinated fluctuations marked a molecular shift from pregnancy progression to pre-labor onset biology 2ā4 weeks before delivery. Our study lays the groundwork for developing blood-based methods for predicting the onset of labor, anchored in mechanisms shared in preterm, term, and postterm pregnancies.
Untargeted urine LC-HRMS metabolomics profiling for bladder cancer binary outcome classification
STUDY_SUMMARY
Two samples cohorts were analysed for bladder cancer biomarkers selection. Untargeted urine RP UPLC-HRMS metabolomics profiling was utilized in SCAN MS mode and positive polarity. Dilute and shoot technique was employed for sample preparation.
A gut microbe-focused metabolomics pipeline enables mechanistic interrogation of microbiome metabolism.
STUDY_SUMMARY
Gut microbes modulate host phenotypes and are associated with numerous health effects in humans, ranging from cancer immunotherapy response to metabolic disease and obesity. However, difficulty in accurate and high-throughput functional analysis of human gut microbes has hindered defining mechanistic connections between individual microbial strains and host phenotypes. One key way the gut microbiome influences host physiology is through the production of small molecules hindered by limited tools calibrated to detect products of anaerobic biochemistry in the gut. Here we construct a microbiome-focused, integrated mass-spectrometry pipeline to accelerate the identification of microbiota-dependent metabolites (MDMs) in diverse sample types. We report the metabolic profiles of 178 gut microbe strains using our library of 833 metabolites. Leveraging this metabolomics resource we establish deviations in the relationships between phylogeny and metabolism, use machine learning to discover novel metabolism in Bacteroides, and employ comparative genomics-based discovery of candidate biochemical pathways. MDMs can be detected in diverse body fluids in gnotobiotic and conventional mice and traced back to corresponding metabolomic profiles of cultured bacteria. Collectively, our microbiome-focused metabolomics pipeline and interactive metabolomics profile explorer are a powerful tool for characterizing microbe and microbe-host interactions.
An overexpression of lipoprotein lipase leads to an alteration in the skeletal muscle metabolome in transgenic rabbits
STUDY_SUMMARY
The purpose of the current study was to characterize the skeletal muscle metabolome in the lipoprotein lipase (LPL) transgenic rabbits. The skeletal muscle metabolite profile was analyzed using capillary electrophoresis time-of flight mass spectrometry in the control rabbits and LPL transgenic rabbits (n = 9, each group).
INSTITUTE
Saga University
LAST_NAME
Nishida
FIRST_NAME
Yuichiro
ADDRESS
Department of Preventive Medicine, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501
Exposure to per- and polyfluoroalkyl substances associates with altered lipid profile of breast milk (Part 2)
STUDY_SUMMARY
In this mother-infant study (n=44) we investigated the levels of PFAS in maternal serum and detailed lipidomic profile in breast milk at birth and at three months using ultra high performance liquid chromatography combined with quadrupole-time-of-flight mass spectrometry.
Exposure to per- and polyfluoroalkyl substances associates with altered lipid profile of breast milk (Part 3)
STUDY_SUMMARY
In this mother-infant study (n=44) we investigated the levels of PFAS in maternal serum and detailed lipidomic profile in breast milk at birth and at three months using ultra high performance liquid chromatography combined with quadrupole-time-of-flight mass spectrometry.
Non-transformed cells respond to fat by inducing glucose metabolism
STUDY_SUMMARY
C57BL/6N mice were obtained from the KU Leuven animal laboratory. 2-week old mice were injected i.p. diethylnitrosamine (DEN, 25 mg/Kg) in phosphate buffered saline (3.17 mg/ml).s At 6-weeks of age, mice were randomized into two groups: control diet (CD, E15742-33 ssniff SpezialdiƤten GmbH) or high fat diet (HFD, S8655-E220 sniff SpezialdiƤten GmbH). At endpoint, mice were sacrificed by injecting approximately 50 Āµl of a 60 mg/ml Nembutal solution (Vetoquinol). Tissues were immediately excised, washed in ice-cold saline, placed into pre-labelled bags and frozen using a liquid nitrogen-cooled biosqueezer. The bags were then placed in liquid nitrogen until all collections were finished and finally stored at -80Ā°C until further processing. Where necessary tumors were rapidly separated from normal tissue prior to freezing with tumor and normal tissue being stored separately.
INSTITUTE
VIB-KU Leuven Center for Cancer Biology
DEPARTMENT
Department of Oncology
LABORATORY
Laboratory of Cellular Metabolism and Metabolic Regulation
Exposure to per- and polyfluoroalkyl substances associates with altered lipid profile of breast milk (Part 1)
STUDY_SUMMARY
In this mother-infant study (n=44) we investigated the levels of PFAS in maternal serum and detailed lipidomic profile in breast milk at birth and at three months using ultra high performance liquid chromatography combined with quadrupole-time-of-flight mass spectrometry.
Untargeted metabolomic analysis of human blood samples via qualitative GC-MS for T1D biomarker identification
STUDY_TYPE
Qualitative GC-MS biomarker identification
STUDY_SUMMARY
"Blood from human subjects at high risk for T1D (and healthy controls; n=4 each) were subjected to parallel unlabeled proteomics, metabolomics, lipidomics, and transcriptomics. The integrated dataset was analyzed using Ingenuity Pathway Analysis (IPA) software for disturbances in the at-risk subjects compared to the controls. The final quadra-omics dataset contained 2292 proteins, 328 miRNAs, 75 metabolites, and 41 lipids that were detected in all samples. Disease/function enrichment analyses consistently indicated increased activation, proliferation, and migration of immune cells, particularly, CD4 T-lymphocytes and macrophages. Integrated molecular network predictions highlighted central involvement and activation of NF-ĪŗB, TGF-Ī², VEGF, arachidonic acid, and arginase, and inhibition of miRNA Let-7a-5p. Parallel multi-omics provided a comprehensive picture of disturbances in high-risk T1D subjects and helped identify an associated integrated biomarker signature, which could ultimately facilitate the classification of T1D progressors from non-progressors."
INSTITUTE
Duke University
DEPARTMENT
Duke Molecular Physiology Institute, School of Medicine
Perfluoroalkyl substances and lipid composition in human milk
STUDY_TYPE
CHEAR Study
STUDY_SUMMARY
PFAS are widely used in commercial products, and so humans have consistent exposure to them via oil- and water-resistant consumer products, fire- fighting foam, and industrial surfactants 1,2. The four PFASs commonly detected in blood, perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS), perfluorononanoic acid (PFNA), and perfluorohexane sulfonate (PFHxS) 3,4, are present in drinking water supplies both in northern New England as well as in 27 states nationally 5-8. Animal models shows that PFASs have can have effects on both the endocrine system and on adiposity 9-12. Epidemiological evidence shows that the presence of PFASs in maternal serum is associated with changes in maternal serum lipid and cholesterol composition 13,14. Similarly, serum levels of PFAS in adolescents have been associated with increases in serum cholesterol 15. These findings raise interesting questions about the association of PFAS and lipids in human milk. Research has shown the PFASs are present in human milk 16-18, and human milk is composed primarily of lipids 19. However, the relation between PFAS in milk and milk composition is unclear. The chemical and compositional profiles of breast milk are important because of the potential effects on the developing infant. The developmental origins of health and disease hypothesis suggests that early life exposures, such as toxins and nutrients via breast milk, have lasting effects on health, particularly obesity outcomes 20. In fact, some studies have shown associations between PFAS in maternal serum and infant birth weight and later childhood BMI 14,21. Our study will help to better illuminate the potential effects of maternal exposure to PFASs on infant exposure, both through direct transmission into breast milk and indirectly via influence on the lipid profiles of milk. To investigate how early life exposure to perfluoroalkyl substances (PFAS) may affect childhood health outcomes as mediated through breast milk, we propose the following specific aims: 1. Characterize the levels of PFAS in breast milk samples (n=495) in the NHBCS; 2. Characterize the lipid profiles of breast milk samples (n=495) in the NHBCS; 3. Test the relation between PFAS concentration and breast milk lipid profiles; and 4. Test the association between PFAS concentrations in maternal plasma collected during pregnancy with paired breast milk samples (n=100).
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Department of Environmental Medicine and Public Health
A cross-sectional study of functional and metabolic changes during aging through the lifespan in male mice (Muscle) part-I
STUDY_SUMMARY
Aging is associated with distinct phenotypical, physiological, and functional changes, leading to the onset of disease and death. The progression of aging-related traits varies widely among individuals, influenced by their environment, lifestyle, and genetics. In this study, we performed physiologic and functional tests cross-sectionally throughout the entire lifespan of male C57BL/6N mice. In parallel, metabolomics analyses in serum, brain, liver, heart, and skeletal muscle were also performed to identify signatures associated with frailty and age-dependent functional decline. Our findings indicate that the decline in gait speed as a function of age and frailty is associated with dramatic increases in the energetic cost of physical activity and decreases in working capacity. Aging and functional decline prompt organs to rewire their substrate selection and metabolism towards redox-related pathways, mainly in liver and heart. Collectively, the data provide a framework to further understand and characterize processes of aging at the individual and organ levels.
INSTITUTE
National Institutes of Health
DEPARTMENT
NIA
LABORATORY
Experimental Gerontology Section and Translational Gerontology Branch
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
A cross-sectional study of functional and metabolic changes during aging through the lifespan in male mice (Serum) part-II
STUDY_SUMMARY
Aging is associated with distinct phenotypical, physiological, and functional changes, leading to the onset of disease and death. The progression of aging-related traits varies widely among individuals, influenced by their environment, lifestyle, and genetics. In this study, we performed physiologic and functional tests cross-sectionally throughout the entire lifespan of male C57BL/6N mice. In parallel, metabolomics analyses in serum, brain, liver, heart, and skeletal muscle were also performed to identify signatures associated with frailty and age-dependent functional decline. Our findings indicate that the decline in gait speed as a function of age and frailty is associated with dramatic increases in the energetic cost of physical activity and decreases in working capacity. Aging and functional decline prompt organs to rewire their substrate selection and metabolism towards redox-related pathways, mainly in liver and heart. Collectively, the data provide a framework to further understand and characterize processes of aging at the individual and organ levels.
INSTITUTE
National Institutes of Health
DEPARTMENT
NIA
LABORATORY
Experimental Gerontology Section and Translational Gerontology Branch
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
A cross-sectional study of functional and metabolic changes during aging through the lifespan in male mice (Liver) part-III
STUDY_SUMMARY
Aging is associated with distinct phenotypical, physiological, and functional changes, leading to the onset of disease and death. The progression of aging-related traits varies widely among individuals, influenced by their environment, lifestyle, and genetics. In this study, we performed physiologic and functional tests cross-sectionally throughout the entire lifespan of male C57BL/6N mice. In parallel, metabolomics analyses in serum, brain, liver, heart, and skeletal muscle were also performed to identify signatures associated with frailty and age-dependent functional decline. Our findings indicate that the decline in gait speed as a function of age and frailty is associated with dramatic increases in the energetic cost of physical activity and decreases in working capacity. Aging and functional decline prompt organs to rewire their substrate selection and metabolism towards redox-related pathways, mainly in liver and heart. Collectively, the data provide a framework to further understand and characterize processes of aging at the individual and organ levels.
INSTITUTE
National Institutes of Health
DEPARTMENT
NIA
LABORATORY
Experimental Gerontology Section and Translational Gerontology Branch
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
A cross-sectional study of functional and metabolic changes during aging through the lifespan in male mice (Heart) part-IV
STUDY_SUMMARY
Aging is associated with distinct phenotypical, physiological, and functional changes, leading to the onset of disease and death. The progression of aging-related traits varies widely among individuals, influenced by their environment, lifestyle, and genetics. In this study, we performed physiologic and functional tests cross-sectionally throughout the entire lifespan of male C57BL/6N mice. In parallel, metabolomics analyses in serum, brain, liver, heart, and skeletal muscle were also performed to identify signatures associated with frailty and age-dependent functional decline. Our findings indicate that the decline in gait speed as a function of age and frailty is associated with dramatic increases in the energetic cost of physical activity and decreases in working capacity. Aging and functional decline prompt organs to rewire their substrate selection and metabolism towards redox-related pathways, mainly in liver and heart. Collectively, the data provide a framework to further understand and characterize processes of aging at the individual and organ levels.
INSTITUTE
National Institutes of Health
DEPARTMENT
NIA
LABORATORY
Experimental Gerontology Section and Translational Gerontology Branch
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
A cross-sectional study of functional and metabolic changes during aging through the lifespan in male mice (Brain) part-V
STUDY_SUMMARY
Aging is associated with distinct phenotypical, physiological, and functional changes, leading to the onset of disease and death. The progression of aging-related traits varies widely among individuals, influenced by their environment, lifestyle, and genetics. In this study, we performed physiologic and functional tests cross-sectionally throughout the entire lifespan of male C57BL/6N mice. In parallel, metabolomics analyses in serum, brain, liver, heart, and skeletal muscle were also performed to identify signatures associated with frailty and age-dependent functional decline. Our findings indicate that the decline in gait speed as a function of age and frailty is associated with dramatic increases in the energetic cost of physical activity and decreases in working capacity. Aging and functional decline prompt organs to rewire their substrate selection and metabolism towards redox-related pathways, mainly in liver and heart. Collectively, the data provide a framework to further understand and characterize processes of aging at the individual and organ levels.
INSTITUTE
National Institutes of Health
DEPARTMENT
NIA
LABORATORY
Experimental Gerontology Section and Translational Gerontology Branch
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
Sclerostin antibody increases trabecular bone and bone mechanical properties by increasing osteoblast activity damaged by whole-body irradiation in mice
STUDY_TYPE
Basic research
STUDY_SUMMARY
Irradiation therapy causes bone deterioration and increased risk for skeletal-related events. Irradiation interferes with trabecular architecture through increased osteoclastic activity, decreased osteoblastic activity, and increased adipocyte expansion in the bone marrow (BM), which further compounds bone-related disease. Neutralizing antibodies to sclerostin (Scl-Ab) increase bone mass and strength by increasing bone formation and reducing bone resorption. We hypothesized that treatment with Scl-Ab would attenuate the adverse effects of irradiation by increasing bone volume and decreasing BM adipose tissue (BMAT), resulting in better quality bone. In this study, 12-week-old female C57BL/6J mice were exposed to 6 Gy whole-body irradiation or were non-irradiated, then administered Scl-Ab (25 mg/kg) or vehicle weekly for 5 weeks. Femoral ĀµCT analysis confirmed that the overall effect of IR significantly decreased trabecular bone volume/total volume (Tb.BV/TV) (2-way ANOVA, p<0.0001) with a -43.8% loss in Tb.BV/TV in the IR control group. Scl-Ab independently increased Tb.BV/TV by 3.07-fold in non-irradiated and 3.6-fold in irradiated mice (2-way ANOVA, p<0.0001). Irradiation did not affect cortical parameters, although Scl-Ab increased cortical thickness and area significantly in both irradiated and non-irradiated mice (2-way ANOVA, p<0.0001). Femoral mechanical testing confirmed Scl-Ab significantly increased bending rigidity and ultimate moment independently of irradiation (2-way ANOVA, p<0.0001). Static and dynamic histomorphometry of the femoral metaphysis revealed osteoblast vigor, not number, was significantly increased in the irradiated mice treated with Scl-Ab. Systemic alterations were assessed through serum lipidomic analysis, which showed that Scl-Ab normalized lipid profiles in the irradiated group. This data supports the theory of sclerostin as a novel contributor to the regulation of osteoblast activity after irradiation. Overall, our data support the hypothesis that Scl-Ab ameliorates the deleterious effects of whole-body irradiation on bone and adipose tissue in a mouse model. Our findings suggest that future research into localized and systemic therapies after irradiation exposure is warranted.
Machine learning-enabled renal cell carcinoma status prediction using multi-platform urine-based metabolomics
STUDY_SUMMARY
Currently, Renal Cell Carcinoma (RCC) is identified through expensive cross-sectional imaging, frequently followed by renal mass biopsy, which is invasive and subject to sampling errors. Hence, there is a critical need for a non-invasive diagnostic assay. RCC is a disease of altered cellular metabolism with the tumor(s) in close proximity to the urine in the kidney suggesting metabolomic profiling would be an excellent choice for assay development. Here, we applied liquid chromatography-mass spectrometry (LC-MS), nuclear magnetic resonance (NMR), and machine learning (ML) for the discovery of candidate metabolic panels for RCC. The study cohort consists of 82 RCC patients and 174 healthy controls, these were separated into two sub-cohorts: model cohort and the test cohort. Discriminatory metabolic features were selected in the model cohort, using univariate, wrapper, and embedded methods of feature selection. Three ML techniques with different induction biases were used for training and hyperparameter tuning. Final assessment of RCC status prediction was made using the test cohort with the selected biomarkers and the tuned ML algorithms. A seven-metabolite panel consisting of endogenous and exogenous metabolites enabled the prediction of RCC with 88% accuracy, 94% sensitivity, and 85% specificity in the test cohort, with an AUC of 0.98.
Machine learning-enabled renal cell carcinoma status prediction using multi-platform urine-based metabolomics NMR (part-II)
STUDY_SUMMARY
Currently, Renal Cell Carcinoma (RCC) is identified through expensive cross-sectional imaging, frequently followed by renal mass biopsy, which is invasive and subject to sampling errors. Hence, there is a critical need for a non-invasive diagnostic assay. RCC is a disease of altered cellular metabolism with the tumor(s) in close proximity to the urine in the kidney suggesting metabolomic profiling would be an excellent choice for assay development. Here, we applied liquid chromatography-mass spectrometry (LC-MS), nuclear magnetic resonance (NMR), and machine learning (ML) for the discovery of candidate metabolic panels for RCC. The study cohort consists of 82 RCC patients and 174 healthy controls, these were separated into two sub-cohorts: model cohort and the test cohort. Discriminatory metabolic features were selected in the model cohort, using univariate, wrapper, and embedded methods of feature selection. Three ML techniques with different induction biases were used for training and hyperparameter tuning. Final assessment of RCC status prediction was made using the test cohort with the selected biomarkers and the tuned ML algorithms. A seven-metabolite panel consisting of endogenous and exogenous metabolites enabled the prediction of RCC with 88% accuracy, 94% sensitivity, and 85% specificity in the test cohort, with an AUC of 0.98.
Lipid Profiling of Mouse Intestinal Organoids for studying APC Mutations
STUDY_SUMMARY
Inactivating mutations including both germline and somatic mutations in the adenomatous polyposis coli (APC) gene drives most familial and sporadic colorectal cancers. Understanding the metabolic implications of this mutation will aid to establish its wider impact on cellular behaviour and potentially inform clinical decisions. However, to date, alterations in lipid metabolism induced by APC mutations remain unclear. Intestinal organoids have gained widespread popularity in studying colorectal cancer and chemotherapies, because their three-dimensional structure more accurately mimics an in vivo environment. Here, we aimed to investigate intra-cellular lipid disturbances induced by APC gene mutations in intestinal organoids using a reversed-phase ultra-high-performance liquid chromatography mass spectrometry (RP-UHPLC-MS)-based lipid profiling method. Lipids of the organoids grown from either wildtype (WT) or mice with Apc mutations (Lgr5āEGFP-IRES-CreERT2 Apcfl/fl) were extracted and analysed using RP-UHPLC-MS. Concentrations of phospholipids (e.g. PC(16:0/16:0), PC(18:1/20:0), PC(38:0), PC(18:1/22:1)), ceramides (e.g. Cer(d18:0/22:0), Cer(d42:0), Cer(d18:1/24:1)) and hexosylceramide (e.g. HexCer(d18:1/16:0), HexCer(d18:1/22:0)) were higher in Apcfl/fl organoids, whereas levels of sphingomyelins (e.g. SM(d18:1/14:0), SM(d18:1/16:0) ) were lower compared to WT. These observations indicate that cellular metabolism of sphingomyelin was upregulated, resulting in the cellular accumulation of ceramides and production of HexCer due to the absence of Apcfl/fl in the organoids. Our observations demonstrated lipid profiling of organoids and provided an enhanced insight into the effects of the APC mutations on lipid metabolism, making for a valuable addition to screening options of the organoid lipidome.
Oxylipin biosynthesis reinforces cellular senescence through a RAS/p53 feedback loop and allows detection of senolysis
STUDY_SUMMARY
Cellular senescence is a stress or damage response that causes a permanent proliferative arrest and secretion of numerous factors with potent biological activities. This senescence-associated secretory phenotype (SASP) has been characterized largely for secreted proteins that participate in embryogenesis, wound healing, inflammation and many age-related pathologies. By contrast, lipid components of the SASP are understudied. We show that senescent cells activate the biosynthesis of several oxylipins that promote segments of the SASP and reinforce the proliferative arrest. Notably, senescent cells synthesize and accumulate an unstudied intracellular prostaglandin, 1a,1b-dihomo-15-deoxy-delta-12,14-prostaglandin J2. Released 15-deoxy-delta-12,14-prostaglandin J2 is a biomarker of senolysis in culture and in vivo. This and other prostaglandin D2-related lipids promote the senescence arrest and SASP by activating RAS signaling. These data identify an important aspect of cellular senescence and a method to detect senolysis
SARS-CoV-2 infection rewires host cell metabolism and is potentially susceptible to mTORC1 inhibition
STUDY_SUMMARY
Viruses hijack host cell metabolism to acquire the building blocks required for viral replication. Understanding how SARS-CoV-2 alters host cell metabolism could lead to potential treatments for COVID-19, the disease caused by SARS-CoV-2 infection. Here we profile metabolic changes conferred by SARS-CoV-2 infection in kidney epithelial cells and lung air-liquid interface cultures and show that SARS-CoV-2 infection increases glucose carbon entry into the TCA cycle via increased pyruvate carboxylase expression. SARS-CoV-2 also reduces host cell oxidative glutamine metabolism while maintaining reductive carboxylation. Consistent with these changes in host cell metabolism, we show that SARS-CoV-2 increases activity of mTORC1, a master regulator of anabolic metabolism, in cell lines and patient lung stem cell-derived airway epithelial cells. We also show evidence of mTORC1 activation in COVID-19 patient lung tissue. Notably, mTORC1 inhibitors reduce viral replication in kidney epithelial cells and patient-derived lung stem cell cultures. This suggests that targeting mTORC1 could be a useful antiviral strategy for SARS-CoV-2 and treatment strategy for COVID-19 patients, although further studies are required to determine the mechanism of inhibition and potential efficacy in patients.
Metabolic signatures of NAFLD - Lipidomics data (part 1 of 3)
STUDY_SUMMARY
Serum samples were randomized and extracted using a modified version of the previously-published Folch procedure, as applied recently [20]. The maternal samples were analysed as one batch and the cord blood samples as a second batch. In short, 10 ĀµL of 0.9% NaCl and, 120 ĀµL of CHCl3: MeOH (2:1, v/v) containing the internal standards (c = 2.5 Āµg/mL) was added to 10 ĀµL of each serum sample. The standard solution contained the following compounds: 1,2-diheptadecanoyl-sn-glycero-3-phosphoethanolamine (PE(17:0/17:0)), N-heptadecanoyl-D-erythro-sphingosylphosphorylcholine (SM(d18:1/17:0)), N-heptadecanoyl-D-erythro-sphingosine (Cer(d18:1/17:0)), 1,2-diheptadecanoyl-sn-glycero-3-phosphocholine (PC(17:0/17:0)), 1-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC(17:0)) and 1-palmitoyl-d31-2-oleoyl-sn-glycero-3-phosphocholine (PC(16:0/d31/18:1)), were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA), and, triheptadecanoylglycerol (TG(17:0/17:0/17:0)) was purchased from Larodan AB (Solna, Sweden). The samples were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 Ć g, 3 min). 60 ĀµL from the lower layer of each sample was then transferred to a glass vial with an insert and 60 ĀµL of CHCl3: MeOH (2:1, v/v) was added to each sample. The samples were stored at -80 Ā°C until analysis. Calibration curves using 1-hexadecyl-2-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine (PC(16:0e/18:1(9Z))), 1-(1Z-octadecenyl)-2-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine (PC(18:0p/18:1(9Z))), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC(18:0)), 1-oleoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC(18:1)), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (PE(16:0/18:1)), 1-(1Z-octadecenyl)-2-docosahexaenoyl-sn-glycero-3-phosphocholine (PC(18:0p/22:6)) and 1-stearoyl-2-linoleoyl-sn-glycerol (DG(18:0/18:2)), 1-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (LPE(18:1)), N-(9Z-octadecenoyl)-sphinganine (Cer(d18:0/18:1(9Z))), 1-hexadecyl-2-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (PE(16:0/18:1)) from Avanti Polar Lipids, 1-Palmitoyl-2-Hydroxy-sn-Glycero-3-Phosphatidylcholine (LPC(16:0)), 1,2,3 trihexadecanoalglycerol (TG(16:0/16:0/16:0)), 1,2,3-trioctadecanoylglycerol (TG(18:0/18:0/18:)) and 3Ī²-hydroxy-5-cholestene-3-stearate (ChoE(18:0)), 3Ī²-Hydroxy-5-cholestene-3-linoleate (ChoE(18:2)) from Larodan, were prepared to the following concentration levels: 100, 500, 1000, 1500, 2000 and 2500 ng/mL (in CHCl3:MeOH, 2:1, v/v) including 1250 ng/mL of each internal standard. The samples were analyzed by ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOFMS). Briefly, the UHPLC system used in this work was a 1290 Infinity II system from Agilent Technologies (Santa Clara, CA, USA). The system was equipped with a multi sampler (maintained at 10 Ā°C), a quaternary solvent manager and a column thermostat (maintained at 50 Ā°C). Injection volume was 1 ĀµL and the separations were performed on an ACQUITY UPLCĀ® BEH C18 column (2.1 mm Ć 100 mm, particle size 1.7 Āµm) by Waters (Milford, MA, USA). The mass spectrometer coupled to the UHPLC was a 6545 QTOF from Agilent Technologies interfaced with a dual jet stream electrospray (Ddual ESI) ion source. All analyses were performed in positive ion mode and MassHunter B.06.01 (Agilent Technologies) was used for all data acquisition. Quality control was performed throughout the dataset by including blanks, pure standard samples, extracted standard samples and control serum samples, including in-house serum and a pooled QC with an aliquot of each sample was collected and pooled and used as quality control sample. Relative standard deviations (% RSDs) for identified lipids in the control serum samples (n = 13) and in the pooled serum samples (n = 54) were on average 22.4% and 17.5%, respectively.
INSTITUTE
Ćrebro University
LAST_NAME
McGlinchey
FIRST_NAME
Aidan
ADDRESS
School of Medical Sciences, Ćrebro, Ćrebro, 70281, Sweden
Metabolic signatures of NAFLD - Polar metabolomics data (part II)
STUDY_SUMMARY
Analysis of polar metabolites Serum samples were randomized and sample preparation was carried out as described previously (Castilloet al. 2011). In summary, 400 ?L of MeOH containing ISTDs (heptadecanoic acid, deuterium-labeled DL-valine, deuterium-labeled succinic acid, and deuterium-labeled glutamic acid, c = 1 Āµg/mL) was added to 30 Āµl of the serum samples which were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 Ć g, 3 min) and 350 ?L of the supernatant was collected after centrifugation. The solvent was evaporated to dryness and 25 ?L of MOX reagent was added and the sample was incubated for 60 min at 45 Ā°C. 25 ?L of MSTFA was added and after 60 min incubation at 45 Ā°C 25 ?L of the retention index standard mixture (n-alkanes, c=10 Āµg/mL) was added. The analyses were carried out on an Agilent 7890B GC coupled to 7200 QTOF MS. Injection volume was 1 ĀµL with 100:1 cold solvent split on PTV at 70 Ā°C, heating to 300 Ā°C at 120 Ā°C/minute. Column: Zebron ZB-SemiVolatiles. Length: 20m, I.D. 0.18mm, film thickness: 0.18 Āµm. With initial Helium flow 1.2 mL/min, increasing to 2.4 mL/min after 16 mins. Oven temperature program: 50 Ā°C (5 min), then to 270Ā°C at 20 Ā°C/min and then to 300 Ā°C at 40 Ā°C/min (5 min). EI source: 250 Ā°C, 70 eV electron energy, 35ĀµA emission, solvent delay 3 min. Mass range 55 to 650 amu, acquisition rate 5 spectra/s, acquisition time 200 ms/spectrum. Quad at 150 Ā°C, 1.5 mL/min N2 collision flow, aux-2 temperature: 280 Ā°C. Calibration curves were constructed using alanine, citric acid, fumaric acid, glutamic acid, glycine, lactic acid, malic acid, 2-hydroxybutyric acid, 3-hydroxybutyric acid, linoleic acid, oleic acid, palmitic acid, stearic acid, cholesterol, fructose, glutamine, indole-3-propionic acid, isoleucine, leucine, proline, succinic acid, valine, asparagine, aspartic acid, arachidonic acid, glycerol-3-phosphate, lysine, methionine, ornithine, phenylalanine, serine and threonine purchased from Sigma-Aldrich (St. Louis, MO, USA) at concentration range of 0.1 to 80 ?g/mL. An aliquot of each sample was collected and pooled and used as quality control samples, together with a NIST SRM 1950 serum sample and an in-house pooled serum sample. Relative standard deviations (% RSDs) of the metabolite concentrations in control serum samples showed % RSDs within accepted analytical limits at averages of 27.2% and 29.2% for in-house QC abd pooled QC samples.
INSTITUTE
Ćrebro University
LAST_NAME
McGlinchey
FIRST_NAME
Aidan
ADDRESS
School of Medical Sciences, Ćrebro, Ćrebro, 70281, Sweden
Metabolomics analysis of plasma from a mouse model of astrocytoma subjected to radiotherapy
STUDY_SUMMARY
Mice were randomized in two groups (n=9 mice/group), one group was subjected to radiotherapy (Monday and Friday for 2 consecutive weeks at 3Gy/session) and the other cohort was the control. Sample were taken approximately each 10 days from the tail vein.
Effects of different planting densities on the metabolism of Panax notoginseng
STUDY_TYPE
Planting density experiment
STUDY_SUMMARY
At the moderate planting density, the primary metabolism (starch and sucrose metabolism) of the plants were significantly enhanced. However, the strong intraspecific competition at the higher planting densities resulted in stress as well as the accumulation of antioxidants (gentiobiose, oxalic acid, dehydroascorbic acid) and other stress resistance-related metabolites. Interestingly, the planting at low densities with low intraspecific competition disturbed normal carbohydrate metabolism by upregulating galactose metabolism.
INSTITUTE
Yunnan Agricultural University
DEPARTMENT
College of Plant Protection
LABORATORY
Key Laboratory for Agro-biodiversity and Pest Control of Ministry of Education
Large-scale enzyme-based xenobiotic identification for exposomics
STUDY_TYPE
Xenobiotic Metabolism
STUDY_SUMMARY
Exposomics methods are limited by low abundance of xenobiotic metabolites and lack of authentic standards, which precludes identification using solely mass spectrometry-based criteria. Here, we validate a method for enzymatic generation of xenobiotic metabolites for use with high-resolution mass spectrometry for chemical identification. Generated xenobiotic metabolites were used to confirm identities of respective metabolites in mice and human samples based upon accurate mass, retention time, and co-occurrence with related xenobiotic metabolites. The data shared here are high-resolution Orbitrap MS data for S9 incubations of 140 xenobiotic compounds with 0 and 24 hour time points for all reactions.
Phospholipid transfer function of PTPIP51 at mitochondria-associated ER membranes
STUDY_SUMMARY
In eukaryotic cells, mitochondria are closely tethered to the endoplasmic reticulum (ER) at sites called mitochondria-associated ER membranes (MAMs). Ca2+ ion and phospholipid transfer occurs at MAMs to support diverse cellular functions. Unlike those in yeast, the protein complexes involved in phospholipid transfer at MAMs in humans have not been identified. Here, we determined the crystal structure of the tetratricopeptide repeat domain of PTPIP51 (PTPIP51_TPR), a mitochondrial protein that interacts with the ER-anchored VAPB protein at MAMs. The structure of PTPIP51_TPR showed an archetypal TPR fold, and an electron density corresponding to an unidentified lipid-like molecule probably derived from the protein expression host was found in the structure. We revealed functions of PTPIP51 in phospholipid binding/transfer, particularly of phosphatidic acid, in vitro. Depletion of PTPIP51 in cells reduced the mitochondrial cardiolipin level. Additionally, we confirmed that the PTPIP51āVAPB interaction is mediated by the FFAT-like motif of PTPIP51 and the MSP domain of VAPB. Our findings suggest that PTPIP51 is a phospholipid transfer protein with a MAM-tethering function similar to the ERMES complex in yeast.
INSTITUTE
Korea Basic Science Institute
DEPARTMENT
Western Seoul Center
LABORATORY
Integrated Metabolomics Research Group
LAST_NAME
Lee
FIRST_NAME
Jueun
ADDRESS
150, Bugahyeon-ro, Seodaemun-gu, Seoul, Republic of Korea (Zip code: 03759)
Commensal intestinal microbiota regulates host luminal proteolytic activity and intestinal barrier integrity through Ī²-glucuronidase activity
STUDY_TYPE
Complex
STUDY_SUMMARY
Proteases constitute the largest enzyme gene family in vertebrates with intracellular and secreted proteases having critical roles in cellular and organ physiology. Intestinal tract contains diverse set of proteases mediating digestion, microbial responses, epithelial and immune signaling. Transit of chyme through the intestinal tract results in significant suppression of proteases. Although endogenous protease inhibitors have been identified, the broader mechanisms underlying protease regulation in the intestinal tract remains unclear. The objective of this study was to determine microbial regulation of proteolytic activity in intestinal tract using phenotype of post-infection irritable bowel syndrome, a condition characterized by high fecal proteolytic activity. Proteases of host pancreatic origin (chymotrypsin like pancreatic elastase 2A, 3B and trypsin 2) drove proteolytic activity. Of the 14 differentially abundant taxa, high proteolytic activity state was characterized by complete absence of the commensal Alistipes putredinis. Germ free mice had very high proteolytic activity (10-fold of specific-pathogen free mice) which dropped significantly upon humanization with microbiota from healthy volunteers. In contrast, high proteolytic activity microbiota failed to inhibit it, a defect that corrected with fecal microbiota transplant as well as addition of A. putredinis. These mice also had increased intestinal permeability similar to that seen in patients. Microbiota Ī²-glucuronidases mediate bilirubin deconjugation and unconjugated bilirubin is an inhibitor of serine proteases. We found that high proteolytic activity patients had lower urobilinogen levels, a product of bilirubin deconjugation. Mice colonized with Ī²-glucuronidase overexpressing E. coli demonstrated significant inhibition of proteolytic activity and treatment with Ī²-glucuronidase inhibitors increased it. The findings establish that specific commensal microbiota mediates effective inhibition of host pancreatic proteases and maintains intestinal barrier function through the production of Ī²-glucuronidases. This suggests an important homeostatic role for commensal intestinal microbiota.
Detecting sex-related changes to the metabolome of a critically endangered freshwater crayfish during the mating season
STUDY_TYPE
LC-MS analysis of crustacean haemolymph
STUDY_SUMMARY
Captive breeding is a vital tool in the conservation of highly endangered species, as it is for the Margaret River hairy marron, Cherax tenuimanus, from the south west of Australia. A close relative, Cherax cainii, has almost completely displaced C. tenuimanus in the wild and is a successful aquaculture species, whereas C. tenuimanus has performed poorly in captivity. We used untargeted liquid chromatography-mass spectrometry to obtain metabolomic profiles of female and male C. tenuimanus held in controlled aquarium conditions during their reproductive period. Using repeated haemolymph sampling we tracked the metabolomic profiles of animals just prior to and for a period of up to 34 days after pairing with a similar sized potential mate. We identified 54 reproducible annotated metabolites including amino acids, fatty acids, biogenic amines, purine and pyrimidine metabolites and excretion metabolites. Hierarchical clustering analysis distinguished five metabolite clusters. Principal component-canonical variate analysis clearly distinguished females from males, both unpaired and paired; similar trends in profile changes in both sexes after pairing; and a striking shift in males upon pairing. We discuss three main patterns of metabolomic responses: differentiation between sexes; reactive responses to the disturbance of pairing; and convergent response to the disturbance of pairing for males. Females generally had higher concentrations of metabolites involved in metabolic rate, mobilisation of energy stores and stress. Responses to the disturbance of pairing were also related to elevated stress. Females were mobilising lipid stores to deposit yolk, whereas males had a rapid and strong response to pairing, with shifts in metabolites associated with gonad development and communication, indicating males could complete reproductive readiness only once paired with a female. The metabolomic profiles support a previously proposed potential mechanism for displacement of C. tenuimanus by C. cainii in the wild and identify several biomarkers for testing hypotheses regarding reproductive success using targeted metabolomics.
INSTITUTE
Edith Cowan University
DEPARTMENT
School of Science
LAST_NAME
Lette
FIRST_NAME
Emily
ADDRESS
270 Joondalup Drive, Joondalup, WA, 6027, Australia
The effects of birth weight and breeding value for protein deposition on the plasma metabolome in growing pigs
STUDY_SUMMARY
An experiment was conducted with growing pigs, to determine the effects of birth weight (BiW) and estimated breeding value for protein deposition (EBV) on the metabolomic profile in plasma samples collected under different dietary regimens: protein adequate (A) or protein restricted (R, 70% of A).
INSTITUTE
Aarhus University
DEPARTMENT
Animal Science
LABORATORY
Metabolomics LC-MS platform Aarhus University Foulum
The effects of birth weight and breeding value for protein deposition on the plasma metabolome in growing pigs (part-II)
STUDY_SUMMARY
An experiment was conducted with growing pigs, to determine the effects of birth weight (BiW) and estimated breeding value for protein deposition (EBV) on the metabolomic profile in plasma samples collected under different dietary regimens: protein adequate (A) or protein restricted (R, 70% of A).
INSTITUTE
Aarhus University
DEPARTMENT
Animal Science
LABORATORY
Metabolomics LC-MS platform Aarhus University Foulum
The effects of birth weight and breeding value for protein deposition on the urine metabolome in growing pigs (part-III)
STUDY_SUMMARY
An experiment was conducted with growing pigs, to determine the effects of birth weight (BiW) and estimated breeding value for protein deposition (EBV) on the metabolomic profile in urine samples collected under different dietary regimens: protein adequate (A) or protein restricted (R, 70% of A).
INSTITUTE
Aarhus University
DEPARTMENT
Animal Science
LABORATORY
Metabolomics LC-MS platform Aarhus University Foulum
LAST_NAME
Hedemann
FIRST_NAME
Mette
ADDRESS
Blichers Alle 20, Tjele, -, 8830, Denmark
EMAIL
Mette.Hedemann@anis.au.dk
PHONE
51448783
TOTAL_SUBJECTS
40
NUM_MALES
40
PUBLICATIONS
The effects of birth weight and breeding value for protein deposition on nitrogen efficiency in growing pigs
Lipidomics dataset of Danio rerio optic nerve regeneration model
STUDY_SUMMARY
The right optic nerve of 1 year old female and male Danio rerio were crushed and collected three days after. Matching controls of uninjured eyes were also collected. The tissue was dissected from euthanized fish and āflashā frozen on dry ice in Eppendorf tubes. Due to the small size of the nerves, for each category (female crush, female control, male crush, male control) n=24 the samples were pooled. The brain from one male fish was also collected for control/calibration. Lipid extraction was done with the Bligh and Dyer [2] method, followed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) lipid profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. The lipids were identified and quantified with LipidSearch 4.2.21 and the statistical analysis was conducted through Metaboanalyst 5.0.
Metabolomics and metagenomics of metformin (Plasma)
STUDY_SUMMARY
Metformin affects the gut microbiome and altered microbiota may contribute to the hypoglycemic effect of metformin. Metabolomic analysis can be useful to elucidate the potential underlying mechanisms of the hypoglycemic effect according to the change in the microbiome and metabolites induced by the administration of metformin.
INSTITUTE
Seoul National University
DEPARTMENT
Department of Clinical Pharmacology and Therapeutics, College of Medicine and Hospital
Metabolomics and metagenomics of metformin (Urine)
STUDY_SUMMARY
Metformin affects the gut microbiome and altered microbiota may contribute to the hypoglycemic effect of metformin. Metabolomic analysis can be useful to elucidate the potential underlying mechanisms of the hypoglycemic effect according to the change in the microbiome and metabolites induced by the administration of metformin.
INSTITUTE
Seoul National University
DEPARTMENT
Department of Clinical Pharmacology and Therapeutics, College of Medicine and Hospital
Metabolomics and metagenomics of metformin (Stool)
STUDY_SUMMARY
Metformin affects the gut microbiome and altered microbiota may contribute to the hypoglycemic effect of metformin. Metabolomic analysis can be useful to elucidate the potential underlying mechanisms of the hypoglycemic effect according to the change in the microbiome and metabolites induced by the administration of metformin.
INSTITUTE
Seoul National University
DEPARTMENT
Department of Clinical Pharmacology and Therapeutics, College of Medicine and Hospital
Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance
STUDY_SUMMARY
Chemotherapy remains the standard of care for most cancers worldwide, however development of chemoresistance due to the presence of the drug-effluxing ABC transporters remains a significant problem. The development of safe and effective means to overcome chemoresistance is critical for achieving durable remissions in many cancer patients. We have investigated the energetic demands of ABC transporters in the context of the metabolic adaptations of chemoresistant cancer cells. Here we show that ABC transporters use mitochondrial-derived ATP as a source of energy to efflux drugs out of cancer cells. We further demonstrate that the loss of MCJ (DnaJC15), an endogenous negative regulator of mitochondrial respiration, in chemoresistant cancer cells boosts their ability to produce ATP from mitochondria and fuel ABC transporters. We have developed novel MCJ mimetics that can attenuate mitochondrial respiration and safely overcome chemoresistance in vitro and in vivo. Administration of MCJ mimetics in combination with standard chemotherapeutic drugs could therefore become an new strategy for treatment of multiple cancers.
INSTITUTE
University of Colorado Denver
DEPARTMENT
Biochemistry and Molecular Genetics
LABORATORY
Angelo D'Alessandro
LAST_NAME
Culp-Hill
FIRST_NAME
Rachel
ADDRESS
12801 E 17th Ave L18-9403D, Aurora, Colorado, 80045, USA
Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance (part-II)
STUDY_SUMMARY
Chemotherapy remains the standard of care for most cancers worldwide, however development of chemoresistance due to the presence of the drug-effluxing ABC transporters remains a significant problem. The development of safe and effective means to overcome chemoresistance is critical for achieving durable remissions in many cancer patients. We have investigated the energetic demands of ABC transporters in the context of the metabolic adaptations of chemoresistant cancer cells. Here we show that ABC transporters use mitochondrial-derived ATP as a source of energy to efflux drugs out of cancer cells. We further demonstrate that the loss of MCJ (DnaJC15), an endogenous negative regulator of mitochondrial respiration, in chemoresistant cancer cells boosts their ability to produce ATP from mitochondria and fuel ABC transporters. We have developed novel MCJ mimetics that can attenuate mitochondrial respiration and safely overcome chemoresistance in vitro and in vivo. Administration of MCJ mimetics in combination with standard chemotherapeutic drugs could therefore become an new strategy for treatment of multiple cancers.
INSTITUTE
University of Colorado Denver
DEPARTMENT
Biochemistry and Molecular Genetics
LABORATORY
Angelo D'Alessandro
LAST_NAME
Culp-Hill
FIRST_NAME
Rachel
ADDRESS
12801 E 17th Ave L18-9403D, Aurora, Colorado, 80045, USA
Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance (part-III)
STUDY_SUMMARY
Chemotherapy remains the standard of care for most cancers worldwide, however development of chemoresistance due to the presence of the drug-effluxing ABC transporters remains a significant problem. The development of safe and effective means to overcome chemoresistance is critical for achieving durable remissions in many cancer patients. We have investigated the energetic demands of ABC transporters in the context of the metabolic adaptations of chemoresistant cancer cells. Here we show that ABC transporters use mitochondrial-derived ATP as a source of energy to efflux drugs out of cancer cells. We further demonstrate that the loss of MCJ (DnaJC15), an endogenous negative regulator of mitochondrial respiration, in chemoresistant cancer cells boosts their ability to produce ATP from mitochondria and fuel ABC transporters. We have developed novel MCJ mimetics that can attenuate mitochondrial respiration and safely overcome chemoresistance in vitro and in vivo. Administration of MCJ mimetics in combination with standard chemotherapeutic drugs could therefore become an new strategy for treatment of multiple cancers.
Understanding systemic and local inflammation induced by nasal polyposis: role of the allergic phenotype (part-I)
STUDY_SUMMARY
Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by persistent symptoms associated to the development of nasal polyps. To this day, the molecular mechanisms involved are still not well defined. However, it has been suggested that a sustained inflammation as allergy is involved in its onset. In this pilot study, we aimed to look into the effect of the allergic status of the patient and in their underlying mechanisms. To achieve this, we recruited 22 patients with CRSwNP and classified them in non-allergic and allergic using ImmunoCAP ISAC molecular diagnosis. Plasma samples were analyzed using liquid chromatography coupled to mass spectrometry (LC-MS). Subsequently, the identified changed metabolites from plasma that were commercially available were then analyzed by targeted analysis in some nasal polyps. Additionally, nasal polyp and mucosa tissue samples were examined for eosinophils and neutrophils. We found that 9 out of the 22 patients were sensitized to some aeroallergens (named as allergic). The other 13 patients had no sensitizations (non-allergic). Regarding metabolomics, we found that bilirubin, cortisol, lysophosphatidylcholines (LPCs) 16:0, 18:0 and 20:4 and lysophosphatidylinositol (LPI) 20:4, metabolites that are usually related to a sustained allergic inflammation, were unexpectedly increased in the plasma of non-allergic patients with CRSwNP compared to allergic patients with CRSwNP. LPC 16:0, LPC 18:0 and LPI 20:4 metabolites followed the same trend in the nasal polyp as they did in plasma. Comparison of nasal polyps with nasal mucosa tissue showed a significant increase in eosinophils (p < 0.001) and neutrophils (p < 0.01) in allergic patients with CRSwNP. There were also more eosinophils in the polyps of non-allergic patients with CRSwNP than in their nasal mucosa (p <0.01). The polyps from non-allergic patients with CRSwNP had less eosinophils than the polyps of allergic patients with CRSwNP (p < 0.05). Our data suggests that there is a systemic inflammatory response associated to CRSwNP in the absence of allergy, which could be accountable for the nasal polyp development. Allergic patients with CRSwNP presented a higher number of eosinophils located in nasal polyps suggesting that eosinophilia might be connected to the development of nasal polyps in these patients.
INSTITUTE
CEMBIO
LAST_NAME
Delgado Dolset
FIRST_NAME
MarĆa Isabel
ADDRESS
Urb. MonteprĆncipe s/n, Ctra. Boadilla del Monte km 5.3, Madrid, Madrid, 28668, Spain
Understanding systemic and local inflammation induced by nasal polyposis: role of the allergic phenotype (part-II)
STUDY_SUMMARY
Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by persistent symptoms associated to the development of nasal polyps. To this day, the molecular mechanisms involved are still not well defined. However, it has been suggested that a sustained inflammation as allergy is involved in its onset. In this pilot study, we aimed to look into the effect of the allergic status of the patient and in their underlying mechanisms. To achieve this, we recruited 22 patients with CRSwNP and classified them in non-allergic and allergic using ImmunoCAP ISAC molecular diagnosis. Plasma samples were analyzed using liquid chromatography coupled to mass spectrometry (LC-MS). Subsequently, the identified changed metabolites from plasma that were commercially available were then analyzed by targeted analysis in some nasal polyps. Additionally, nasal polyp and mucosa tissue samples were examined for eosinophils and neutrophils. We found that 9 out of the 22 patients were sensitized to some aeroallergens (named as allergic). The other 13 patients had no sensitizations (non-allergic). Regarding metabolomics, we found that bilirubin, cortisol, lysophosphatidylcholines (LPCs) 16:0, 18:0 and 20:4 and lysophosphatidylinositol (LPI) 20:4, metabolites that are usually related to a sustained allergic inflammation, were unexpectedly increased in the plasma of non-allergic patients with CRSwNP compared to allergic patients with CRSwNP. LPC 16:0, LPC 18:0 and LPI 20:4 metabolites followed the same trend in the nasal polyp as they did in plasma. Comparison of nasal polyps with nasal mucosa tissue showed a significant increase in eosinophils (p < 0.001) and neutrophils (p < 0.01) in allergic patients with CRSwNP. There were also more eosinophils in the polyps of non-allergic patients with CRSwNP than in their nasal mucosa (p <0.01). The polyps from non-allergic patients with CRSwNP had less eosinophils than the polyps of allergic patients with CRSwNP (p < 0.05). Our data suggests that there is a systemic inflammatory response associated to CRSwNP in the absence of allergy, which could be accountable for the nasal polyp development. Allergic patients with CRSwNP presented a higher number of eosinophils located in nasal polyps suggesting that eosinophilia might be connected to the development of nasal polyps in these patients.
INSTITUTE
CEMBIO
LAST_NAME
Delgado Dolset
FIRST_NAME
MarĆa Isabel
ADDRESS
Urb. MonteprĆncipe s/n, Ctra. Boadilla del Monte km 5.3, Madrid, Madrid, 28668, Spain
The COVIDome Explorer Researcher Portal (Red Blood Cells)
STUDY_SUMMARY
COVID-19 pathology involves dysregulation of diverse molecular, cellular, and physiological processes. In order to expedite integrated and collaborative COVID-19 research, we completed multi-omics analysis of hospitalized COVID-19 patients including matched analysis of the whole blood transcriptome, plasma proteomics with two complementary platforms, cytokine profiling, plasma and red blood cell metabolomics, deep immune cell phenotyping by mass cytometry, and clinical data annotation. We refer to this multidimensional dataset as the COVIDome. We then created the COVIDome Explorer, an online researcher portal where the data can be analyzed and visualized in real time. We illustrate here the use of the COVIDome dataset through a multi-omics analysis of biosignatures associated with C-reactive protein (CRP), an established marker of poor prognosis in COVID-19, revealing associations between CRP levels and damage-associated molecular patterns, depletion of protective serpins, and mitochondrial metabolism dysregulation. We expect that the COVIDome Explorer will rapidly accelerate data sharing, hypothesis testing, and discoveries worldwide.
COVID-19 pathology involves dysregulation of diverse molecular, cellular, and physiological processes. In order to expedite integrated and collaborative COVID-19 research, we completed multi-omics analysis of hospitalized COVID-19 patients including matched analysis of the whole blood transcriptome, plasma proteomics with two complementary platforms, cytokine profiling, plasma and red blood cell metabolomics, deep immune cell phenotyping by mass cytometry, and clinical data annotation. We refer to this multidimensional dataset as the COVIDome. We then created the COVIDome Explorer, an online researcher portal where the data can be analyzed and visualized in real time. We illustrate here the use of the COVIDome dataset through a multi-omics analysis of biosignatures associated with C-reactive protein (CRP), an established marker of poor prognosis in COVID-19, revealing associations between CRP levels and damage-associated molecular patterns, depletion of protective serpins, and mitochondrial metabolism dysregulation. We expect that the COVIDome Explorer will rapidly accelerate data sharing, hypothesis testing, and discoveries worldwide.
1H HRMAS NMR Spectroscopy based Metabolomics of Urinary Bladder Tissues from NMIBC Patients
STUDY_TYPE
NMR Based Metabolomics
STUDY_SUMMARY
Application of 1H HRMAS NMR Spectroscopy to study malignancy induced metabolomic changes in urinary bladder tissues from 26 NMIBC patients. Predict the possible biomarker of NMIBC in urinary bladder tissues.
INSTITUTE
Centre of Biomedical Research, Lucknow, India
DEPARTMENT
Department of Radiology
LABORATORY
NMR Based Metabolomics
LAST_NAME
Roy
FIRST_NAME
Raja
ADDRESS
Centre of Biomedical Research, Lucknow, India 226014
AdipoAtlas: A Reference Lipidome for Human White Adipose Tissue
STUDY_SUMMARY
Obesity, characterized by expansion and metabolic dysregulation of white adipose tissue (WAT), has reached pandemic proportions and acts as a primer for a wide range of metabolic disorders. Remodelling of WAT lipidome in obesity and associated comorbidities can explain disease etiology and provide valuable diagnostic and prognostic markers. To support understanding of WAT lipidome remodelling at molecular level, we performed in-depth lipidomics profiling of human subcutaneous and visceral WAT of lean and obese individuals. Tissue-tailored preanalytical and analytical workflows allowed accurate identification and semi-absolute quantification of 1636 and 737 lipid molecular species, respectively, and summarized here in a form of human WAT reference lipidome. Deep lipidomic profiling allowed to identify main lipid (sub)classes undergoing depot/phenotype specific remodelling. Furthermore, previously unanticipated diversity of WAT ceramides was uncovered. AdipoAtlas reference lipidome will serve as a data-rich resource for development of WAT-specific high-throughput methods and as a scaffold for systems medicine data integration.
INSTITUTE
University of Leipzig
DEPARTMENT
Faculty for Chemistry and Mineralogy, Biotechnological-Biomedical Center
LABORATORY
Fedorova Lab
LAST_NAME
Fedorova
FIRST_NAME
Maria
ADDRESS
Deutscher Platz 5
EMAIL
maria.fedorova@bbz.uni-leipzig.de
PHONE
03419731336
NUM_GROUPS
4
STUDY_COMMENTS
Pools of subcutaneous and visceral white adipose tissue were generated from lean patients (BMI < 25; n=5) and obese (BMI > 40; n=81)
Differential Metabolites and Disturbed Metabolic Pathways Associated with chlorpromazine Poisoning
STUDY_SUMMARY
Perturbed pathways and differential metabolites analysis for lethal chlorpromzaine poisoning relative to control and non-lethal chlorpromazine poisoning group.
Mice homozygous for the Scd1ab-2J allele have a defect Scd1 gene with an in-frame stop codon in exon 2. To identify SCD1-derived phospholipid species, we analysed PI and PC species in organs and tissues that highly express SCD1 and are considered as targets for intervention with SCD1 inhibitors, i.e., liver, skin, hind leg skeletal muscle, and white abdominal fat.
Blood was collected from diabetic dogs and healthy control dogs. Diabetic dogs and control dogs were breed matched where possible. Timing of blood collection after a meal was matched as best as posible. Serum was frozen at -80C.
The Role of Intestinal-derived FGF15 and Vertical Sleeve Gastrectomy on Plasma Bile Acid Composition in Mice
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Bariatric surgeries such as the Vertical Sleeve Gastrectomy (VSG) are invasive but provide the most effective long-term metabolic improvements in individuals with obesity and/or Type 2 diabetes. These powerful effects of manipulating the gastrointestinal tract point to an important role of gastrointestinal signals in regulating both energy balance and metabolism. To that end, we have used mouse models of VSG to identify key gut signals that mediate these beneficial effects. Previous data from our rodent model of VSG led us to hypothesize a potential role for the hormone Fibroblast-Growth Factor15/19 (mouse/human ortholog) which pharmacologically can regulate many aspects of energy homeostasis and glucose handling. FGF15 is expressed in ileal enterocytes of the small intestine and is released postprandially. Like many other gut hormones, postprandial plasma concentrations of the human ortholog FGF19 and ileal FGF15 expression in mice increase after VSG. We generated intestinal-specific FGF15 knock out (VilCreERT2; Fgf15f/f) mice and controls, which were maintained on 60% high-fat diet. VSG resulted in increased plasma bile acid levels. However, intestinal-specific FGF15 knock out mice had considerably higher levels of circulating total and hydrophobic bile acids after VSG. Unlike what we had predicted, intestinal-specific FGF15 knock out mice lost more weight after VSG as a result of increased lean tissue loss compared to control mice. Further, the loss of bone mineral density and bone marrow adipose tissue observed after VSG in control mice was even greater in intestinal-specific FGF15 knock out mice, perhaps secondary to anemia and elevated erythropoietin/FGF23. Finally the effect of VSG to improve glucose tolerance and to reduce hepatic cholesterol was also absent in intestinal-specific FGF15 knock out mice. These data point to an important role for intestinal FGF15 to protect the organism from deleterious effects of bile acid toxicity after VSG.
X13CMS: Global Tracking of Isotopic Labels in Untargeted Metabolomics
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Studies of isotopically labeled compounds have been fundamental to understanding metabolic pathways and fluxes. They have traditionally, however, been used in conjunction with targeted analyses that identify and quantify a limited number of labeled downstream metabolites. Here we describe an alternative workflow that leverages recent advances in untargeted metabolomic technologies to track the fates of isotopically labeled metabolites in a global, unbiased manner. This untargeted approach can be applied to discover novel biochemical pathways and characterize changes in the fates of labeled metabolites as a function of altered biological conditions such as disease. To facilitate the data analysis, we introduce X13CMS, an extension of the widely used mass spectrometry-based metabolomic software package XCMS. X13CMS uses the XCMS platform to detect metabolite peaks and perform retention-time alignment in liquid chromatography/mass spectrometry (LC/MS) data. With the use of the XCMS output, the program then identifies isotopologue groups that correspond to isotopically labeled compounds. The retrieval of these groups is done without any a priori knowledge besides the following input parameters: (i) the mass difference between the unlabeled and labeled isotopes, (ii) the mass accuracy of the instrument used in the analysis, and (iii) the estimated retention-time reproducibility of the chromatographic method. Despite its name, X13CMS can be used to track any isotopic label. Additionally, it detects differential labeling patterns in biological samples collected from parallel control and experimental conditions. We validated the ability of X13CMS to accurately retrieve labeled metabolites from complex biological matrices both with targeted LC/MS/MS analysis of a subset of the hits identified by the program and with labeled standards spiked into cell extracts. We demonstrate the full functionality of X13CMS with an analysis of cultured rat astrocytes treated with uniformly labeled (U-)13C-glucose during lipopolysaccharide (LPS) challenge. Our results show that out of 223 isotopologue groups enriched from U-13C-glucose, 95 have statistically significant differential labeling patterns in astrocytes challenged with LPS compared to unchallenged control cells. Only two of these groups overlap with the 32 differentially regulated peaks identified by XCMS, indicating that X13CMS uncovers different and complementary information from untargeted metabolomic studies. Like XCMS, X13CMS is implemented in R. It is available from our laboratory website at http://pattilab.wustl.edu/x13cms.php.
INSTITUTE
Washington University, St. Louis
LAST_NAME
Cho
FIRST_NAME
Kevin
ADDRESS
1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA
Metabolomic profiling of the rat hippocampus across developmental ages and after learning
STUDY_TYPE
Developmental study
STUDY_SUMMARY
Little is known about how the hippocampal metabolomic profile changes across development and in response to learning at different ages. To fill this knowledge gap, we employed an untargeted metabolomic analyses in rats to determine how the hippocampal metabolome changes over the course of post-natal development under basal conditions and following inhibitory avoidance (IA) training, an aversive episodic event. We found that unique metabolomic profiles accompany learning at different ages. Subsequent biochemical and behavioral studies based on unique metabolomic regulations in the infant hippocampus established that infantile learning selectively recruits the glutathione-mediated antioxidant defenses for the formation of infantile memory.
Lung metabolomics after ischemic acute kidney injury reveals increased oxidative stress, altered energy production, and ATP depletion
STUDY_SUMMARY
Acute kidney injury (AKI) is a complex disease associated with increased mortality that may be due to deleterious distant organ effects. AKI associated with respiratory complications, in particular, has a poor outcome. In murine models, AKI is characterized by increased circulating cytokines, lung chemokine upregulation, and neutrophilic infiltration, similar to other causes of indirect acute lung injury (ALI)(e.g., sepsis). Many causes of lung inflammation are associated with a lung metabolic profile characterized by increased oxidative stress, a shift towards the use of other forms of energy production, and/or a depleted energy state. To our knowledge, there are no studies that have evaluated pulmonary energy production and metabolism after AKI. We hypothesized that based on the parallels between inflammatory acute lung injury and AKI-mediated lung injury, a similar metabolic profile would be observed. Lung metabolomics and ATP levels were assessed 4 hours, 24 hours, and 7 days after ischemic AKI in mice. Numerous novel findings regarding the effect of AKI on the lung were observed including 1) increased oxidative stress, 2) a shift toward alternate methods of energy production, and 3) depleted levels of ATP. The findings in this report bring to light novel characteristics of AKI-mediated lung injury and provide new leads into the mechanisms by which AKI in patients predisposes to pulmonary complications.
Rationally designed bacterial consortia to treat chronic immune-mediated colitis and restore intestinal homeostasis
STUDY_TYPE
Targeted metabolomics
STUDY_SUMMARY
GUT103 and GUT108, live biotherapeutic products rationally designed to complement missing or underrepresented functions in the dysbiotic microbiome of IBD patients; they address upstream targets, rather than targeting a single cytokine to block downstream inflammation responses. Systematic colonization experiments in colitis mouse models were performed to test their therapeutic effects. Targeted fecal metabolomics data uploaded here of bile acids, short-chain fatty acids, and tryptophan metabolites provides a unique metabolome perspective for evaluation of the therapeutic potential of GUT103 and GUT108.
INSTITUTE
University of North Carolina at Chapel Hill
LABORATORY
Gusto Global LLC.
LAST_NAME
Lai
FIRST_NAME
Yunjia
ADDRESS
1104 MHRC, 135 Dauer Drive, Chapel Hill, NC 27599, USA
Increasing evidence indicates that physical activity and exercise training may delay or prevent the onset of Alzheimerās disease (AD). However, systemic biomarkers that can measure exercise effects on brain function and that link to relevant metabolic responses are lacking. This study utilized blood samples of 23 asymptomatic late middle-aged adults with familial and genetic risk for AD who underwent 26 weeks of supervised treadmill training. Metabolomic profiles were evaluated using MS.
INSTITUTE
University of Wisconsin - Madison
DEPARTMENT
Medicine
LABORATORY
Wisconsin Alzheimer's Disease Research Center
LAST_NAME
GaitƔn
FIRST_NAME
Julian
ADDRESS
600 Highland Ave. J5/1M CSC MC2420, Madison, WI 53792
Gut microbiome influence on metabolic 1 disease in HIV and high-risk populations
STUDY_SUMMARY
Poor metabolic health, characterized by insulin resistance and dyslipidemia, is higher in people living with HIV (PLWH) and has been linked with inflammation, anti-retroviral therapy (ART) drugs, and ART-associated lipodystrophy (LD). Metabolic disease is associated with gut microbiome composition outside the context of HIV but has not been deeply explored in HIV infection nor in high-risk men who have sex with men (HR-MSM), who have a highly altered gut microbiome composition. Furthermore, the contribution of increased bacterial translocation and associated systemic inflammation that has been described in HIV-positive and HR-MSM individuals has not been explored. We used a multi-omic approach to explore relationships between gut microbes, immune phenotypes, diet, and metabolic health across ART-treated PLWH with and without .D untreated PLWH; and HR-MSM. For PLWH on ART, we further explored associations with the plasma metabolome.
Dual RNA regulator VcdRP in V. cholerae modulates central metabolism
STUDY_SUMMARY
Bacterial small RNAs (sRNAs) are well-known to modulate gene expression by base-pairing with trans-coded transcripts and are typically considered to be non-coding. However, several sRNAs have been reported to also contain an open reading frame and thus are considered dual-function regulators. We discovered a dual-function regulator from Vibrio cholerae, called VcdRP, harboring a 29 amino acid protein (VcdP), as well as a base-pairing sequence. In this study, we measured the metabolite abundance of glycolytic and citric acid cycle intermediates using LC-MS.
Modifying Chromatography Conditions for Improved Unknown Feature Identification in Untargeted Metabolomics
STUDY_SUMMARY
Project represents an effort to modify chromatographic conditions for improved compound identification in untargeted metabolomics. Two different modes of chromatograph (HILIC and RPLC) and multiple run conditions (sample loading, gradient duration, iterative acquisition) were evaluated. All relevant data from different conditions are contained within the raw data archive file attached to this submission. Metadata associated with this Metabolomics Workbench submission reflects only the manually reviewed identifications obtained using modified HILIC conditions. See protocol file "Mod_vs_Con_Chrom_IDs_Protocol.pdf" for details.
INSTITUTE
University of Michigan
DEPARTMENT
Chemistry/Internal Medicine/CCMB/Biomedical Research Core Facilities
LABORATORY
Michigan Compound Identification Development Core/BRCF Metabolomics Core
Blood was collected from diabetic dogs and healthy control dogs. Diabetic dogs and control dogs were breed matched where possible. Timing of blood collection after a meal was matched as best as posible. Serum was frozen at -80C.
A reductionist approach using primary and metastatic cell-derived extracellular vesicles reveals hub proteins associated with oral cancer prognosis
STUDY_SUMMARY
Oral squamous cell carcinoma (OSCC) has high mortality rates that are largely associated with lymph node metastasis. However, the molecular mechanisms that drive OSCC metastasis are unknown. Extracellular vesicles (EVs) are membrane-bound particles that play a role in intercellular communication and impact cancer development and progression. Thus, profiling EVs would be of great significance to decipher the role of EV cargo in OSCC metastasis. For that purpose, we used a reductionist approach to map the proteomic, miRNA, metabolomic, and lipidomic profiles of extracellular vesicles (EVs) derived from human primary tumor (SCC-9) cells and matched lymph node metastases (LN1) cells. Distinct omics profiles were associated with the metastatic phenotype, including 670 proteins, 217 miRNAs, 26 metabolites, and 64 lipids differentially abundant between LN1- and SCC-9-derived EVs. A multi-omics integration identified 11 āhub proteinsā significantly decreased at the metastatic site compared to primary tumor-derived EVs. We confirmed the validity of these findings with analysis of data from multiple public databases, and found that low abundance of seven hub proteins in metastatic EVs is correlated with reduced survival and tumor aggressiveness in cancer patients. In summary, this multi-omics approach identified proteins transported by EVs that are associated with metastasis, and which may potentially serve as prognostic markers in OSCC.
INSTITUTE
National Center for Research in Energy and Materials
Metabolomics of mouse feces comparing GF and CONV-R mice
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
Using high coverage metabolomics, we profiled feces, blood sera and cerebral cortical brain tissues of germ-free C57BL/6 mice and their age-matched conventionally raised counterparts. Results revealed for all three sample matrices metabolomic signatures owing to microbiota, yielding hundreds of identified metabolites including 533 altered for feces, 231 for sera and 58 for brain tissues with numerous significantly enriched pathways involving aromatic amino acids and neurotransmitters.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Department of Environmental Sciences and Engineering
Metabolomics of mouse blood sera comparing GF and CONV-R mice
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
Using high coverage metabolomics, we profiled feces, blood sera and cerebral cortical brain tissues of germ-free C57BL/6 mice and their age-matched conventionally raised counterparts. Results revealed for all three sample matrices metabolomic signatures owing to microbiota, yielding hundreds of identified metabolites including 533 altered for feces, 231 for sera and 58 for brain tissues with numerous significantly enriched pathways involving aromatic amino acids and neurotransmitters.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Department of Environmental Sciences and Engineering
Metabolomics of mouse cerebral cortical brain tissues comparing GF and CONV-R mice
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
Using high coverage metabolomics, we profiled feces, blood sera and cerebral cortical brain tissues of germ-free C57BL/6 mice and their age-matched conventionally raised counterparts. Results revealed for all three sample matrices metabolomic signatures owing to microbiota, yielding hundreds of identified metabolites including 533 altered for feces, 231 for sera and 58 for brain tissues with numerous significantly enriched pathways involving aromatic amino acids and neurotransmitters.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Department of Environmental Sciences and Engineering
Application of the redox metabolite detection method for mouse liver
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on ZIC-pHILIC chromatography. This study is an independent replicate.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mouse kidney
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on ZIC-pHILIC chromatography. This study is an independent replicate
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mouse biofluids
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian biofluids. Three different extraction conditions were compared, including derivatization of glutathione. This study was with mouse cerebrospinal fluid.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mouse kidney (part II)
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on ZIC-pHILIC chromatography. This study is an independent repeat.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mouse liver (part II)
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on ZIC-pHILIC chromatography. This study is an independent repeat.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for profiling redox state following pharmacologic perturbation with methotrexate
STUDY_SUMMARY
This study aimed to test methods for detection of redox metabolites from mammalian cells upon metabolism perturbation by methotrexate. Three time points and three extraction conditions are explored
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Optimization of redox metabolite detection in mammalian cells (part I)
STUDY_SUMMARY
Conditions were tested to optimize number of cells and extraction buffer for the detection of redox reactive metabolites from mammalian cells. Four different extraction buffers were compared. Derivatization of glutathione was explored as a condition as well. This is an independent repeat.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mammalian tissues (part I)
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on LUNA NH2 HILIC chromatography
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mammalian tissues (part II)
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on Accucore Amide HILIC chromatography
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mammalian tissues (part III)
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian tissues. Three different chromatographic conditions were compared as well as three different extraction conditions. This study was run on ZIC-pHILIC chromatography.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for profiling redox state following pharmacologic perturbations of redox balance in cells (part I)
STUDY_SUMMARY
This study aimed to test our optimized method for detection of redox metabolites from mammalian cells upon redox stress and metabolism perturbations. Redox balance was perturbed using H2O2 and diamide, metabolism was perturbed by methotrexate or oligomycin.
Application of the redox metabolite detection method for profiling redox state following pharmacologic perturbations of redox balance in cells (part II)
STUDY_SUMMARY
This study aimed to test our optimized method for detection of redox metabolites from mammalian cells upon redox stress and metabolism perturbations. Redox balance was perturbed using H2O2 and diamide, metabolism was perturbed by methotrexate or oligomycin. This experiment is an independent repeat.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for profiling redox state following pharmacologic perturbations of redox balance in cells (part III)
STUDY_SUMMARY
This study aimed to test our optimized method for detection of redox metabolites from mammalian cells upon redox stress and metabolism perturbations. Redox balance was perturbed using H2O2 and diamide, metabolism was perturbed by methotrexate or oligomycin. This experiment is an independent repeat.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Optimization of redox metabolite detection in mammalian cells (part II)
STUDY_SUMMARY
Conditions were tested to optimize number of cells and extraction buffer for the detection of redox reactive metabolites from mammalian cells. Four different extraction buffers were compared. Derivatization of glutathion was explored as a condition as well.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Application of the redox metabolite detection method for mouse biofluids (part II)
STUDY_SUMMARY
This study was aimed at optimizing redox metabolites detection from mammalian biofluids. Three different extraction conditions were compared, including derivatization of glutathione. This study was with mouse plasma.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Timecourse on primary human highly differentiated airway epithelial cells infected with HRV-C15
STUDY_SUMMARY
At each time point (4, 12, 24 h), ALI basolateral media was collected and incubated 1:1 in ice-cold 100% methanol for 30 minutes on ice, vortexing every 10 minutes. Basolateral media was centrifuged at 20,000 Ć š for 10 minutes at 4Ā°C, and further diluted in 50% methanol prior to mass spectrometry plate loading.
Plasma metabolomics of diverse mouse strains infected with Plasmodium chabaudi
STUDY_SUMMARY
To uncover links between metabolism and disease severity in murine malaria, we performed plasma metabolomics via Metabolon on eight inbred, Plasmodium chabaudi-infected mouse strains with diverse disease phenotypes. We sacrificed and collected plasma from >=3 mice per strain per day of acute infection alongside uninfected control mice (approximately days 5-12 depending on mouse strain). We collected disease severity data, e.g. weight loss, liver enzymes, and anemia, concurrently. Together, these data enable 1) a picture of strain-specific and conserved metabolic responses during acute malaria, and 2) a comparison between metabolic responses and disease severity.
Study on Metabolic Response of HEK 293 Cells Exposed to Methylmercury
STUDY_SUMMARY
HEK 293 cells were treated with 7.5 Ī¼M methylmercury (HgMe) for 48 h. Metabolites were extracted and subjected to LC-HRMS based metabolomics. LC-HRMS data were processed by SIEVE2.2 software. Metabolites associated with HgMe toxicity were screened and identified.
Comparison of High-Resolution Fourier Transform Mass Spectrometry Platforms for Metabolite Annotation - C. elegans
STUDY_SUMMARY
Fourier transform ion cyclotron resonance (FT-ICR) and Orbitrap mass spectrometry (MS) are among the highest-performing analytical platforms in metabolomics. Their high mass measurement accuracy and mass resolving power enable detailed investigation of biological metabolomes. Non-targeted MS experiments, however, yield extremely complex datasets that make metabolite annotation very challenging, if not impossible. High-resolution accurate mass measurements greatly facilitate this process by reducing mass errors and spectral overlaps. When applied together with relative isotopic abundance (RIA) measurements, heuristic rules, and constraints during searches, the number of candidate elemental formula(s) can be significantly reduced. Here, we evaluate the performance of two leading analytical MS platforms, Orbitrap ID-X and 12T solariX FT-ICR mass spectrometers, in terms of mass accuracy and RIA measurements, and how these factors affect the assignment of the correct elemental formulae in metabolite annotation. Quality of the mass measurements was evaluated under various experimental conditions (resolution: 120 K, 240 K, 500 K; automatic gain control: 5e4, 1e5, 5e5) for the Orbitrap MS platform.
Comparison of High-Resolution Fourier Transform Mass Spectrometry Platforms for Metabolite Annotation
STUDY_SUMMARY
Fourier transform ion cyclotron resonance (FT-ICR) and Orbitrap mass spectrometry (MS) are among the highest-performing analytical platforms in metabolomics. Their high mass measurement accuracy and mass resolving power enable detailed investigation of biological metabolomes. Non-targeted MS experiments, however, yield extremely complex datasets that make metabolite annotation very challenging, if not impossible. High-resolution accurate mass measurements greatly facilitate this process by reducing mass errors and spectral overlaps. When applied together with relative isotopic abundance (RIA) measurements, heuristic rules, and constraints during searches, the number of candidate elemental formula(s) can be significantly reduced. Here, we evaluate the performance of two leading analytical MS platforms, Orbitrap ID-X and 12T solariX FT-ICR mass spectrometers, in terms of mass accuracy and RIA measurements, and how these factors affect the assignment of the correct elemental formulae in metabolite annotation.
Untargeted Metabolomics analysis of A549 treated with 0.5 mM extracellular ATP and 10 ng/ml TGF-beta
STUDY_SUMMARY
Control, 0.5 mM extracellular ATP and 10 ng/ml TGF-beta were used to treated 5 million A549 lung cancer cells in vitro for 2, 6 and 12 hours. The untargeted metabolomics analysis was performed on the cell lysates. The main objective of the study was to determine changes in metabolite abundances in lung cancer after treatment with extracellular ATP and TGF-beta (a known EMT inducer).
INSTITUTE
Ohio University
DEPARTMENT
Biological Sciences
LABORATORY
Dr. Xiaozhuo Chen, Edison biotechnology Institute
LAST_NAME
Shriwas
FIRST_NAME
Pratik
ADDRESS
Room 425, Parks Hall, College of Pharmacy, Ohio State University, Columbus Ohio. 43210
EMAIL
ps774614@ohio.edu
NUM_GROUPS
7
STUDY_TYPE
Untargeted metabolomics analysis in lung cancer cells
Comparative analysis of metabolomic profiles in cerebrospinal fluid before and after endurance exercise
STUDY_SUMMARY
This study compared the metabolomic differences in the cerebrospinal fluid of young adult volunteers between before (Pre), and 60 minutes after the 90-min run (Post).
INSTITUTE
UCSD School of Medicine
LAST_NAME
Naviaux
FIRST_NAME
Robert
ADDRESS
214 Dickinson St., Room C-107, San Diego, CA, 92103, USA
The study population includes 844 healthy volunteers of which 183 women and 661 men, with a median age of 43 Ā± 12 yrs and 40 Ā± 11 yrs, respectively. The participants in this study were selected from the Tus-cany section of the Italian Association of Blood Donors (AVIS) in the Transfusion Service of the Pistoia Hospital. Plasma samples were obtained according to the Italian guidelines for blood donations. How age and sex influence the human metabolome and lipidome were investigated.
Multi-omics analysis of glucose-mediated signaling by a moonlighting GĪ² protein Asc1/RACK1
STUDY_TYPE
Untargeted UPLC-MS Metabolomics Analysis
STUDY_SUMMARY
While much is known about glucose metabolism in yeast, less is known about the receptors and signaling pathways that indicate glucose availability. We obtained metabolic profiles for wildtype and 16 mutants affecting the yeast glucose sensing pathway, comparing 0.05% glucose vs 10 min after glucose addition to 2%. First, we determined that the G protein-coupled receptor (Gpr1/Gpa2) directs early events in glucose utilization while the transceptors (Snf3/Rgt2) regulate subsequent processes and downstream products of glucose metabolism. Whereas the large G protein transmits the signal from its cognate receptor, Ras2 (but not Ras1) integrates responses from both receptor pathways. Second, we determined the relative contributions of the G protein Ī± (Gpa2) and Ī² (Asc1) subunits to glucose-initiated processes. We determined that Gpa2 is primarily involved in regulating carbohydrate metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Collectively, our analysis reveals the molecular basis for glucose detection and the earliest events of glucose-dependent signal transduction in yeast.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Nutrition
LABORATORY
Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel Hill
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
500 Laureate Way, Nutrition Research Institute, UNC Chapel Hill
GC-XLE method development: dSPE and MgSO4 as clean-up for sample preparation
STUDY_TYPE
Untargeted MS anlaysis
STUDY_SUMMARY
Compared to using dispersive SPE (dSPE) based on the QuEChERS procedure, we found similar reproducibility using high purity MgSO4 to analyze standard reference material (SRM) of human serum and human plasma samples and slightly higher recovery of targeted chemicals using MgSO4. To avoid contamination by environmental chemicals in solvents and reagents used for QuEChERS, we chose to use high purity MgSO4 to remove water-soluble interferences.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
Ī²-Adrenergic regulation of metabolism in macrophages (part-IV)
STUDY_SUMMARY
Macrophages have important roles in the immune system including clearing pathogens and wound healing. Metabolic phenotypes have been associated with functional phenotypes, where pro-inflammatory macrophages have an increased rate of glycolysis and anti-inflammatory macrophages primarily use oxidative phosphorylation. Ī²-adrenoceptor (Ī²AR) signalling in macrophages has been implicated in disease states such as cancer, atherosclerosis and rheumatoid arthritis. The impact of Ī²-adrenoceptor signalling on macrophage metabolism has not been defined. Here we expand on defining the phenotype of macrophages treated with isoprenaline and describe the impact that Ī²AR signalling has on the metabolome and proteome. We found that Ī²AR signalling alters proteins involved in cytoskeletal rearrangement and redox control of the cell. We showed that Ī²AR signalling in macrophages shifts glucose metabolism from glycolysis towards the tricarboxylic acid cycle and pentose phosphate pathways. We also show that Ī²AR signalling perturbs purine metabolism by accumulating adenylate pools. Taken together these results indicate that Ī²AR signalling shifts metabolism to support redox perturbations and upregulate proteins involved in cytoskeletal changes that may impact migration and phagocytosis processes.
INSTITUTE
Monash University
LAST_NAME
Peterson
FIRST_NAME
Amanda
ADDRESS
Drug delivery, disposition and dynamics, Pharmacy and Pharmaceutical Sciences, 381 Royal Parade, Parkville, Victoria, 3052, Australia
Acute metabolomic changes of plasma in response to endurance exercise
STUDY_SUMMARY
This study compared the metabolomic differences in plasma of young adult volunteers between before (Pre), immediately after the run (Time 0) and 60 minutes after the 90-min run (Time 60).
Effect of external low-dose rate radiation on mouse biofluid metabolomic signatures
STUDY_SUMMARY
An important component of ionizing radiation (IR) exposure after a radiological incident may include low-dose rate (LDR) exposures either externally or internally, such as from 137Cs deposition. LDR exposures can have different effects compared to acute high-dose rate exposures from a health and biodosimetry perspective. In this study, a novel irradiation system, VAriable Dose-rate External 137Cs irradiatoR (VADER), was used to expose male and female mice to a variable LDR over a 30-day time span to cumulative doses of 1 (only in males), 2, 2.8, 4.1, 8.8 (only in males), or 9.7 Gy to simulate fall-out type exposures. Urine and serum from mice exposed to an acute dose (~0.8 Gy/min) of x-rays were collected in parallel. Radiation markers were identified by global mass spectrometry based metabolomics and the machine learning algorithm Random Forests.
Effect of external low-dose rate radiation on mouse biofluid metabolomic signatures
STUDY_SUMMARY
An important component of ionizing radiation (IR) exposure after a radiological incident may include low-dose rate (LDR) exposures either externally or internally, such as from 137Cs deposition. LDR exposures can have different effects compared to acute high-dose rate exposures from a health and biodosimetry perspective. In this study, a novel irradiation system, VAriable Dose-rate External 137Cs irradiatoR (VADER), was used to expose male and female mice to a variable LDR over a 30-day time span to cumulative doses of 1 (only in males), 2, 2.8, 4.1, 8.8 (only in males), or 9.7 Gy to simulate fall-out type exposures. Urine and serum from mice exposed to an acute dose (~0.8 Gy/min) of x-rays were collected in parallel. Radiation markers were identified by global mass spectrometry based metabolomics and the machine learning algorithm Random Forests.
Effect of external low-dose rate radiation on mouse biofluid metabolomic signatures
STUDY_SUMMARY
An important component of ionizing radiation (IR) exposure after a radiological incident may include low-dose rate (LDR) exposures either externally or internally, such as from 137Cs deposition. LDR exposures can have different effects compared to acute high-dose rate exposures from a health and biodosimetry perspective. In this study, a novel irradiation system, VAriable Dose-rate External 137Cs irradiatoR (VADER), was used to expose male and female mice to a variable LDR over a 30-day time span to cumulative doses of 1 (only in males), 2, 2.8, 4.1, 8.8 (only in males), or 9.7 Gy to simulate fall-out type exposures. Urine and serum from mice exposed to an acute dose (~0.8 Gy/min) of x-rays were collected in parallel. Radiation markers were identified by global mass spectrometry based metabolomics and the machine learning algorithm Random Forests.
Effect of external low-dose rate radiation on mouse biofluid metabolomic signatures
STUDY_SUMMARY
An important component of ionizing radiation (IR) exposure after a radiological incident may include low-dose rate (LDR) exposures either externally or internally, such as from 137Cs deposition. LDR exposures can have different effects compared to acute high-dose rate exposures from a health and biodosimetry perspective. In this study, a novel irradiation system, VAriable Dose-rate External 137Cs irradiatoR (VADER), was used to expose male and female mice to a variable LDR over a 30-day time span to cumulative doses of 1 (only in males), 2, 2.8, 4.1, 8.8 (only in males), or 9.7 Gy to simulate fall-out type exposures. Urine and serum from mice exposed to an acute dose (~0.8 Gy/min) of x-rays were collected in parallel. Radiation markers were identified by global mass spectrometry based metabolomics and the machine learning algorithm Random Forests.
Metabolomics Analysis of Time-Series Gastrointestinal Lumen Samples
STUDY_SUMMARY
Samples were retrieved from a human small intestine samples over 8 hours. Metabolomics analysis followed resulting in many annotated metabolites. Intensity profiles gives insight into gastrointestinal functions.
Changes in mesenteric lymph lipid profile of mice upon high-fat diet with and without Celecoxib (part I)
STUDY_SUMMARY
Untargeted lipid profiling of mesenteric mice lymph from mice fed with CHOW, HFD and HFD supplemented with COx-2 inhibitor drug Celecoxib. It is proposed that Celecoxib can protect from deleterious morphological changes in lymphatic system caused by obesity/HFD.
INSTITUTE
Monash Institute of Pharmaceutical Sciences
DEPARTMENT
Drug Delivery, Disposition and Dynamics
LABORATORY
Trevaskis Lab, Creek Lab
LAST_NAME
Anderson
FIRST_NAME
Dovile
ADDRESS
6 Anderson
EMAIL
dovile.anderson@gmail.com
NUM_GROUPS
3
TOTAL_SUBJECTS
26
NUM_MALES
26
PUBLICATIONS
Manuscript NATMETAB-A20022406A Mesenteric lymphatic dysfunction promotes insulin resistance and represents a potential novel treatment target in obesity
Changes in mesenteric lymph lipid profile of mice upon high-fat diet with and without Celecoxib
STUDY_TYPE
Untargeted lipidomics analysis
STUDY_SUMMARY
Untargeted lipid profiling of mesenteric mice lymph from mice fed with CHOW, HFD and HFD supplemented with COx-2 inhibitor drug Celecoxib. It is proposed that Celecoxib can protect from deleterious morphological changes in lymphatic system caused by obesity due to HFD.
INSTITUTE
Monash Institute of Pharmaceutical Sciences
DEPARTMENT
Drug Delivery, Disposition and Dynamics
LABORATORY
Trevaskis Lab, Creek Lab
LAST_NAME
Anderson
FIRST_NAME
Dovile
ADDRESS
6 Anderson
EMAIL
dovile.anderson@gmail.com
PHONE
8671141
NUM_GROUPS
4
TOTAL_SUBJECTS
20
NUM_MALES
20
PUBLICATIONS
Manuscript NATMETAB-A20022406A Mesenteric lymphatic dysfunction promotes insulin resistance and represents a potential novel treatment target in obesity
We tested XLE quantification of chemicals in a non-fortified reference material: SRM-1957. Results show that XLE detected 29 out of 32 chemicals with certified or estimated reference values in the ng/kg range in SRM-1957; 16 out of 29 chemicals were quantified at >65% of the reference levels.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
We evaluated quantification using XLE by testing 68 different chemicals (PCB, PBDEs, chlorinated pesticides) in SRM-1958 using external calibration curves (0.05 to 2 ng/mL) and comparing measured values to the reference concentrations reported for SRM. We identified all 40 PCBs that are reported with a reference mass fraction (including certified values and non-certified estimates) in the range of 46.6 to 490 ng/kg in SRM-1958 certificate of analysis (issue date: 11 October 2018). Quantification without adjustment for recovery was reproducible with 29 PCB qualifications at >70% and 35 PCBs at >65% of the reference levels. Eleven out of 13 PBDE/PBBs and all 17 organochlorine pesticides were identifiable and reproducibly quantified in this experiment. Therefore, XLE provides sufficient recovery to support accurate absolute quantification of a broad range of environmental chemicals. Overall, XLE supported measurement of 68 out of the 70 chemicals that are in the ng/kg range in SRM-1958.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
Validation of XLE quantification using standard reference material. High recovery of [13C] labelled chemicals was obtained for important classes of environmental chemicals (PCB, PBDE, PAH, chlorinated pesticides) in NIST SRM-1957. Recoveries ranged from 110Ā±7% for [13C10]mirex to 91 to 105% for congeners of universally [13C] labeled PCBs, PBDEs and chlorinated pesticides, with only [13C12]p,pā-dichlorodiphenyldichloroethylene (p,pā-DDE) having low recovery of 65Ā±6%. Therefore, the simplified extraction procedure provides an efficient recovery of environment chemicals in an organic phase.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
We analyzed 80 archival samples from individuals (57 females, 23 males; aged 41 to 68 y) without known disease or occupational or environmental exposures of concern as a pilot to test the utility of XLE in large-scale human biomonitoring studies. Using a requirement for at least 3 co-eluting accurate mass m/z features (ļ± 5 ppm) within 30 s of database retention time, we identified 49 chemicals belonging to various environmental chemical classes. An unsupervised 2-way hierarchical cluster analysis (HCA) of log transformed intensity showed clustering according to chemical class. In particular, persistent chemicals were highly correlated with each other (all raw P < 0.001), including p,pā-DDE, PCBs 153, 180, 138, 118 and 74, PBDE-47, hexachlorobenzene (HCB) and trans-nonachlor. Results showed a general increase of chemical levels with increasing age quartiles (Q3 and Q4 : 53 to 68 versus Q1 and Q2: 41 to 52) using unsupervised clustering, a trend particularly evident for the cluster of p,pā-DDE, PCBs 153, 180, 138, 118 and 74, PBDE-47, HCB and trans-nonachlor. Examination of data according to body mass index (BMI) showed that individuals with BMI ā„ 40 had lower levels of environmental chemicals, which may be attributed to high lipophilicity and propensity to distribute in adipose tissue versus plasma. Quantification with reference standardization showed that use of two SRM samples with differing environmental chemical concentrations can overcome variable batch effects in quantification for large-scale studies. Examples of the most frequently detected chemicals shows that overall distributions were positively skewed by a small subset of individuals with high concentrations.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
We analyzed 80 archival samples from individuals (57 females, 23 males; aged 41 to 68 y) without known disease or occupational or environmental exposures of concern as a pilot to test the utility of XLE in large-scale human biomonitoring studies. Using a requirement for at least 3 co-eluting accurate mass m/z features (ļ± 5 ppm) within 30 s of database retention time, we identified 49 chemicals belonging to various environmental chemical classes. An unsupervised 2-way hierarchical cluster analysis (HCA) of log transformed intensity showed clustering according to chemical class. In particular, persistent chemicals were highly correlated with each other (all raw P < 0.001), including p,pā-DDE, PCBs 153, 180, 138, 118 and 74, PBDE-47, hexachlorobenzene (HCB) and trans-nonachlor. Results showed a general increase of chemical levels with increasing age quartiles (Q3 and Q4 : 53 to 68 versus Q1 and Q2: 41 to 52) using unsupervised clustering, a trend particularly evident for the cluster of p,pā-DDE, PCBs 153, 180, 138, 118 and 74, PBDE-47, HCB and trans-nonachlor. Examination of data according to body mass index (BMI) showed that individuals with BMI ā„ 40 had lower levels of environmental chemicals, which may be attributed to high lipophilicity and propensity to distribute in adipose tissue versus plasma. Quantification with reference standardization showed that use of two SRM samples with differing environmental chemical concentrations can overcome variable batch effects in quantification for large-scale studies. Examples of the most frequently detected chemicals shows that overall distributions were positively skewed by a small subset of individuals with high concentrations.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
We tested the general utility of XLE in a variety of human biological samples by analyzing human lung and thyroid tissues and stool samples. We quantified 32 environmental chemicals in 11 human lungs, with HCB, PCB-28 and PCB-18 being most frequently detected (10 out of 11). The commonly detected chemicals in human plasma were detected less frequently in the lung. For the 11 lungs, p,pā-DDE was detected in eight, PCB-153 in five, PBDE-47 and PCB-138 in four and PCB-180 in three. Although the plasma samples were from non-diseased individuals and the lungs were both diseased and non-diseased individuals, HCA results suggest that environmental chemical profiles in human lung may be very different from plasma.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
In the small number of thyroids that was analyzed with XLE, 14 environmental chemicals were quantified. The most prevalent was p,pā-DDE, detected in 4 out of 5 thyroid samples, with median concentration (2.20 ng/g). The amounts of individual chemicals were highly variable among the individuals, and the small number of samples precludes any generalization. Nevertheless, HCA of correlation matrix showed high correlation of chemicals measured in the thyroid samples was similar to that in the lung and plasma.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
Human stool samples, as a noninvasive matrix, have unique value in exposome research but have not been extensively studied for environmental chemical exposures. For lipophilic and unabsorbed dietary environmental chemicals, stool is a primary route of elimination and can therefore provide useful information on body burden and clearance of chemicals. In a pilot analysis of six human stool samples, we detected 52 and quantified 21 environmental chemicals, with HCB found in all samples. Quantification of HCB showed a median concentration of 0.057 ng/g. HCA of correlation matrix showed co-exposures of chemicals are likely as shown in the plasma, lung and thyroid. The high correlations of these persistent chemicals are not surprising as they likely derive from similar environmental exposure events.
INSTITUTE
Emory University
DEPARTMENT
Medicine/Pulmonary
LABORATORY
Dean Jones
LAST_NAME
Hu
FIRST_NAME
Xin
ADDRESS
Emory University Whitehead building (Rm 225), 615 Michael Street
Metabolic responses of two pioneer wood decay fungi to diurnally cycling temperature
STUDY_SUMMARY
1. Decomposition of lignin-rich wood by fungi drives nutrient recycling in woodland ecosystems. Fluctuating abiotic conditions are known to promote the functioning of ecological communities and ecosystems. In the context of wood decay, fluctuating temperature increases decomposition rates. Metabolomics, in tandem with other āomics tools, can highlight the metabolic processes affected by experimental treatments, even in the absence of genome sequences and annotations. Globally, natural wood decay communities are dominated by the phylum Basidiomycota. We examined the metabolic responses of Mucidula mucida, a dominant constituent of pioneer communities in beech branches in British woodlands, and Exidia glandulosa, a stress-selected constituent of the same communities, in response to constant and diurnally cycling temperature. 2. We applied untargeted metabolomics and proteomics to beech wood blocks, colonised by M. mucida or E. glandulosa and exposed to either diurnally cycling (mean 15 Ā± 10Ā°C) or constant (15Ā°C) temperature, in a fully factorial design. 3. Metabolites and proteins linked to lignin breakdown, the citric acid cycle, pentose phosphate pathway, carbohydrate metabolism, fatty acid metabolism and protein biosynthesis and turnover were under-enriched in fluctuating, compared to stable temperatures, in the generalist M. mucida. Conversely E. glandulosa showed little differential response to the experimental treatments. 4. Synthesis. By demonstrating temperature dependant metabolic signatures related to nutrient acquisition in a generalist wood decay fungus, we provide new insights into how abiotic conditions can affect community-mediated decomposition and carbon turnover in forests. We show that mechanisms underpinning important biogeochemical processes can be highlighted using untargeted metabolomics and proteomics in the absence of well-annotated genomes.
INSTITUTE
Swansea University
DEPARTMENT
Biosciences
LABORATORY
Fungal Molecular Ecology
LAST_NAME
Eastwood
FIRST_NAME
Daniel
ADDRESS
Wallace 102, Biosciences, College of Science, Swansea University, Swansea, SA2 8PP
Effect of external low-dose rate radiation on mouse biofluid metabolomic signatures (part V)
STUDY_SUMMARY
An important component of ionizing radiation (IR) exposure after a radiological incident may include low-dose rate (LDR) exposures either externally or internally, such as from 137Cs deposition. LDR exposures can have different effects compared to acute high-dose rate exposures from a health and biodosimetry perspective. In this study, a novel irradiation system, VAriable Dose-rate External 137Cs irradiatoR (VADER), was used to expose male and female mice to a variable LDR over a 30-day time span to cumulative doses of 1 (only in males), 2, 2.8, 4.1, 8.8 (only in males), or 9.7 Gy to simulate fall-out type exposures. Urine and serum from mice exposed to an acute dose (~0.8 Gy/min) of x-rays were collected in parallel. Radiation markers were identified by global mass spectrometry based metabolomics and the machine learning algorithm Random Forests.
Untargeted metabolomics of Daphnia magna exposed to a lithium cobalt oxide nanomaterial
STUDY_TYPE
Untargeted MS
STUDY_SUMMARY
The goal of this project was to determine lithium cobalt oxide (LCO)ās effects on pathways in the model organism Daphnia magna through untargeted metabolomics.
Impact of high intensity and moderate exercise on genomic and metabolic remodeling with age in male mice
STUDY_SUMMARY
How skeletal muscle adapts to different types of exercise intensity with age is not known. Young and old C57BL/6 male mice were assigned to either a sedentary or two types of exercise regimes consisting of daily high-intensity intermittent (HIIT) or moderate intensity continuous (MICT) training for 4 weeks, compatible with the older groupās exercise capacity. Body composition, fasting blood glucose levels, and muscle strength were improved in exercised old mice compared to sedentary controls, while the exercise benefits were absent in younger animals. Skeletal muscle exhibited structural and functional adaptations in response to exercise, as revealed by electron microscopy, OXPHOS assays, respirometry, and PGC-1ļ” and LC3-II protein levels. Transcriptomics analysis of gastrocnemius muscle combined with liver and serum metabolomics unveiled an age-dependent metabolic remodeling provoked by exercise through mitochondrial biogenesis, energy metabolism, and cellular plasticity. These results are supportive of a tailored exercise prescription approach with the goal of improving health and ameliorating age-associated loss of muscle mass, strength and function in the elderly.
INSTITUTE
National Institutes of Health
DEPARTMENT
Experimental Gerontology Section and Translational Gerontology Branch, NIA
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
The Metabolic Benefits of Short Cycles of Very Low Caloric Intake are Dependent on Diet Composition in Middle-Aged Mice
STUDY_SUMMARY
Diet composition, calories, and fasting times contribute to maintenance of health. Here, middle-aged mice were maintained for 5 months on 4:10 feeding cycles, consisting of 4 days of very low-calorie intake (VLCI) achieved with either standard laboratory chow (SD) or a fasting mimicking diet (FMD), followed by 10 days of ad libitum access to SD. Fat and lean mass loss was accompanied with improved performance, glucoregulation, and metabolic flexibility independent of diet composition. However, only the 4:10/SD cycles elicited a long-lasting metabolomic reprograming in serum and liver that was preserved six days after refeeding. Challenged with an obesogenic diet, cycles of VLCI achieved with either high-fat diet (HFD) or FMD during the low-calorie period did not prevent diet-induced obesity nor did they elicited a long-lasting metabolic memory, despite achieving modest metabolic flexibility. Our results highlight the importance of diet composition in mediating the metabolic benefits of short cycles of VLCI.
INSTITUTE
National Institutes of Health
DEPARTMENT
Experimental Gerontology Section and Translational Gerontology Branch, NIA
LAST_NAME
de Cabo
FIRST_NAME
Rafael
ADDRESS
251 Bayview Blvd. Suite 100/Room 5C214. Baltimore, MD 21224
Metabolomics-driven identification of biochemical mechanisms underlying the neuroprotective effects of pleiotrophin in a mouse model of Parkinsonās disease
STUDY_SUMMARY
Pleiotrophin (PTN) is a cytokine involved in nerve tissue repair processes, neuroinflammation and neuronal survival. PTN expression levels are upregulated in the nigrostriatal pathway of Parkinsonās Disease (PD) patients. We aimed to characterize the dopaminergic injury and glial activation in the nigrostriatal pathway of mice with transgenic Ptn overexpression in the brain (Ptn-Tg) after intrastriatal injection of the Parkinsonian toxin 6-hydroxydopamine (6-OHDA). The injection of 6-OHDA induced a significant decrease of the number of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra and of the striatal TH contents in Wild type (Wt) mice. In contrast, these effects of 6-OHDA were blocked in Ptn-Tg mice. 6-OHDA injection did not cause robust changes in microglia but induced an exacerbated astrocytic response in Wt mice compared with Ptn-Tg mice. In metabolomics studies, we detected interesting metabolites that significantly discriminate the more injured 6-OHDA-injected Wt striatum and the more protected 6-OHDA-injected Ptn-Tg striatum. Particularly, we detected groups of metabolites, mostly corresponding to phospholipids, whose trends were opposite in both groups. In summary, the data confirm the neuroprotective effect of brain PTN overexpression in this mouse model of PD. New lipid-related PD drug candidates emerge from this study and the data presented here support the increasingly recognized ālipid cascadeā in PD.
Evidence that class I glutamine amidotransferase, GAT1_2.1, acts as a glutaminase in roots of Arabidopsis thaliana
STUDY_TYPE
Targeted Metabolite Quantification
STUDY_SUMMARY
In this study, we use a targeted metabolite quantification approach to demonstrate the difference in quantities of pathway intermediates between wild type Arabidopsis roots and gat1_2.1 mutants using glutamine as organic nitrogen treatment and KNO3 and Glu treatments as negative and positive controls, respectively.
Evidence that class I glutamine amidotransferase, GAT1_2.1, acts as a glutaminase in roots of Arabidopsis thaliana
STUDY_TYPE
Targeted Metabolite Quantification
STUDY_SUMMARY
In this study, we used Arabidopsis root extracts, spiked with amide nitrogen labeled (15N1) Glutamine and a purified recombinant protein, both full length and glutaminase domain only versions, to determine the amido group acceptor, if any, in the glutamine amidotransferase reaction.
Associations between the gut microbiome and metabolome in early life
STUDY_SUMMARY
The infant intestinal microbiome plays an important role in metabolism and immune development with impacts on lifelong health. The linkage between the taxonomic composition of the microbiome and its metabolic phenotype is undefined and complicated by redundancies in the taxon-function relationship within microbial communities. To inform a more mechanistic understanding of the relationship between the microbiome and health, we performed an integrative statistical and machine learning-based analysis of microbe taxonomic structure and metabolic function (using untargeted (binned) NMR and relative concentration data) in order to characterize the taxa-function relationship in early life.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Nutrition
LABORATORY
Metabolomics and Exposome Laboratory at UNC CH Nutrition Research Institute
LAST_NAME
Sumner
FIRST_NAME
Susan
ADDRESS
500 Laureate Way, Kannapolis, NC, 28081, USA
EMAIL
susan_sumner@unc.edu
TOTAL_SUBJECTS
440
STUDY_TYPE
Untargeted and semi-targeted metabolomics analysis
Searching for prognostic biomarkers of ParkinsonĀ“s Disease development in the Spanish EPIC cohort through a multiplatform metabolomics approach
STUDY_SUMMARY
The lack of knowledge about the onset and progression of Parkinsonās disease (PD) hampers its early diagnosis and treatment. We used a multiplatform untargeted metabolomics-based approach to uncover the biochemical remodeling induced by PD in a really early and pre-symptomatic stage and unveiling early potential diagnostic biomarkers. Baseline pre-clinical plasma samples (Pre-PD n=39; control group n=39) were obtained from the European Prospective Study on Nutrition and Cancer (EPIC) cohort, which healthy volunteers were followed for around 15 years to ascertain incident PD. Our finding revealed alterations in fatty acids metabolism, mitochondrial dysfunction, oxidative stress, and gut-brain axis dysregulation. This study is of inestimable value since this is the first study conducted with samples collected many years before the disease development.
INSTITUTE
Universidad CEU San Pablo
LAST_NAME
Gonzalez-Riano
FIRST_NAME
Carolina
ADDRESS
km 0, Universidad CEU-San Pablo UrbanizaciĆ³n MonteprĆncipe. M-501, 28925 AlcorcĆ³n
Metabolic Markers of Methotrexate Response in Juvenile Idiopathic Arthritis
STUDY_TYPE
Clinical
STUDY_SUMMARY
Plasma from children with juvenile idiopathic arthritis collected pre-treatment and following 3 months of treatment with methotrexate were submitted for metabolomic profiling to the NIH West Coast Metabolomics Center.
Phosphoinositides (PIPs) species, differing in phosphorylation at hydroxyls of the inositol head group, play roles in various cellular events. Here we developed a supercritical fluid chromatography-mass spectrometry (SFC-MS) method that allows us to quantify molecular species of all seven PIP regioisomers in culture cells and tissues. Using this methodology, we quantified PIPs species in 13 mouse tissues.
Quantification of PIPs species in LPIAT1 KO mouse embryonic fibroblasts (MEFs).
STUDY_SUMMARY
Using a supercritical fluid chromatography-mass spectrometry (SFC-MS)-based methodology, we quantified phosphoinositides (PIPs) species in LPIAT1 KO mouse embryonic fibroblasts (MEFs).
Quantification of PIPs species in mouse embryonic fibroblasts (MEFs) during autophagosome formation.
STUDY_SUMMARY
Using a supercritical fluid chromatography-mass spectrometry (SFC-MS)-based methodology, we quantified phosphoinositides (PIPs) species in mouse embryonic fibroblasts (MEFs) from WT or FIP200 KO mice during autophagosome formation.
The RNA-binding protein RBP42 regulates cellular energy metabolism in mammalian-infective Trypanosoma brucei
STUDY_SUMMARY
Metabolic changes following two days of RBP42 knockdown was investigated using a targeted metabolomics approach, designed to capture intermediary metabolites in central carbon metabolism including glycolytic intermediates, TCA compounds, amino acids, nucleotides and derivatives, were obtained using hydrophilic interaction liquid chromatography (HILIC) separation method coupled with mass spectrometry run in negative ionization mode
INSTITUTE
Rutgers University
LAST_NAME
Das
FIRST_NAME
Anish
ADDRESS
Department of Microbiology, Biochemistry and Molecular Genetics, Rutgers-New Jersey Medical School, Newark, NJ 07103
Alterations in the fecal microbiome and metabolome of horses with antimicrobial-associated diarrhea compared to antibiotic-treated and non-treated healthy case controls
STUDY_SUMMARY
Horses receiving antimicrobials may develop diarrhea due to changes in the gastrointestinal microbiome and metabolome. This matched, case-controlled study compared the fecal microbiome and metabolome in hospitalized horses on antibiotics that developed diarrhea (AAD), hospitalized horses on antibiotics that did not develop diarrhea (ABX) and a healthy, non-hospitalized control population (CON). Naturally-voided fecal samples were collected from AAD horses (n=17) the day that diarrhea developed and matched to ABX (n=15) and CON (n=31) horses for diet, antimicrobial agent and duration of antimicrobial therapy (< 5 days or > 5 days). Illumina sequencing of 16S rRNA genes on fecal DNA was performed. Alpha and beta diversity metrics were generated using QIIME 2.0. A Kruskal-Wallis with Dunnās post-test and ANOSIM testing was used for statistical analysis. Microbiome composition in AAD was significantly different from CON (ANOSIM, R= 0.568, p=0.001) and ABX (ANOSIM, R=0.121, p=0.0012). Fecal samples were lyophilized and extracted using a solvent-based method. Untargeted metabolomics using gas chromatography-mass spectrometry platforms was performed. Metabolomic data was analyzed using Metaboanalyst 4.0 and Graphpad Prism v 7. Principal component analysis plots (PCA) were used to visualize the distribution of metabolites between groups. Heat maps were used to identify the relative concentrations amongst the most abundant 25 metabolites. A one-way ANOVA was used to compare differences in metabolites amongst the three groups of horses. Only named metabolites were included in the analysis. The microbiome of AAD and ABX horses had significantly decreased richness and evenness than CON horses (p<0.05). Actinobacteria (q=0.0192) and Bacteroidetes (q=0.0005) were different between AAD and CON. Verrucomicrobia was markedly decreased in AAD compared to ABX and CON horses (q=0.0005). Horses with AAD have a dysbiosis compared to CON horses, and show minor differences in bacterial community composition to ABX horses. Metabolite profiles of horses with AAD clustered separately from those with AAD or CON. Ten metabolites were found to be significantly different between groups (P<0.05) and are listed according to their metabolic pathway: amino acid metabolism (R-equol, L-tyrosine, kynurenic acid, xanthurenic acid, 5-hydroxyindole-3-acetic acid ) lipid metabolism (docosahexaenoic acid ethyl ester), biosynthesis of secondary metabolites (daidzein, isoquinoline) and two metabolites with unidentified pathways (1,3-divinyl-2-imidazolidinone, N-acetyltyramine).
INSTITUTE
Texas A&M University
LAST_NAME
Arnold
FIRST_NAME
Carolyn
ADDRESS
4475 TAMU College of Veterinary Medicine and Biomedical Sciences College Station, Texas 77843-4475
Metabolomic and lipidomic profiles of CKD in obese patients in serum and urine (part 1 of 3)
STUDY_TYPE
Human nephropathy in CKD obese patients
STUDY_SUMMARY
Obesity is a global pandemic with an increase prevalence over the years. This condition elevates the risk of developing cardiovascular diseases, hypertension and renal pathologies, like chronic kidney disease (CKD). In the present study, the metabolomic and the lipidomic profiles of CKD obese patients were analyzed comparing with obese subjects without CKD. Subsequently, CKD obese patients underwent bariatric surgery and the effect of surgery in the CKD progression of these subjects was evaluated. Serum and urine were measured by LC-MS and GC-HRAM equipment.
INSTITUTE
University Rey Juan Carlos
DEPARTMENT
Basics Science of Health
LAST_NAME
Lanzon
FIRST_NAME
Borja
ADDRESS
Avenida de Atenas S/N
EMAIL
borja.lanzon@urjc.es
PHONE
663692554
NUM_GROUPS
3
TOTAL_SUBJECTS
36
STUDY_COMMENTS
Serum LC-MS data: part 1 of 3. Samples were analyzed per duplicated.
Metabolomic and lipidomic profiles of CKD in obese patients in serum and urine (part 2 of 3)
STUDY_SUMMARY
Obesity is a global pandemic with an increase prevalence over the years. This condition elevates the risk of developing cardiovascular diseases, hypertension and renal pathologies, like chronic kidney disease (CKD). In the present study, the metabolomic and the lipidomic profiles of CKD obese patients were analyzed comparing with obese subjects without CKD. Subsequently, CKD obese patients underwent bariatric surgery and the effect of surgery in the CKD progression of these subjects was evaluated. Serum and urine were measured by LC-MS and GC-HRAM equipment.
INSTITUTE
University Rey Juan Carlos
DEPARTMENT
Basics Science of Health
LAST_NAME
Lanzon
FIRST_NAME
Borja
ADDRESS
Avenida de Atenas S/N, AlcorcĆ³n, Madrid, 28922, Spain
Metabolomic and lipidomic profiles of CKD in obese patients in serum and urine (part 3 of 3)
STUDY_SUMMARY
Obesity is a global pandemic with an increase prevalence over the years. This condition elevates the risk of developing cardiovascular diseases, hypertension and renal pathologies, like chronic kidney disease (CKD). In the present study, the metabolomic and the lipidomic profiles of CKD obese patients were analyzed comparing with obese subjects without CKD. Subsequently, CKD obese patients underwent bariatric surgery and the effect of surgery in the CKD progression of these subjects was evaluated. Serum and urine were measured by LC-MS and GC-HRAM equipment.
INSTITUTE
University Rey Juan Carlos
DEPARTMENT
Basics Science of Health
LAST_NAME
Lanzon
FIRST_NAME
Borja
ADDRESS
Avenida de Atenas S/N, AlcorcĆ³n, Madrid, 28922, Spain
The pregnancy metabolome from a multi-ethnic pregnancy cohort
STUDY_SUMMARY
The PRogramming of Intergenerational Stress Mechanisms (PRISM) study is an urban, ethnically diverse pregnancy cohort that was designed to study a range of chemical and non-chemical stressors in relation to maternal health, pregnancy outcomes, and child development. Pregnant women were enrolled from Boston and New York City hospitals and affiliated prenatal clinics beginning in 2011. Eligibility criteria included English or Spanish-speaking, over 18 years of age at enrollment, and singleton pregnancy. Exclusion criteria included HIV+ status or self-reported drinking ā„7 alcoholic drinks per week before pregnancy or any alcohol after pregnancy recognition
Nested caseācontrol study of environmental exposure within the ongoing Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) pregnancy cohort.
STUDY_SUMMARY
Preterm birth is the leading cause of infant mortality worldwide, and also greatly increases the risk of developing serious health complications in childhood and throughout life. Conditions that contribute to preterm birth remain largely unclear, though an influence by environmental exposures is suspected but poorly understood. In our preliminary work, we found that exposure to common chemicals, such as phthalates and bisphenol A (BPA), may contribute to this serious but understudied public health problem. We also observed strong associations between phthalates/BPA and general markers of oxidative stress, and in turn between these general markers of oxidative stress and risk of preterm birth. In this nested case-control study within an ongoing cohort of pregnant women in Puerto Rico (PROTECT), we will explore untargeted lipidomics, as well as levels of eicosanoids specific to arachidonic acid oxidation pathways, in relation to both environmental exposures and preterm birth.
Analysis and annotation of oxidized PCs generated in vitro.
STUDY_SUMMARY
Oxidized PC16:0/PUFA (18:2, 20:4 and 22:6) generated in vitro were analyzed by LC/HRMS/MS analysis.All mass spectrometry raw data obtained in this study were deposited.
Exhaustive analysis of endogenous oxPCs in APAP-treated mice.
STUDY_SUMMARY
A single dose of APAP at 300 mg/kg body weight was intraperitoneally administered into eight-week-old C57BL/6J male mice. After extraction of hepatic lipids, oxPCs were analyzed by LC/HRMS/MS. All mass spectrometry raw data obtained in this study were deposited.
INSTITUTE
Kyushu university
LAST_NAME
Matsuoka
FIRST_NAME
Yuta
ADDRESS
3-1-1 Maidashi Higashi-ku, Fukuoka, Not USCanada, 812-8582, Japan
Nested caseācontrol study of environmental exposure within the ongoing Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) pregnancy cohort (part II)
STUDY_SUMMARY
Preterm birth is the leading cause of infant mortality worldwide, and also greatly increases the risk of developing serious health complications in childhood and throughout life. Conditions that contribute to preterm birth remain largely unclear, though an influence by environmental exposures is suspected but poorly understood. In our preliminary work, we found that exposure to common chemicals, such as phthalates and bisphenol A (BPA), may contribute to this serious but understudied public health problem. We also observed strong associations between phthalates/BPA and general markers of oxidative stress, and in turn between these general markers of oxidative stress and risk of preterm birth. In this nested case-control study within an ongoing cohort of pregnant women in Puerto Rico (PROTECT), we will explore untargeted lipidomics, as well as levels of eicosanoids specific to arachidonic acid oxidation pathways, in relation to both environmental exposures and preterm birth.
A metabolomics comparison of plant-based meat and grass-fed meat indicates large nutritional differences despite comparable nutrition facts labels
STUDY_SUMMARY
A new generation of plant-based meat alternativesāformulated to mimic the taste and nutritional composition of red meatāhave attracted considerable consumer interest, research attention, and media coverage. This has raised questions of whether plant-based meat alternatives represent proper nutritional replacements to animal meat. Given that food sources have considerable complexity and contain a wide variety of nutrients (e.g., phenols, anti-oxidants, peptides, amino acids, fatty acids, and other carboxylic acids), the majority of which do not appear on nutrition labels, it is important to explore expanded nutrient profiles when determining whether beef and plant-based meat alternatives are nutritionally interchangeable. Important nutritional differences may exist between beef and novel plant-based alternatives, given their materials origin; however, this has not been thoroughly assessed. Given the scientific and commercial interest in plant-based meat alternatives, the goal of our study was to use untargeted metabolomics to provide an in-depth comparison of the metabolite profiles of grass-fed ground beef and a popular plant-based meat alternative.
Use of Integrated Metabolomics, Transcriptomics, and Signal Protein Profile to Characterize the Effector Function and Associated Metabotype of Polarized Macrophage Phenotypes
STUDY_SUMMARY
Macrophages (MĪ¦s) display remarkable plasticity and the ability to activate diverse responses to a host of intracellular and external stimuli. Despite extensive characterization of M1 MĪ¦s and a broad set of M2 MĪ¦s, comprehensive characterization of functional phenotype and associated metabotype driving this diverse MĪ¦ activation remains. Herein, we utilized an ex vivo model to produce six MĪ¦ functional phenotypes. Isolated CD14+ PBMCs were differentiated into resting M0 MĪ¦s, and then polarized into M1 (IFN-Ī³/LPS), M2a (IL-4/IL-13), M2b (IC/LPS), M2c (IL-10), and M2d (IL-6/LIF) MĪ¦s. The MĪ¦s were profiled using a bioanalyte matrix of four cell surface markers, ~50 secreted proteins, ~800 expressed myeloid genes, and ~450 identified metabolites relative to M0 MĪ¦s. Signal protein and expressed gene profiles grouped the MĪ¦s into inflammatory (M1 and M2b) and wound resolution (M2a, M2c, and M2d) phenotypes; however, each had a unique metabolic profile. While both M1 and M2b MĪ¦s shared metabotype profiles consistent with an inflammatory signature; key differences were observed in the TCA cycle, FAO, and OXPHOS. Additionally, M2a, M2c, and M2d MĪ¦s all profiled as tissue repair MĪ¦s; however, metabotype differences were observed in multiple pathways including hexosamine, polyamine, and fatty acid metabolism. These metabolic and other key functional distinctions suggest phagocytic and proliferative functions for M2a MĪ¦s, and angiogenesis and ECM assembly capabilities for M2b, M2c, and M2d MĪ¦s. By integrating metabolomics into a systems analysis of MĪ¦ phenotypes, we provide the most comprehensive map of MĪ¦ diversity to date, along with the global metabolic shifts that correlate to MĪ¦ functional plasticity in these phenotypes.
INSTITUTE
Idaho Veterans Research and Education Foundation
DEPARTMENT
Research
LABORATORY
Ammons
LAST_NAME
Ammons
FIRST_NAME
Mary Cloud
ADDRESS
Mail Stop 151, Bldg 117, 500 W. Fort St. Boise, Idaho 83702
Reversing Epigenetic Gene Silencing to Overcome Immune Evasion in CNS Malignancies
STUDY_SUMMARY
Glioblastoma is an aggressive brain malignancy with a dismal prognosis. With emerging evidence that disproves the immune privileged environment in the brain, there is much interest in examining various immunotherapy strategies to treat these incurable cancers. Unfortunately, to date, clinical studies investigating immunotherapy regimens have not provided much evidence of efficacy, leading to questions about the suitability of immunotherapy strategies for these tumors. Inadequate inherent populations of lymphocytes in tumor (TILs) and limited trafficking of systemic circulating T cells into the central nervous system (CNS) likely contribute to the poor response to immunotherapy treatment for primary CNS cancers. This paucity of TILs is in concert with the finding of epigenetic silencing of genes that promote immune cell movement (chemotaxis) to the tumor. In this study we evaluated the ability of GSK126, a blood-brain barrier permeable small molecule inhibitor of EZH2, to reverse the epigenetic silencing of chemokines like CXCL9 and CXCL10. When combined with anti-PD-1 treatment, these IFN driven chemokines promote T cell infiltration, resulting in decreased tumor growth and enhanced survival in immunocompetent murine sub-cutaneous and intracranial tumor syngeneic models of GBM. Examination of the tumor micro-environment revealed that the decrease in tumor growth in the mice treated with the drug combination was accompanied by increased tumor CD8 T cell infiltration along with higher IFN expression. Additionally, a significant increase in CXCR3+ T cells in the draining lymph nodes was also found. Taken together, our data suggests that in glioblastoma, epigenetic modulation using GSK126 could improve current immunotherapy strategies by reversing the epigenetic changes that enable immune cell evasion leading to enhanced immune cell trafficking to the tumor.
Metabolite profiling of Malaysian Gracilaria edulis reveals Eplerenone as novel antibacterial compound for drug repurposing against MDR Bacteria
STUDY_SUMMARY
The current study re-defines a method to reveal bioactive compounds from the crude extracts of Malaysian red seaweed Gracilaria edulis, having promising antibacterial activities against selected bacterial species. Three species of Gram-positive and - negative characters were remarkably inhibited by the sequential and direct extracts of ethyl acetate and acetone. These were further separated through chromatographic methods to reveal a plethora of chemical constituents to be considered for a downstream virtual screening against selected crucial proteins of the six bacteria.
INSTITUTE
Sunway University
DEPARTMENT
Biological Sciences, Sunway University, Selangor, Malaysia
LABORATORY
Disease Complexity
LAST_NAME
Lahiri
FIRST_NAME
Chandrajit
ADDRESS
Sunway University, No. 5, Jalan Universiti, Bandar Sunway, Petaling Jaya 47500, Selangor, Malaysia
Metabolomics of lung microdissections reveals region- and sex-specific metabolic effects of acute naphthalene exposure in mice (part I)
STUDY_SUMMARY
Naphthalene is a ubiquitous environmental contaminant produced by combustion of fossil fuels and is a primary constituent of both mainstream and side stream tobacco smoke. Naphthalene elicits region-specific toxicity in airway club cells through cytochrome P450 (P450)-mediated bioactivation, resulting in depletion of glutathione and subsequent cytotoxicity. While effects of naphthalene in mice have been extensively studied, few experiments have characterized global metabolomic changes in the lung. In individual lung regions, we found metabolomic changes in microdissected mouse lung conducting airways and parenchyma obtained from animals sacrificed 2, 6, and 24 hours following naphthalene treatment. Data on 577 unique identified metabolites were acquired by accurate mass spectrometry-based assays focusing on lipidomics and non-targeted metabolomics of hydrophilic compounds. Statistical analyses revealed distinct metabolite profiles between the two major lung regions. In addition, the number and magnitude of statistically significant exposure-induced changes in metabolite abundance were different between lung airways and parenchyma for unsaturated lysophosphatidylcholines (LPCs), dipeptides, purines, pyrimidines, and amino acids. Importantly, temporal changes were found to be highly distinct for male and female mice, with males exhibiting predominant treatment-specific changes only at two hours post-exposure. In females, metabolomic changes persisted until six hours post-naphthalene treatment, which may explain the previously characterized higher susceptibility of female mice to naphthalene toxicity. In both males and females, treatment-specific changes corresponding to lung remodeling, oxidative stress response, and DNA damage were observed, which may provide insights into potential mechanisms contributing to the previously reported effects of naphthalene exposure in the lung.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome Center
LABORATORY
Fiehn Lab
LAST_NAME
Stevens
FIRST_NAME
Nathanial C.
ADDRESS
451 Health Sciences Drive University of California Davis Davis, CA 95616
Metabolomics of lung microdissections reveals region- and sex-specific metabolic effects of acute naphthalene exposure in mice (part II)
STUDY_SUMMARY
Naphthalene is a ubiquitous environmental contaminant produced by combustion of fossil fuels and is a primary constituent of both mainstream and side stream tobacco smoke. Naphthalene elicits region-specific toxicity in airway club cells through cytochrome P450 (P450)-mediated bioactivation, resulting in depletion of glutathione and subsequent cytotoxicity. While effects of naphthalene in mice have been extensively studied, few experiments have characterized global metabolomic changes in the lung. In individual lung regions, we found metabolomic changes in microdissected mouse lung conducting airways and parenchyma obtained from animals sacrificed 2, 6, and 24 hours following naphthalene treatment. Data on 577 unique identified metabolites were acquired by accurate mass spectrometry-based assays focusing on lipidomics and non-targeted metabolomics of hydrophilic compounds. Statistical analyses revealed distinct metabolite profiles between the two major lung regions. In addition, the number and magnitude of statistically significant exposure-induced changes in metabolite abundance were different between lung airways and parenchyma for unsaturated lysophosphatidylcholines (LPCs), dipeptides, purines, pyrimidines, and amino acids. Importantly, temporal changes were found to be highly distinct for male and female mice, with males exhibiting predominant treatment-specific changes only at two hours post-exposure. In females, metabolomic changes persisted until six hours post-naphthalene treatment, which may explain the previously characterized higher susceptibility of female mice to naphthalene toxicity. In both males and females, treatment-specific changes corresponding to lung remodeling, oxidative stress response, and DNA damage were observed, which may provide insights into potential mechanisms contributing to the previously reported effects of naphthalene exposure in the lung.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome Center
LABORATORY
Fiehn Lab
LAST_NAME
Stevens
FIRST_NAME
Nathanial C.
ADDRESS
451 Health Sciences Drive University of California Davis Davis, CA 95616
Identification of unique metabolite networks between Latino and Caucasian patients with nonalcoholic fatty liver disease (NAFLD) (part II)
STUDY_SUMMARY
Nonalcoholic fatty liver disease (NAFLD) is a spectrum of liver pathology ranging from simple steatosis to nonalcoholic steatohepatitis (NASH); the latter is characterized by inflammation and fibrosis. Risk factors for NALFD include obesity, diabetes, hyperlipidemia, and hypertensionāall of which are features of metabolic syndrome. NAFLD is a very heterogeneous disease, as it presents in different patterns in males and females and in patients from different ethnicities, with unclear predictors for development and severity of disease. Previous studies have shown that NAFLD is 1.4 times more frequent in Hispanics than in Caucasians. One of the major challenges in NAFLD is the lack of accurate, noninvasive biomarkers for the detection of the most aggressive presentation, NASH. The gold standard for the diagnosis is liver biopsy, which is an invasive procedure associated with possible complications. Noninvasive diagnosis of NASH is a major unmet medical need and there are no ethnicity-specific biomarkers that can diagnose this condition and predict its progression. Therefore, the main gap in knowledge that this proposal and line of research will address is the characterizing the different plasma and liver metabolomics profile of patients with fatty liver from two ethnicities (Latinos vs. Caucasians) and of both sexes. The overall hypothesis of the present study is that the higher incidence of nonalcoholic fatty liver (NAFL) in Latino patients is reflected in a different plasma and liver metabolomics profile compared to Caucasian patients with further sex-related differences. Characterization of metabolite networks can aid in identifying the mechanistic underpinnings of sex and ethnic driven differences in NAFL which could help diagnose and establish a prognosis of this condition, especially in the critical transition from NAFL to the more aggressive nonalcoholic steatohepatitis (NASH).To address this hypothesis, plasma metabolomics profile of samples from male and female Latino and Caucasian bariatric surgery patients with NAFL and from healthy subjects will be compared. Metabolomics findings will be related with liver pathology and liver transcriptome profiles from intraoperatively obtained liver biopsies using correlation, network, and pathway analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Department of Internal Medicine, Division of Gastroenterology and Hepatology
LABORATORY
Medici Lab
LAST_NAME
Medici
FIRST_NAME
Valentina
ADDRESS
4150 V Street - PSSB Suite 3500 - 95817 Sacramento CA
Identification of unique metabolite networks between Latino and Caucasian patients with nonalcoholic fatty liver disease (NAFLD) (part III)
STUDY_SUMMARY
Nonalcoholic fatty liver disease (NAFLD) is a spectrum of liver pathology ranging from simple steatosis to nonalcoholic steatohepatitis (NASH); the latter is characterized by inflammation and fibrosis. Risk factors for NALFD include obesity, diabetes, hyperlipidemia, and hypertensionāall of which are features of metabolic syndrome. NAFLD is a very heterogeneous disease, as it presents in different patterns in males and females and in patients from different ethnicities, with unclear predictors for development and severity of disease. Previous studies have shown that NAFLD is 1.4 times more frequent in Hispanics than in Caucasians. One of the major challenges in NAFLD is the lack of accurate, noninvasive biomarkers for the detection of the most aggressive presentation, NASH. The gold standard for the diagnosis is liver biopsy, which is an invasive procedure associated with possible complications. Noninvasive diagnosis of NASH is a major unmet medical need and there are no ethnicity-specific biomarkers that can diagnose this condition and predict its progression. Therefore, the main gap in knowledge that this proposal and line of research will address is the characterizing the different plasma and liver metabolomics profile of patients with fatty liver from two ethnicities (Latinos vs. Caucasians) and of both sexes. The overall hypothesis of the present study is that the higher incidence of nonalcoholic fatty liver (NAFL) in Latino patients is reflected in a different plasma and liver metabolomics profile compared to Caucasian patients with further sex-related differences. Characterization of metabolite networks can aid in identifying the mechanistic underpinnings of sex and ethnic driven differences in NAFL which could help diagnose and establish a prognosis of this condition, especially in the critical transition from NAFL to the more aggressive nonalcoholic steatohepatitis (NASH).To address this hypothesis, plasma metabolomics profile of samples from male and female Latino and Caucasian bariatric surgery patients with NAFL and from healthy subjects will be compared. Metabolomics findings will be related with liver pathology and liver transcriptome profiles from intraoperatively obtained liver biopsies using correlation, network, and pathway analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Department of Internal Medicine, Division of Gastroenterology and Hepatology
LABORATORY
Medici Lab
LAST_NAME
Medici
FIRST_NAME
Valentina
ADDRESS
4150 V Street - PSSB Suite 3500 - 95817 Sacramento CA
Identification of unique metabolite networks between Latino and Caucasian patients with nonalcoholic fatty liver disease (NAFLD) (part IV)
STUDY_SUMMARY
Nonalcoholic fatty liver disease (NAFLD) is a spectrum of liver pathology ranging from simple steatosis to nonalcoholic steatohepatitis (NASH); the latter is characterized by inflammation and fibrosis. Risk factors for NALFD include obesity, diabetes, hyperlipidemia, and hypertensionāall of which are features of metabolic syndrome. NAFLD is a very heterogeneous disease, as it presents in different patterns in males and females and in patients from different ethnicities, with unclear predictors for development and severity of disease. Previous studies have shown that NAFLD is 1.4 times more frequent in Hispanics than in Caucasians. One of the major challenges in NAFLD is the lack of accurate, noninvasive biomarkers for the detection of the most aggressive presentation, NASH. The gold standard for the diagnosis is liver biopsy, which is an invasive procedure associated with possible complications. Noninvasive diagnosis of NASH is a major unmet medical need and there are no ethnicity-specific biomarkers that can diagnose this condition and predict its progression. Therefore, the main gap in knowledge that this proposal and line of research will address is the characterizing the different plasma and liver metabolomics profile of patients with fatty liver from two ethnicities (Latinos vs. Caucasians) and of both sexes. The overall hypothesis of the present study is that the higher incidence of nonalcoholic fatty liver (NAFL) in Latino patients is reflected in a different plasma and liver metabolomics profile compared to Caucasian patients with further sex-related differences. Characterization of metabolite networks can aid in identifying the mechanistic underpinnings of sex and ethnic driven differences in NAFL which could help diagnose and establish a prognosis of this condition, especially in the critical transition from NAFL to the more aggressive nonalcoholic steatohepatitis (NASH).To address this hypothesis, plasma metabolomics profile of samples from male and female Latino and Caucasian bariatric surgery patients with NAFL and from healthy subjects will be compared. Metabolomics findings will be related with liver pathology and liver transcriptome profiles from intraoperatively obtained liver biopsies using correlation, network, and pathway analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Department of Internal Medicine, Division of Gastroenterology and Hepatology
LABORATORY
Medici Lab
LAST_NAME
Medici
FIRST_NAME
Valentina
ADDRESS
4150 V Street - PSSB Suite 3500 - 95817 Sacramento CA
Identification of unique metabolite networks between Latino and Caucasian patients with nonalcoholic fatty liver disease (NAFLD) (part V)
STUDY_SUMMARY
Nonalcoholic fatty liver disease (NAFLD) is a spectrum of liver pathology ranging from simple steatosis to nonalcoholic steatohepatitis (NASH); the latter is characterized by inflammation and fibrosis. Risk factors for NALFD include obesity, diabetes, hyperlipidemia, and hypertensionāall of which are features of metabolic syndrome. NAFLD is a very heterogeneous disease, as it presents in different patterns in males and females and in patients from different ethnicities, with unclear predictors for development and severity of disease. Previous studies have shown that NAFLD is 1.4 times more frequent in Hispanics than in Caucasians. One of the major challenges in NAFLD is the lack of accurate, noninvasive biomarkers for the detection of the most aggressive presentation, NASH. The gold standard for the diagnosis is liver biopsy, which is an invasive procedure associated with possible complications. Noninvasive diagnosis of NASH is a major unmet medical need and there are no ethnicity-specific biomarkers that can diagnose this condition and predict its progression. Therefore, the main gap in knowledge that this proposal and line of research will address is the characterizing the different plasma and liver metabolomics profile of patients with fatty liver from two ethnicities (Latinos vs. Caucasians) and of both sexes. The overall hypothesis of the present study is that the higher incidence of nonalcoholic fatty liver (NAFL) in Latino patients is reflected in a different plasma and liver metabolomics profile compared to Caucasian patients with further sex-related differences. Characterization of metabolite networks can aid in identifying the mechanistic underpinnings of sex and ethnic driven differences in NAFL which could help diagnose and establish a prognosis of this condition, especially in the critical transition from NAFL to the more aggressive nonalcoholic steatohepatitis (NASH).To address this hypothesis, plasma metabolomics profile of samples from male and female Latino and Caucasian bariatric surgery patients with NAFL and from healthy subjects will be compared. Metabolomics findings will be related with liver pathology and liver transcriptome profiles from intraoperatively obtained liver biopsies using correlation, network, and pathway analysis.
INSTITUTE
University of California, Davis
DEPARTMENT
Department of Internal Medicine, Division of Gastroenterology and Hepatology
LABORATORY
Medici Lab
LAST_NAME
Medici
FIRST_NAME
Valentina
ADDRESS
4150 V Street - PSSB Suite 3500 - 95817 Sacramento CA
Proteasome Inhibitor Related Cardiotoxicity (part I)
STUDY_SUMMARY
BALB/c mice were injected with CFZ (16 mg/kg, iv), BTZ (1 mg/kg, iv), CFZ/atRA (CFZ: 16 mg/kg iv, atRA: 1 mg/kg iv) or Captisol (Vehicle, iv) (6 mice per cohort). ECGs were recorded under anesthesia at the end of 1h treatment and the mice were sacrificed by cervical dislocation. Hearts were extracted and were frozen and stored at -80Ā°C until analysis
Standardized gnotobiotic mouse model for NMR based phenotyping
STUDY_SUMMARY
Germ-free mice were inoculated with 15 representative bacterial strains selected from specific pathogen-free mice. All three microbiota mouse models were generated in two different facility sites and collected plasma sample phenotyped by NMR. 58 mice from Facility 1 (32 males and 26 females) and 59 mice from Facility 2 (29 males and 30 females) were used for analysis.
Untargeted mass spectrometry reveals impact of high fat diet on peripheral amino acid regulation in a mouse model of Alzheimerās Disease
STUDY_SUMMARY
APPSwe/PS1ĪE9 (APP/PSEN1) transgenic mice (to represent familial or early-onset AD) and wild-type litter mater controls were fed either a high-fat diet (HFD, 60% kcal from lard), low-fat diet (LFD, 10% kcal from lard) from 2 months of age, or reversal diet (REV, high-fat followed by low-fat from 9.5 months).
Longitudinal Metabolomics of Human Plasma Reveals Robust Prognostic Markers of COVID-19 Disease Severity (part I)
STUDY_SUMMARY
There is an urgent need to identify which COVID-19 patients will develop life-threatening illness so that medical resources can be optimally allocated and rapid treatment can be administered early in the disease course, when clinical management is most effective. To aid in the prognostic classification of disease severity, we perform untargeted metabolomics on plasma from 339 patients, with samples collected at six longitudinal time points. Using the temporal metabolic profiles and machine learning, we build a predictive model of disease severity. We discover that a panel of metabolites measured at the time of study entry successfully determine disease severity. Through analysis of longitudinal samples, we confirm that the majority of these markers are directly related to disease progression and that their levels are restored to baseline upon disease recovery. Finally, we validate that these metabolites are also altered in a hamster model of COVID-19. Our results indicate that metabolic changes associated with COVID-19 severity can be effectively used to stratify patients and inform resource allocation during the pandemic.
INSTITUTE
Washington University in St. Louis
DEPARTMENT
Chemistry
LABORATORY
Patti
LAST_NAME
Patti
FIRST_NAME
Gary
ADDRESS
McMillen Chemistry Laboratory Washington University 1 Brookings Dr @ Throop Drive Rm 102 St. Louis, MO 63130-4899
Unbiased LC-MS-based metabolomics analysis for both whole cell and mitochondria metabolites to gain an insight into the role of Tug1/PGC1 axis on metabolite profiles in podocytes
STUDY_SUMMARY
unbiased LC-MS-based metabolomics analysis for both whole cell and mitochondria metabolites to gain an insight into the role of Tug1/PGC1 axis on metabolite profiles in podocytes
Quantitative analysis of bile acids in fecal samples from centenarians, elderly and young subjects.
STUDY_SUMMARY
Fecal samples from centenarians (>100 yo), elderly (85-89 yo) and young (21-55) subjects were analysed using LC-MS/MS. 48 bile acids were measured by targeted metabolomics.
Screening of unique bile acid metabolizing bacteria
STUDY_SUMMARY
We incubated individual bacterial strains at pH 7 or pH 9 with either CDCA, LCA, or 3-oxo-Ī4-LCA as starting substrates. Culture supernatants were collected after 48 hours and 14 bile acids were measured by targeted metabolomics.
Longitudinal Metabolomics of Human Plasma Reveals Robust Prognostic Markers of COVID-19 Disease Severity
STUDY_SUMMARY
There is an urgent need to identify which COVID-19 patients will develop life-threatening illness so that medical resources can be optimally allocated and rapid treatment can be administered early in the disease course, when clinical management is most effective. To aid in the prognostic classification of disease severity, we perform untargeted metabolomics on plasma from 339 patients, with samples collected at six longitudinal time points. Using the temporal metabolic profiles and machine learning, we build a predictive model of disease severity. We discover that a panel of metabolites measured at the time of study entry successfully determine disease severity. Through analysis of longitudinal samples, we confirm that the majority of these markers are directly related to disease progression and that their levels are restored to baseline upon disease recovery. Finally, we validate that these metabolites are also altered in a hamster model of COVID-19. Our results indicate that metabolic changes associated with COVID-19 severity can be effectively used to stratify patients and inform resource allocation during the pandemic.
INSTITUTE
Washington University, St. Louis
DEPARTMENT
Chemistry
LABORATORY
Patti
LAST_NAME
Patti
FIRST_NAME
Gary
ADDRESS
McMillen Chemistry Laboratory, Washington University 1 Brookings Dr @ Throop Drive, Rm 102, St. Louis, MO 63130-4899
Metabolic profiling of Rafflesia-infected Tetrastigma and applications for propagation
STUDY_SUMMARY
Endemic to the forests of Southeast Asia, Rafflesia (Rafflesiaceae) is a genus of holoparasitic plants producing the largest flowers in the world, yet completely dependent on its host, the tropical grape vine, Tetrastigma. Rafflesia species are threatened with extinction, making them an iconic symbol of plant conservation. Thus far, propagation has proved challenging, greatly decreasing efficacy of conservation efforts. This study compared the metabolites in the shoots of Rafflesia-infected and non-infected Tetrastigma loheri to examine how Rafflesia infection affects host metabolomics and elucidate the Rafflesia infection process. Results from LC-MS-based untargeted metabolomics analysis showed benzylisoquinoline alkaloids were significantly elevated in non-infected shoots and are here reported for the first time in the genus Tetrastigma, and in the grape family, Vitaceae. These metabolites have been implicated in plant defense mechanisms and may prevent a Rafflesia infection. In Rafflesia-infected shoots, oxygenated fatty acids, or oxylipins, and a flavonoid, previously shown involved in plant immune response, were abundant. This study provides a preliminary assessment of metabolites that differ between Rafflesia-infected and non-infected Tetrastigma hosts and may have applications in Rafflesia propagation to meet conservation goals.
The metabolomic resetting effect of FG4592 in AKI to CKD transition-day 21
STUDY_SUMMARY
C57BL/6 mice were anesthetized using isoflurane. UIR was induced by clamping the right renal pedicle for 45 minutes and then releasing it to allow reperfusion, leaving the left kidney intact. Sham treated mice served as controls. Each mouse was located supine on a thermostatic pad (37 Ā°C) to maintain its body temperature throughout the whole process. After 3 days of recovery, the mice received a daily intraperitoneal (i.p.) injection of FG4592 (10 mg/kg) or vehicle for 18 consecutive days. After treatment, the mice were sacrificed. Non-target metabolomics analysis was carried out using the kidney tissues of mice sacrificed at day 21 after UIR.
INSTITUTE
Childrenās Hospital of Nanjing Medical University
The metabolomic resetting effect of FG4592 in AKI to CKD transition-day 21
STUDY_SUMMARY
C57BL/6 mice were anesthetized using isoflurane. UIR was induced by clamping the right renal pedicle for 45 minutes and then releasing it to allow reperfusion, leaving the left kidney intact. Sham treated mice served as controls. Each mouse was located supine on a thermostatic pad (37 Ā°C) to maintain its body temperature throughout the whole process. After 3 days of recovery, the mice received a daily intraperitoneal (i.p.) injection of FG4592 (10 mg/kg) or vehicle for 7 consecutive days. After treatment, the mice were sacrificed.Non-target metabolomics analysis was carried out using the kidney tissues of mice sacrificed at day 10 after UIR.
Proteasome Inhibitor Related Cardiotoxicity (part II)
STUDY_SUMMARY
BALB/c mice were injected with CFZ (16 mg/kg, iv), BTZ (1 mg/kg, iv), CFZ/atRA (CFZ: 16 mg/kg iv, atRA: 1 mg/kg iv) or Captisol (Vehicle, iv) (6 mice per cohort). ECGs were recorded under anesthesia at the end of 1h treatment and the mice were sacrificed by cervical dislocation. Plasma was obtained after centrifugation by heart puncture. Samples were immediately frozen and stored at -80Ā°C until analysis
dTor affects the fat body lipidome via Nep1r1, Ctdnep1 and Lipin
STUDY_SUMMARY
Quantitative MS analysis was performed on ten 4 day-old Drosophila larval fat bodies homogenized in 50Āµl D-PBS (Dulbeccoās Phosphate Buffered Saline without Mg2+ and Ca2+) by Lipotype using previously described methods (Grillet et al, 2016).
INSTITUTE
VIB KULeuven
DEPARTMENT
Dept. of Neurosciences, KU Leuven, 3000 Leuven, Belgium
LAST_NAME
Jacquemyn
FIRST_NAME
Julie
ADDRESS
ON 4, 6e verd Campus Gasthuisberg, Herestraat 49, bus 602, Leuven, NA, 3000, Belgium
Spontaneous hydrolysis and spurious metabolic properties of Ī±-ketoglutarate esters
STUDY_SUMMARY
8988T cells treated with methyl acetate or 1 mM of alpha-ketoglutarate disodium salt or 1 mM of dimethyl-alpha-ketoglutarate for 3 hours prior to rapid quenching of metabolism and extraction of metabolites in 80% methanol (-80Ā°C) containing internal QC standards.
INSTITUTE
University of British Columbia
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, 2099, Vancouver, British Columbia, Canada V6H 0B3
Parallelized multidimensional analytic framework, PAMAF, applied to mammalian cells uncovers novel regulatory principles in EMT
STUDY_SUMMARY
Painting a holistic picture of disease etiology will require longitudinal systems-scale reconstruction of the multitiered architecture of eukaryotic signaling. As opposed to āone omic at a timeā, which provides an incomplete view on disease mechanisms, here we developed an experimental and analytics framework, PAMAF, to simultaneously acquire and analyze twelve omic modalities from the same set of samples, i.e., protein abundance from whole-cells, nucleus, exosomes, secretome and membrane; peptidome; N-glycosylation, phosphorylation; metabolites; mRNA, miRNA; and, in parallel, single-cell transcriptomes. We applied PAMAF in a well-studied in vitro model of TGFĪ²-induced EMT to generate the EMT-ExMap dataset, cataloguing >61,000 expression profiles (>10,000 differential) over 12 days. PAMAF revealed that EMT is more complex than currently understood and identified numerous stage-specific mechanisms and vulnerabilities not captured in literature. Broad application of PAMAF will provide unprecedented insights into multifaceted biological processes relevant to human health and disease.
Cross-feeding between intestinal pathobionts promotes their overgrowth during undernutrition
STUDY_SUMMARY
Child undernutrition is a global health issue associated with a high burden of infectious disease. Undernourished children display an overabundance of intestinal pathogens and pathobionts, and these bacteria induce enteric dysfunction in undernourished mice; however, the cause of their overgrowth remains poorly defined. Here, we show that disease-inducing human isolates of Enterobacteriaceae and Bacteroidales spp. are capable of multi-species symbiotic cross-feeding, resulting in synergistic growth of a mixed community in vitro. Growth synergy occurs uniquely under malnourished conditions limited in protein and iron: in this context, Bacteroidales spp. liberate diet- and mucin-derived sugars and Enterobacteriaceae spp. enhance the bioavailability of iron. Analysis of human microbiota datasets reveals that Bacteroidaceae and Enterobacteriaceae are strongly correlated in undernourished children, but not in adequately nourished children, consistent with a diet-dependent growth synergy in the human gut. Together these data suggest that dietary cross-feeding fuels the overgrowth of pathobionts in undernutrition.
Targeting host glycolysis as a strategy for antimalarial development
STUDY_SUMMARY
Glycolysis controls cellular energy, redox balance, and biosynthesis. Antiglycolytic therapies are under investigation for treatment of obesity, cancer, aging, autoimmunity, and microbial diseases. Interrupting glycolysis is highly valued as a therapeutic strategy, because glycolytic disruption is generally tolerated in mammals. Unfortunately, anemia is a known dose-limiting side effect of these inhibitors and presents a major caveat to development of antiglycolytic therapies. We developed specific inhibitors of enolase ā a critical enzyme in glycolysis ā and validated their metabolic and cellular effects on human erythrocytes. Enolase inhibition increases erythrocyte susceptibility to oxidative damage and induces rapid and premature erythrocyte senescence, rather than direct hemolysis. We apply our model of red cell toxicity to address questions regarding erythrocyte glycolytic disruption in the context of Plasmodium falciparum malaria pathogenesis. Our study provides a framework for understanding red blood cell homeostasis under normal and disease states and clarifies the importance of erythrocyte reductive capacity in malaria parasite growth.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Prolonged cellular hypoxia leads to energetic failure and death. However, sublethal hypoxia can trigger an adaptive response called hypoxic preconditioning. While prolyl-hydroxylase (PHD) enzymes and hypoxia inducible factors (HIFs) have been identified as key elements of oxygen sensing machinery, the mechanisms by which hypoxic preconditioning protects against insults remain unclear. Here, we perform serum metabolomic profiling to assess alterations induced by hypoxic preconditioning. We discover that hypoxic preconditioning increases serum kynurenine levels and enhance kynurenine biotransformation leading to preservation of NAD+ in the post-ischemic kidney. Furthermore, we show that Indoleamine 2,3-dioxygenase 1 (Ido1) deficiency abolishes the systemic increase of kynurenine and the subsequent renoprotection generated by hypoxic preconditioning. Importantly, exogenous administration of kynurenine restores the hypoxic preconditioning in the context of Ido1 deficiency. Collectively, our findings demonstrate a critical role of Ido1/kynurenine axis in mediating hypoxic preconditioning
INSTITUTE
Northwestern University
DEPARTMENT
Medicine/Nephrology
LABORATORY
Kapitsinou
LAST_NAME
Kapitsinou
FIRST_NAME
Pinelopi
ADDRESS
303 East Superior Street
EMAIL
pinelopi.kapitsinou@northwestern.edu
NUM_GROUPS
2
TOTAL_SUBJECTS
16
NUM_MALES
16
STUDY_COMMENTS
N/A
PUBLICATIONS
Accepted in Cell Reports
STUDY_TYPE
Comparative metabolomic analysis of serum metabolites detected by untargeted LC/MS and GC/MS platform
Prolonged cellular hypoxia leads to energetic failure and death. However, sublethal hypoxia can trigger an adaptive response called hypoxic preconditioning. While prolyl-hydroxylase (PHD) enzymes and hypoxia inducible factors (HIFs) have been identified as key elements of oxygen sensing machinery, the mechanisms by which hypoxic preconditioning protects against insults remain unclear. Here, we perform serum metabolomic profiling to assess alterations induced by hypoxic preconditioning. We discover that hypoxic preconditioning increases serum kynurenine levels and enhance kynurenine biotransformation leading to preservation of NAD+ in the post-ischemic kidney. Furthermore, we show that Indoleamine 2,3-dioxygenase 1 (Ido1) deficiency abolishes the systemic increase of kynurenine and the subsequent renoprotection generated by hypoxic preconditioning. Importantly, exogenous administration of kynurenine restores the hypoxic preconditioning in the context of Ido1 deficiency. Collectively, our findings demonstrate a critical role of Ido1/kynurenine axis in mediating hypoxic preconditioning
INSTITUTE
Northwestern University
DEPARTMENT
Medicine/Nephrology
LABORATORY
Kapitsinou
LAST_NAME
Kapitsinou
FIRST_NAME
Pinelopi
ADDRESS
303 East Superior Street
EMAIL
pinelopi.kapitsinou@northwestern.edu
NUM_GROUPS
2
TOTAL_SUBJECTS
14
NUM_MALES
14
STUDY_COMMENTS
N/A
PUBLICATIONS
Accepted in Cell Reports
STUDY_TYPE
Comparative metabolomic analysis of serum metabolites detected by untargeted LC/MS and GC/MS platform
Sodium dichloroacetate stimulates cardiac mitochondrial metabolism and improves cardiac conduction in the ovine fetus during labor (part I)
STUDY_SUMMARY
Previous studies in our laboratory have suggested that the increase in stillbirth in pregnancies complicated by chronic maternal stress or hypercortisolemia is associated with cardiac dysfunction in late stages of labor and delivery. Transcriptomics analysis of the overly represented differentially expressed genes in the fetal heart of hypercortisolemic ewes indicated involvement of mitochondrial function. Sodium dichloroacetate (DCA) has been used to improve mitochondrial function in several disease states. We hypothesized that administration of DCA to laboring ewes would improve both cardiac mitochondrial activity and cardiac function in their fetuses. Four groups of ewes and their fetuses were studied: control, cortisol-infused (1 g/kg/d from 115 to term; CORT), DCA-treated (over 24h) or DCA+CORT-treated; oxytocin was delivered starting 48h before the DCA treatment. DCA significantly decreased cardiac lactate, alanine and glucose/glucose-6-phosphate and increased acylcarnitine/isobutyryl-carnitine. DCA increased mitochondrial activity, increasing oxidative phosphorylation (PCI, PCI+II)) per tissue weight or per unit of citrate synthase. DCA also decreased the duration of the QRS, attenuating the prolongation of the QRS observed in CORT fetuses. The effect to reduce QRS duration with DCA treatment correlated with increased glycerophosphocholine and serine and decreased phophocholine after DCA treatment. There were negative correlations of acylcarnitine/isobutyryl-carnitine to both HR and MAP. These results suggest that improvements in mitochondrial respiration with DCA produced changes in the cardiac lipid metabolism that favor improved conduction in the heart. DCA may therefore be an effective treatment of fetal cardiac metabolic disturbances in labor that can contribute to impairments of fetal cardiac conduction.
INSTITUTE
University of Georgia
DEPARTMENT
Biochemistry and Molecular Biology and Complex Carbohydrate Research Center, Department of Pharmacodynamics (University of Florida), Department of Physiology and Functional Genomics (University of Florida)
LABORATORY
Edison Lab, Keller-Wood Lab, and Wood Lab
LAST_NAME
Zhang
FIRST_NAME
Sicong
ADDRESS
315 Riverbend Road, Complex Carbohydrate Research Center
Sodium dichloroacetate stimulates cardiac mitochondrial metabolism and improves cardiac conduction in the ovine fetus during labor (part II)
STUDY_TYPE
untargeted NMR analysis-noesy
STUDY_SUMMARY
Previous studies in our laboratory have suggested that the increase in stillbirth in pregnancies complicated by chronic maternal stress or hypercortisolemia is associated with cardiac dysfunction in late stages of labor and delivery. Transcriptomics analysis of the overly represented differentially expressed genes in the fetal heart of hypercortisolemic ewes indicated involvement of mitochondrial function. Sodium dichloroacetate (DCA) has been used to improve mitochondrial function in several disease states. We hypothesized that administration of DCA to laboring ewes would improve both cardiac mitochondrial activity and cardiac function in their fetuses. Four groups of ewes and their fetuses were studied: control, cortisol-infused (1 g/kg/d from 115 to term; CORT), DCA-treated (over 24h) or DCA+CORT-treated; oxytocin was delivered starting 48h before the DCA treatment. DCA significantly decreased cardiac lactate, alanine and glucose/glucose-6-phosphate and increased acylcarnitine/isobutyryl-carnitine. DCA increased mitochondrial activity, increasing oxidative phosphorylation (PCI, PCI+II)) per tissue weight or per unit of citrate synthase. DCA also decreased the duration of the QRS, attenuating the prolongation of the QRS observed in CORT fetuses. The effect to reduce QRS duration with DCA treatment correlated with increased glycerophosphocholine and serine and decreased phophocholine after DCA treatment. There were negative correlations of acylcarnitine/isobutyryl-carnitine to both HR and MAP. These results suggest that improvements in mitochondrial respiration with DCA produced changes in the cardiac lipid metabolism that favor improved conduction in the heart. DCA may therefore be an effective treatment of fetal cardiac metabolic disturbances in labor that can contribute to impairments of fetal cardiac conduction.
INSTITUTE
University of Georgia
DEPARTMENT
Biochemistry and Molecular Biology and Complex Carbohydrate Research Center, Department of Pharmacodynamics (University of Florida), Department of Physiology and Functional Genomics (University of Florida)
LABORATORY
Edison Lab, Keller-Wood Lab, and Wood Lab
LAST_NAME
Zhang
FIRST_NAME
Sicong
ADDRESS
315 Riverbend Road, Complex Carbohydrate Research Center
Effects of GP130 Antagonism on Right Ventricular Metabolism in Monocrotaline Rats
STUDY_SUMMARY
We used global metabolomics profiling to evaluate right ventricular metabolism in control, monocrotaline rats treated with vehicle, and monocrotaline rats treated with SC144 (GP130 antagonists).
Untargeted LC-MS metabolomics analysis of cecal content of mice treated with TCDD vs. vehicle control (part I)
STUDY_SUMMARY
Six-week-old female wildtype (WT) C57BL/6 mice were administered a single 100Āµl intraperitoneal injection containing sterile corn oil (VEH group) or an intraperitoneal injection of 10Āµg/kg TCDD suspended within sterile corn oil (TCDD group). At the 72h time point following TCDD or VEH exposure, the mice were humanely euthanized by an overdose of inhaled isoflurane. During necropsy, cecal content and blood serum samples were collected for untargeted metabolomics profiling.
Untargeted LC-MS metabolomics analysis of cecal content of mice treated with TCDD vs. vehicle control (part II)
STUDY_SUMMARY
Six-week-old female wildtype (WT) C57BL/6 mice were administered a single 100Āµl intraperitoneal injection containing sterile corn oil (VEH group) or an intraperitoneal injection of 10Āµg/kg TCDD suspended within sterile corn oil (TCDD group). At the 72h time point following TCDD or VEH exposure, the mice were humanely euthanized by an overdose of inhaled isoflurane. During necropsy, cecal content and blood serum samples were collected for untargeted metabolomics profiling.
Metabolomics analysis of multiple samples on Agilent 6546-Part 1
STUDY_SUMMARY
Metabolomics analysis of multiple samples from human, trying to annotate the metabolites in them. Agilent 6546 LC-QTOF was used for metabolomics detection.
INSTITUTE
Dalian Institute Of Chemical Physics
LABORATORY
Laboratory of High Resolution Separation/Analysis and Metabonomics
LAST_NAME
Zheng
FIRST_NAME
Fujian
ADDRESS
No. 457 Zhongshan Road, Shahekou District, Dalian, Liaoning Province, China
Metabolomics analysis of multiple samples on Agilent 6546-Part 2
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Metabolomics analysis of multiple samples from mouse, trying to annotate the metabolites in them. Agilent 6546 LC-QTOF was used for metabolomics detection.
INSTITUTE
Dalian Institute Of Chemical Physics
LABORATORY
Laboratory of High Resolution Separation/Analysis and Metabonomics
LAST_NAME
Zheng
FIRST_NAME
Fujian
ADDRESS
No. 457 Zhongshan Road, Shahekou District, Dalian, Liaoning Province, China
Targeted Sphingolipid analysis of HeLa knockout for expression of GRASP55
STUDY_SUMMARY
Golgi apparatus, the main glycosylation station of the cell, consists of a stack of discontinuous cisternae. Glycosylation enzymes are usually concentrated in one or two specific cisternae along the cis-trans axis of the organelle. How such compartmentalized localization of enzymes is achieved and how it contributes to glycosylation are not clear. Here we show that the Golgi matrix protein GRASP55 directs the compartmentalized localization of key enzymes involved in glycosphingolipid (GSL) biosynthesis. GRASP55 acts by binding to these enzymes and preventing their entry to COPI derived retrograde transport vesicles thus concentrating them in the trans-Golgi. In genome edited cells lacking GRASP55 the enzymes relocate to cis-Golgi. Here we evaluated the impact of deleting GRASP55 on sphingolipid composition of HeLa cells by targeted lipid analysis.
INSTITUTE
IBBC, CNR
LAST_NAME
parashuraman
FIRST_NAME
seetharaman
ADDRESS
Via Pietro Castellino 111, Napoli, NA, 80131, Italy
The study comprehends two consecutive LC-QqQ/MS analyses of Babesia divergens merozoite extracts isolated from B. divergens infected red blood cell cultures performed under identical chromatographic conditions and targeting distinct transitions corresponding to metabolites from specific pathways including the glycolysis, the TCA cycle, the pentose phosphate pathway, purine and pyrimidine biosynthesis and amino acid metabolism.
Proteomics reveals an increase in the abundance of glycolytic and ethanolic fermentation enzymes in developing sugarcane culms during sucrose accumulation
STUDY_SUMMARY
Sugarcane is an economically important crop contributing to the worldās sugar and ethanol production with 80% and 40%, respectively. Metabolites from I5-4M and I9-4M were extracted from six biological samples of four-month-old plants. Following the removal of leaves, the internodes were identified and cut, and the bark removed, and the remaining tissue was immediately frozen in liquid nitrogen.
1H NMR Metabolomics to elucidate signaling pathway
STUDY_SUMMARY
Metabolomics investigation of the phytoplankton Alexandrium minutum with and without copepod cues in order to explore cell signaling involved in toxin induction.
INSTITUTE
Georgia Institute of Technology
DEPARTMENT
School of Biological Sciences, School of Chemistry and Biochemistry, Center for Microbial Dynamics and Infection, Parker H. Petit Institute for Bioengineering and Bioscience
LABORATORY
Kubanek Lab
LAST_NAME
Brown
FIRST_NAME
Emily
ADDRESS
950 Atlantic Dr Atlanta, Georgia USA 30332
EMAIL
julia.kubanek@biosci.gatech.edu
PHONE
404-894-8424
NUM_GROUPS
2
TOTAL_SUBJECTS
40
STUDY_COMMENTS
Part 1 of 3. This part includes NMR analysis of polar metabolites using oPLSDA. Parts 2 and 3 inlcude NMR analysis of nonpolar metabolites and the corresponding mass spectrometry metabolomics for both polar and non-polar metabolites.
1H NMR Metabolomics to elucidate signaling pathway
STUDY_SUMMARY
Metabolomics investigation of the phytoplankton Alexandrium minutum with and without copepod cues in order to explore cell signaling involved in toxin induction.
INSTITUTE
Georgia Institute of Technology
DEPARTMENT
School of Biological Sciences, School of Chemistry and Biochemistry, Center for Microbial Dynamics and Infection, Parker H. Petit Institute for Bioengineering and Bioscience
LABORATORY
Kubanek Lab
LAST_NAME
Brown
FIRST_NAME
Emily
ADDRESS
950 Atlantic Dr Atlanta, Georgia USA 30332
EMAIL
julia.kubanek@biosci.gatech.edu
PHONE
404-894-8424
NUM_GROUPS
2
TOTAL_SUBJECTS
40
STUDY_COMMENTS
Part 2 of 3. This part includes NMR analysis of non-polar metabolites using oPLSDA. Parts 1 and 3 inlcude NMR analysis of polar metabolites and the corresponding mass spectrometry metabolomics for both polar and non-polar metabolites.
LC-MS for Predator Cues Target Signaling Pathways in Toxic Algal Metabolome Protocol
STUDY_SUMMARY
Metabolomics investigation of the phytoplankton Alexandrium minutum with and without copepod cues in order to explore cell signaling involved in toxin induction.
MS Differentiating toxic and nontoxic cogeneric harmful algae using the non-polar metabolome.
STUDY_SUMMARY
Recognition and rejection of chemically defended prey is critical to maximizing fitness for predators. Paralytic shellfish toxins (PSTs) which strongly inhibit voltage-gated sodium channels in diverse animal taxa are produced by several species of the bloom-forming algal genus Alexandrium where they appear to function as chemical defenses against grazing copepods. Despite PSTs being produced and localized within phytoplankton cells, some copepods distinguish toxic from non-toxic prey, selectively ingesting less toxic cells, in ways that suggest cell surface recognition perhaps associated with non-polar metabolites. In this study LC/MS and NMR-based metabolomics revealed that the non-polar metabolomes of two toxic species (Alexandrium catenella and Alexandrium pacificum) vary considerably from their non-toxic congener Alexandrium tamarense despite all three being very closely related. Toxic and non-toxic Alexandrium spp. were distinguished from each other by metabolites belonging to seven lipid classes. Of these, 17 specific metabolites were significantly more abundant in both toxic A. catenella and A. pacificum compared to non-toxic A. tamarense suggesting that just a small portion of the observed metabolic variability is associated with toxicity. Future experiments aimed at deciphering chemoreception mechanisms of copepod perception of Alexandrium toxicity should consider these metabolites, and the broader lipid classes phosphatidylcholines and sterols, as potential candidate cues.
Untargeted metabolomics of hypertrophic cardiomyopathy (part I)
STUDY_SUMMARY
Hypertrophic cardiomyopathy (HCM) is a complex disease partly explained by the effects of individual gene variants on sarcomeric protein biomechanics. At the cellular level, HCM mutations most commonly enhance force production, leading to higher energy demands. Despite significant advances in elucidating sarcomeric structure-function relationships, there is still much to be learned about the mechanisms that link altered cardiac energetics to HCM phenotypes. In this work, we test the hypothesis that changes in cardiac energetics represent a common pathophysiologic pathway in HCM.
Untargeted lipidomics of hypertrophic cardiomyopathy (part II)
STUDY_SUMMARY
Hypertrophic cardiomyopathy (HCM) is a complex disease partly explained by the effects of individual gene variants on sarcomeric protein biomechanics. At the cellular level, HCM mutations most commonly enhance force production, leading to higher energy demands. Despite significant advances in elucidating sarcomeric structure-function relationships, there is still much to be learned about the mechanisms that link altered cardiac energetics to HCM phenotypes. In this work, we test the hypothesis that changes in cardiac energetics represent a common pathophysiologic pathway in HCM.
A Metabolome Atlas of the Aging Mouse Brain (Study part II)
STUDY_SUMMARY
The mammalian brain relies on neurochemistry to fulfill its functions. Yet, the complexity of the brain metabolome and its changes during diseases or aging remains poorly understood. To start bridging this gap, we generated a metabolome atlas of the aging wildtype male and female mouse brain from 10 anatomical regions spanning from adolescence to old age. We combined data from three chromatography-based mass spectrometry assays and structurally annotated 1,547 metabolites to reveal the underlying architecture of aging-induced changes in the brain metabolome. Overall differences between sexes were minimal. We found 99% of all metabolites to significantly differ between brain regions in at least one age group. We also discovered that 97% of the metabolome showed significant changes with respect to age groups. For example, we identified a shift in sphingolipid patterns during aging that is related to myelin remodeling in the transition from adolescent to aging brains. This shift was accompanied by large changes in overall signature in a range of other metabolic pathways. We found clear metabolic similarities in brain regions that were functionally related such as brain stem, cerebrum and cerebellum. In cerebrum, metabolic correlation patterns got markedly weaker in the transition from adolescent to adulthood, whereas the overall correlation patterns between all regions reflected a decreased brain segregation at old age. We were also able to map metabolic changes to gene and protein brain atlases to link molecular changes to metabolic brain phenotypes. Metabolic profiles can be investigated via https://mouse.atlas.metabolomics.us/. This new resource enables brain researchers to link new metabolomic studies to a foundation data set.
INSTITUTE
University of California, Davis
DEPARTMENT
Genome Center
LABORATORY
West Coast Metabolomics Center
LAST_NAME
Ding
FIRST_NAME
Jun
ADDRESS
451 East Health Science Drive, Davis, CA, 95616, USA
Multiomics Longitudinal Modeling of Preeclamptic Pregnancies (part I)
STUDY_SUMMARY
Preeclampsia is a complex disease of pregnancy whose physiopathology remains unclear and that poses a threat to both mothers and infants. Specific complex changes in women's physiology precede a diagnosis of preeclampsia. Understanding multiple aspects of such a complex changes at different levels of biology can be enabled by simultaneous application of multiple assays. We developed prediction models for preeclampsia risk by analyzing six omics datasets from a longitudinal cohort of pregnant women. A machine learning-based multiomics model had high accuracy (area under the receiver operating characteristics curve (AUC) of 0.94, 95% confidence intervals (CI): [0.90, 0.99]). A prediction model using only ten urine metabolites provided an accuracy of the whole metabolomic dataset and was validated using an independent cohort of 16 women (AUC=0.87, 95% CI: [0.76, 0.99]). Integration with clinical variables further improved prediction accuracy of the urine metabolome model (AUC=0.90, 95% CI: [0.80, 0.99], urine metabolome, validated). We identified several biological pathways to be associated with preeclampsia. The findings derived from models were integrated with immune system cytometry data, confirming known physiological alterations associated with preeclampsia and suggesting novel associations between the immune and proteomic dynamics. While further validation in larger populations is necessary, these encouraging results will serve as a basis for a simple, early diagnostic test for preeclampsia.
Multiomics Longitudinal Modeling of Preeclamptic Pregnancies (part II)
STUDY_SUMMARY
Preeclampsia is a complex disease of pregnancy whose physiopathology remains unclear and that poses a threat to both mothers and infants. Specific complex changes in women's physiology precede a diagnosis of preeclampsia. Understanding multiple aspects of such a complex changes at different levels of biology can be enabled by simultaneous application of multiple assays. We developed prediction models for preeclampsia risk by analyzing six omics datasets from a longitudinal cohort of pregnant women. A machine learning-based multiomics model had high accuracy (area under the receiver operating characteristics curve (AUC) of 0.94, 95% confidence intervals (CI): [0.90, 0.99]). A prediction model using only ten urine metabolites provided an accuracy of the whole metabolomic dataset and was validated using an independent cohort of 16 women (AUC=0.87, 95% CI: [0.76, 0.99]). Integration with clinical variables further improved prediction accuracy of the urine metabolome model (AUC=0.90, 95% CI: [0.80, 0.99], urine metabolome, validated). We identified several biological pathways to be associated with preeclampsia. The findings derived from models were integrated with immune system cytometry data, confirming known physiological alterations associated with preeclampsia and suggesting novel associations between the immune and proteomic dynamics. While further validation in larger populations is necessary, these encouraging results will serve as a basis for a simple, early diagnostic test for preeclampsia.
Small molecule signatures of mice lacking T-cell p38 alternate activation, a model for immunosuppression conditions, after exposure to total body radiation (part I)
STUDY_SUMMARY
Introduction Novel biodosimetry assays are needed in the event of radiological/nuclear emergencies for both immediate triage and identifying delayed effects of acute radiation exposure. Genetically engineered mouse models are used to assess how genotypic variation in the general population may affect post-irradiation classification performance. Here, we used a mouse model that lacks the T-cell receptor specific alternative p38 pathway (p38Ī±Ī²Y323F, double knock-in [DKI] mice) to determine how attenuated autoimmune and inflammatory responses may affect dose reconstruction. Objectives To determine if deficient alternative p38 activation differentially affects biofluid metabolic signatures post-irradiation compared to wild-type (WT). Methods Untargeted global metabolomics was used to assess biofluid signatures between WT and DKI mice (8 ā 10 weeks old) after exposure to total body radiation (0, 2, or 7 Gy). Urine was analyzed in the first week (1, 3, and 7 d) and serum at 1 d. Spectral features of interest were identified using the machine learning algorithm Random Forests and MetaboLyzer. Validated metabolite panels were constructed and classification performance was assessed by determining the area under the receiver operating characteristic curve (AUROC). Results A multidimensional scaling plot showed excellent separation of IR exposed groups in WT with slightly dampened responses in DKI mice. For both urine and serum, excellent sensitivity and specificity (AUROC > 0.90) was observed for 0 Gy vs. 7 Gy groups irrespective of genotype using identical metabolite panels. Similarly, excellent to fair classification (AUROC > 0.75) was observed for ā¤ 2 Gy vs. 7 Gy post-irradiation mice for both genotypes, however, model performance declined (AUROC < 0.75) between genotypes post-irradiation. Conclusion Overall, these results suggest less influence of the alternative p38 activation pathway for dose reconstruction compared to other radiosensitive genotypes.
Small molecule signatures of mice lacking T-cell p38 alternate activation, a model for immunosuppression conditions, after exposure to total body radiation (part II)
STUDY_SUMMARY
Introduction Novel biodosimetry assays are needed in the event of radiological/nuclear emergencies for both immediate triage and identifying delayed effects of acute radiation exposure. Genetically engineered mouse models are used to assess how genotypic variation in the general population may affect post-irradiation classification performance. Here, we used a mouse model that lacks the T-cell receptor specific alternative p38 pathway (p38Ī±Ī²Y323F, double knock-in [DKI] mice) to determine how attenuated autoimmune and inflammatory responses may affect dose reconstruction. Objectives To determine if deficient alternative p38 activation differentially affects biofluid metabolic signatures post-irradiation compared to wild-type (WT). Methods Untargeted global metabolomics was used to assess biofluid signatures between WT and DKI mice (8 ā 10 weeks old) after exposure to total body radiation (0, 2, or 7 Gy). Urine was analyzed in the first week (1, 3, and 7 d) and serum at 1 d. Spectral features of interest were identified using the machine learning algorithm Random Forests and MetaboLyzer. Validated metabolite panels were constructed and classification performance was assessed by determining the area under the receiver operating characteristic curve (AUROC). Results A multidimensional scaling plot showed excellent separation of IR exposed groups in WT with slightly dampened responses in DKI mice. For both urine and serum, excellent sensitivity and specificity (AUROC > 0.90) was observed for 0 Gy vs. 7 Gy groups irrespective of genotype using identical metabolite panels. Similarly, excellent to fair classification (AUROC > 0.75) was observed for ā¤ 2 Gy vs. 7 Gy post-irradiation mice for both genotypes, however, model performance declined (AUROC < 0.75) between genotypes post-irradiation. Conclusion Overall, these results suggest less influence of the alternative p38 activation pathway for dose reconstruction compared to other radiosensitive genotypes.
Involvement of Mieap in Cardiolipin metabolism (part I)
STUDY_SUMMARY
Quantitative assessment of total cardiolipin (CL) and comparison of CL species conducted with A549 (Ad-Mieap infected vs. non-infected) and LS174T cells (LS174T-cont vs. Mieap-KD). The A549 cells were harvested 24 hr after infection with Ad-Mieap and were compared with non-infected cells by mass spectrometric analysis. The LS174T-cont and Mieap-KD cells incubated under a normal condition were harvested and subjected to mass spectrometric analysis.
NMR Differentiating toxic and nontoxic congeneric harmful algae using the non-polar metabolome (Experiment 1)
STUDY_TYPE
1H NMR Metabolomics to compare toxic and non-toxic species
STUDY_SUMMARY
Metabolomics comparison of toxic and non-toxic species of phytoplankton from the genus Alexandrium.This study was carried out using 2 pairwise experiments, A. catenella compared to A. tamarense (Experiment 1) and A. pacificum compared to A. tamarense (Experiment 2). This study includes the NMR data from Experiment 1.
INSTITUTE
Georgia Institute of Technology
DEPARTMENT
School of Biological Sciences, School of Chemistry and Biochemistry, Center for Microbial Dynamics and Infection, Parker H. Petit Institute for Bioengineering and Bioscience
LABORATORY
Kubanek Lab
LAST_NAME
Brown
FIRST_NAME
Emily
ADDRESS
950 Atlantic Dr Atlanta, Georgia USA 30332
EMAIL
julia.kubanek@biosci.gatech.edu
PHONE
404-894-8424
NUM_GROUPS
2
TOTAL_SUBJECTS
30
STUDY_COMMENTS
Part 1 of 3. This part includes NMR analysis of non-polar metabolites from Experiment 1 using oPLSDA. Parts 2 and 3 inlcude NMR analysis of non-polar metabolites from Experiment 2 and the corresponding mass spectrometry metabolomics for both Experiments 1 and 2.
NMR Differentiating toxic and nontoxic congeneric harmful algae using the non-polar metabolome (Experiment 2)
STUDY_TYPE
1H NMR Metabolomics to compare toxic and non-toxic species
STUDY_SUMMARY
Metabolomics comparison of toxic and non-toxic species of phytoplankton from the genus Alexandrium.This study was carried out using 2 pairwise experiments, A. catenella compared to A. tamarense (Experiment 1) and A. pacificum compared to A. tamarense (Experiment 2). This study includes the NMR data from Experiment 2.
INSTITUTE
Georgia Institute of Technology
DEPARTMENT
School of Biological Sciences, School of Chemistry and Biochemistry, Center for Microbial Dynamics and Infection, Parker H. Petit Institute for Bioengineering and Bioscience
LABORATORY
Kubanek Lab
LAST_NAME
Brown
FIRST_NAME
Emily
ADDRESS
950 Atlantic Dr Atlanta, Georgia USA 30332
EMAIL
julia.kubanek@biosci.gatech.edu
PHONE
404-894-8424
NUM_GROUPS
2
TOTAL_SUBJECTS
30
STUDY_COMMENTS
Part 2 of 3. This part includes NMR analysis of non-polar metabolites from Experiment 2 using oPLSDA. Parts 1 and 3 inlcude NMR analysis of non-polar metabolites from Experiment 1 and the corresponding mass spectrometry metabolomics for both Experiments 1 and 2.
A local source of insulin in the eye governed by phagocytosis and starvation
STUDY_SUMMARY
Untargeted metabolite analysis was performed on a Thermo Orbitrap IDX Tribrid MS to understand changes in metabolites in eye retina and RPE tissue during starvation. This study also probes the role of certain metabolites during phagocytosis.
INSTITUTE
University of Virginia
DEPARTMENT
Center of Cell Clearance , Microbiology, Immunology, and Cancer Biology
LABORATORY
Kodi Ravichandraan Lab
LAST_NAME
Etchegaray
FIRST_NAME
Iker
ADDRESS
Center of Cell Clearance , Microbiology, Immunology, and Cancer Biology
Evolution of diapause in the African killifish by remodeling ancient gene regulatory landscape
STUDY_SUMMARY
Suspended animation (e.g. hibernation, diapause) allows organisms to survive extreme environments. But the mechanisms underlying the evolution of suspended animation states are unknown. The African turquoise killifish has evolved diapause as a form of suspended development to survive the complete drought that occurs every summer. Here, we show that gene duplicates ā paralogs ā exhibit specialized expression in diapause compared to normal development in the African turquoise killifish. Surprisingly, paralogs with specialized expression in diapause are evolutionarily very ancient and are present even in vertebrates that do not exhibit diapause. To determine if evolution of diapause is due to the regulatory landscape rewiring at ancient paralogs, we assessed chromatin accessibility genome-wide in fish species with or without diapause. This analysis revealed an evolutionary recent increase in chromatin accessibility at very ancient paralogs in African turquoise killifish. The increase in chromatin accessibility is linked to the presence of new binding sites for transcription factors, likely due to de novo mutations and transposable element (TE) insertion. Interestingly, accessible chromatin regions in diapause are enriched for lipid metabolism genes, and our lipidomics studies uncover a striking difference in lipid species in African turquoise killifish diapause, which could be critical for long-term survival. Together, our results show that diapause likely originated by repurposing pre-existing gene programs via recent changes in the regulatory landscape. This work raises the possibility that suspended animation programs could be reactivated in other species for long-term preservation via transcription factor remodeling and suggests a mechanism for how complex adaptations evolve in nature.
Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites (part I)
STUDY_SUMMARY
Previous reports suggest that the maturation rate of malaria parasites within red blood cells (RBC) is not constant for a given species in vivo. For instance, maturation can be influenced by host nutrient status or circadian rhythm. Here we observed in mice that systemic host inflammation, induced by lipopolysaccharide (LPS) conditioning or ongoing acute malaria infection, slowed the progression of a single cohort of parasites from one generation of RBC to the next. LPS-conditioning and acute infection both triggered substantial changes to the metabolomic composition of plasma in which parasites circulated. This altered plasma directly slowed parasite maturation in a manner that could not be rescued by supplementation, consistent with the presence of inhibitory factors. Single-cell transcriptomic assessment of mixed parasite populations, exposed to a short period of systemic host inflammation in vivo, revealed specific impairment in the transcriptional activity and translational capacity of trophozoites compared to rings or schizonts. Thus, we provide in vivo evidence of transcriptomic and phenotypic plasticity of asexual blood-stage Plasmodium parasites when exposed to systemic host inflammation
Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites (part II)
STUDY_SUMMARY
Previous reports suggest that the maturation rate of malaria parasites within red blood cells (RBC) is not constant for a given species in vivo. For instance, maturation can be influenced by host nutrient status or circadian rhythm. Here we observed in mice that systemic host inflammation, induced by lipopolysaccharide (LPS) conditioning or ongoing acute malaria infection, slowed the progression of a single cohort of parasites from one generation of RBC to the next. LPS-conditioning and acute infection both triggered substantial changes to the metabolomic composition of plasma in which parasites circulated. This altered plasma directly slowed parasite maturation in a manner that could not be rescued by supplementation, consistent with the presence of inhibitory factors. Single-cell transcriptomic assessment of mixed parasite populations, exposed to a short period of systemic host inflammation in vivo, revealed specific impairment in the transcriptional activity and translational capacity of trophozoites compared to rings or schizonts. Thus, we provide in vivo evidence of transcriptomic and phenotypic plasticity of asexual blood-stage Plasmodium parasites when exposed to systemic host inflammation
INSTITUTE
QIMR Berghofer Medical Research Institute
DEPARTMENT
Cell & Molecular Biology Department
LABORATORY
Precision & Systems Biomedicine
LAST_NAME
Stoll
FIRST_NAME
Thomas
ADDRESS
300 Herston Road
EMAIL
thomas.stoll@qimrberghofer.edu.au
NUM_GROUPS
5
TOTAL_SUBJECTS
30
STUDY_TYPE
Study part 2 of 2 (independent experiment 2; replication experiment of Study part 1)
Mitochondrial-Derived Compartments Facilitate Cellular Adaptation to Amino Acid Stress
STUDY_SUMMARY
Amino acids are essential building blocks of life. However, increasing evidence suggests that elevated amino acids cause cellular toxicity associated with numerous metabolic disorders. How cells cope with elevated amino acids remains poorly understood. Here, we show that a previously identified cellular structure, the mitochondrial-derived compartment (MDC), functions to protect cells from amino acid stress. In response to amino acid elevation, MDCs are generated from mitochondria, where they selectively sequester and deplete SLC25A nutrient carriers and their associated import receptor Tom70 from the organelle. Generation of MDCs promotes amino acid catabolism, and their formation occurs simultaneously with transporter removal at the plasma membrane via the multi-vesicular body (MVB) pathway. Combined loss of vacuolar amino acid storage, MVBs and MDCs renders cells sensitive to high amino acid stress. Thus, we propose that MDCs operate as part of a coordinated cell network that facilitates amino acid homoeostasis through post-translational nutrient transporter remodeling.
INSTITUTE
University of Utah School of Medicine
DEPARTMENT
Biochemistry
LABORATORY
Hughes Lab
LAST_NAME
Hughes
FIRST_NAME
Adam
ADDRESS
15 N Medical Drive East, RM 4100, Salt Lake City, UT, 84112, USA
Metabolomics analysis of AsPC-1 PDAC cells treated with Porcupine inhibitor (LGK974)
STUDY_SUMMARY
WNT signaling promotes pancreatic ductal adenocarcinoma (PDAC) through diverse effects on proliferation, differentiation, survival, and stemness. A subset of PDAC with inactivating mutations in ring finger protein 43 (RNF43) have growth dependency on autocrine WNT ligand signaling, which renders them susceptible to porcupine inhibitors (PORCNi) that block WNT ligand acylation and secretion. For this study, non-targeted metabolomic analyses were performed to explore the therapeutic response of RNF43-mutant PDAC to the PORCNi LGK974. AsPC-1 (RNF43-mutant) PDAC cells were treated with 25 nM LGK974 to explore stable isotope-resolved metabolomics with uniform 1, D-glucose [U13-C6] labeling.
Effect of ketogenic diet on the plasma and tumor metabolome of melanoma-bearing mice
STUDY_SUMMARY
Growing evidence supports the use of low-carbohydrate/high-fat ketogenic diets (KDs) together with standard therapies to improve cancer treatment outcomes. However, conflicting data exist regarding the efficacy of KDs in melanoma, the deadliest skin cancer. Here, we show that two different KD formulations effectively reduced tumor growth in immunocompromised mice bearing genetically and metabolically heterogeneous human melanoma xenografts. Furthermore, the KDs exerted a metastasis-reducing effect in an immunocompetent syngeneic melanoma mouse model. Ketone bodies did not directly influence melanoma cell proliferation; therefore, we performed an in-depth metabolomics analysis using the MxPĀ® Quant 500 kit combined with a acylcarnitine assay (biocrates life sciences ag)to elucidate potential anti-tumor mechanisms in vivo. Targeted analysis of plasma and tumor metabolomes revealed distinct changes in amino acid metabolism induced by the KDs. Moreover, the KDs increased sphingomyelin synthesis and hydroxylation of certain lipids. Thus, KDs simultaneously affect multiple metabolic pathways to create an unfavorable environment for melanoma cell proliferation, supporting their potential as a complementary nutritional approach to melanoma therapy.
INSTITUTE
University Hospital of the Paracelsus Medical University Salzburg
Lipidomics analysis of outer membrane vesicles and elucidation of the ceramide phosphoinositol biosynthetic pathway in Bacteroides thetaiotaomicron
STUDY_SUMMARY
In this work, we characterized the lipid composition of membranes and OMV from Bacteroides thetaiotaomicron VPI-5482. LC-MS analysis indicate that OMV carry sphingolipids, glycerophospholipids and serine-dipeptide lipids. Sphingolipid species represent more than 50% of the total lipid content of OMV. The most abundant sphingolipids in OMV are ceramide phosphoethanolamine (CerPE) and ceramide phosphoinositol (CerPI). Bioinformatic analysis allowed the identification of the BT1522-1526 operon putatively involved in CerPI synthesis. Mutagenesis studies revealed BT1522-1526 are essential for synthesis of PI and CerPI, confirming the role of this operon in biosynthesis of CerPI. BT1522-1526 mutant strains lacking CerPI produced OMV that were indistinguishable from the wild-type strain, indicating that CerPI sphingolipid species are not involved in OMV biogenesis. Bacteroides sphingolipids are thought to modulate host-commensal interactions, and based on our data, we propose that OMV could act as long distance delivery vehicles for these molecules.
Metabolomic profiling of saliva in diabetes patients
STUDY_SUMMARY
We performed comprehensive profiling of plasma and salivary metabolomes, and investigated multivariate covariations with clinical markers of oral and cardiometabolic health in healthy subjects and type 2 diabetes patients. The key findings highlight the potential utility of salivary metabolites for reflecting cardiometabolic changes, including hyperglycemia and dyslipidemia. Our study shows that analysis of panels of salivary metabolites may become an attractive alternative to blood testing for screening of metabolic disorders.
Metabolomic profiling of plasma in diabetes patients
STUDY_SUMMARY
We performed comprehensive profiling of plasma and salivary metabolomes, and investigated multivariate covariations with clinical markers of oral and cardiometabolic health in healthy subjects and type 2 diabetes patients. The key findings highlight the potential utility of salivary metabolites for reflecting cardiometabolic changes, including hyperglycemia and dyslipidemia. Our study shows that analysis of panels of salivary metabolites may become an attractive alternative to blood testing for screening of metabolic disorders.
Training-induced bioenergetic improvement in human skeletal muscle is associated with non-stoichiometric changes in the mitochondrial proteome without reorganisation of respiratory chain content
STUDY_SUMMARY
Lipidomic analysis of muscle mitochondrial isolates. 10 men with repeated measures.
Post Acute Myocardial Infarction Left Ventricular Remodeling Bio marker Analysis (PAMILA)
STUDY_SUMMARY
Patients with acute myocardial infarction (a condition classified under coronary heart disease, including STEMI and NSTEMI) are at high risk for recurrent ischemic events, but the pathways and factors which contribute to this elevated risk are incompletely understood. This study aims to identify biomarkers associated with acute myocardial infarction through various omics strategies. For the identified biomarkers, we aim to demonstrate prognostic value, and predict/stratify the risks of adverse cardiovascular events (e.g., stroke, heart failure, death).
INSTITUTE
National University of Singapore
LAST_NAME
Lim
FIRST_NAME
Si Ying
ADDRESS
Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore 117543
ATF3 is a common stress sensor, and its expression can be induced by serine deprivation. The goal of this project is to determine whether ATF3 regulates serine biosynthesis. ATF3-wildtype and āknockout cells are cultured in complete or serine-free medium supplemented with 13C-6-glucose for 24 h for stable isotope tracing. The results show that ATF3 appears to promote serine biosynthesis.
Impact of host dietary BCAA to gut symbiont-derived lipid profile
STUDY_SUMMARY
We analyzed mouse feces samples of C57BL/6 mice from diet intervention experiment. Briefly, mice were fed with BCAA-sufficient formulated diet (Testdiet 5CC7) for seven days, followed by BCAA-deficient formulated diet (Testdiet 58ZX) for the following seven days. Fecal sample were collected from mice at baseline (n=11), at day 7 (n=10) and at day 14 (n=10).
A high-fat diet induced obesity murine model treated with bitter melon (Momordica charantia)
STUDY_TYPE
A 40-day Intervention experiment
STUDY_SUMMARY
Metabolic effects of lyophilized bitter melon juice (BMJ) extract (oral gavage 200mg/kg/body weight-daily for 40 days) intake were evaluated in diet-induced obese C57BL/6J male mice [fed-high fat diet (HFD), 60 kcal% fat]. Changes in metabolite abundance levels in lipid-phase plasma [liquid chromatography mass spectrometry (LC-MS)-based metabolomics] after BMJ intervention were assessed.
INSTITUTE
University of Colorado Anschutz Medical Campus
DEPARTMENT
Pharmaceutical Sciences
LABORATORY
Dr. Rajesh Agarwal's Laboratory
LAST_NAME
Agarwal
FIRST_NAME
Rajesh
ADDRESS
12850 E. Montview Blvd, C238, Room V20-2118, Aurora, CO 80045, USA
Identification of effector metabolites using exometabolite profiling of diverse microalgae
STUDY_SUMMARY
We conducted an untargeted metabolomic analysis of non-polar exometabolites from four phylogenetically and ecologically diverse eukaryotic microalgal strains grown in the laboratory: freshwater Chlamydomonas reinhardtii, brackish Desmodesmus sp., marine Phaeodactylum tricornutum, and marine Microchloropsis salina. We analyzed both exometabolomes and cell pellet metabolomes to identify released metabolites based on relative enrichment in the exometabolomes.
Stool metabolomics in the New Hampshire Birth Cohort Study
STUDY_TYPE
Untargeted and semi-targeted metabolomics analysis
STUDY_SUMMARY
This is a study of data collected from fecal samples from larger New Hampshire Birth Cohort Study (NHBCS). 1H NMR metabolomic profiling of 524 infant stool samples collected at 6 week - 3 year age was performed and spectra were binned (untargeted metabolomics). A set of host-microbiome co-metabolites were library matched in individual sample spectra and their relative concentrations were determined. This study investigated associations of the functional metabolic response of the microbial milieu of the infant gut with environmental and other factors.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Nutrition
LABORATORY
Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel Hill
Fecal Metabolomics Reveals Products of Dysregulated Proteolysis and Altered Microbial Metabolism in Obesity-Related Osteoarthritis
STUDY_TYPE
C18 Untargeted UPLC-MS Metabolomics Analysis
STUDY_SUMMARY
Objective. The objective of this study was to determine if perturbations in gut microbial composition and the gut metabolome could be linked to individuals with obesity and osteoarthritis (OA). Methods. Fecal samples were collected from obese individuals diagnosed with radiographic hand plus knee OA (n=59), defined as involvement of at least 3 joints across both hands, and a Kellgren-Lawrence (KL) grade 2-4 (or total knee replacement) in at least one knee. Controls (n=33) were without hand OA and with KL grade 0-1 knees. Fecal metabolomes were analyzed by a UHPLC/Q Exactive HFx mass spectrometer. Microbiome composition was determined in fecal samples by 16S ribosomal RNA amplicon sequencing (rRNA-seq). Stepwise logistic regression models were built to determine microbiome and/or metabolic characteristics of OA. Results. Untargeted metabolomics analysis indicated that OA cases had significantly higher levels of di- and tri-peptides and significant perturbations in microbial metabolites including propionic acid, indoles and other tryptophan metabolites. Pathway analysis revealed several significantly perturbed pathways associated with OA including leukotriene metabolism, amino acid metabolism and fatty acid utilization. Logistic regression models selected metabolites associated with the gut microbiota and leaky gut syndrome as significant predictors of OA status, particularly when combined with the rRNA-seq data. Conclusions. Adults with obesity and OA have distinct fecal metabolomes characterized by increased products of proteolysis, perturbations in leukotriene metabolism, and changes in microbial metabolites compared with controls. These metabolic perturbations indicate a possible role of dysregulated proteolysis in OA.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Nutrition
LABORATORY
Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel Hill
Agonists and antagonists of nuclear receptor Rev-erbĪ±/Ī², key components of the circadian clock, can benefit the heart. Here, we show that mice with cardiomyocyte-specific knockout (KO) of Rev-erbĪ±/Ī² display progressive cardiac dilation and lethal heart failure. Inducible ablation of Rev-erbĪ±/Ī² in adult hearts causes similar phenotypes. Impaired fatty acid oxidation in the KO myocardium, particularly in the light cycle, precedes contractile dysfunctions with a reciprocal overreliance on carbohydrate utilization, particularly in the dark cycle. These findings delineate temporal coordination between clock-mediated anticipation and nutrient-induced response in myocardial metabolism.
Agonists and antagonists of nuclear receptor Rev-erbĪ±/Ī², key components of the circadian clock, can benefit the heart. Here, we show that mice with cardiomyocyte-specific knockout (KO) of Rev-erbĪ±/Ī² display progressive cardiac dilation and lethal heart failure. Inducible ablation of Rev-erbĪ±/Ī² in adult hearts causes similar phenotypes. Impaired fatty acid oxidation in the KO myocardium, particularly in the light cycle, precedes contractile dysfunctions with a reciprocal overreliance on carbohydrate utilization, particularly in the dark cycle. These findings delineate temporal coordination between clock-mediated anticipation and nutrient-induced response in myocardial metabolism.
The snap-frozen mouse heart ventricles that was harvested at ZT6 or ZT18 from both WT and Rev-erb cardiomyocytes specific KO mice were used for metabolomics study.
Metabolome-wide association study of occupational exposure to benzene
STUDY_SUMMARY
Benzene is a recognized hematotoxin and leukemogen; however, its mechanism of action in humans remain unclear. To provide insight into the processes underlying benzene hematotoxicity, we performed high-resolution metabolomic (HRM) profiling of plasma collected from a cross-sectional study of 33 healthy workers exposed to benzene (median 8-hr time-weighted average exposure; 20 ppma), and 25 unexposed controls in Shanghai, China. Metabolic features associated with benzene were identified using a metabolome-wide association study (MWAS) that tested for the relationship between feature intensity and benzene exposure. MWAS identified 478 mass spectral features associated with benzene exposure at FDR<20%. Comparison to a list of 13 known benzene metabolites and metabolites predicted using a multi-component biotransformation algorithm showed five metabolites were detected, which included the known metabolites phenol and benzene diolepoxide. Metabolic pathway enrichment identified 41 pathways associated with benzene exposure, with altered pathways including carnitine shuttle, fatty acid metabolism, sulfur amino acid metabolism, glycolysis, gluconeogenesis, and branched chain amino acid metabolism. These results suggest disruption to fatty acid uptake, energy metabolism and increased oxidative stress, and point towards pathways related to mitochondrial dysfunction, which has previously been linked to benzene exposure in animal models and human studies. Taken together, these results suggest benzene exposure is associated with disruption of mitochondrial pathways, and provide promising, systems biology biomarkers for risk assessment of benzene-induced hematotoxicity in humans.
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Environmental Medicine and Public Health
LABORATORY
High Resolution Exposomics
LAST_NAME
Walker
FIRST_NAME
Douglas
ADDRESS
Atran Building RM AB3-39, 1428 Madison Ave, New York, NY, 10029, USA
EMAIL
douglas.walker@mssm.edu
NUM_GROUPS
3
TOTAL_SUBJECTS
58
NUM_MALES
28
NUM_FEMALES
30
PUBLICATIONS
N Rothman, R Vermeulen, L Zhang, W Hu, S Yin, SM Rappaport, MT Smith, DP Jones, M Rahman, Qing Lan, DI Walker. (2021). Metabolome-wide association study of occupational exposure to benzene. Carcinogenesis. In Review
Exposure to environmental contaminants is associated with alterations in hepatic lipid metabolism in non-alcoholic fatty liver disease
STUDY_SUMMARY
Background & aims: Recent experimental models and epidemiological studies suggest that specific environmental contaminants (ECs) contribute to the initiation and pathology of NAFLD. However, the underlying mechanisms linking EC exposure with NAFLD remain poorly understood and there is no data on their impact on the human liver metabolome. Herein, we hypothesized that exposure to ECs, particularly perfluorinated alkyl substances (PFAS), impacts liver metabolism, specifically bile acid metabolism. Methods: In a well-characterized human NAFLD cohort of 105 individuals, we investigated the effects of EC exposure on liver metabolism. We characterized the liver (via biopsy) and circulating metabolomes using four mass spectrometry-based analytical platforms, and measured PFAS and other ECs in serum. We subsequently compared these results with an exposure study in a PPARa-humanized mouse model. Results: PFAS exposure appears associated with perturbation of key hepatic metabolic pathways previously found altered in NAFLD, particularly as regards bile acid metabolism. Specifically, we identified stronger associations between the liver metabolome, chemical exposure and NAFLD-associated clinical variables in female subjects versus males. The murine exposure study further corroborated our findings, vis-Ć -vis a sex-specific association between PFAS exposure and NAFLD-associated lipid changes. Conclusions: Females may be more sensitive to the harmful impacts of PFAS. Lipid-related changes subsequent to PFAS exposure may be secondary to the interplay between PFAS and bile acid metabolism.
INSTITUTE
Ćrebro University
DEPARTMENT
Department of Medical Sciences
LAST_NAME
McGlinchey
FIRST_NAME
Aidan
ADDRESS
School of Medical Sciences, Ćrebro, Ćrebro, 70281, Sweden
Metabolic and lipidomic characterization of radioresistant MDA-MB-231 human breast cancer cells to investigate potential therapeutic targets
STUDY_SUMMARY
To provide preliminary insights into metabolic and lipidomic characteristics in radioresistant triple-negative breast cancer (TNBC) cells and suggest potential therapeutic targets, we performed a comprehensive metabolic and lipidomic profiling of radioresistant MDA-MB-231 (MDA-MB-231/RR) TNBC cells and their parental cells using gas chromatography-mass spectrometry and nano electrospray ionization-mass spectrometry, followed by multivariate statistical analysis. Buthionine sulfoximine (BSO) and radiation were co-treated to radioresistant TNBC cells. The level of glutathione (GSH) was significantly increased, and the levels of GSH synthesis-related metabolites, such as cysteine, glycine, and glutamine were also increased in MDA-MB-231/RR cells. In contrast, the level of lactic acid was significantly reduced. In addition, reactive oxygen species (ROS) level was decreased in MDA-MB-231/RR cells. In the lipidomic profiles of MDA-MB-231/RR cells, the levels of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were significantly increased, whereas those of most of the phosphatidylinositol species were significantly decreased. BSO sensitized MDA-MB-231/RR cells to radiotherapy, which resulted in decreased GSH level and increased ROS level and apoptosis. Radioresistant TNBC cells showed distinct metabolic and lipidomic characteristics compared to their parental cells. We suggested activated GSH, PC, and PE biosynthesis pathways as potential targets for treating radioresistant TNBC cells. Particularly, enhanced radiosensitivity was achieved by inhibition of GSH biosynthesis in MDA-MB-231/RR cells.
INSTITUTE
ChungAng University
DEPARTMENT
College of Pharmacy
LABORATORY
Natural product biotechnology and Metabolomics
LAST_NAME
Lee
FIRST_NAME
Hwanhui
ADDRESS
College of Pharmacy, Chung-Ang University, Seoul 06974, Republic of Korea
An Airway Organoid-Based Screen Identifies a Role for the HIF1Ī±-Glycolysis Axis in SARS-CoV-2 Infection
STUDY_SUMMARY
SARS-CoV-2 causes the COVID-19 pandemic. It is urgent to develop disease models to dissect mechanisms regulating SARS-CoV-2 infection. Here, we derive airway organoids from human pluripotent stem cells (hPSC-AOs). The hPSC-AOs, particularly ciliated-like cells, are permissive to SARS-CoV-2 infection. Using this platform, we perform a high content screen and identify GW6471, which blocks SARS-CoV-2 infection. GW6471 can also block infection of the B.1.351 SARS-CoV-2 variant. RNA-seq analysis suggests that GW6471 blocks SARS-CoV-2 infection at least in part by inhibiting HIF1Ī±, which is further validated by chemical inhibitor and genetic perturbation targeting HIF1Ī±. Metabolic profiling identifies decreased rates of glycolysis upon GW6471 treatment, consistent with transcriptome profiling. Finally, xanthohumol, 5-(Tetradecyloxy)-2-furoic acid, and ND-646, three compounds that suppress fatty acid biosynthesis, also block SARS-CoV-2 infection. Together, a high content screen coupled with transcriptome and metabolic profiling reveals a key role of the HIF1Ī±-glycolysis axis in mediating SARS-CoV-2 infection of human airway epithelium.
Sublytic membrane attack complex drives glycolysis and mitochondrial dysfunction with inflammatory consequences in human monocyte-derived macrophages
STUDY_SUMMARY
The terminal stage in the complement activation pathways, the membrane attack complex (MAC), is upregulated in diabetic and rheumatoid arthritis patients, contributing pathologically by increasing inflammation. Previous research has highlighted that a sublytic dose of MAC can initiate NLRP3 inflammasome activation via calcium influx and loss of mitochondrial membrane potential. Here, we show that sublytic concentrations of MAC mediate a previously undescribed perturbation in cellular energy metabolism in human monocyte-derived macrophages, by phenotypic skewing towards glycolysis and upregulation of glycolysis-promoting genes. Sublytic MAC concentrations drive mitochondrial dysfunction, characterised by a fragmented mitochondrial morphology, loss of maximal respiratory response, depleted mitochondrial membrane potential as well as increased mitochondrial reactive oxygen species production. The consequences of these alterations in glycolytic metabolism and mitochondrial dysfunction lead to NLRP3 inflammasome activation, driving gasdermin D formation and IL-18 release. This novel link between sublytic MAC and immunometabolism, with direct consequences for downstream inflammatory processes, is important for development of novel therapeutics for areas where MAC may mediate disease.
Urine-Based Metabolomics and Machine Learning Reveals Metabolites Associated with Renal Cell Carcinoma Progression
STUDY_SUMMARY
Every year, hundreds of thousands of cases of renal carcinoma (RCC) are reported worldwide. Accurate staging of the disease is important for treatment and prognosis purposes; however, contemporary methods such as computerized tomography (CT) and biopsies are expensive and prone to sampling errors, respectively. As such, a non-invasive diagnostic assay for staging would be beneficial. This study aims to investigate urine metabolites as potential biomarkers to stage RCC. In the study, we identified a panel of such urine metabolites with machine learning techniques.
Urine-Based Metabolomics and Machine Learning Reveals Metabolites Associated with Renal Cell Carcinoma Progression NMR (part-I)
STUDY_SUMMARY
Every year, hundreds of thousands of cases of renal carcinoma (RCC) are reported worldwide. Accurate staging of the disease is important for treatment and prognosis purposes; however, contemporary methods such as computerized tomography (CT) and biopsies are expensive and prone to sampling errors, respectively. As such, a non-invasive diagnostic assay for staging would be beneficial. This study aims to investigate urine metabolites as potential biomarkers to stage RCC. In the study, we identified a panel of such urine metabolites with machine learning techniques.
Multiple Myeloma patients were treated with the proteasome inhibitor carfilzomib in different protocols according to their clinical stage. Paired serum samples were obtained before i.v injection and after 1-8h post treatment and snap frozen. Samples were stored at -80Ā°C until analysis.
In this study, we look at both targeted and untargeted metabolomic data from both sexes and 11 species of Drosophila. For the targeted analysis, we also looked at two ages to understand conserved changes with age in the Drosophila genus.
INSTITUTE
University of Washington
LAST_NAME
Promislow
FIRST_NAME
Daniel
ADDRESS
1959 NE Pacific Street, Seattle, Washington, 98195, USA
Fungal consortium of two Beauveria bassiana strains increases their virulence, growth, and resistance to stress: a metabolomic approach.
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
Entomopathogenic fungi have been successfully used to control agricultural pests. They infect insects by coming into direct contact with their cuticle or when feeding on contaminated leaves or fruits. After contact with the insect, the entomopathogenic fungus penetrates its body cavity, where it grows and colonizes it from within, causing its death The use of two or more microorganisms in a microbial consortium has been increasingly applied in the biological control of diseases and pests. Beauveria bassiana is one of the most widely studied fungal species in biological control, yet little is known about its role in fungal consortiums. In a previous study, our group found that a consortium formed by two strains of B. bassiana had significantly greater biocontrol potential against the polyphagous caterpillars Duponchelia fovealis (Lepidoptera: Crambidae) than either strain on its own. Despite recent developments and growing efforts to better understand fungal metabolism and metabolites, much remains unknown. Metabolomics therefore represents an important field for evaluating the metabolites produced or modified by an organism or its relationship with the environment. In the present study, we aim to use untargeted metabolomics with gas and liquid chromatography coupled to mass spectrometers (GC-MS and LC-MS/MS) to evaluate the metabolic alterations caused by the co-cultivation of these strains and to correlate the metabolites produced by this consortium with the increased mortality in D. fovealis observed previosly.
INSTITUTE
Universidade Federal do ParanĆ”
DEPARTMENT
Patologia BƔsica
LABORATORY
LaboratĆ³rio de Microbiologia e Biologia Molecular
Metabolomics profiles of premenopausal women are different based on O-desmethylangolensin metabotype
STUDY_SUMMARY
Urinary O-desmethylangolensin (ODMA) concentrations provide a functional gut microbiome marker of dietary isoflavone daidzein metabolism to ODMA. Individuals who do not have gut microbial environments that produce ODMA have less favorable cardiometabolic and cancer risk profiles. Urinary metabolomics profiles were evaluated in relation to ODMA metabotypes within and between individuals over time. Secondary analysis was conducted of data from the BEAN2 trial, which was a cross-over study of premenopausal women consuming six months on a high- and a low-soy diet, each separated by a 1-month washout period. In all of the 672 samples in the study, 66 of the 84 women had the same ODMA metabotype at seven or all eight time points. Two or four urine samples per woman were selected based on temporal metabotypes in order to compare within and across individuals. Metabolomics assays for primary metabolism and biogenic amines were conducted in 60 urine samples from 20 women. Partial least-squares discriminant analysis was used to compare metabolomics profiles.
Metabolomics profiles of premenopausal women are different based on O-desmethylangolensin metabotype (Part 2)
STUDY_SUMMARY
Urinary O-desmethylangolensin (ODMA) concentrations provide a functional gut microbiome marker of dietary isoflavone daidzein metabolism to ODMA. Individuals who do not have gut microbial environments that produce ODMA have less favorable cardiometabolic and cancer risk profiles. Urinary metabolomics profiles were evaluated in relation to ODMA metabotypes within and between individuals over time. Secondary analysis was conducted of data from the BEAN2 trial, which was a cross-over study of premenopausal women consuming six months on a high- and a low-soy diet, each separated by a 1-month washout period. In all of the 672 samples in the study, 66 of the 84 women had the same ODMA metabotype at seven or all eight time points. Two or four urine samples per woman were selected based on temporal metabotypes in order to compare within and across individuals. Metabolomics assays for primary metabolism and biogenic amines were conducted in 60 urine samples from 20 women.
Integrated molecular response of exposure to traffic-related pollutants in the US trucking industry
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
Exposure to traffic-related pollutants, including diesel exhaust, is associated with increased risk of cardiopulmonary disease and mortality; however, the precise biochemical pathways underlying these effects are not known. To investigate biological response mechanisms underlying exposure to traffic related pollutants, we used an integrated molecular response approach that included high-resolution metabolomic profiling and peripheral blood gene expression to identify biological responses to diesel exhaust exposure. Plasma samples were collected from 73 non-smoking males employed in the US trucking industry between February 2009 and October 2010 and analyzed using untargeted high-resolution metabolomics to characterize association with shift- and week-averaged levels of elemental carbon (EC), organic carbon (OC) and particulate matter with diameter ā¤ 2.5 Ī¼m (PM2.5). Annotated metabolites associated with exposure were then tested for relationships with the peripheral blood transcriptome using multivariate selection and network correlation. Week-averaged EC and OC levels, which were averaged across multiple shifts during the workweek, resulted in the greatest exposure-associated metabolic alterations compared to shift-averaged exposure levels. Metabolic changes associated with EC exposure suggest increased lipid peroxidation products, biomarkers of oxidative stress, thrombotic signaling lipids, and metabolites associated with endothelial dysfunction from altered nitric oxide metabolism, while OC exposures were associated with antioxidants, oxidative stress biomarkers and critical intermediates in nitric oxide production. Correlation with whole blood RNA gene expression provided additional evidence of changes in processes related to endothelial function, immune response, inflammation, and oxidative stress. We did not detect metabolic associations with PM2.5. This study provides an integrated molecular assessment of human exposure to traffic-related air pollutants that includes diesel exhaust. Metabolite and gene expression changes associated with exposure to EC and OC are consistent with increased risk of cardiovascular diseases and the adverse health effects of traffic-related air pollution.
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Environmental Medicine and Public Health
LABORATORY
High Resolution Exposomics
LAST_NAME
Walker
FIRST_NAME
Douglas
ADDRESS
Atran Building RM AB3-39, 1428 Madison Ave, New York, NY, 10029, USA
EMAIL
douglas.walker@mssm.edu
PHONE
1-212-241-4392
NUM_GROUPS
1
TOTAL_SUBJECTS
95
NUM_MALES
94
NUM_FEMALES
1
PUBLICATIONS
DI Walker, JE Hart, CJ Patel, R Rudel, J Chu, E Garshick, KD Pennel, F Laden, DP Jones. Integrated molecular response of exposure to traffic-related pollutants in the US trucking industry. Environment International. In review
Pesticides, Perfluorinated compounds and the development of adolescents in agricultural communities
STUDY_SUMMARY
This project aims to evaluate the developmental effects of pesticides and perfluroalkyl and polyfluroralkyl substances (PFAS) in adolescent participants of the study of Secondary Pesticide Exposure on Children and Adolescents (ESPINA) living in floricultural communities in Pedro Moncayo County, Ecuador. This prospective cohort study of children examines pesticide and PFAS exposures and the association with neurobehavioral performance, including anxiety and depression, and endocrine changes in adrenal and sex hormones in children aged 12 to 17 years. Analysis of adolescent serum metabolome changes aim to characterize metabolic and biological effects and their association to neuro behavioral outcomes and hormonal changes related to the developmental effects of PFAS from agricultural sources.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Tran
FIRST_NAME
ViLinh
ADDRESS
615 Michael St, suite 225
EMAIL
vtran6@emory.edu
PHONE
4047275091
TOTAL_SUBJECTS
527
STUDY_COMMENTS
Both CHEAR pooled plasma samples and Clinical Biomarker Laboratory pooled plasma samples were used
Perfluorinated compounds and high fat diet in relation to CVD-relevant metabolomic pathways in the SEARCH for Diabetes in Youth study
STUDY_SUMMARY
This project aims to evaluate the association between environmental exposure to perfluorinated alkyl substances (PFCs) and the development of risk factors for cardiometabolic disease in youth diagnosed with diabetes in the SEARCH Cohort Study. The longitudinal study of newly diagnosed cases of type 1 and type 2 diabetes examines serum metabolome changes at baseline and follow-up at approximately 5 years (all >3 years from baseline). Exposures to PFCs and biological effects characterized by serum metabolome changes will be associated with known cardiometabolic risk factors in youth diagnosed with diabetes.
INSTITUTE
Emory University
DEPARTMENT
School of Medicine
LABORATORY
Clincal Biomarkers Laboratory
LAST_NAME
Tran
FIRST_NAME
ViLinh
ADDRESS
615 Michael St, suite 225
EMAIL
vtran6@emory.edu
PHONE
4047275091
TOTAL_SUBJECTS
1796
STUDY_COMMENTS
Both CHEAR pooled plasma samples and Clinical Biomarker Laboratory pooled plasma samples were used
Absolute quantification of plasma cytokines and metabolome reveals the glycylproline regulating antibody-fading in convalescent COVID-19 patients
STUDY_SUMMARY
COVID-19 pandemic has caused tremendous costs worldwide and is still threatening public health in the ānew normalā. The association between neutralizing antibody levels and metabolic alterations in convalescent patients with COVID-19 is still poorly understood. In the present work, we conducted absolutely quantitative approach to profile the metabolomes in the plasma of the ordinary convalescent patients with antibody (CA), the convalescents of rapidly faded antibodies (CO) as well as the healthy subjects.
INSTITUTE
Hong Kong Baptist University
LAST_NAME
Yang
FIRST_NAME
Zhu
ADDRESS
OEW901, Oen Hall Building West Wing,, Ho Sin Hang Campus, Hong Kong Baptist University, Kowloon Tong, Kowloon, 999077, Hong Kong
Differential Accumulation of Metabolites and Transcripts Related to Flavonoid, Styrylpyrone, and Galactolipid Biosynthesis in Equisetum Species and Tissue Types
STUDY_SUMMARY
Members of the genus Equisetum are often referred to as āliving fossilsā, partly because they are the only extant representatives of the Equisetidae, a subclass that was once prominent in late Paleozoic forests. Several classes of specialized metabolites have been reported to occur in the genus Equisetum. However, while steady progress is being made with identifying individual novel metabolites of Equisetum, few if any analyses have focused on assessing the chemical diversity across the genus. The present study focused on three species: E. hyemale subsp. affine (rough horsetail or scouring rush), which is native to the temperate to artic portions of North America; E. arvense (common horsetail), which is endemic to the arctic and temperate regions of the northern hemisphere; and Equisetum telmateia subsp. braunii (Milde) Hauke (giant horsetail), which is native to western North America. Both below-ground rhizome and above-ground shoot material was harvested from each species, extracted with aqueous methanol, and subjected to non-targeted HPLC-QTOF-MS analysis. This research project was designed to lay the foundation for continued research to capture the metabolic capabilities in the ferns and fern allies.
INSTITUTE
Washington State University
DEPARTMENT
Institute of Biological Chemistry
LABORATORY
Lange
LAST_NAME
Lange
FIRST_NAME
Mark
ADDRESS
Plant Sciences Building, Pullman, Washington 99164
Metabolomic profiling of spontaneous macaque model for diabetes mellitus
STUDY_SUMMARY
The prevalence of diabetes mellitus has been increasing for decades worldwide. To develop safe and potent therapeutics, insights into the mechanisms underlying its pathogenesis are urgently needed. We reported the multi-omics profiling of the liver and sera of both peripheral blood and hepatic portal vein blood from Macaca fascicularis with spontaneous diabetes mellitus with a chow diet. The other two groups of the monkeys fed with chow diet and high-sugar high-fat (HSHF) diet, respectively, were included for comparison. These multi-omics datasets can provide a comprehensive picture of the molecular changes caused by diabetes in primates. Analyses of various omics datasets revealed the alterations of high consistency. Correlation between transcripts and proteins derived from the same genes was observed across individuals for most genes, especially the ones of differential expression. As a result, we found that distinct patterns of the metabolome, proteome, and transcriptome between spontaneous diabetes and HSHF diet-induced diabetes compared with healthy individuals.
INSTITUTE
Xiamen University
LAST_NAME
Yang
FIRST_NAME
Zhu
ADDRESS
OEW901, Oen Hall Building West Wing,, Ho Sin Hang Campus, Hong Kong Baptist University
Pseudoexfoliation aqueous humor lipidome suggests enrichment of specific pathways
STUDY_SUMMARY
Pseudoexfoliation syndrome (PEX) is systemic disorder that manifests as white, fluffy, proteinaceous fibrillar material throughout the body. In the eye such deposits result in Pseudoexfoliation glaucoma (PEXG), due to impeding aqueous humor outflow. Serum lipid alterations and increased lipid peroxidation have been reported in PEX. We report first ever comprehensive lipid profiling of the aqueous humor (AH) of PEXG. Our untargeted lipidomic analysis of 23 non-glaucomatous control, 19 primary open angle glaucoma, 9 PEX, and 14 PEXG AH with 13 deuterated lipid internal standards for normalization among the lipid classes resulted in the combined identification of 489 lipid species within 26 lipid classes across PEX, PEXG, POAG, and control AH. The mean total lipid content in the AH across samples showed that control AH (mean peak area 13.54 Ā± 56.1) had, on average, greater total lipid content than PEX (4.21 Ā± 10.90), PEXG (9.08 Ā± 25.97), and POAG (5.66 Ā± 15.75) samples. Multiple cholesterol esters (ChE), phosphatidylcholines (PC), triglycerides (TG), and ceramides (Cer) were present in higher concentrations for the PEXG AH samples. Some of the lipids found in high concentrations in the PEXG samples are ChE(16:0), ChE(20:3), ChE(18:1), ChE(18:3), ChE(22:6), ChE(18:2), ChE(20:4), PC(16:0/16:0), PC(16:0/18:2), TG(18:1/18:1/20:4), and Cer(t18:0/24:0). The CerG2GNAc1(d34:1) was enriched in control samples and depleted both in PEX and PEXG samples. The PC (18:0/18:2), PC (36:2), and PC (34:1e) are in low concentrations for PEX but highly concentrated in PEXG, despite both having similar material deposits, suggesting they are fundamentally different in composition. Elevations in Apolipoprotein A-I (APOA1) correlated to increase abundance of PC lipid species in the AH of patients with PEXG. Machine learning prediction with three supervised logistic regression binary classification tasks showed 1) POAG vs control, with 86% accuracy 2) PEXG vs control, with 71% accuracy and 3) PEX vs control, with 86% accuracy, respectively.
Comprehensive plasma metabolomics and lipidomics based management of benign and malignant solitary pulmonary nodules
STUDY_SUMMARY
This study found evidence of early metabolic alterations that can distinguish SPNs from healthy controls, but not for benign and malignant SPNs (lung cancer in stage I), highlighting that malignant SPNs less than 3 cm in diameter are most likely lung cancer at some early stage that does not affect blood circulation. Benign SPNs seldom require excessive treatment, and the strategy to detecting and managing malignant SPNs is to perform periodic radiographic examinations and, if operable, bring patients to surgery as quickly as feasible to prevent cancer cells from entering the circulation.
INSTITUTE
China Pharmaceutical University
LAST_NAME
Zhou
FIRST_NAME
Wei
ADDRESS
NO.639 Longmian avenue, Jiangning District, Nanjing city, Jiangsu Province, China
Metabolomics characterized concentration-dependent metabolic influence of magnesium on biofilm formation in Escherichia coli (Part1)
STUDY_SUMMARY
Biofilms are broadly formed by a diversity of microorganisms that enable them to adapt stressful environments. Biofilms often impose harmful influences in many niches, as they can cause food contamination, antibiotics resistance, and environmental issues. However, eradicating biofilms remains difficult since the formation mechanism of biofilms are still incompletely clarified. In this study, we aimed at exploring the regulatory role of magnesium (Mg2+) on biofilm formation in Escherichia coli (E. coli) using phenotype visualization combined with targeted metabolomics method. We found that Mg2+ could exert significant influence on biofilm formation in a concentration-dependent manner by regulating phenotypic morphology and triggering metabolic modifications of biofilm. Phenotypic imaging revealed that increasing concentration of Mg2+ gradually inhibited biofilm formation, Mg2+ was observed to restore the microstructure of E. coli strain in biofilms to that in the relevant planktonic cells. In addition, our metabolomics analysis characterized 20 differential metabolites and associated 2 metabolic pathways including nucleotide metabolism and amino acid metabolism that were notably modified during biofilm formation under the treatments of different concentrations of Mg2+. Altogether, our work provides a novel insight into the influence of Mg2+ on biofilm formation at a metabolic level, which are implicated in the novel solution to disturb biofilm formation through the regulation of Mg2+ and functional metabolite interaction, then biofilms associated harmful impacts in different niches could be well tangled accordingly.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Metabolomics characterized concentration-dependent metabolic influence of magnesium on biofilm formation in Escherichia coli (Part 2)
STUDY_SUMMARY
Biofilms are broadly formed by a diversity of microorganisms that enable them to adapt stressful environments. Biofilms often impose harmful influences in many niches, as they can cause food contamination, antibiotics resistance, and environmental issues. However, eradicating biofilms remains difficult since the formation mechanism of biofilms are still incompletely clarified. In this study, we aimed at exploring the regulatory role of magnesium (Mg2+) on biofilm formation in Escherichia coli (E. coli) using phenotype visualization combined with targeted metabolomics method. We found that Mg2+ could exert significant influence on biofilm formation in a concentration-dependent manner by regulating phenotypic morphology and triggering metabolic modifications of biofilm. Phenotypic imaging revealed that increasing concentration of Mg2+ gradually inhibited biofilm formation, Mg2+ was observed to restore the microstructure of E. coli strain in biofilms to that in the relevant planktonic cells. In addition, our metabolomics analysis characterized 20 differential metabolites and associated 2 metabolic pathways including nucleotide metabolism and amino acid metabolism that were notably modified during biofilm formation under the treatments of different concentrations of Mg2+. Altogether, our work provides a novel insight into the influence of Mg2+ on biofilm formation at a metabolic level, which are implicated in the novel solution to disturb biofilm formation through the regulation of Mg2+ and functional metabolite interaction, then biofilms associated harmful impacts in different niches could be well tangled accordingly.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Cognitive Behavioral Therapy for Irritable Bowel Syndrome Induces Bidirectional Alterations in the Brain-Gut-Microbiome Axis Associated with Gastrointestinal Symptom Improvement
STUDY_SUMMARY
34 Rome III-diagnosed IBS patients receiving CBT were drawn from the Irritable Bowel Syndrome Outcome Study (IBSOS; ClinicalTrials.gov NCT00738920). Fecal samples were collected at baseline and post-treatment for 16S rRNA gene sequencing, untargeted metabolomics, and measurement of short chain fatty acids. Multimodal neuroimaging was performed at baseline and post-treatment.
Untargeted metabolomics of breast cell lines in the presence or the absence of CtBP inhibitors
STUDY_SUMMARY
Experiments to test the effect of CtBP2 inhibition on metabolism of breast cell lines. In particular, experiment 1 involves comparison between a normal breast cell line (MCF102A) and a triple-negative breast cancer cell line (MDA-MB231). Experiment 2 is a study between MDA-MB231 silenced for CtBP2 by stable RNA interference (shCtBP2 cells) compared to scramble (shCTRL cells). Experiment 3 is a comparison between a normal breast cell line (MCF102A) and a triple-negative breast cancer cell line (MDA-MB231)in the presence of the absence of small-molecule CtBP inhibitors: HIPP (400 Ī¼M) or P4 (300 Ī¼M)for 48 hours.
Urinary signature of chronic kidney disease in patients with severe obesity by CE-MS
STUDY_TYPE
Human nephropathy in CKD obese patients
STUDY_SUMMARY
Urine metabolomic characterization of severe obese patients with and without chronic kidney disease (CKD) by CE-MS. Analysis was performed in patients before and after bariatric surgery. In the present studio, samples obtained from CKD patients with severe obesity before bariatric surgery will be referred as OD (obese disease). Samples obtained from CKD patients with severe obesity after bariatric surgery will be referred as ODBS (Obese disease bariatric surgery). Patients with severe obesity without CKD will be referenced as O (obese) and after BS, OBS (obese bariatric surgery). In healthy group, when results refer to the first void urine samples, the acronym will be Healthy1V, and the acronym for urine samples collected at 24-hour will be Healthy24h.
Phytoplankton-derived metabolites fuel a large fraction of heterotrophic bacterial production in the global ocean, yet methodological challenges have limited our knowledge of organic molecules transferred between these two microbial groups. In an experimental bloom study in which the diatom Thalassiosira pseudonana was co-cultured with three heterotrophic marine bacteria, we concurrently measured diatom endometabolites (i.e., potential exometabolite supply) by nuclear magnetic resonance (NMR) spectroscopy and bacterial gene expression (i.e., potential exometabolite uptake) by metatranscriptomic sequencing.
INSTITUTE
University of Georgia
DEPARTMENT
Department of Marine Sciences; Complex Carbohydrate Research Center
The largest living rodent dwelling Pantanal wetlands and Amazon basin, capybara, can efficiently depolymerize and utilize lignocellulosic biomass through microbial symbiotic mechanisms yet elusive. Herein, combining multi-meta-omics approaches, carbohydrate enzymology and X-ray crystallography, we elucidated the microbial community composition and structure, enzymatic systems and metabolic pathways involved in the conversion of recalcitrant dietary fibers into short-chain fatty acids, a main energy source for the host. The high efficiency of this microbiota in the deconstruction of plant polysaccharides is underpinned on the combination of unique enzymatic mechanisms from Fibrobacteres to degrade cellulose with a broad arsenal of Carbohydrate-Active enZymes (CAZymes) organized in polysaccharide utilization loci (PULs) from Bacteroidetes, to tackle with complex hemicelluloses typically found in gramineous and aquatic plants. Exploring the genomic dark matter of this community, two novel CAZy families were unveiled including a glycoside hydrolase family of Ī²-galactosidases and a carbohydrate-binding module family involved in xylan binding that establishes an unprecedented three-dimensional fold among associated modules to CAZymes. Together, these results demonstrate at community and molecular levels how the capybara gut microbiota orchestrates the deconstruction and utilization of dietary fibers, representing an untapped reservoir of new and intricate enzymatic mechanisms to overcome the lignocellulose recalcitrance, a central challenge toward a bio-based and sustainable economy.
INSTITUTE
Brazilian Center for Research in Energy and Materials (CNPEM)
LAST_NAME
Persinoti
FIRST_NAME
Gabriela
ADDRESS
Rua Giuseppe MƔximo Scolfaro, 10.000, Polo II de Alta Tecnologia de Campinas, Campinas, Sao Paulo, 13083-100, Brazil
Bipolar disorder metabolomics analysis using FiehnLib and GMD for curation
STUDY_TYPE
Untargeted GC analysis
STUDY_SUMMARY
In this study we analyzed the blood serum from 14 controls and 14 BD patients using GC-MS under the conditions required for the use of FiehnLib library with GMD as secondary library for curation on a untargeted metabolomics approach
INSTITUTE
new
DEPARTMENT
Chemistry Institute, Department of Analytical Chemistry
LABORATORY
Laboratory of Bioanalytics and Integrated Omics (LaBIOmics)
Metabolite profiling of shoots of juvenile maize plants
STUDY_TYPE
Metabolic diversity analysis
STUDY_SUMMARY
Profiles of primary metabolites in the shoots of juvenile maize inbred lines in the Goodman association panel were analyzed by GC-TOFMS to identify genetic components associated with metabolic control and plant performance. The samples also include those from landrace lines and maize wild relatives.
Metabolites Associated with Gestational Diabetes in Plasma
STUDY_TYPE
Case:Control ancillary analysis of RCT
STUDY_SUMMARY
"Gestational diabetes mellitus (GDM) significantly increases maternal and fetal health risks, but factors predictive of GDM are poorly understood. Plasma metabolomics analyses were conducted in early pregnancy to identify potential biomarkers for early prediction of Gestational Diabetes Mellitus (GDM). Sixty-eight pregnant women with overweight/obesity from a clinical trial of a lifestyle intervention were included. Participants who developed GDM (n=34; GDM group) were matched on treatment group, age, body mass index, and ethnicity with those who did not develop GDM (n=34; Non-GDM group). Blood draws were completed early in pregnancy (10-16 weeks). Plasma samples were analyzed by UPLC-MS using three metabolomics assays. "
INSTITUTE
California Polytechnic State University
DEPARTMENT
Food Science and Nutrition
LABORATORY
Cal Poly Metabolomics Service Center
LAST_NAME
La Frano
FIRST_NAME
Michael
ADDRESS
CALIFORNIA POLYTECHNIC STATE UNIVERSITY, 1 GRAND AVE
Plasma Metabolome Normalization in Rheumatoid Arthritis following initiation of Methotrexate and the Identification of Metabolic Biomarkers of Efficacy
STUDY_TYPE
Clinical
STUDY_SUMMARY
Methotrexate (MTX) efficacy in the treatment of rheumatoid arthritis (RA) is variable and unpredictable, resulting in a need to identify biomarkers to guide drug therapy. This study evaluates changes in the plasma metabolome associated with response to MTX in RA with the goal of understanding the metabolic basis for MTX efficacy towards the identification of potential metabolic biomarkers of MTX response.
Lipidome Alterations Following Mild Traumatic Brain Injury.
STUDY_TYPE
Untargeted Lipidomics
STUDY_SUMMARY
Traumatic brain injury (TBI) poses a major health challenge, with tens of millions of new cases reported globally every year. Brain damage resulting from TBI can vary significantly due to factors including injury severity, diffusivity, modality, time delay relative to impact, and exposure to repeated injury events. Untargeted lipidomic analysis of Sprague-Dawley rat serum within 24 hours of mild single and repeat controlled cortical impact (CCI) injury events led to the discovery of biomarker candidates of TBI. Lipid biomarkers have a unique potential to serve as objective molecular measures of the bodyās response to injury as their alteration in brain tissue can be more freely observed than for larger protein markers. Animal serum was analyzed via ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) in positive and negative ion modes. Known lipid species were identified through matching to in-house tandem MS databases. Machine learning and feature selection approaches were used to construct lipid panels capable of distinguishing serum from injured and uninjured animals across a range of injury severities and timepoints within the first day of injury. The best multivariate lipid panels had over 90% cross-validated sensitivity, selectivity, and accuracy and consisted of species from nine different lipid classes. These mapped onto sphingolipid signaling, autophagy, necroptosis and glycerophospholipid metabolism pathways, with FDR corrected p-values better than 0.05.
Quantification of Ļ-3 fatty acids and their derivatives in lungs from hypoxia-induced pulmonary hypertension (PH) mice.
STUDY_SUMMARY
Using a liquid chromatography tandem mass spectrometry (LC-MS/MS) system, we quantified the Ļ-3 fatty acid (EPA and DHA) metabolites in the lung tissues of mice with PH induced by chronic hypoxia (10% oxygen concentration).
Test the effect of GLS2 knockout in primary mouse hepatocytes. We isolated hepatocytes from GLS2 knockout and wild-type mice, and briefly applied media lacking L-glutamine (2 hours). Sixty minutes after resupplying Gln, metabolites were extracted and analyzed with the Mixed Mode method. Data were processed through XCMS, and features were filtered for p<0.01, fold change >2, and a minimum intensity of 1x10^6. Sorting by smallest p value, the first extracted ion chromatogram (EIC) with good chromatographic peak shape corresponded to 188.0567m/z at 24.41 min. Six putative IDs were within 3ppm of the experimentally observed m/z, representing two chemical formulas, none of which had documented retention times in training or test sets. Amongst these potential IDs, the Message Passing Neural Network (MPNN) model correctly predicted N-acetyl-L-glutamic acid as the most likely candidate, as verified by injection of purchased standards. The next most significant difference between GLS2KO vs WT was 117.0196m/z observed at 20.07 minutes. The model had been trained on 2/6 of the putative IDs. Despite the four additional isomers suggested, the model correctly selected succinate as reduced by GLS2 KO (Figure 5B). The third most significant hit corresponds to 171.0068m/z at 23.11 min. Although glycerol 1-P and 2-P are both potential hits, almost indistinguishable by the model, the large gap in retention times between these top hits and the Cl- adducts of threonate (+ isomers) is apparent, further supporting the correct identification as glycerol mono-phosphate. Expansion of the list to include p<0.05 leads to the identification of Glutamine, Glutamate, and other downstream metabolites known to be altered by GLS2 KO.
Identifying metabolite changes in human islets treated with Phospho-BAD mimicry and Inflammatory cytokines
STUDY_SUMMARY
The goal of this study was to associate metabolite changes with protection of human islets from cell death induced by the diabetogenic stress of inflammatory cytokines. Protection of human islet viability was accomplished via enhanced glucose metabolism using phospho-BAD mimicry peptide treatment.
A pathogenic role for histone H3 copper reductase activity in a yeast model of Friedreichās Ataxia
STUDY_SUMMARY
Disruptions to iron-sulfur (Fe-S) clusters, essential cofactors for a broad range of proteins, cause widespread cellular defects resulting in human disease. An underappreciated source of damage to Fe-S clusters are cuprous (Cu1+) ions. Since histone H3 enzymatically produces Cu1+ to support copper-dependent functions, we asked whether this activity could become detrimental to Fe-S clusters. Here, we report that histone H3-mediated Cu1+ toxicity is a major determinant of cellular functional pool of Fe-S clusters. Inadequate Fe-S cluster supply, either due to diminished assembly as occurs in Friedreichās Ataxia or defective distribution, causes severe metabolic and growth defects in S. cerevisiae. Decreasing Cu1+ abundance, through attenuation of histone cupric reductase activity or depletion of total cellular copper, restored Fe-S cluster-dependent metabolism and growth. Our findings reveal a novel interplay between chromatin and mitochondria in Fe-S cluster homeostasis, and a potential pathogenic role for histone enzyme activity and Cu1+ in diseases with Fe-S cluster dysfunction.
Metabonomics analysis reveals the physiological mechanism of promoting maize shoots growth under negative pressure to stabilize soil water content
STUDY_SUMMARY
The purpose of this study is to analyze maize shoots growth under negative pressure to stabilize soil water contentļ¼Maize plants were subjected to two irrigation treatments. The first treatment was soil moisture dry-wet cycles, which was obtained using drip irrigation (control, DW). The second treatment was negative pressure to stabilize soil water content treatment (SW), which was obtained using the negative pressure irrigation (NPI) system.
INSTITUTE
Heilongjiang Bayi Agricultural University
LAST_NAME
Zhang
FIRST_NAME
Jili
ADDRESS
High tech Zone, Longfei District, Daqing, Heilongjiang, 163319, China
Timecourse exometabolome analysis of glucose grown Rubrivivax benzoatilyticus cells
STUDY_TYPE
Timecourse experiment
STUDY_SUMMARY
Bacterial cells were grown on glucose under photoheterotrophic conditions for 18 days. Spent media of cells, harvested at 3rd, 9th and 18th day of growth, was vacuum dried and the metabolome was extracted in methanol. The extracted metabolites were derivatized and analyzed using GC-MS.
Untargeted Mass Spectrometry Metabolomic Profiles of iPSC-derived Dopaminergic Neurons from Clinically Discordant Brothers with Identical PRKN Deletions
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
We have previously reported on two brothers, PM and SM, who carry identical compound heterozygous PRKN mutations but present with very different clinical Parkinsonās disease (PD) phenotypes, with PM, but not SM having been diagnosed with early onset disease. The occurrence of juvenile cases demonstrates that PD is not necessarily an age-associated disease, indeed evidence is accumulating that there is a developmental component to PD pathogenesis. We hypothesize that additional genetic modifiers, potentially including genetic loci relevant to mesencephalic dopamine neuron development may play a role. We differentiated human-induced pluripotent stem cells (hiPSCs) derived from SM and PM into mitotically active mesencephalic neural precursor cells and early postmitotic dopaminergic neurons and performed whole exome sequencing, transcriptomic- and metabolomic analyses. No significant differences in canonical markers of differentiation were observed between SM and PM. Yet our transcriptomic analysis revealed a significant down regulation of three neurodevelopmentally relevant cell adhesion molecules, CNTN6, CNTN4 and CHL1 in PM- compared to SM cultures on days 11 and 25 of differentiation. In addition, several HLA genes, known to play a role in neurodevelopment, independent of their well-established function in immunity, were differentially regulated in PM and SM developing dopamine neurons. EN2, a transcription factor crucial for mesencephalic dopamine neuron development, was also differentially regulated. We further observed differences in cellular processes relevant to dopamine homeostasis. Lastly, our whole exome sequencing, transcriptomics and metabolomics data of SM and PM neurons revealed differences in GSH homeostasis, the dysregulation of which has been associated with PD.
Data on changes in lipid profiles during differentiation and maturation of human subcutaneous white adipocytes analyzed using chromatographic and bioinformatics tools
STUDY_SUMMARY
Three cell lines of Caucasian-derived subcutaneous preadipocytes were divided into five stages (stage-1 to stage-5) from subcutaneous preadipocytes to mature subcutaneous adipocytes filled with many lipid droplets. Lipids were extracted from cells in each stage and processed using untargeted liquid chromatography and Q-Exactive Orbitrap tandem mass spectrometry. The lipids were identified using LipidSearch 4.2.13.
INSTITUTE
Hamamatsu University School of Medicine
LAST_NAME
Kitamoto
FIRST_NAME
Takuya
ADDRESS
1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan
Investigation of serum metabolites in the AMPK intestinal KO mice
STUDY_SUMMARY
Conducted serum untargeted metabolomics analysis in AMP-activated protein kinase (AMPK) intestinal KO mice and control mice under high-fat diet (HFD) conditions
Lipidomics of brown adipocytes treated with d9-choline
STUDY_TYPE
targeted LC-QTOF/MS lipidomic profiling
STUDY_SUMMARY
Brown adipose tissue (BAT) was initially characterised as a thermogenic organ, and recent studies have suggested it plays a crucial role in maintaining systemic metabolic health. In this project, we demonstrated that alteration of BAT function contributes to development of heart failure through disorientation in choline metabolism. To analyze the detail effect of choline accumulation on brown adipocytes, we conducted the LC-QTOF/MS analysis using cultured brown adipocytes treated with d9-choline. In brown adipocytes treated with d9-choline, we found increase of phosphatidylcholine and lysophosphatidylcholine, suggesting that choline was metabolized in healthy brown adipocytes.
INSTITUTE
Juntendo University
DEPARTMENT
Department of Cardiovascular Biology and Medicine
LAST_NAME
Yoshida
FIRST_NAME
Yohko
ADDRESS
2-1-1, Hongo, Bunkyo-ku, Tokyo, Tokyo, 1138421, Japan
Metabolomics of brown adipose tissue in murine heart failure model
STUDY_TYPE
untargeted CE-TOF/MS metabolomic profiling
STUDY_SUMMARY
Brown adipose tissue (BAT) was initially characterised as a thermogenic organ, and recent studies have suggested it plays a crucial role in maintaining systemic metabolic health. In this project, we demonstrated that alteration of BAT function contributes to development of heart failure through disorientation in choline metabolism. To analyze the changes of metabolites, we conducted the CE-TOF/MS analysis using BAT from TAC (thoracic aortic constriction) or sham-operated mice. In BAT from TAC-operated mice, we found increase of choline and glycerophosphorylcholine and a decrease of phosphorylcholine, suggesting that BAT dysfunction induces the disorientation of choline metabolism.
INSTITUTE
Juntendo University
DEPARTMENT
Department of Cardiovascular Biology and Medicine
LAST_NAME
Yoshida
FIRST_NAME
Yohko
ADDRESS
2-1-1, Hongo, Bunkyo-ku, Tokyo, Tokyo, 1138421, Japan
THEM6-mediated lipid remodelling sustains stress resistance in cancer
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Despite the clinical benefit of androgen-deprivation therapy (ADT), the majority of patients with advanced prostate cancer (PCa) ultimately develop lethal castration-resistant prostate cancer (CRPC). In this study, we identified thioesterase superfamily member 6 (THEM6) as a marker of ADT resistance in PCa. In patients, THEM6 expression correlates with progressive disease and is associated with poor survival. THEM6 deletion reduces in vivo tumour growth and restores castration sensitivity in orthograft models of CRPC. Mechanistically, THEM6 is located at the endoplasmic reticulum (ER) membrane and controls lipid homeostasis by regulating intracellular levels of ether lipids. As a consequence, THEM6 loss in CRPC cells significantly alters ER function, preventing lipid-mediated induction of ATF4 and reducing de novo sterol biosynthesis. Finally, we show that THEM6 is required for the establishment of the MYC-induced stress response. Thus, similar to PCa, THEM6 loss significantly impairs tumorigenesis in the MYC-dependent subtype of triple negative breast cancer. Altogether our results highlight THEM6 as a novel component of the treatment-induced stress response and a promising target for the treatment of CRPC and MYC-driven cancer.
Quantitative genome-scale analysis of human liver reveals dysregulation of glycosphingolipid pathways in progressive nonalcoholic fatty liver disease
STUDY_SUMMARY
Nonalcoholic fatty liver disease (NAFLD) is a well-defined chronic liver diseases closely related with metabolic disorders. The prevalence of NAFLD is rapidly increasing worldwide, while the pathology and the underlying mechanisms driving NAFLD are not fully understood. In NAFLD, a series of metabolic changes takes place in the liver. However, the alteration of the metabolic pathways in the human liver along the progression of NAFLD, i.e., the transition from nonalcoholic steatosis (NAFL) to steatohepatitis (NASH) through cirrhosis remains to be discovered. Here, we sought to examine the metabolic pathways of the human liver across the full histological spectrum of NAFLD. We analyzed the whole liver tissue transcriptomic (RNA-Seq) and serum metabolomics data obtained from a large, prospectively enrolled cohort of histologically characterized patients derived from the European NAFLD Registry (n=206), and developed genome-scale metabolic models (GEMs) of human hepatocytes at different stages of NAFLD. The integrative approach employed in this study has enabled us to understand the regulation of the metabolic pathways of human liver in NAFL, and with progressive NASH-associated fibrosis (F0āF4). Our study identified several metabolic signatures in the liver and blood of these patients, specifically highlighting the alteration of vitamins (A, E) and glycosphingolipids (GSLs), and their link with complex glycosaminoglycans (GAGs) in advanced fibrosis. The study provides insights into the underlying pathways of the progressive fibrosing steatohepatitis. Furthermore, by applying genome-scale metabolic modeling (GSMM), we were able to identify the metabolic differences among carriers of widely validated genetic variants associated with NAFLD / NASH disease severity in three genes (PNPLA3, TM6SF2 and HSD17B13).
INSTITUTE
University of Turku
LAST_NAME
Sen
FIRST_NAME
Partho
ADDRESS
Systems Medicine group, Turku Bioscience, University of Turku (UTU), Tykistƶkatu 6B, P.O. Box 123 FIN-20521 Turku, Finland
Integration of Metabolomics and Proteomics to Unveil Orchestration of Photorespiration and Central Carbon Pathway in Microchloropsis gaditana NIES 2587
STUDY_TYPE
Time Course VLC HC Metabolome
STUDY_SUMMARY
Photosynthetic organisms have evolved and adapted strategies to overcome the limiting concentrations of CO2. In this regard, the CO2-concentrating mechanism (CCM) developed by microalgae implies an efficient machinery to acquire CO2 in limiting environment. Inorganic carbon transporters channelize CO2 towards Rubisco, however, there are significant differences in the CCM of some species and it is obscurely understood. In the present study, we performed qualitative metabolomics and proteomics on Microchloropsis gaditana, under the influence of very-low CO2 (VLC; 300 ppm, or 0.03%) and high CO2 (HC; 30,000 ppm, or 3% v/v) at the time intervals of 0, 6, 12 and 24 hrs. Our results demonstrate that HC supplementation channelizes the carbon flux towards enhancing the biomass yield, increasing up to 1.7-fold. Cyclic electron flow driven (CEF) by PSI confers energy to the cells in the case of VLC in the initial acclimatization stage. Our qualitative metabolomic analyses has identified nearly 35 essential metabolites among which significant fold-change was observed as a photorespiratory by-product, glycolate, in VLC resulting in delayed growth and lower biomass. Whole cell proteomics study was performed in M. gaditana in both VLC and HC conditions and a total of 998 proteins were identified. Cells in VLC, undergoes dynamic changes to activate biophysical CCM with the help of bicarbonate transporters. In conclusion, comprehensive changes occur inside the cell that consequently mediate the assimilation and regulation of carbon metabolic loadout such that it favours fatty acid biosynthesis in HC. In conclusion, our emphasis is to delineate carbon assimilation in M. gaditana with the help of advanced multi-omics tools and provide translational approach for the enhanced production of biofuels and biorenewables.
INSTITUTE
International Centre for Genetic Engineering and Biotechnology
DEPARTMENT
Integrative Biotechnology
LABORATORY
Omics of Algae
LAST_NAME
Jutur
FIRST_NAME
Pavan
ADDRESS
Omics of Algae Lab, 2nd Floor, New Building, ICGEB, Aruna Asaf Ali Marg, New Delhi
NMR Hydrophilic Metabolomic Analysis of Bacterial Resistance Pathways using Multivalent Antimicrobials with Challenged and Unchallenged Wild Type and Mutated Gram Positive Bacteria
STUDY_TYPE
NMR Hydrophilic Metabolomics
STUDY_SUMMARY
Multivalent membrane disruptors are a relatively new antimicrobial scaffold that are difficult for bacteria to develop resistance to and can act on both gram-positive and gram-negative bacteria. Nuclear Magnetic Resonance (NMR) metabolomics is an important method for studying resistance development in bacteria since it is both a quantitative and qualitative method to study and identify phenotypes by changes in metabolic pathways. Determine the likely metabolic differences between antimicrobially challenged and unchallenged growth and wild type and antimicrobially mutated Bacillus cereus (B. cereus) samples by using NMR hydrophilic metabolomics. Proton (1H) NMR hydrophilic metabolite analysis was conducted using B. cereus wild type and B. cereus that was mutated with C16-DABCO and mannose functionalized poly(amidoamine) dendrimers (DABCOMD). Both the wild type and the mutated sample types were grown in low levels of DABCOMD (challenged samples) or without the addition of DABCOMD to the growth media (unchallenged samples) for sample collection at the mid log and stationary phases and for growth curve procurement. Hierarchical clustering of only the challenged sample type showed that both the stationary phase sample types (mutant and wild type) clustered together while the both the mid log phase sample types were distinct. Hierarchical clustering of the unchallenged samples showed complete separation of all sample types. There were statistically significant (p-value and fold change) changes in the concentrations of metabolites in both energy related pathways and peptidoglycan synthesis between all sample types, especially with mutants and especially the challenged sample types have more N-acetylglucosamine (as much as a 94.2-fold increase). The mid log phase sample types showed a larger difference between sample types than their stationary phase counter parts. The challenged and unchallenged mutant samples showed a larger difference between sample types in comparison to the differences between the challenged and unchallenged wild type sample types. There was a larger metabolite difference when comparing the challenged mutant samples to the challenged wild type samples than when comparing the unchallenged mutant samples to the unchallenged wild type samples. The metabolomic analysis of wild type and multivalent DABCOMD mutated B. cereus under both challenged and unchallenged conditions indicated that the mutants, especially the challenged mutants, are likely changing their peptidoglycan layer to protect themselves from the high positive charge on the membrane disrupting DABCOMD. This membrane fortification most likely led to the slow growth curve of the mutated and especially the challenged mutant samples. The association of these sample types with metabolites associated with energy expenditure is attributed to the increased energy required for these changes to occur as well as to the decreased diffusion of nutrients across the membrane.
Analytical methodology for a metabolome atlas of goatās plas-ma, milk and feces using 1H-NMR and UHPLC-HRMS
STUDY_SUMMARY
Metabolomics has been increasingly used in animal and food sciences. Animal health is one of the most important factor that can also alter animal integrity and welfare. Some studies have al-ready investigated the link between health and metabolic profile of dairy animals. These studies in metabolomics often consider a single type of sample using a single analytical platform (Nu-clear Magnetic Resonance or Mass Spectrometry). Only few studies with multi-platform ap-proaches are also used with a single or a multi type of sample, but they mainly consider dairy cows metabolome although dairy goats present similar diseases, that it could be interesting to detect early to preserve animal health and milk production. This study aims to create a metabol-ic atlas of goat plasma, milk and feces, based on healthy animals. Our study describes a Standard Operating Procedure for three goat matrices: blood plasma, milk, and feces using multiple plat-forms (NMR (1H), UHPLC (RP)-MS and UHPLC (HILIC)-MS) that follows a unique sample prep-aration procedure for each sample type to be analyzed on multi-platforms basis. Our method was evaluated for its robustness and allowed a better characterization of goat metabolic profile in healthy conditions.
Analytical methodology for a metabolome atlas of goatās plas-ma, milk and feces using 1H-NMR and UHPLC-HRMS
STUDY_SUMMARY
Metabolomics has been increasingly used in animal and food sciences. Animal health is one of the most important factor that can also alter animal integrity and welfare. Some studies have al-ready investigated the link between health and metabolic profile of dairy animals. These studies in metabolomics often consider a single type of sample using a single analytical platform (Nu-clear Magnetic Resonance or Mass Spectrometry). Only few studies with multi-platform ap-proaches are also used with a single or a multi type of sample, but they mainly consider dairy cows metabolome although dairy goats present similar diseases, that it could be interesting to detect early to preserve animal health and milk production. This study aims to create a metabol-ic atlas of goat plasma, milk and feces, based on healthy animals. Our study describes a Standard Operating Procedure for three goat matrices: blood plasma, milk, and feces using multiple plat-forms (NMR (1H), UHPLC (RP)-MS and UHPLC (HILIC)-MS) that follows a unique sample prep-aration procedure for each sample type to be analyzed on multi-platforms basis. Our method was evaluated for its robustness and allowed a better characterization of goat metabolic profile in healthy conditions.
Analytical methodology for a metabolome atlas of goatās plas-ma, milk and feces using 1H-NMR and UHPLC-HRMS
STUDY_SUMMARY
Metabolomics has been increasingly used in animal and food sciences. Animal health is one of the most important factor that can also alter animal integrity and welfare. Some studies have al-ready investigated the link between health and metabolic profile of dairy animals. These studies in metabolomics often consider a single type of sample using a single analytical platform (Nu-clear Magnetic Resonance or Mass Spectrometry). Only few studies with multi-platform ap-proaches are also used with a single or a multi type of sample, but they mainly consider dairy cows metabolome although dairy goats present similar diseases, that it could be interesting to detect early to preserve animal health and milk production. This study aims to create a metabol-ic atlas of goat plasma, milk and feces, based on healthy animals. Our study describes a Standard Operating Procedure for three goat matrices: blood plasma, milk, and feces using multiple plat-forms (NMR (1H), UHPLC (RP)-MS and UHPLC (HILIC)-MS) that follows a unique sample prep-aration procedure for each sample type to be analyzed on multi-platforms basis. Our method was evaluated for its robustness and allowed a better characterization of goat metabolic profile in healthy conditions.
Analytical methodology for a metabolome atlas of goatās plasma, milk and feces using 1H-NMR and UHPLC-HRMS:MS/milk
STUDY_SUMMARY
Metabolomics has been increasingly used in animal and food sciences. Animal health is one of the most important factors that can also alter animal integrity and welfare. Some studies have already investigated the link between health and metabolic profile of dairy animals. These studies in metabolomics often consider a single type of sample using a single analytical platform (Nuclear Magnetic Resonance or Mass Spectrometry). Only few studies with multi-platform approaches are also used with a single or a multi type of sample, but they mainly consider dairy cows metabolome although dairy goats present similar diseases, that it could be interesting to detect early to preserve animal health and milk production. This study aims to create a metabolic atlas of goat plasma, milk, and feces, based on healthy animals. Our study describes a Standard Operating Procedure for three goat matrices: blood plasma, milk, and feces using multiple platforms (NMR (1H), UHPLC (RP)-MS and UHPLC (HILIC)-MS) that follows a unique sample preparation procedure for each sample type to be analyzed on multi-platforms basis. Our method was evaluated for its robustness and allowed a better characterization of goat metabolic profile in healthy conditions.
Lipid Profiling in African-American Men with Prostate Cancer
STUDY_SUMMARY
we examined the lipidomes from cancer-benign matched prostate tissues in PCa patients of AA and EA men to determine potential alterations in lipid metabolism which might explain the observed disparity in tumor progression.
Analytical methodology for a metabolome atlas of goatās plasma, milk and feces using 1H-NMR and UHPLC-HRMS:NMR/feces
STUDY_SUMMARY
Metabolomics has been increasingly used in animal and food sciences. Animal health is one of the most important factors that can also alter animal integrity and welfare. Some studies have already investigated the link between health and metabolic profile of dairy animals. These studies in metabolomics often consider a single type of sample using a single analytical platform (Nuclear Magnetic Resonance or Mass Spectrometry). Only few studies with multi-platform approaches are also used with a single or a multi type of sample, but they mainly consider dairy cows metabolome although dairy goats present similar diseases, that it could be interesting to detect early to preserve animal health and milk production. This study aims to create a metabolic atlas of goat plasma, milk, and feces, based on healthy animals. Our study describes a Standard Operating Procedure for three goat matrices: blood plasma, milk, and feces using multiple platforms (NMR (1H), UHPLC (RP)-MS and UHPLC (HILIC)-MS) that follows a unique sample preparation procedure for each sample type to be analyzed on multi-platforms basis. Our method was evaluated for its robustness and allowed a better characterization of goat metabolic profile in healthy conditions.
Analytical methodology for a metabolome atlas of goatās plasma, milk and feces using 1H-NMR and UHPLC-HRMS:MS/feces
STUDY_SUMMARY
Metabolomics has been increasingly used in animal and food sciences. Animal health is one of the most important factors that can also alter animal integrity and welfare. Some studies have already investigated the link between health and metabolic profile of dairy animals. These studies in metabolomics often consider a single type of sample using a single analytical platform (Nuclear Magnetic Resonance or Mass Spectrometry). Only few studies with multi-platform approaches are also used with a single or a multi type of sample, but they mainly consider dairy cows metabolome although dairy goats present similar diseases, that it could be interesting to detect early to preserve animal health and milk production. This study aims to create a metabolic atlas of goat plasma, milk, and feces, based on healthy animals. Our study describes a Standard Operating Procedure for three goat matrices: blood plasma, milk, and feces using multiple platforms (NMR (1H), UHPLC (RP)-MS and UHPLC (HILIC)-MS) that follows a unique sample preparation procedure for each sample type to be analyzed on multi-platforms basis. Our method was evaluated for its robustness and allowed a better characterization of goat metabolic profile in healthy conditions.
Anti-oxidative metabolism measurement in mammalian cells and tissues by quantitative LC/MS method (I)
STUDY_SUMMARY
Anti-oxidative metabolism measurement in mammalian cells and tissues by quantitative LC/MS method to establish baseline metabolism in mouse controls 0-48hrs in CSF and hippocampus.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Anti-oxidative metabolism measurement in mammalian cells and tissues by quantitative LC/MS method (II)
STUDY_SUMMARY
Anti-oxidation metabolism measurement in mouse CSF by quantitative LC/MS method to establish MTX effects on mouse metabolism in mouse controls 0-48hrs in CSF (repeat of 20200124 ChP-MTX-Anti-oxidative-study-test)
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Anti-oxidation metabolism measurement in mammalian cells and tissues by quantitative LC/MS method (III)
STUDY_SUMMARY
Anti-oxidative metabolism measurement in mouse CSF by quantitative LC/MS method of mouse CSF at 24H of MTX treatment, for either control GFP or SOD3-overexpressing ChP mice.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Metabolomic profiles in S. mutans, S. gordonii, and S. oralis cells treated with D-tagatose
STUDY_SUMMARY
Recent studies have shown phenotypic and metabolic heterogeneity in related species including Streptococcus oralis, a typical oral commensal bacterium, Streptococcus mutans, a cariogenic bacterium, and Streptococcus gordonii, which functions as an accessory pathogen in periodontopathic biofilm. In this study, metabolites characteristically contained in the saliva of individuals with good oral hygiene were determined, after which the effects of an identified prebiotic candidate, D-tagatose, on phenotype, gene expression, and metabolic profiles of those three key bacterial species were investigated. Examinations of the saliva metabolome of 18 systemically healthy volunteers identified salivary D-tagatose as associated with lower dental biofilm abundance in the oral cavity (Spearmanās correlation coefficient; r = -0.603, p = 0.008), then the effects of D-tagatose on oral streptococci were analyzed in vitro. In chemically defined medium (CDM) containing D-tagatose as the sole carbohydrate source, S. mutans and S. gordonii each showed negligible biofilm formation, whereas significant biofilms were formed in cultures of S. oralis. Furthermore, even in the presence of glucose, S. mutans and S. gordonii showed growth suppression and decreases in the final viable cell count in a D-tagatose concentration-dependent manner. In contrast, no inhibitory effects of D-tagatose on the growth of S. oralis were observed. To investigate species-specific inhibition by D-tagatose, the metabolomic profiles of D-tagatose-treated S. mutans, S. gordonii, and S. oralis cells were examined. The intracellular amounts of pyruvate-derived amino acids in S. mutans and S. gordonii, but not in S. oralis, such as branched-chain amino acids and alanine, tended to decrease in the presence of D-tagatose. This phenomenon indicates that D-tagatose inhibits growth of those bacteria by affecting glycolysis and its downstream metabolism. In conclusion, the present study provides evidence that D-tagatose is abundant in saliva of individuals with good oral health. Additionally, experimental results demonstrated that D-tagatose selectively inhibits growth of the oral pathogens S. mutans and S. gordonii. In contrast, the oral commensal S. oralis seemed to be negligibly affected, thus highlighting the potential of administration of D-tagatose as an oral prebiotic for its ability to manipulate the metabolism of those targeted oral streptococci.
Non-destructive characterization of Mesenchymal stem cells
STUDY_SUMMARY
Culture media from the growth of three different MSC cell lines (two bone marrow, one iPSC) were sampled daily for NMR metabolomics analysis. T cell proliferation and IDO assays were used as surrogates of anti-inflammatory function. Linear regression was used to assess the media metabolic changes over time, and partial least squares regression (PLSR) was then used to obtain predictive media markers (PMMs) based on variable importance in projection (VIP) scores. In addition, pathway analysis was performed to show the relations between media metabolites (MMs) and cell metabolites (CMs).
Lipidomic characterization of Candida albicans in response to Aureobasidin treatment in vitro.
STUDY_SUMMARY
Candida albicans is an opportunistic yeast pathogen that causes a wide range of infections especially amongst immunocompromised patients. Aureobasidin A (AbA) has been shown to inhibit inositolphosphoryl ceramide synthase (IPCS), a key enzyme responsible for sphingolipid biosynthesis. There are limited studies exploring IPCS as a target molecule for antifungal treatment. It is hypothesized that the mechanism of AbA inhibition involves alteration of C. albicans phospholipid and sphingolipid profiles. The profiling of C. albicans phospholipid and sphingolipid upon exposure to 0.5-4 Āµg/ml of AbA were determined using Liquid chromatography-mass spectrometry (LC-MS).
INSTITUTE
University of Malaya
LAST_NAME
Hamdan
FIRST_NAME
Nur Wahida
ADDRESS
Jalan Profesor Diraja Ungku Aziz, 50603 Kuala Lumpur, Wilayah Persekutuan Kuala Lumpur, Malaysia
Metabolomic Fingerprinting of Human High Grade Serous Ovarian Carcinoma Cell Lines
STUDY_SUMMARY
Focusing on defining the metabolomic basis of intratumoral heterogeneity in ovarian cancer, the metabolic diversity of a panel of high grade serous ovarian carcinoma (HGSOC) cell-lines we investigated using a metabolomics platform that interrogate 731 compounds. Metabolic fingerprinting followed by 2-dimensional and 3-dimensional principal component analysis defined the heterogeneity of the HGSOC cells and clustered them into five distinct metabolic groups. An overall increase in the metabolites associated with aerobic glycolysis and phospholipid metabolism were observed in the majority of the cancer cells. A preponderant increase in the levels of metabolites involved in trans-sulfuration and glutathione synthesis was also observed. Subsets of HGSOC cells showed an increase in the levels of 5-Hydroxytryptamine, gamma-aminobutyric acid, or glutamate, pointing to their potential role as oncometabolites. In summary, our results identify increased glycolysis, phospholipid metabolism and amino acid metabolism with the resultant increase in the levels of 5-Hydoxytryptamine, GABA, and Glutamate as metabolomic correlates underlying the heterogeneity of ovarian cancer cell lines.
INSTITUTE
University of Oklahoma Health Sciences Center
DEPARTMENT
Cell Biology
LABORATORY
Danny N Dhanasekaran
LAST_NAME
Jayaraman
FIRST_NAME
Muralidharan
ADDRESS
975 NE 10th street, BRC1470, Oklahoma City, Oklahoma, 73104, USA
Metabolic adaptations in an endocrine-related breast cancer mouse model unveil potential markers of tumor response to hormonal therapy
STUDY_TYPE
case - control study
STUDY_SUMMARY
Breast cancer (BC) is the most common type of cancer in women and, in most cases, it is hormone-dependent (HD), thus relying on ovarian hormone activation of intracellular receptors to stimulate tumor growth. Endocrine therapy (ET) aimed at preventing hormone receptor activation is the primary treatment strategy, however, about half of the patients, develop resistance in time. This involves the development of hormone independent tumors that initially are ET-responsive (HI), which may subsequently become resistant (HIR). The mechanisms that promote the conversion of HI to HIR tumors are varied and not completely understood. The aim of this work was to characterize the metabolic adaptations accompanying this conversion through the analysis of the polar metabolomes of tumor tissue and non-compromised mammary gland from mice implanted subcutaneously with HD, HI and HIR tumors from a medroxyprogesterone acetate (MPA)-induced BC mouse model. This was carried out by nuclear magnetic resonance (NMR) spectroscopy of tissue polar extracts and data mining through multivariate and univariate statistical analysis. Initial results unveiled marked changes between global tumor profiles and non-compromised mammary gland tissues, as expected. More importantly, specific metabolic signatures were found to accompany progression from HD, through HI and to HIR tumors, impacting on amino acids, nucleotides, membrane percursors and metabolites related to oxidative stress protection mechanisms. For each transition, sets of polar metabolites are advanced as potential markers of progression, including acquisition of resistance to ET. Putative biochemical interpretation of such signatures are proposed and discussed.
INSTITUTE
University of Aveiro
DEPARTMENT
Chemistry
LABORATORY
Metabolomics
LAST_NAME
Silva
FIRST_NAME
Ana
ADDRESS
Campus UniversitƔrio de Santiago, 3810-193
EMAIL
anarita.asilva@ua.pt
PHONE
234370200
NUM_GROUPS
8
TOTAL_SUBJECTS
48
NUM_FEMALES
48
STUDY_COMMENTS
For this study 48 female 2-month old Balb/c mice were used
Profiling Plasmodium falciparum parasites and human red blood cells after treatment with MMV693183
STUDY_SUMMARY
Compound MMV693183 was added to either Plasmodium falciparum or uninfected red blood cells. The concentrations used were 240 nM, 24 nM, and 2.4 nM. Untreated parasites and red blood cells were also used as controls.
INSTITUTE
Pennsylvania State University
LAST_NAME
LlinƔs
FIRST_NAME
Manuel
ADDRESS
W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA
Fbxo7 promotes Cdk6 activity to inhibit PFKP and glycolysis in T cells
STUDY_SUMMARY
Deregulated Fbxo7 expression is associated with many pathologies, including anaemia, male sterility, cancer, and Parkinsonās disease, demonstrating its critical role in a variety of cell types. Although Fbxo7 is an F-box protein that recruits substrates for SCF-type E3 ubiquitin ligases, it also promotes the formation of cyclin D/Cdk6/p27 complexes in an E3-ligase independent fashion. We discovered PFKP, the major gatekeeper of glycolysis, in a screen for Fbxo7 substrates. PFKP has been previously shown to be a critical substrate of Cdk6 for the viability of T-ALL cells experiencing high levels of reactive oxygen species. We investigated the molecular relationships between Fbxo7, Cdk6 and PFKP, and the functional effect Fbxo7 has on T cell metabolism, viability, and activation. Fbxo7 promotes Cdk6-independent ubiquitination and Cdk6-dependent phosphorylation of PFKP. Importantly Fbxo7-deficient cells have reduced Cdk6 activity, and haematopoietic and lymphocytic cell lines show a significant dependency on Fbxo7. CD4+ T cells with reduced Fbxo7 have increased glycolysis, and lower cell viability and activation levels. Metabolomic studies of activated CD4+ T cells confirm increased glycolytic flux in Fbxo7-deficient cells, as well as altered nucleotide biosynthesis and arginine metabolism. We show Fbxo7 expression is glucose-responsive, and we propose Fbxo7 inhibits PFKP and glycolysis via its activation of Cdk6.
Transcriptomic and lipidomic analysis unravels the response of Faecalibacterium prausnitzii to calcium palmitate
STUDY_SUMMARY
Infant formula is a suggested alternative to human milk if breastfeeding is not an option; vegetable oil blends are commonly used in infant formula (IF) to replace dairy fat, which can induce the formation of the poorly soluble soap calcium palmitate (CP) in the infantās gut. Previously, we observed that CP at a low concentration of 0.01 mg/ml inhibits the growth of dominant infant bacteria such as Faecalibacterium prausnitzii both during the exponential phase as well as in the stationary phase. Here, we investigate the underlying mechanism of the CP inhibition on infant-gut bacteria using F. prausnitzii as a model by analysing its growth at a transcriptomic and lipidomic level.
THEM6-mediated lipid remodelling sustains stress resistance in cancer (Part 2)
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Despite the clinical benefit of androgen-deprivation therapy (ADT), the majority of patients with advanced prostate cancer (PCa) ultimately develop lethal castration-resistant prostate cancer (CRPC). In this study, we identified thioesterase superfamily member 6 (THEM6) as a marker of ADT resistance in PCa. In patients, THEM6 expression correlates with progressive disease and is associated with poor survival. THEM6 deletion reduces in vivo tumour growth and restores castration sensitivity in orthograft models of CRPC. Mechanistically, THEM6 is located at the endoplasmic reticulum (ER) membrane and controls lipid homeostasis by regulating intracellular levels of ether lipids. As a consequence, THEM6 loss in CRPC cells significantly alters ER function, preventing lipid-mediated induction of ATF4 and reducing de novo sterol biosynthesis. Finally, we show that THEM6 is required for the establishment of the MYC-induced stress response. Thus, similar to PCa, THEM6 loss significantly impairs tumorigenesis in the MYC-dependent subtype of triple negative breast cancer. Altogether our results highlight THEM6 as a novel component of the treatment-induced stress response and a promising target for the treatment of CRPC and MYC-driven cancer.
THEM6-mediated lipid remodelling sustains stress resistance in cancer (Part 3)
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Despite the clinical benefit of androgen-deprivation therapy (ADT), the majority of patients with advanced prostate cancer (PCa) ultimately develop lethal castration-resistant prostate cancer (CRPC). In this study, we identified thioesterase superfamily member 6 (THEM6) as a marker of ADT resistance in PCa. In patients, THEM6 expression correlates with progressive disease and is associated with poor survival. THEM6 deletion reduces in vivo tumour growth and restores castration sensitivity in orthograft models of CRPC. Mechanistically, THEM6 is located at the endoplasmic reticulum (ER) membrane and controls lipid homeostasis by regulating intracellular levels of ether lipids. As a consequence, THEM6 loss in CRPC cells significantly alters ER function, preventing lipid-mediated induction of ATF4 and reducing de novo sterol biosynthesis. Finally, we show that THEM6 is required for the establishment of the MYC-induced stress response. Thus, similar to PCa, THEM6 loss significantly impairs tumorigenesis in the MYC-dependent subtype of triple negative breast cancer. Altogether our results highlight THEM6 as a novel component of the treatment-induced stress response and a promising target for the treatment of CRPC and MYC-driven cancer.
Metabolomics of the interaction between a consortium of entomopathogenic fungi and their target insect: mechanisms of attack and survival
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
One of the most concerning pests that attack strawberries in Brazil is Duponchelia fovealis, a non-native moth with no registered control methods to date. Our group recently observed that a fungal consortium formed by two strains of Beauveria bassiana increased the mortality of D. fovealis more than inoculation with each strain on its own. However, the molecular interaction between the fungal consortium and the caterpillars is unknown, raising several questions about the enhanced pest control observed. Furthermore, concerns over the emergency of resistance and the selection for resistance to chemical and biological products that are constantly applied in agriculture highlight the need for careful examination of novel pest control methods. Thus, in this work, we sought to pioneer the evaluation of the molecular interaction between a fungal consortium of B. bassiana and D. fovealis caterpillars. We aimed to understand the biocontrol process involved in this interaction and the defense system of the caterpillar. Therefore, seven days after D. fovealis caterpillars were inoculated with the B. bassiana consortium, the dead and surviving caterpillars were analyzed using GC-MS and LC-MS/MS.
INSTITUTE
Universidade Federal do ParanĆ”
DEPARTMENT
Patologia BƔsica
LABORATORY
LaboratĆ³rio de Microbiologia e Biologia Molecular
LAST_NAME
Katiski da Costa Stuart
FIRST_NAME
Andressa
ADDRESS
Av. Cel. Francisco HerƔclito dos Santos, 100, Curitiba, ParanƔ, 81530-000, Brazil
Dynamics of bile acid metabolism between the host and gut microbiome in progression to islet autoimmunity (Blood)
STUDY_SUMMARY
Previous studies suggest that the human gut microbiome is dysregulated in islet autoimmunity, preceding the clinical onset of type 1 diabetes (T1D). The Gut microbiota of the gut plays an important role in the regulation of bile acid (BA) metabolism. However, not much is known about the regulation of BAs during progression to T1D. Here, we analyzed BAs in a longitudinal series of serum (n= 333) and stool (n= 304) samples, collected at 3, 6, 12, 18, 24 and 36 months of age, from children who developed a single islet autoantibody (P1Ab), multiple islet autoantibodies (P2Ab), and controls (CTRs) who remained autoantibody (AAb) negative during the follow-up. In addition, we analyzed the stool microbiome by shotgun metagenomics in a subgroup of these children (n=111). Factor analysis showed that age had the strongest impact on BA and microbiome profiles. We found that, at an early age, the systemic BA (including taurine and glycine conjugates) and microbial secondary BA pathways were altered in the P2Ab group as compared to the P1Ab or CTR groups. Our findings thus suggest that dysregulated BA metabolism in early life may contribute to the risk and pathogenesis of T1D.
Dynamics of bile acid metabolism between the host and gut microbiome in progression to islet autoimmunity (Feces)
STUDY_SUMMARY
Previous studies suggest that the human gut microbiome is dysregulated in islet autoimmunity, preceding the clinical onset of type 1 diabetes (T1D). The Gut microbiota of the gut plays an important role in the regulation of bile acid (BA) metabolism. However, not much is known about the regulation of BAs during progression to T1D. Here, we analyzed BAs in a longitudinal series of serum (n= 333) and stool (n= 304) samples, collected at 3, 6, 12, 18, 24 and 36 months of age, from children who developed a single islet autoantibody (P1Ab), multiple islet autoantibodies (P2Ab), and controls (CTRs) who remained autoantibody (AAb) negative during the follow-up. In addition, we analyzed the stool microbiome by shotgun metagenomics in a subgroup of these children (n=111). Factor analysis showed that age had the strongest impact on BA and microbiome profiles. We found that, at an early age, the systemic BA (including taurine and glycine conjugates) and microbial secondary BA pathways were altered in the P2Ab group as compared to the P1Ab or CTR groups. Our findings thus suggest that dysregulated BA metabolism in early life may contribute to the risk and pathogenesis of T1D.
CE-MS based metabolomics to study plasma samples that reveal new pathways implicated in SARS-CoV-2 pathogenesis
STUDY_SUMMARY
CE-MS based metabolomics was used in this study to analize COVID-19 disease and the susceptibility to SARS-CoV-2. In total, 63 plasma samples were analyzed and different comparisons were performed. To our knowledge, CE-MS has never been used to study COVID-19 and it is considered in this study an appropriate approach to extend the polar metabolome beyond what has been obtained by LC-MS and GC-MS
Mutasynthetic production and antimicrobial characterisation of Darobactin darobactin analogs (NMR analysis)
STUDY_SUMMARY
There is great need for therapeutics against multi-drug resistant, Gram-negative bacterial pathogens. Recently, darobactin A, a novel bicyclic heptapeptide that selectively kills Gram-negative bacteria by targeting the outer-membrane protein BamA, was discovered. Its efficacy was proven in animal infection models of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, thus promoting darobactin A as a promising lead compound. Originally discovered from members of the nematode symbiotic genus Photorhabdus, the biosynthetic gene cluster (BGC) encoding for the synthesis of darobactin A can also be found in other Ī³-proteobacterial families. Therein, the precursor peptides DarB-F, which differ in their core sequence from darobactin A, were identified in silico. Even though production of these analogs was not observed in the putative producer strains, we were able to generate them by mutasynthetic derivatization of a heterologous expression system. The generated analogs were isolated and tested for their bioactivity. The most potent compound, darobactin B, was used for co-crystallization with the target BamA, revealing an identical binding site to darobactin A. Besides its potency, darobactin B did not exhibit cytotoxicity and was slightly more active against Acinetobacter baumanii isolates than darobactin A. Furthermore, we evaluated the plasma protein binding of darobactin A and B, indicating their different pharmacokinetic properties. This is the first report on new members of this new antibiotics class, which is likely to expand to several promising therapeutic candidates
Mutasynthetic production and antimicrobial characterisation of Darobactin darobactin analogs (MS analysis)
STUDY_SUMMARY
There is great need for therapeutics against multi-drug resistant, Gram-negative bacterial pathogens. Recently, darobactin A, a novel bicyclic heptapeptide that selectively kills Gram-negative bacteria by targeting the outer-membrane protein BamA, was discovered. Its efficacy was proven in animal infection models of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, thus promoting darobactin A as a promising lead compound. Originally discovered from members of the nematode symbiotic genus Photorhabdus, the biosynthetic gene cluster (BGC) encoding for the synthesis of darobactin A can also be found in other Ī³-proteobacterial families. Therein, the precursor peptides DarB-F, which differ in their core sequence from darobactin A, were identified in silico. Even though production of these analogs was not observed in the putative producer strains, we were able to generate them by mutasynthetic derivatization of a heterologous expression system. The generated analogs were isolated and tested for their bioactivity. The most potent compound, darobactin B, was used for co-crystallization with the target BamA, revealing an identical binding site to darobactin A. Besides its potency, darobactin B did not exhibit cytotoxicity and was slightly more active against Acinetobacter baumanii isolates than darobactin A. Furthermore, we evaluated the plasma protein binding of darobactin A and B, indicating their different pharmacokinetic properties. This is the first report on new members of this new antibiotics class, which is likely to expand to several promising therapeutic candidates.
Polyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells (Part 1)
STUDY_SUMMARY
Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naĆÆve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1ļ¢ļ or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Polyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells (Part 2)
STUDY_SUMMARY
Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naĆÆve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1ļ¢ļ or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Polyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells (Part 3)
STUDY_SUMMARY
Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naĆÆve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Polyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells (Part 4)
STUDY_SUMMARY
Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naĆÆve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
In order to identify metabolites descriptive of alterations of the working themperature during the process of anaerobic digestion, we performed untargeted metabolomics on samples of sewage sludge collected from two reactors working in parallel but with different temperature settings.
INSTITUTE
INRAE
LAST_NAME
Chapleur
FIRST_NAME
Olivier
ADDRESS
1 rue Pierre-Gilles de Gennes, 92761 Antony Cedex, FRANCE
Isotope tracing analysis of stress erythroid progenitors
STUDY_SUMMARY
Isotope tracing analysis to study the intracellular metabolic changes of progenitors during the expansion stage of stress erythropoiesis and assess the effect of 1400w treatment.
INSTITUTE
Pennsylvania State University
DEPARTMENT
Veterinary and Biomedical Sciences
LABORATORY
Paulson Lab
LAST_NAME
Ruan
FIRST_NAME
Baiye
ADDRESS
228 AVBS Building Shortlidge Road University Park, PA 16802
Glycine betaine uptake and metabolism in marine microbial communities
STUDY_TYPE
Quantitative and qualitative exploration of isotope-labeled glycine betaine uptake and use in natural marine microbial communities
STUDY_SUMMARY
Glycine betaine (GBT) is a component of labile dissolved organic matter and a compatible solute in high concentrations in marine microbial populations. GBT has complex biochemical potential, but, once taken up from the environment, the cellular fate of the carbon and nitrogen from GBT is unknown. Here we determine the uptake kinetics and metabolism of GBT in two natural microbial communities characterized by different nitrate concentrations in the North Pacific transition zone. Dissolved GBT had maximum uptake rates of 0.36 and 0.56 nM hr -1 and half-saturation constants of 79 and 11 nM in the high nitrate and low nitrate stations, respectively. GBT taken into cells was predominantly retained as an untransformed compatible solute. A portion of GBT was transformed into other metabolites, through characterized and uncharacterized pathways. Where nitrate was scarce, GBT was primarily catabolized via the demethylation to glycine. Resulting metabolites were used to build protein biomass, and remineralized ammonia was re-assimilated into cells. Gene expression data from this region show that bacteria, especially SAR11, are the dominant organisms expressing the demethylation genes. Where nitrate concentrations were higher, more GBT was used for choline synthesis. Our data highlight undiscussed metabolic pathways and potential routes of microbial metabolite exchange.
Chemoresistance remains the major barrier to effective ovarian cancer treatment. The molecular features and associated biological functions of this phenotype remain poorly understood. We developed carboplatin resistant cell line models using OVCAR5 and CaOV3 cell lines with the aim of identifying chemoresistance-specific molecular features. Mass spectrometry analysis was used to analyse the metabolome of these cell lines and was able to separate these populations based on their molecular features. It revealed signaling and metabolic perturbations in chemoresistant cell lines. A comprehensive analysis of a larger patient cohort, including advanced in vitro and in vivo models, promises to help better understand the molecular mechanisms of chemo-resistance and associated enhancement of migration and invasion.
INSTITUTE
The University of South Australia
LAST_NAME
Acland
FIRST_NAME
Mitchell
ADDRESS
Cnr North Terrace and Morphett Street, Adelaide SA 5000
The anticancer human mTOR inhibitor MLN0128/Sapanisertib with potent multistage in vitro antiplasmodium activity and in vivo antimalarial efficacy in a humanised mouse model is an inhibitor of multiple Plasmodium falciparum kinases.
STUDY_SUMMARY
Here we interrogated the in vitro metabolic effects of 6 drugs using ultra-high performance liquid chromatography mass-spectrometry (UHPLC-MS). The resulting metabolic fingerprints provide information on the parasite biochemical pathways affected by pharmacologic intervention and offer a critical blueprint for selecting and advancing lead compounds as next-generation antimalarial drugs. Our results reveal several distinctions between compounds with polypharmacological effects.
INSTITUTE
Pennsylvania State University
DEPARTMENT
Department of Biochemistry and Molecular Biology
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millenium Science Complex, University Park, PA 16802
Untargeted primary metabolite profiling in Arabidopsis thaliana
STUDY_SUMMARY
The goal of this project is to compare the primary metabolite profile in different tissue types of the model plant Arabidopsis thaliana. Specifically, plants were grown hydroponically under the long-day (16hr light/day) condition at 21C. Tissue samples, including leaves, inflorescences, and roots were harvest 4 1/2 weeks post sowing. Untargeted primary metabolites profiling was carried out using GCTOF.
Impact of microcin J25 on the porcine microbiome in a continuous culture model
STUDY_SUMMARY
The increased prevalence of Salmonella spp. resistance in swine spurs the search for alternatives to antibiotics. Microcin J25 (MccJ25), a bacteriocin produced by Escherichia coli, is a potent inhibitor of several pathogenic bacteria including Salmonella enterica. In this study, we aimed to evaluate in vitro the impact of MccJ25 on the metabolic activity of the swine colonic microbiota. The PolyFermS in vitro continuous fermentation model was used with modified Macfarlane medium to simulate the porcine proximal colon. During 35 days of fermentation, a first-stage reactor containing immobilized swine fecal microbiota fed two second-stage control and test reactors operated in parallel and used to test the effectsof MccJ25 on the composition and the metabolic activity of the microbiota. Reuterin, a broad spectrum antimicrobial produced by Limosilactobacillus reuteri and the antibiotic rifampicin were tested for comparison. LC-MS analysis of the cell extracts was used to assess the bacteriocin/antibiotic degradation products and monitor changes in the swine colonic microbiota metabolome.
Dysregulated Alanine as a Potential Predictive Marker of GliomaāAn Insight from Untargeted HRMAS-NMR and Machine Learning Data
STUDY_TYPE
Untargeted NMR
STUDY_SUMMARY
Metabolic alterations play a crucial role in glioma development and progression and can be detected even before the appearance of the fatal phenotype. We have compared the circulating metabolic fingerprints of glioma patients versus healthy controls, for the first time, in a quest to identify a panel of small, dysregulated metabolites with potential to serve as a predictive and/or diagnostic marker in the clinical settings. High-resolution magic angle spinning nuclear magnetic resonance spectroscopy (HRMAS-NMR) was used for untargeted metabolomics and data acquisition followed by a machine learning (ML) approach for the analyses of large metabolic datasets. Crossvalidation of ML predicted NMR spectral features was done by statistical methods (Wilcoxon-test) using JMP-pro16 software. Alanine was identified as the most critical metabolite with potential to detect glioma with precision of 1.0, recall of 0.96, and F1 measure of 0.98. The top 10 metabolites identified for glioma detection included alanine, glutamine, valine, methionine, N-acetylaspartate (NAA), Ī³-aminobutyric acid (GABA), serine, Ī±-glucose, lactate, and arginine. We achieved 100% accuracy for the detection of glioma using ML algorithms, extra tree classifier, and random forest, and 98% accuracy with logistic regression. Classification of glioma in low and high grades was done with 86% accuracy using logistic regression model, and with 83% and 79% accuracy using extra tree classifier and random forest, respectively. The predictive accuracy of our ML model is superior to any of the previously reported algorithms, used in tissue- or liquid biopsy-based metabolic studies. The identified top metabolites can be targeted to develop early diagnostic methods as well as to plan personalized treatment strategies.
INSTITUTE
University of the Punjab
DEPARTMENT
School of Biochemistry and Biotechnology
LABORATORY
Biopharmaceuticals and Biomarkers Discovery Lab
LAST_NAME
Firdous
FIRST_NAME
Safia
ADDRESS
Quaid e Azam Campus, University of the Punjab, Lahore.
EMAIL
saima.ibb@pu.edu.pk
PHONE
+924299231098
NUM_GROUPS
2
TOTAL_SUBJECTS
42
NUM_MALES
25
NUM_FEMALES
17
PUBLICATIONS
Dysregulated Alanine as a Potential Predictive Marker of GliomaāAn Insight from Untargeted HRMAS-NMR and Machine Learning Data
Untargeted metabolite analysis was performed on a Thermo Orbitrap IDX Tribrid MS to understand changes in metabolites due to COVID severity.
INSTITUTE
University of Virginia
DEPARTMENT
1Department of Biochemistry & Molecular Genetics; School of Medicine Core Facilities; Department of Microbiology, Immunology, and Cancer Biology; Department of Biomedical Engineering
LABORATORY
Biomolecular Analysis Facility, Univ of Virginia School of Medicine
LAST_NAME
Wase
FIRST_NAME
Nishikant
ADDRESS
Biomolecular Analysis Facility, Pinn Hall Room No 1105B
Multi-omic analysis of the microbiome and metabolome in healthy subjects (blood)
STUDY_SUMMARY
We conducted multi-omic phenotyping of healthy individuals, in order to investigate the interaction between diet, the microbiome, and the metabolome in a cross-sectional sample. We applied metabolomic profiling (at Metabolon Inc.) to plasma and stool samples in a subset of individuals (N=75).
INSTITUTE
Vanderbilt University Medical Center
LAST_NAME
Ferguson
FIRST_NAME
Jane
ADDRESS
2220 Pierce Ave, PRB 354, Nashville, TN, 37232, USA
Multi-omic analysis of the microbiome and metabolome in healthy subjects (feces)
STUDY_SUMMARY
We conducted multi-omic phenotyping of healthy individuals, in order to investigate the interaction between diet, the microbiome, and the metabolome in a cross-sectional sample. We applied metabolomic profiling (at Metabolon Inc.) to plasma and stool samples in a subset of individuals (N=75).
INSTITUTE
Vanderbilt University Medical Center
LAST_NAME
Ferguson
FIRST_NAME
Jane
ADDRESS
2220 Pierce Ave, PRB 354, Nashville, TN, 37232, USA
A prospective cohort of 38 European (EA) and African American (AA) omnivorous females were recruited. Samples were collected pre-intervention while subjects consumed habitual animal-based diet, and following 4 days of interventional vegetarian diet.
INSTITUTE
Vanderbilt University Medical Center
LAST_NAME
Ferguson
FIRST_NAME
Jane
ADDRESS
2220 Pierce Ave, PRB 354, Nashville, TN, 37232, USA
A prospective cohort of 38 European (EA) and African American (AA) omnivorous females were recruited. Samples were collected pre-intervention while subjects consumed habitual animal-based diet, and following 4 days of interventional vegetarian diet.
INSTITUTE
Vanderbilt University Medical Center
LAST_NAME
Ferguson
FIRST_NAME
Jane
ADDRESS
2220 Pierce Ave, PRB 354, Nashville, TN, 37232, USA
An integrated-omics approach reveals specific bacterial and fungal taxa associated with roots of Alkanna tinctoria L. Tausch correlating with medicinally relevant alkannin derivatives and other secondary metabolites
STUDY_SUMMARY
Plants are naturally associated with diverse microbial communities, which play significant roles in plant performance, such as growth promotion or fending off pathogens. The roots of Alkanna tinctoria L. are rich in naphthoquinones, particularly the medicinally used chiral compounds alkannin, shikonin and their derivatives. Former studies already have shown that microorganisms may modulate plant metabolism. To further investigate the potential interaction between A. tinctoria and associated microorganisms we performed a greenhouse experiment, in which A. tinctoria plants were grown in the presence of three distinct soil microbiomes. At four defined plant developmental stages we made an in-depth assessment of bacterial and fungal root-associated microbiomes as well as all primary and secondary metabolites. Our results showed that the plant developmental stage was the most important driver influencing the plant metabolite content, revealing peak contents of alkannin/shikonin at the fruiting stage. In contrast, the soil microbiome had the biggest impact on the plant root microbiome. Correlation analyses performed on the measured metabolite content and the abundance of individual bacterial and fungal taxa suggested a dynamic, at times positive or negative relationship between root-associated microorganisms and root metabolism. In particular, the bacterial Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium group and the fungal species Penicillium jensenii were found to be positively correlated with higher content of alkannins.
INSTITUTE
Aristotle University of Thessaloniki, MICROMETABOLITE project
A Sentinel Serum Quality Management Program for NMR Metabolomics
STUDY_SUMMARY
Pooled human serum (Sentinel Serum Samples) was aliquoted for storage at -80C and subsequently had NMR spectra acquired annually for three years. This was to determine sentinel serum sample utility for a quality management program to monitor NMR reproducibility. Sentinel serum quality control samples allowed the quantification of variance between different profilers within a project. They also demonstrated declines in metabolite concentrations over time. A sentinel program is useful for tracking changes in QC serum samples from long-term storage. Separately, QC samples show negligible variance introduced by multiple profilers of NMR spectra within a project.
INSTITUTE
University of Michigan
DEPARTMENT
Clinical Pharmacy
LABORATORY
UMich NMR Metabolomics Laboratory
LAST_NAME
Stringer
FIRST_NAME
Kathleen
ADDRESS
428 Church Street
EMAIL
stringek@med.umich.edu; stringek@umich.edu
NUM_GROUPS
3
TOTAL_SUBJECTS
1
STUDY_COMMENTS
Samples were aliquots of a batch of pooled human serum, collected from multiple different individuals
A targeted metabolomics study for assessing rodent thyroid toxicity
STUDY_SUMMARY
The thyroid gland regulates various physiological mechanisms in mammals/humans, such as individual development, cell proliferation and differentiation. Thus, disorders can lead to diseases. In this study, we aimed to apply targeted metabolomics approach to investigate rodent thyroid toxicity. For this purpose, male Wistar rats have been exposed to a direct (6-propyl-2-thiouracil, PTU) and an indirect (phenytoin) thyroid toxicant, respectively. Thereby, two doses (low:5ppm for PTU and 300ppm for phenytoin , high: 50ppm for PTU and 2400ppm for phenytoin) and three exposure time phase (short: 2 weeks, long:4 weeks, and long+recovery: 4weeks+2weeks) were investigated, allowing insights into the modes of action during thyroid toxicity. Targeted metabolomics were applied to both liver and thyroid gland tissue.
Plasmodium falciparum stable-isotope carbon labeling to explore metabolic consequences of ketoāacid dehydrogenase disruption
STUDY_SUMMARY
Plasmodium falciparum cells in culture were treated with respective universally labelled carbon-13 metabolites for 2.5 hours at standard culture concentrations (glucose or glutamine) or 5 mM (acetate). Metabolites were isolated using 90% methanol, dried, reconstituted in HPLC-grade water, and analyzed by HPLC/MS. Resulting data were analyzed and compiled to generate study data.
INSTITUTE
Pennsylvania State University
DEPARTMENT
Chemistry
LABORATORY
LlinƔs Laboratory
LAST_NAME
LlinƔs
FIRST_NAME
Manuel
ADDRESS
W126 Millennium Science Complex, University Park, PENNSYLVANIA, 16802, USA
Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Serum/GC)
STUDY_SUMMARY
Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Stool/GC)
STUDY_SUMMARY
Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Serum/HILIC)
STUDY_SUMMARY
: Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Stool/HILIC)
STUDY_SUMMARY
: Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Serum/Bile acids)
STUDY_SUMMARY
Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Metabolomics Analysis of Blood Plasma and Stool from Six Week Flaxseed Dietary Intervention in Postmenopausal Women (Stool/Bile acids)
STUDY_SUMMARY
Samples were taken before and after 6 weeks of flaxseed dietary intervention. Serum and fecal metabolomics were performed to characterize the metabolomic profiles of serum and stool metabolites pre- and post-intervention.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Whole blood)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Blood plasma)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
INSTITUTE
University of Colorado Anschutz School of Medicine
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Prestool)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
INSTITUTE
University of Colorado Anschutz School of Medicine
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Poststool)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Heart)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Kidney)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Liver)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Duodenum)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Brain)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Colon)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
INSTITUTE
University of Colorado Anschutz School of Medicine
Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Spleen)
STUDY_SUMMARY
Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.
This project seeks to understand the metabolic consequences of gestational hypoxia on fetal, newborn, and adult plasma, arteries and other tissues using a sheep model of fetal growth restriction. Specifically we are interested testing the hypothesis that gestational hypoxia will result in discernable differences in glucose and lipid metabolism in tissues and plasma as well influence indicators of oxidative stress and inflammation. These studies aim to delineate pathways and biomarkers that help explain how hypoxia leads to the development of neonatal as well as adult-onset diseases associated with chronic hypoxia that are inter-related with fetal growth restriction. From a vascular perspective this includes cerebrovascular hemorrhage and pulmonary hypertension in the newborn, but more broadly it includes development of diseases later in life including diabetes, hypertension, and coronary artery disease.
INSTITUTE
LOMA LINDA UNIVERSITY | School of Medicine
DEPARTMENT
Lawrence D. Longo, MD Center for Perinatal Biology
LABORATORY
Sean Wilson, Center for Perinatal Biology
LAST_NAME
Wilson
FIRST_NAME
Sean
ADDRESS
11234 Anderson Street, MC A582, Loma Linda, California 92350
A Taguchi Design of Experiments Approach for Untargeted Metabolomics Sample Preparation Optimization
STUDY_TYPE
Design of Experiments - Extraction Optimization
STUDY_SUMMARY
We describe an efficient Taguchi method design of experiments (DOE) approach to optimize the extraction solvent and volume, extraction time, and LC reconstitution solvent for a sequential non-polar and polar Caenorhabditis elegans extraction. DOE is rarely used in metabolomics yet provides a systematic approach for optimizing sample preparation while simultaneously decreasing the number of experiments required to obtain high-quality data.
Lyso-lipid induced oligodendrocytes maturation underlie restoration of optic nerve function
STUDY_SUMMARY
Protein hyper-deimination and deficiency of lyso-phospholipids (LPC 18:1) has been associated with the pathology of demyelinating disease in both humans and mice. We uncovered interesting biology of LPC 18:1, in which LPC 18:1 induced optic nerve function restoration through oligodendrocyte maturation and remyelination in mouse model systems. Our in vitro studies show LPC 18:1 protection against neuron-ectopic hyper-deimination and stimulation of oligodendrocyte maturation, while in vivo investigations recorded optic nerve function improvement following optic nerve injections of LPC 18:1, in contrast to LPC 18:0. Thus just a change in a single bond renders a dramatic alternation in biological function. The incorporation of isobaric C13-histidine in newly synthesized myelin proteins and quantitative proteome shifts are consistent with remyelination underlying restoration in optic nerve function. These results suggest that exogenous LPC 18:1 may provide a therapeutic avenue for stemming vision loss in demyelinating diseases.
Perfluoroalkyl Compounds and Child Metabolic Health - Healthy Start Cohort
STUDY_TYPE
Untargeted LC-MS Metabolomics Study
STUDY_SUMMARY
Healthy Start is a prospective, pre-birth cohort study that recruited pregnant participants from outpatient prenatal clinics at the University of Colorado Hospital between 2009 and 2014. Eligible participants were 16 years or older with singleton pregnancies, no history of stillbirth or extremely preterm birth (<25 weeks of gestation) and no serious medical conditions, and had not yet completed 24 weeks of gestation at the time of enrollment. Mothers completed two study visits during pregnancy (median gestational ages 17 and 27 weeks). Mother-child pairs were thereafter assessed at birth, and for the childās follow up, in mid-childhood (median age 4.8 years). For the present project, we will use data from 523 mother-child pairs of the Healthy Start cohort with available information on prenatal PFAS concentrations, available cord plasma samples at delivery, and outcomes of interest. Please contact Wei Perng at wei.perng@cuanschutz.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Healthy Start is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Parker CB; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Perfluoroalkyl Compounds and Child Metabolic Health (Project Viva)
STUDY_SUMMARY
Project Viva: Pregnant women were enrolled in Project Viva between 1999 and 2002 at their first prenatal visit at one of 8 obstetric clinics of Atrius Harvard Vanguard Medical Associates, a multispecialty group practice in eastern Massachusetts. Eligible mothers were fluent in English, had singleton gestations, were <22 weeks gestation, and had no plans to move away from the study area. Research staff performed in-person study visits with participating mothers in the first (median gestational age 9.9 weeks) and second (median gestational age 28.1 weeks) trimesters of pregnancy, and with mothers and children during the first few days after delivery, during infancy (median age 6.3 months), in early childhood (median age 3.3 years), mid-childhood (median age 7.7 years), and adolescence (median age13 years). In this analysis, we will use data from 188 mother-child pairs in Project Viva with available information on prenatal PFAS concentrations, available umbilical cord serum samples at delivery, and outcomes of interest. Please contact Nicole Bornkamp at Nicole_Bornkamp@harvardpilgrim.org for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Project Viva is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Parker CB; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
The apicomplexan parasite Toxoplasma gondii forms bradyzoite-containing tissue cysts that cause chronic and drug-tolerant infections.
STUDY_SUMMARY
The apicomplexan parasite Toxoplasma gondii forms bradyzoite-containing tissue cysts that cause chronic and drug-tolerant infections. Here, we developed a human myotube-based in vitro culture model of functionally mature tissue cysts. Metabolomic characterization of purified cysts reveals global changes that comprise increased levels of amino acids and decreased abundance of nucleobase- and tricarboxylic acid cycle-associated metabolites. In contrast to fast replicating tachyzoite forms of T. gondii these tissue cysts tolerate exposure to the aconitase inhibitor sodium fluoroacetate.
Multi-omic Attributes and Unbiased Computational Modeling for the Prediction of Immunomodulatory Potency of Mesenchymal Stromal Cells
STUDY_SUMMARY
Mesenchymal stromal cells (MSCs) are āliving medicinesā that continue to be evaluated in clinical trials to treat various clinical indications, yet remain unapproved. Because these cell therapies can be harvested from different tissue sources, are manufactured ex vivo, and are composed of highly responsive cells from donors of varying demographics, significant complexities limit the current understanding and advancements to clinical practice. However, we propose a model workflow used to overcome challenges by identifying multi-omic features that can serve as predictive therapeutic outcomes of MSCs. Here, features were identified using unbiased symbolic regression and machine learning models that correlated multi-omic datasets to results from in vitro functional assays based on putative mechanisms of action of MSCs. Together, this study provides a compelling framework for achieving the identification of candidate CQAs specific to MSCs that may help overcome current challenges, advancing MSCs to broad clinical use. This upload contain the metabolomic datasets, which were correlated with quality metrics, such as potency.
Resistance to NaFAc of in vitro maturated Toxoplasma gondii bradyzoites in human myotubes.
STUDY_SUMMARY
The apicomplexan parasite Toxoplasma gondii forms bradyzoite-containing tissue cysts that cause chronic and drug-tolerant infections. Here, we developed a human myotube-based in vitro culture model of functionally mature tissue cysts. In contrast to fast replicating tachyzoite forms of T. gondii these tissue cysts tolerate exposure to the aconitase inhibitor sodium fluoroacetate. These data characterize bradyzoites and tachyzoites treated with NaFAc.
Metabolomic Profiling of Human Pluripotent Stem Cell Differentiation into Lung Progenitors
STUDY_SUMMARY
Metabolism is vital to cellular function and tissue homeostasis during human lung development. In utero, embryonic pluripotent stem cells undergo endodermal differentiation towards a lung progenitor cell fate that can be mimicked in vitro using induced human pluripotent stem cells (hiPSCs) to study genetic mutations. To identify differences between wild type and surfactant protein B (SFTPB)-deficient cell lines during endoderm specification towards lung, we used an untargeted metabolomics approach to evaluate the developmental changes in metabolites. We found that the metabolites most enriched during the differentiation from pluripotent stem cell to lung progenitor cell, regardless of cell line, were sphingomyelins and phosphatidylcholines, two important lipid classes in fetal lung development. The SFTPB mutation had no metabolic impact on early endodermal lung development. The identified metabolite signatures during lung progenitor cell differentiation may be utilized as biomarkers for normal embryonic lung development.
Integrated Multilayer Omics Reveals the Genomic, Proteomic and Metabolic Influences of the Histidyl Dipeptides on Heart
STUDY_SUMMARY
Histidyl dipeptides, such as carnosine, present in a micro-millimolar ranges in the heart, are synthesized via the enzyme carnosine synthase (Carns). These dipeptides facilitate glycolysis by proton buffering, form conjugates with reactive aldehydes, such as acrolein, and attenuate ischemia and reperfusion injury. While these dipeptides exhibit multifunctional properties, a composite understanding of their roles in myocardium is lacking. To identify the landscape of histidyl dipeptide mediated responses in the heart, we used a triomics approach of genome wide RNA sequencing, global proteomics and unbiased metabolomics in the cardio specific Carns transgenic (Tg) mice and integrated the three data sets. Our result show higher myocardial levels of histidyl dipeptides lead to extensive changes in several microRNAs, which could target the expression of contractile proteins, beta-fatty acid oxidation and citric acid cycle (TCA) enzymes. Global proteomics shows, expression of contractile proteins, enzymes of beta-fatty acid oxidation and TCA cycle, were enriched in the CarnsTg heart. Under aerobic conditions, the CarnsTg hearts had lower levels of short and long-chain fatty acids and TCA cycle intermediate-succinic acid, whereas, under ischemic conditions the accumulation of fatty acids and TCA cycle intermediates were significantly attenuated in the CarnsTg heart. Integration of multiple data sets suggests that beta-fatty acid oxidation and TCA cycle pathways exhibited correlative changes in the CarnsTg hearts at all three levels. Our triomics approach shows histidyl dipeptides are critical regulators of myocardial structure, function and energetics.
Distinct Human Hepatocyte Lipidomics Profiles for Nonalcoholic Steatohepatitis and In Vitro-Induced Steatosis
STUDY_SUMMARY
Nonalcoholic steatohepatitis (NASH) is a severe form of steatotic liver injury that can be caused by a variety of stimuli and has a significant mortality rate. A common technique to induce in vitro steatosis involves culturing primary human hepatocytes (PHH) in a fatty acid-enriched media. This study compared the lipidome of PHH cultured in a fatty acid-enriched media to hepatocytes from patients with NASH and healthy controls to determine whether such culture techniques could generate a hepatocellular lipid profile similar to that observed in NASH patients. LC-MS lipidomics analysis of hepatocytes from patients with NASH revealed increases in the total cellular abundance of glycerolipids, phosphatidylcholines, phosphatidylethanolamines, phosphatidylglycerols, phosphatidylinositols and phosphatidylserines compared to healthy control hepatocytes. PHH cultured in a fatty acid-enriched environment demonstrated an increase in total lipid abundance, however, changes were limited to glycerolipids; in contrast to NASH hepatocytes, increases in the abundance of phospholipids were not observed.
Skeletal muscle (SkM, tibialis anterior and upper arm) and lung samples analyzed by LC-MS metabolomics. These tissue types are analyzed as healthy control, Mcherry+ tumor, or adjacent tissue.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Nemkov
FIRST_NAME
Travis
ADDRESS
12801 East 17th Ave. Research 1 South Rm 9121 Aurora CO 80045
Pollen metabolomics using Arabidopsis thaliana: Comparison of pollen at mature, hydration and germination stage
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
In this study, we investigated the differential metabolic pathway enrichment among mature, hydrated, and germinated pollen using untargeted metabolomics analysis. Integration of publicly available transcriptome with presented metabolome data revealed starch and sucrose metabolism was significantly increased during pollen hydration and germination. The alterations in central metabolism focusing on sugar, fatty acids, and lipids were analyzed in detail. Several metabolites, including palmitic acid, oleic acid, linolenic acid, quercetin, luteolin/kaempferol, and Ī³-aminobutyric acid (GABA), were elevated in the hydrated pollen, suggesting a potential role in activating pollen tube emergence. The metabolite levels of mature, hydrated, and germinated pollen, presented in this work provide insights on the molecular basis of pollen germination.
Glutamine flux in macrophages treated with stable-isotope labeled analog 4 mM (U-13C5) glutamine
STUDY_TYPE
Glutamine flux
STUDY_SUMMARY
107 BMDMs per group (Sirt3 WT and Sirt3 K223R)were seeded in 10cm plates and incubated in RPMI-1640 cell culture medium with 10% FBS. Prior to isotopic labeling, the medium was replaced with RPMI-1640 without glutamine for 4 hrs. Then stable-isotope labeled analog 4 mM (U-13C5) glutamine (Cambridge Isotope) was added together with IL-4 (20ng/ml) for 4 hrs.
INSTITUTE
Shanghai Jiao Tong University affiliated Renji Hospital
Endophytic bacteria are key players in the modulation of the secondary metabolome of Lithospermum officinale L.
STUDY_SUMMARY
Endophytic bacteria influence plant growth and development and therefore are an attractive resource for applications in agriculture. However, little is known about the impact of these microorganisms on secondary metabolite (SM) production by medicinal plants. Here we assessed, for the first time, the effects of root endophytic bacteria on the modulation of SMs in the medicinal plant Lithospermum officinale (Boraginaceae family), with a focus on the naphthoquinones alkannin/shikonin (A/S). The study was conducted using a newly developed in vitro system as well as in the greenhouse. Targeted and non-targeted metabolomics approaches were used and supported by expression analysis of the gene PGT, encoding a key enzyme in the A/S biosynthesis pathway. Three bacterial strains, Chitinophaga sp. R-73072, Xanthomonas sp. R-73098 and Pseudomonas sp. R-71838 induced a significant increase of diverse SMs, including A/S, in L. officinale in both systems, demonstrating the strength of our approach for screening A/S derivative-inducing bacteria. Our results highlight the impact of root-endophytic bacteria on secondary metabolism in plants and indicate that production of A/S derivatives in planta likely involves cross-modulation of different metabolic pathways that can be manipulated by bacterial endophytes.
Intravenous lipopolysaccharide infusion and the bovine metabolome
STUDY_SUMMARY
The effects of lipopolysaccharides (i.e., endotoxin; LPS) on metabolism are poorly defined in lactating dairy cattle experiencing hyperlipidemia. Our objective was to explore the effects of acute intravenous LPS administration on metabolism in late-lactation Holstein cows experiencing hyperlipidemia. Ten non-pregnant lactating Holstein cows (273 Ā± 35 d in milk) were administered a single bolus of saline (3 mL of saline; n = 5) or LPS (0.375 Ī¼g of LPS/kg of body weight; n = 5). Simultaneously, cows were intravenously infused a triglyceride emulsion and fasted for 16 h to induce hyperlipidemia in an attempt to model the periparturient period. Blood was sampled at routine intervals. Changes in circulating total fatty acid concentrations and inflammatory parameters were measured. Plasma samples were analyzed using untargeted lipidomics and metabolomics. Endotoxin increased circulating serum amyloid A, LPS-binding protein, and cortisol concentrations. Endotoxin administration decreased plasma lysophosphatidylcholine (LPC) concentrations and increased select plasma ceramide concentrations. These outcomes suggest modulation of the immune response and insulin action. Lipopolysaccharide decreased the ratio of phosphatidylcholine to phosphatidylethanomanine, which potentially indicate a decrease in the hepatic activation of phosphatidylethanolamine N-methyltransferase and triglyceride export. Endotoxin administration also increased plasma concentrations of pyruvic and lactic acids, and decreased plasma citric acid concentrations, which implicate the upregulation of glycolysis and downregulation of the citric acid cycle (i.e., the Warburg effect), potentially in leukocytes. Acute intravenous LPS administration decreased circulating LPC concentrations, modified ceramide and glycerophospholipid concentrations, and influenced intermediary metabolism in dairy cows experiencing hyperlipidemia.
Metabolic impact of anticancer drugs Pd2Spermine and Cispla-tin on the brain of healthy mice
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Palladium (Pd(II))-complexes are alternatives due to similar metal coordination and promising cytotoxic properties. Metabolomics can measure the metabolic response of drug-exposed tissues, unveiling insight into drug mechanisms and new markers of drug efficacy/toxicity. The present 1H NMR metabolomics study aims to characterize the in vivo response of the impact of a Pd(II)-complex with polyamine spermine (Pd2Spm), compared to cDDP, on polar metabolism of brain from healthy mice at 1, 12 and 48 h post-injection times.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Metabolic impact of anticancer drugs Pd2Spermine and Cisplatin on the brain of healthy mice
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Palladium (Pd(II))-complexes are alternatives due to similar metal coordination and promising cytotoxic properties. Metabolomics can measure the metabolic response of drug-exposed tissues, unveiling insight into drug mechanisms and new markers of drug efficacy/toxicity. The present 1H NMR metabolomics study aims to characterize the in vivo response of the impact of a Pd(II)-complex with polyamine spermine (Pd2Spm), compared to cDDP, on nonpolar metabolism of brain from healthy mice at 1, 12 and 48 h post-injection times.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Glutaminase inhibition impairs CD8 T cell activation in STK11/Lkb1 deficient lung cancer
STUDY_TYPE
Biomedical research
STUDY_SUMMARY
To characterize the impact of KRAS co-mutations KEAP1 and STK11/Lkb1 on the metabolic and immune microenvironment of lung adenocarcinoma in immune-intact models, we generated a cohort of genetically engineered mouse models (GEMMs) to reflect the diverse tumor suppressor landscape seen in patients. Mice carrying the KrasG12D allele (K)21 were crossed with Keap1flox/flox (KK)22 and/or Lkb1flox/flox (KKL, KL)23 mice and genetic recombination induced by intranasal inhalation of Ad5-CMV-Cre adenovirus. We interrogated the metabolites present in lung tumor nodules collected from cohorts of KK, KL and KKL mice.
INSTITUTE
The Walter and Eliza Hall Institute of Medical Research
LABORATORY
Kate Sutherland
LAST_NAME
Sarah
FIRST_NAME
Best
ADDRESS
1G, Royal Parade, Parkville VIC 3052, Australia
EMAIL
best@wehi.edu.au
PHONE
+61-3-9345-2452
NUM_GROUPS
5
TOTAL_SUBJECTS
25
NUM_MALES
17
NUM_FEMALES
8
STUDY_COMMENTS
L lobe of mice lung
PUBLICATIONS
Glutaminase inhibition impairs CD8 T cell activation in STK11/Lkb1 deficient lung cancer
Time-Resolved Metabolomics of a Mouse Model of Ovarian High-Grade Serous Carcinoma (LC-MS)
STUDY_SUMMARY
The dismally-low survival rate of ovarian cancer patients diagnosed with high-grade serous carcinoma (HGSC) emphasizes the lack of effective screening strategies. One major obstacle is the limited knowledge of the underlying mechanisms of HGSC pathogenesis at very early stages. Here, we present the first 10-month time-resolved serum metabolic profile of a triple mutant (TKO) HGSC mouse model, along with the spatial lipidome profile of its entire reproductive system. A high-coverage liquid chromatography mass spectrometry-based metabolomics approach was applied to longitudinally-collected serum samples from both TKO and TKO control mice, tracking metabolome and lipidome changes from disease onset until death. Spatial lipid distributions within the reproductive system were also mapped via ultrahigh-resolution matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, and compared with serum lipid profiles for various lipid classes. Altogether, our results show that the remodeling of lipid and fatty acid metabolism, amino acid biosynthesis, TCA cycle and ovarian steroidogenesis are critical components of HGSC onset and development. These metabolic alterations are accompanied by changes in energy metabolism, mitochondrial and peroxisomal function, redox homeostasis, and inflammatory response, collectively supporting tumorigenesis.
Mutant CHCHD10 causes an extensive metabolic rewiring that precedes OXPHOS dysfunction in a murine model of mitochondrial cardiomyopathy
STUDY_SUMMARY
Mitochondrial cardiomyopathies are fatal diseases, with no effective treatment. Alterations of heart mitochondrial function activate the mitochondrial integrated stress response (ISRmt), a transcriptional program affecting cell metabolism, mitochondrial biogenesis, and proteostasis. In humans, mutations in CHCHD10, a mitochondrial protein with unknown function, were recently associated with dominant multi-system mitochondrial diseases, whose pathogenic mechanisms remain to be elucidated. Here, in CHCHD10 knock-in mutant mice, we identify an extensive cardiac metabolic rewiring triggered by proteotoxic ISRmt. The stress response arises early on, before the onset of bioenergetic impairments, triggering a switch from oxidative to glycolytic metabolism, enhancement of transsulfuration and one carbon (1C) metabolism, and widespread metabolic imbalance. In parallel, increased NADPH oxidases elicit antioxidant responses leading to heme depletion. As the disease progresses, the adaptive metabolic stress response fails, resulting in fatal cardiomyopathy. Our findings suggest that early interventions to counteract metabolic imbalance could ameliorate mitochondrial cardiomyopathy associated with proteotoxic ISRmt.
Systemic impact of the expression of the mitochondrial alternative oxidase on Drosophila development
STUDY_SUMMARY
Despite the beneficial effects shown when the mitochondrial alternative oxidase AOX from Ciona intestinalis (Tunicata: Ascidiacea) is xenotopically expressed in mammalian and insect models, important detrimental outcomes have also been reported, raising concerns regarding its envisioned deployment as a therapy enzyme for human mitochondrial and related diseases. We have shown previously that AOX-expressing flies present a dramatic drop in pupal viability when the larvae are cultured on a low nutrient (LN) diet, indicating that AOX interferes with normal developmental metabolism. Here, we applied the metabolomics approach to gain insights into the molecular basis of the fatal developmental interaction between AOX expression and LN diet. We investigated the whole-body metabolome of AOX-expressing and control larvae cultured on SD and LN diets.
Lipidomic Comparison of 2D and 3D Colon Cancer Cell Culture Models
STUDY_TYPE
Untargeted Lipidomics
STUDY_SUMMARY
Altered lipid metabolism is one of the hallmarks of cancer. Cellular proliferation and de novo synthesis of lipids are related to cancer progression. In this study, we evaluated the lipidomic profile of two-dimensional (2D) monolayer and multicellular tumor spheroids from the HCT 116 colon carcinoma cell line. We utilized serial trypsinization on the spheroid samples to generate three cellular populations representing the proliferative, quiescent, and necrotic regions of the spheroid. This analysis enabled a comprehensive identification and quantification of lipids produced in each of the spheroid layer and 2D cultures. We show that lipid subclasses associated with lipid droplets form in oxygen-restricted and acidic regions of spheroids and are produced at higher levels than in 2D cultures. Additionally, sphingolipid production, which is implicated in cell death and survival pathways, is higher in spheroids relative to 2D cells. Finally, we show that increased numbers of lipids comprised of polyunsaturated fatty acids (PUFAs) are produced in the quiescent and necrotic regions of the spheroid. The lipidomic signature for each region and cell culture type highlights the importance of understanding the spatial aspects of cancer biology. These results provide additional lipid biomarkers in the tumor microenvironment that can be further studied during potential therapeutic studies which target pivotal lipid production pathways.
A non-dividing population with high pyruvate dehydrogenase kinase activity drives metabolic heterogeneity and tumorigenesis in the intestine
STUDY_SUMMARY
Although reprogramming of cellular metabolism is a hallmark of cancer, little is known about how metabolic reprogramming contributes to early stages of transformation. Here, we show that the histone deacetylase SIRT6 regulates tumor initiation during intestinal cancer by controlling glucose metabolism. Loss of SIRT6 results in increased number of intestinal stem cells (ISCs), which translates into enhanced tumor initiating potential in APCmin mice. More importantly, we found a metabolic compartmentalization within the intestinal epithelium and adenomas, where a rare population of cells exhibit features of Warburg-like metabolism characterized by high pyruvate dehydrogenase kinase (PDK) activity. Our results show that these cells are quiescent cells expressing +4 ISCs and enteroendocrine markers. Active glycolysis in these cells suppresses ROS accumulation and enhances their stem cell and tumorigenic potential. Our studies reveal that aerobic glycolysis represents a highly heterogeneous feature of cancer, and more importantly, they indicate that this metabolic adaptation occurs in non-dividing cells, suggesting a role for the Warburg effect beyond biomass production in tumors.
Profiling of the human intestinal microbiome and bile acids under physiologic conditions using an ingestible sampling device
STUDY_SUMMARY
15 human subjects were sample using an ingestible sampling device to target specific regions of the small intestine by using different capsule types (capsule types 1 to 4). Stool was also analyzed.
Metabolomic study of Escherichia coli K-12 MG1655 WT and its transcriptional regulator double mutants under anaerobic fermentation conditions
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Metabolomic analysis of Wildtype, fnr_arcA (FA), arcA_ihf (AI) and fnr_ihf (FI) mutants in glucose minimal media under anaerobic fermentation conditions during its exponential phase of growth. Three biological and two technical replicate samples (n=6) were harvested for each of the strains while growing in a bioreactor anaerobically at 37 degrees Celsius and 150 rpm. This study aims to characterize and compare the metabolic profiles of all these strains.
INSTITUTE
IIT Bombay
DEPARTMENT
Department of Chemical Engineering
LABORATORY
Systems Biology and Metabolic Engineering Laboratory (SBMEL)
LAST_NAME
Pal
FIRST_NAME
Ankita
ADDRESS
Department of Chemical Engineering, IIT Bombay, Mumbai, Maharashtra, 400076, India
Algal bacterial interactions in phycosphere microbial communities have important implications for the stability and productivity of algal biofuel systems, and algal metabolites are important mediators of those interactions. We characterized exometabolites and cell associated metabolites from the model diatom Phaeodactylum tricornutum across different growth stages.
Multiple modes of interfering with the activity of Plasmodium falciparum cytoplasmic isoleucyl-tRNA synthetase illustrate the enzyme is a promising antimalarial target.
STUDY_SUMMARY
Here we interrogated the in vitro metabolic effects of 6 drugs using ultra-high performance liquid chromatography mass-spectrometry (UHPLC-MS). The resulting metabolic fingerprints provide information on the parasite biochemical pathways affected by pharmacologic intervention. We interrogate the metabolic effects on the 3D7 parasite strain. The metabolic effects were compared to their parent strain. The metabolic fingerprints provided show that certain biochemical pathways are affected by the drug inhibitory effect.
INSTITUTE
Pennsylvania State University
DEPARTMENT
Department of Biochemistry and Molecular Biology
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millenium Science Complex, University Park, PA 16802
Defining the mammalian coactivation of hepatic 12-hour clock and lipid metabolism
STUDY_SUMMARY
The 12-hour clock coordinates lipid homeostasis, energy metabolism and stress rhythms via the transcriptional regulator XBP1. However, the biochemical and physiological basis for integrated control of the 12-hour clock and diverse metabolic pathways remains unclear. Here, we show that steroid receptor coactivator SRC-3 coactivates XBP1 transcription and regulates hepatic 12-hour cistrome and gene rhythmicity. Mice lacking SRC-3 show abnormal 12-hour rhythms in hepatic transcription, metabolic functions, systemic energetics, and rate-limiting lipid metabolic processes including triglyceride, phospholipid and cardiolipin pathways. Notably, 12-hour clock coactivation is not only preserved, with its cistromic activation priming ahead of the zeitgeber cue of light, but concomitant with rhythmic remodeling in the absence of food. These findings reveal that SRC-3 integrates the mammalian 12-hour clock, energy metabolism, and membrane and lipid homeostasis, and demonstrates a role for the 12-hour clock machinery as an active transcriptional mechanism in anticipating physiological and metabolic energy needs and stresses.
Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of Glutaminolysis
STUDY_TYPE
GCMS
STUDY_SUMMARY
Ferroptosis is a novel iron-dependent regulated cell death mechanism that affects cell metabolism; however, a detailed metabolomic analysis of ferroptotic cells is not yet available. Here, we elucidated the metabolome of H9c2 cardioblasts by gas chromatography-mass spectrometry during ferroptosis induced by RSL3, a GPX4 inhibitor, in the presence of ferrostatin-1 (a ferroptosis inhibitor), XJB-5-131 (a mitochondrial-targeted ROS scavenger), or TSM-1005-44 (a newly developed cellular ROS scavenger). Results demonstrated that RSL3 decreased the levels of amino acids involved in glutathione synthesis more than two-fold. In contrast, saturated fatty acids levels were markedly increased in RSL3-challenged cells, with no effects on unsaturated fatty acids. RSL3 significantly altered the levels of mitochondrial tricarboxylic acid cycle intermediates; isocitrate and 2-oxoglutarate were found to increase, whereas succinate was significantly decreased in RSL3-challenged cells. Ferrostatin-1, XJB-5-131, and TSM-1005-44 prevented RSL3-induced cell death and conserved the metabolomic profile of the cells. Since 2-oxoglutarate is involved in the regulation of ferroptosis, particularly through glutamine metabolism, we further assessed the role of glutaminolysis in ferroptosis in H9c2 cardioblasts. Genetic silencing of GLS1, which encodes the K-type mitochondrial glutaminase (glutaminase C), protected against ferroptosis in the early stage. In conclusion, our study demonstrates that RSL3-induced ferroptosis impairs the metabolome of H9c2 cardioblasts.
INSTITUTE
University of Puerto Rico, School of Medicine
DEPARTMENT
Physiology
LABORATORY
Cardiovascular Physiology, DR. Sabzali Javadov's Lab
LAST_NAME
Rodriguez-Graciani
FIRST_NAME
Keishla M
ADDRESS
Medical Sciences Campus, Main Building, 6th Floor, Department of Physiology, San Juan, Puerto Rico, 00936-5067, USA
Dynamic Lipidome Alterations Associated with Human Health, Disease, and Aging
STUDY_SUMMARY
Lipids comprise a complex mixture of molecules that play central but undercharacterized roles across a wide range of functions such as cell membrane maintenance, energy management, and cell signaling. Here, we describe a comprehensive longitudinal lipidomic profiling approach aiming to provide new physiological insights into aging, diabetes, inflammation, and cytokine regulations. By profiling the plasma lipidome to a depth of more than 800 lipid species across 1,546 samples collected from 109 subjects spanning up to 9 years (3.2 average), we identified a myriad of dysregulated lipid species highly associated with transitions from health to disease. Our data suggest distinct physiological roles of complex lipid subclasses including large and small triacylglycerols (TAG), ester- and ether-linked phosphatidylethanolamines (PE), lysophosphatidylcholines (LPC), and lysophosphatidylethanolamine (LPE). The dynamic changes in the plasma lipidome under the conditions of respiratory viral infections, insulin resistance (IR), and aging indicate a putative role of these different lipids in regulating immune homeostasis in health as well as in acute and chronic inflammation. Moreover, metabolically unhealthy subjects diagnosed with IR show (1) disturbed immune homeostasis and differences in specific lipid-clinical measure associations, (2) altered dynamics for particular complex lipids including TAGs, LPCs PEs, and PCs in response to acute infections, and (3) elevated levels of complex lipids such as TAGs and CEs and accelerated aging, highlighting the importance of context specific interpretation of lipid profiles. Overall, our study exemplifies the power of deep and quantitative lipidomics profiling in conjunction with other omics measures to provide new insights into lipidome dynamics in health and disease.
INSTITUTE
Stanford University
LAST_NAME
Hornburg
FIRST_NAME
Daniel
ADDRESS
Department of Genetics, Stanford University School of Medicine, Stanford, CA, 94305, USA
Predicting dying: a study of the metabolic changes and the dying process in patients with lung cancer
STUDY_TYPE
Observational study
STUDY_SUMMARY
Background: Accurately recognising that a person may be dying is central for improving their experience of care. Yet recognising dying is difficult and predicting dying frequently inaccurate. Methods: Serial urine samples from patients (n=112) with lung cancer were analysed using high resolution untargeted mass spectrometry. ANOVA and volcano plot analysis demonstrated metabolites that changed in the last weeks of life. Further analysis identified potential biological pathways affected. Cox lasso logistic regression was engaged to develop a multivariable model predicting the probability of survival within the last 30 days of life. Results: In total 124 metabolites changed. ANOVA analysis identified 93 metabolites and volcano plot analysis 85 metabolites. 53 metabolites changed using both approaches. Pathways altered in the last weeks included those associated with decreased oral intake, muscle loss, decreased RNA and protein synthesis, mitochondrial dysfunction, disrupted Ī²-oxidation and one carbon metabolism. Epinephrine and cortisol increased in the last 2 weeks and week respectively. A model predicting time to death using 7 metabolites had excellent accuracy (AUC= 0.86 at day 30, 0.88 at day 20 and 0.85 at day 10) and enabled classification of patients at low, medium and high risk of dying on a Kaplan-Meier survival curve. Conclusions: Metabolomic analysis identified metabolites and their associated pathways that change in the last weeks and days of life in patients with lung cancer. Prognostic tests based on the metabolites identified have the potential to change clinical practice and improve the care of dying patients.
INSTITUTE
University of Liverpool Institute of Life Course & Medical Sciences
LAST_NAME
Norman
FIRST_NAME
Brendan
ADDRESS
William Henry Duncan Building, 6 West Derby Street, Liverpool, UK. L7 8TX
Time-Resolved Metabolomics of a Mouse Model of High-Grade Serous Ovarian Cancer (MSI)
STUDY_SUMMARY
The dismally low survival rate of ovarian cancer patients diagnosed with high-grade serous carcinoma (HGSC) emphasizes the lack of effective screening strategies. One major obstacle is the limited knowledge of the underlying mechanisms of HGSC pathogenesis at very early stages. Here, we present the first 10-month time-resolved serum metabolic profile of a triple mutant (TKO) HGSC mouse model, along with the spatial lipidome profile of its entire reproductive system. A high-coverage liquid chromatography mass spectrometry-based metabolomics approach was applied to longitudinally collected serum samples from both TKO and TKO control mice, tracking metabolome and lipidome changes from disease onset until mouse death. Spatial lipid distributions within the reproductive system were also mapped via ultrahigh-resolution matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and compared with serum lipid profiles for various lipid classes. Altogether, our results show that the remodeling of lipid and fatty acid metabolism, amino acid biosynthesis, TCA cycle and ovarian steroidogenesis are critical components of HGSC onset and development. These metabolic alterations are accompanied by changes in energy metabolism, mitochondrial and peroxisomal function, redox homeostasis, and inflammatory response, collectively supporting tumorigenesis.
INSTITUTE
Georgia Institute of Technology
DEPARTMENT
School of Chemistry & Biochemistry
LABORATORY
Facundo M. Fernandez
LAST_NAME
Sah
FIRST_NAME
Samyukta
ADDRESS
School of Chemistry & Biochemistry, 901 Atlantic Dr
A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy
STUDY_SUMMARY
Chimeric antigen receptor (CAR)-T cell-based immunotherapy for cancer and immunological diseases has made great strides, but it still faces multiple hurdles. Finding the right molecular targets to engineer T cells toward a desired function has broad implications for the armamentarium of T cell-centered therapies. Here, we developed a dead-guide RNA (dgRNA)-based CRISPR activation screen in primary CD8+ T cells, and identified gain-of-function (GOF) targets for CAR-T engineering. Targeted knock-in or overexpression of a lead target, PRODH2, enhanced CAR-T-based killing and in vivo efficacy in multiple cancer models. Transcriptomics and metabolomics in CAR-T cells revealed that augmenting PRODH2 expression re-shaped broad and distinct gene expression and metabolic programs. Mitochondrial, metabolic and immunological analyses showed that PRODH2 engineering enhances the metabolic and immune functions of CAR-T cells against cancer. Together these findings provide a system for identification of GOF immune boosters, and demonstrate PRODH2 as a target to enhance CAR-T efficacy.
A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy(Part 1)
STUDY_SUMMARY
Chimeric antigen receptor (CAR)-T cell-based immunotherapy for cancer and immunological diseases has made great strides, but it still faces multiple hurdles. Finding the right molecular targets to engineer T cells toward a desired function has broad implications for the armamentarium of T cell-centered therapies. Here, we developed a dead-guide RNA (dgRNA)-based CRISPR activation screen in primary CD8+ T cells, and identified gain-of-function (GOF) targets for CAR-T engineering. Targeted knock-in or overexpression of a lead target, PRODH2, enhanced CAR-T-based killing and in vivo efficacy in multiple cancer models. Transcriptomics and metabolomics in CAR-T cells revealed that augmenting PRODH2 expression re-shaped broad and distinct gene expression and metabolic programs. Mitochondrial, metabolic and immunological analyses showed that PRODH2 engineering enhances the metabolic and immune functions of CAR-T cells against cancer. Together these findings provide a system for identification of GOF immune boosters, and demonstrate PRODH2 as a target to enhance CAR-T efficacy.
Serum NMR profiling reveals differential alterations in the lipoproteome induced by Pfizer-BioNTech vaccine in COVID-19ārecovered subjects and naĆÆve subjects
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
1H NMR spectra of sera have been used to define the changes induced by vaccination with Pfizer-BioNTech vaccine (2 shots, 21 days apart) in 10 COVID-19-recovered subjects and 10 COVID-19-naĆÆve subjects at different time points, starting from before vaccination, then weekly until 7 days after second injection, and finally 1 month after the second dose. The data show that vaccination does not induce any significant variation in the metabolome, whereas it causes changes at the level of lipoproteins. The effects are different in the COVID-19-recovered subjects with respect to the naĆÆve subjects, suggesting that a previous infection reduces the vaccine modulation of the lipoproteome composition.
Profiling metabolites and lipoproteins in COMETA, an Italian cohort of COVID-19 patients
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
1H NMR spectra of EDTA-plasma from 246 COVID-19-positive subjects in the acute phase of infection were compared to those of 94 COVID-19-recovered subjects. The two cohorts are largely different (discrimination accuracy > 93%) due to a pool of 16 metabolites and 74 lipoprotein parameters significantly up- or down-regulated in the patients and within the healthy range in the recovered subjects. In 28 post-acute COVID-19-positive patients, the metabolites levels are reverted back to normality whereas the lipoprotein parameters are still altered. Therefore, the metabolite biomarkers might be used as the timeliest sign of the individual response to treatment or spontaneous healing.
Plasma Metabolomic signatures of COPD in a SPIROMICS cohort
STUDY_TYPE
Observational Cohort
STUDY_SUMMARY
Plasma Metabolomic signatures of COPD in a SPIROMICS cohort The Subpopulations and Intermediate Outcome Measures in COPD Study (SPIROMICS) (ClinicalTrials.gov Identifier: NCT01969344) includes 2,771 subjects, aged 40-80 years with at least 20 pack-years of smoking. An additional 202 subjects were never smokers. Fasting blood drawn at the enrollment visit using a p100 tube. The first 648 subjects who returned for a 5-7 year visit (Visit 5) were selected for this study. The blood profiled were from the year 1 visit.
The NIH sponsored multicenter Genetic Epidemiology of COPD (COPDGene (ClinicalTrials.gov Identifier: NCT01969344) study was approved and reviewed by the institutional review board at all participating centers (1). All study participants provided written informed consent. This study enrolled 10,198 non-Hispanic white (NHW) and African American (AA) individuals from January 2008 until April 2011 (Phase 1) who were aged 45-80 with ā„10 pack-year smoking history and no exacerbations for greater than 30 days. In addition, 465 age and gender matched healthy individuals with no history of smoking were enrolled as controls (mostly at Phase 2). From July 2013 to July 2017, 5,697 subjects returned for an in-person 5-year visit. Each in-person visit included spirometry before and after albuterol, quantitative CT imaging of the chest, and blood sampling. From two clinical centers (National Jewish Health and University of Iowa) 1,136 subjects (1,040 NHW, 96 AA) participated in an ancillary study in which they provided fresh frozen plasma collected using an 8.5 ml p100 tube (Becton Dickinson) at Phase 2.
Commensal intestinal microbiota regulates host luminal proteolytic activity and intestinal barrier integrity through Ī²-glucuronidase activity (Part 2)
STUDY_SUMMARY
Proteases constitute the largest enzyme gene family in vertebrates with intracellular and secreted proteases having critical roles in cellular and organ physiology. Intestinal tract contains diverse set of proteases mediating digestion, microbial responses, epithelial and immune signaling. Transit of chyme through the intestinal tract results in significant suppression of proteases. Although endogenous protease inhibitors have been identified, the broader mechanisms underlying protease regulation in the intestinal tract remains unclear. The objective of this study was to determine microbial regulation of proteolytic activity in intestinal tract using phenotype of post-infection irritable bowel syndrome, a condition characterized by high fecal proteolytic activity. Proteases of host pancreatic origin (chymotrypsin like pancreatic elastase 2A, 3B and trypsin 2) drove proteolytic activity. Of the 14 differentially abundant taxa, high proteolytic activity state was characterized by complete absence of the commensal Alistipes putredinis. Germ free mice had very high proteolytic activity (10-fold of specific-pathogen free mice) which dropped significantly upon humanization with microbiota from healthy volunteers. In contrast, high proteolytic activity microbiota failed to inhibit it, a defect that corrected with fecal microbiota transplant as well as addition of A. putredinis. These mice also had increased intestinal permeability similar to that seen in patients. Microbiota Ī²-glucuronidases mediate bilirubin deconjugation and unconjugated bilirubin is an inhibitor of serine proteases. We found that high proteolytic activity patients had lower urobilinogen levels, a product of bilirubin deconjugation. Mice colonized with Ī²-glucuronidase overexpressing E. coli demonstrated significant inhibition of proteolytic activity and treatment with Ī²-glucuronidase inhibitors increased it. The findings establish that specific commensal microbiota mediates effective inhibition of host pancreatic proteases and maintains intestinal barrier function through the production of Ī²-glucuronidases. This suggests an important homeostatic role for commensal intestinal microbiota.
Serum lipids are associated with nonalcoholic fatty liver disease: a pilot case-control study in Mexico
STUDY_SUMMARY
A nested case-control study was conducted with a sample of 98 NAFLD cases and 100 healthy controls who are participating in an on-going, longitudinal study in Mexico. NAFLD cases were clinically confirmed using elevated liver enzyme tests and liver ultrasound or liver ultrasound elastography, after excluding alcohol abuse, and 100 controls were identified as having at least two consecutive normal alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (<ā40āU/L) results in a 6-month period, and a normal liver ultrasound elastography result in January 2018. Samples were analyzed on the Sciex Lipidyzer Platform and quantified with normalization to serum volume. As many as 1100 lipid species can be identified using the Lipidyzer targeted multiple-reaction monitoring list. The association between serum lipids and NAFLD was investigated using analysis of covariance, random forest analysis, and by generating receiver operator characteristic (ROC) curves.
INSTITUTE
University of California, Los Angeles
DEPARTMENT
Biological Chemistry
LABORATORY
UCLA Lipidomics
LAST_NAME
Williams
FIRST_NAME
Kevin
ADDRESS
BSRB 257, 615 Charles E. Young Drive S., Los Angeles, CA, 90095
Addressing batch effects in large-scale metabolomics with augmented experimental design and meta-analysis
STUDY_SUMMARY
Untargeted LC-MS study conducted using RP and HILIC chromatography on three groups of C. elegans: natural isolates, central metabolism mutants, and UDP-glucuronosyltransferase mutants. An augmented study design, rank transformation of the raw data, strict QA/QC followed by a meta-analysis was used for data analysis.
INSTITUTE
University of Georgia - Complex Carbohydrate Research Center
Obesity exacerbates inflammation upon lung injury; however, the mechanisms by which obesity primes pulmonary dysregulation prior to injury are not well studied. Notably, little is known about how obesity dysregulates pulmonary polyunsaturated fatty acid (PUFA) metabolism that is central to inflammation initiation and resolution. Herein, we first show with mass spectrometry that a high fat diet (HFD) administered to C57BL/6J mice increases the relative abundance of pulmonary PUFA-containing triglycerides and the concentration of PUFA-derived oxylipins, independent of an increase in total pulmonary PUFAs. Experiments with a genetic model of obesity did not recapitulate the effects of the HFD on the pulmonary oxylipin signature, suggesting a diet-driven effect. Subsequent pulmonary next-generation RNA sequencing showed complex and unique transcriptional regulation with a HFD. The HFD increased pathways related to glycerophospholipid metabolism, innate immunity, and inflammation including an elevation in B cell differentiation and signaling. Finally, computational integration of lipidomic with transcriptomic data revealed novel networks with the HFD between glycerophospholipid metabolism and B cell receptor signaling with specific oxylipins. Collectively, these data show obesity dysregulates pulmonary PUFA metabolism prior to lung injury, which may be a mechanism by which obesity primes the lungs to respond poorly upon infectious and/or inflammatory challenges.
Commensal intestinal microbiota regulates host luminal proteolytic activity and intestinal barrier integrity through Ī²-glucuronidase activity (Part 1)
STUDY_TYPE
MS
STUDY_SUMMARY
Proteases constitute the largest enzyme gene family in vertebrates with intracellular and secreted proteases having critical roles in cellular and organ physiology. Intestinal tract contains diverse set of proteases mediating digestion, microbial responses, epithelial and immune signaling. Transit of chyme through the intestinal tract results in significant suppression of proteases. Although endogenous protease inhibitors have been identified, the broader mechanisms underlying protease regulation in the intestinal tract remains unclear. The objective of this study was to determine microbial regulation of proteolytic activity in intestinal tract using phenotype of post-infection irritable bowel syndrome, a condition characterized by high fecal proteolytic activity. Proteases of host pancreatic origin (chymotrypsin like pancreatic elastase 2A, 3B and trypsin 2) drove proteolytic activity. Of the 14 differentially abundant taxa, high proteolytic activity state was characterized by complete absence of the commensal Alistipes putredinis. Germ free mice had very high proteolytic activity (10-fold of specific-pathogen free mice) which dropped significantly upon humanization with microbiota from healthy volunteers. In contrast, high proteolytic activity microbiota failed to inhibit it, a defect that corrected with fecal microbiota transplant as well as addition of A. putredinis. These mice also had increased intestinal permeability similar to that seen in patients. Microbiota Ī²-glucuronidases mediate bilirubin deconjugation and unconjugated bilirubin is an inhibitor of serine proteases. We found that high proteolytic activity patients had lower urobilinogen levels, a product of bilirubin deconjugation. Mice colonized with Ī²-glucuronidase overexpressing E. coli demonstrated significant inhibition of proteolytic activity and treatment with Ī²-glucuronidase inhibitors increased it. The findings establish that specific commensal microbiota mediates effective inhibition of host pancreatic proteases and maintains intestinal barrier function through the production of Ī²-glucuronidases. This suggests an important homeostatic role for commensal intestinal microbiota.
Addressing batch effects in large-scale metabolomics with augmented experimental design and meta-analysis (Part 1)
STUDY_SUMMARY
Untargeted NMR study conducted using a NEO 800āMHz Bruker NMR spectrometer where polar extracts were collected on three groups of C. elegans: natural isolates, central metabolism mutants, and UDP-glucuronosyltrasnferase mutants. An augmented design, rank transformation of the raw data, strict QA/QC followed by a meta-analysis was performed.
INSTITUTE
University of Georgia - Complex Carbohydrate Research Center
LABORATORY
Edison Lab
LAST_NAME
Garcia
FIRST_NAME
Brianna
ADDRESS
315 Riverbend Road, Athens, GA, 30602, USA
EMAIL
brianna.garcia@uga.edu
NUM_GROUPS
3
TOTAL_SUBJECTS
116
STUDY_COMMENTS
Three study groups of C. elegans strains were used: central metabolism mutants (CMM), UDP-glucuronosyltransferase (UGT) mutants, and natural isolates.
Addressing batch effects in large-scale metabolomics with augmented experimental design and meta-analysis
STUDY_SUMMARY
Untargeted NMR study conducted using a NEO 800āMHz Bruker NMR spectrometer where non-polar extracts were collected on three groups of C. elegans: natural isolates, central metabolism mutants, and UDP-glucuronosyltrasnferase mutants. An augmented design, rank transformation of the raw data, strict QA/QC followed by a meta-analysis was performed.
INSTITUTE
University of Georgia - Complex Carbohydrate Research Center
LABORATORY
Edison Lab
LAST_NAME
Garcia
FIRST_NAME
Brianna
ADDRESS
315 Riverbend Road, Athens, GA, 30602, USA
EMAIL
brianna.garcia@uga.edu
PHONE
7065424401
NUM_GROUPS
3
TOTAL_SUBJECTS
116
STUDY_COMMENTS
Three study groups of C. elegans strains were used: central metabolism mutants (CMM), UDP-glucuronosyltransferase (UGT) mutants, and natural isolates.
Functional metabolomics-based molecular profiling of acute and chronic hepatitis (Liver Metabolomics)
STUDY_SUMMARY
Non-alcoholic steatohepatitis (NASH) is a metabolic dysregulation triggered by an overload disrupting the hepatic tolerance to external molecules. With the complexity and diversity of hepatitis triggers, no effective clinical classification and treatment are available, and even using the same strategies or approaches for acute and chronic hepatitis. For us, it is really difficult to precisely diagnose and treat hepatitis accordingly. To overcome this challenge, we integrated metabolomic, lipidomics, transcriptomics and other life science frontier technologies for functional metabolomics studies, and pioneered the redefinition of hepatitis at the molecular level. Our findings suggested that acute hepatitis mainly interferes with purine metabolism and amino acids metabolism, while chronic hepatitis mainly causes disruption of hepatic bile acids and lipidome, especially glycerolipids. Based on the liver-gut axis, we also found that the metabolic regulation of the gut microbiota is another key factor for chronic hepatitis development. In conclusion, functional metabolomics enables the cognition of disease occurrence, development and regression from small molecule metabolic modifications and modulations, realizing the ultimate goal of treating diseases and improving population health through regulation of dysregulated metabolism
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Functional metabolomics-based molecular profiling of acute and chronic hepatitis (Serum Metabolomics)
STUDY_SUMMARY
Non-alcoholic steatohepatitis (NASH) is a metabolic dysregulation triggered by an overload disrupting the hepatic tolerance to external molecules. With the complexity and diversity of hepatitis triggers, no effective clinical classification and treatment are available, and even using the same strategies or approaches for acute and chronic hepatitis. For us, it is really difficult to precisely diagnose and treat hepatitis accordingly. To overcome this challenge, we integrated metabolomic, lipidomics, transcriptomics and other life science frontier technologies for functional metabolomics studies, and pioneered the redefinition of hepatitis at the molecular level. Our findings suggested that acute hepatitis mainly interferes with purine metabolism and amino acids metabolism, while chronic hepatitis mainly causes disruption of hepatic bile acids and lipidome, especially glycerolipids. Based on the liver-gut axis, we also found that the metabolic regulation of the gut microbiota is another key factor for chronic hepatitis development. In conclusion, functional metabolomics enables the cognition of disease occurrence, development and regression from small molecule metabolic modifications and modulations, realizing the ultimate goal of treating diseases and improving population health through regulation of dysregulated metabolism
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Functional metabolomics-based molecular profiling of acute and chronic hepatitis (Stool Metabolomics)
STUDY_SUMMARY
Non-alcoholic steatohepatitis (NASH) is a metabolic dysregulation triggered by an overload disrupting the hepatic tolerance to external molecules. With the complexity and diversity of hepatitis triggers, no effective clinical classification and treatment are available, and even using the same strategies or approaches for acute and chronic hepatitis. For us, it is really difficult to precisely diagnose and treat hepatitis accordingly. To overcome this challenge, we integrated metabolomic, lipidomics, transcriptomics and other life science frontier technologies for functional metabolomics studies, and pioneered the redefinition of hepatitis at the molecular level. Our findings suggested that acute hepatitis mainly interferes with purine metabolism and amino acids metabolism, while chronic hepatitis mainly causes disruption of hepatic bile acids and lipidome, especially glycerolipids. Based on the liver-gut axis, we also found that the metabolic regulation of the gut microbiota is another key factor for chronic hepatitis development. In conclusion, functional metabolomics enables the cognition of disease occurrence, development and regression from small molecule metabolic modifications and modulations, realizing the ultimate goal of treating diseases and improving population health through regulation of dysregulated metabolism
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Functional metabolomics-based molecular profiling of acute and chronic hepatitis (Human Serum Metabolomics)
STUDY_SUMMARY
Non-alcoholic steatohepatitis (NASH) is a metabolic dysregulation triggered by an overload disrupting the hepatic tolerance to external molecules. With the complexity and diversity of hepatitis triggers, no effective clinical classification and treatment are available, and even using the same strategies or approaches for acute and chronic hepatitis. For us, it is really difficult to precisely diagnose and treat hepatitis accordingly. To overcome this challenge, we integrated metabolomic, lipidomics, transcriptomics and other life science frontier technologies for functional metabolomics studies, and pioneered the redefinition of hepatitis at the molecular level. Our findings suggested that acute hepatitis mainly interferes with purine metabolism and amino acids metabolism, while chronic hepatitis mainly causes disruption of hepatic bile acids and lipidome, especially glycerolipids. Based on the liver-gut axis, we also found that the metabolic regulation of the gut microbiota is another key factor for chronic hepatitis development. In conclusion, functional metabolomics enables the cognition of disease occurrence, development and regression from small molecule metabolic modifications and modulations, realizing the ultimate goal of treating diseases and improving population health through regulation of dysregulated metabolism
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Functional metabolomics-based molecular profiling of acute and chronic hepatitis (Liver Lipidomics)
STUDY_SUMMARY
Non-alcoholic steatohepatitis (NASH) is a metabolic dysregulation triggered by an overload disrupting the hepatic tolerance to external molecules. With the complexity and diversity of hepatitis triggers, no effective clinical classification and treatment are available, and even using the same strategies or approaches for acute and chronic hepatitis. For us, it is really difficult to precisely diagnose and treat hepatitis accordingly. To overcome this challenge, we integrated metabolomic, lipidomics, transcriptomics and other life science frontier technologies for functional metabolomics studies, and pioneered the redefinition of hepatitis at the molecular level. Our findings suggested that acute hepatitis mainly interferes with purine metabolism and amino acids metabolism, while chronic hepatitis mainly causes disruption of hepatic bile acids and lipidome, especially glycerolipids. Based on the liver-gut axis, we also found that the metabolic regulation of the gut microbiota is another key factor for chronic hepatitis development. In conclusion, functional metabolomics enables the cognition of disease occurrence, development and regression from small molecule metabolic modifications and modulations, realizing the ultimate goal of treating diseases and improving population health through regulation of dysregulated metabolism
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Volatilomic compound identification in several Indonesian underutilized Zingiberaceae spices using SPME-GC/MS
STUDY_SUMMARY
To identify compounds in 12 minor Zingiberaceae spices grown in Indonesia using SPME-GC/MS
INSTITUTE
IPB University
DEPARTMENT
Departement of Food Science and Technology
LABORATORY
Food Chemistry
LAST_NAME
Yuliana
FIRST_NAME
Nancy Dewi
ADDRESS
Fakultas Teknologi Pertanian (FATETA) Institut Pertanian Bogor. Kampus IPB Darmaga Jl. Lingkar Akademik, Jawa Barat 16680, Telp./faksimili : (0251) 8626725
NC HHEAR Hub Pilot Study within the HHEAR Consortium
STUDY_TYPE
C18 Reversed-Phase Broad Spectrum Metabolomics
STUDY_SUMMARY
This data generation, and subsequent data analysis was conducted by the NC HHEAR Hub as part of a larger HHEAR Consortium Pilot Study. A goal of this study to compare the metabolite identifications and annotations provided by HHEAR Consortium Laboratories as well as non-HHEAR Consortium Laboratories. Ā Urine samples were provided by the Duke HHEAR Hub.Ā This included pools of 3 spot urines collected over 48 hours, which were selected based on maximum overlap of collected data on other exposure matrices (targeted PFAS panel (serum), passive air sampling, wristband data). Dr. Yuan Li of the NC HHEAR Hub had primary responsibility for the generation of the untargeted data.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Nutrition Research Institute
LABORATORY
Untargeted Resource Laboratory for the NC HHEAR Hub
LAST_NAME
Li
FIRST_NAME
Yuan
ADDRESS
Nutrition Research Institute, UNC-CH, 500 Laureate Way, Kannapolis, NC 28081
Chemoresistant Cancer Cell Lines are Characterized by Migratory, Amino Acid Metabolism, Protein Catabolism and IFN1 Signalling Perturbations
STUDY_TYPE
Biomedical research
STUDY_SUMMARY
Our analysis was able to separate chemoresistant cells from their parental cells based on their metabolomic and proteomic features and identified altered biological processes and pathways which are of further interest. Preliminary investigation of patient-derived cells highlighted the need to perform broad biological and molecular analyses, compre-hensive in vitro and in vivo studies, using a larger patient cohort to achieve a deeper and clinically relevant characterization of the molecular drivers of chemoresistance.
INSTITUTE
Future Industries Institute
LABORATORY
Manuela Klingler-Hoffmann
LAST_NAME
Acland
FIRST_NAME
Mitchell
ADDRESS
X Building, Mawson Lakes South Australia 5095
EMAIL
mitch.acland@gmail.com
PHONE
0448671141
NUM_GROUPS
4
TOTAL_SUBJECTS
12
NUM_MALES
NA
NUM_FEMALES
NA
STUDY_COMMENTS
OVCAR5 and CaOV3 cell lines and their carboplatin resistant cell lines
PUBLICATIONS
Chemoresistant Cancer Cell Lines are Characterized by Migra-tory, Amino Acid Metabolism, Protein Catabolism and IFN1 Signalling Perturbations
Genetic and chemical validation of Plasmodium falciparum aminopeptidase PfA-M17 as a drug target in the hemoglobin digestion pathway (Part 1)
STUDY_SUMMARY
Plasmodium falciparum, the causative agent of malaria, continues to remain a global health threat since these parasites are now resistant to all anti-malaria drugs used throughout the world. Accordingly, drugs with novel modes of action are desperately required to combat malaria. P. falciparum parasites infect human red blood cells where they digest the hosts main protein constituent, hemoglobin. Leucine aminopeptidase PfA-M17 is one of several aminopeptidases that have been implicated in the last step of this digestive pathway. Here we utilize both reverse genetics and a compound specifically designed to inhibit the activity of PfA-M17 to show that PfA-M17 is essential for P. falciparum survival as it provides parasites with free amino acids for growth, many of which are highly likely to originate from hemoglobin. We further show that our inhibitor is on-target for PfA-M17 and has the ability to kill parasites at nanomolar concentrations. Thus, in contrast to other hemoglobin-degrading proteases that have overlapping redundant functions, we validate PfA-M17 as a potential novel drug target.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Genetic and chemical validation of Plasmodium falciparum aminopeptidase PfA-M17 as a drug target in the hemoglobin digestion pathway (Part 2)
STUDY_SUMMARY
Plasmodium falciparum, the causative agent of malaria, continues to remain a global health threat since these parasites are now resistant to all anti-malaria drugs used throughout the world. Accordingly, drugs with novel modes of action are desperately required to combat malaria. P. falciparum parasites infect human red blood cells where they digest the hosts main protein constituent, hemoglobin. Leucine aminopeptidase PfA-M17 is one of several aminopeptidases that have been implicated in the last step of this digestive pathway. Here we utilize both reverse genetics and a compound specifically designed to inhibit the activity of PfA-M17 to show that PfA-M17 is essential for P. falciparum survival as it provides parasites with free amino acids for growth, many of which are highly likely to originate from hemoglobin. We further show that our inhibitor is on-target for PfA-M17 and has the ability to kill parasites at nanomolar concentrations. Thus, in contrast to other hemoglobin-degrading proteases that have overlapping redundant functions, we validate PfA-M17 as a potential novel drug target.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Genetic and chemical validation of Plasmodium falciparum aminopeptidase PfA-M17 as a drug target in the hemoglobin digestion pathway (Part 3)
STUDY_SUMMARY
Plasmodium falciparum, the causative agent of malaria, continues to remain a global health threat since these parasites are now resistant to all anti-malaria drugs used throughout the world. Accordingly, drugs with novel modes of action are desperately required to combat malaria. P. falciparum parasites infect human red blood cells where they digest the hosts main protein constituent, hemoglobin. Leucine aminopeptidase PfA-M17 is one of several aminopeptidases that have been implicated in the last step of this digestive pathway. Here we utilize both reverse genetics and a compound specifically designed to inhibit the activity of PfA-M17 to show that PfA-M17 is essential for P. falciparum survival as it provides parasites with free amino acids for growth, many of which are highly likely to originate from hemoglobin. We further show that our inhibitor is on-target for PfA-M17 and has the ability to kill parasites at nanomolar concentrations. Thus, in contrast to other hemoglobin-degrading proteases that have overlapping redundant functions, we validate PfA-M17 as a potential novel drug target.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Towards a mechanistic understanding of patient response to neoadjuvant SBRT with anti-PDL1 in human HPV-unrelated locally advanced HNSCC: Phase I/Ib trial results (Part 1)
STUDY_SUMMARY
Five-year survival for HPV-unrelated head and neck squamous cell carcinomas (HNSCC) remains below 50%. We assessed the safety of administering combination hypofractionated stereotactic body radiation therapy (SBRT) with anti-PDL-1 neoadjuvantly followed by adjuvant anti-PDL-1 with standard of care therapy (n=21). The primary endpoint of the study was safety, which was met. Secondary endpoints included radiographic, pathologic, and objective response, locoregional control (LRC), progression-free survival (PFS), and overall survival (OS). Among evaluable patients at early median follow-up of 16 months (448 days), OS was 83.3%, LRC and PFS were 83.3%, and major pathological response (MPR) or complete response (CR) was 75%. Circulating CD8/Treg ratio, CD4 effector memory T cells, and TCR repertoire emerged as biologic correlates of response to therapy. Using high-dimensional multi-omics and spatial data as well as biological correlatives pre- and post-treatment, three major changes were noted in responders within the tumor microenvironment (TME) (and within the blood) post-treatment: 1) an increase in effector T cells; 2) a decrease in immunosuppressive cells; and 3) an increase in antigen presentation. Non-responders appeared to fail due to a lack of one of these three identified steps needed for priming and maintaining activation of T cells. Multiple correlates for response, along with subsets of non-responders that may benefit from additional or alternative immunotherapies, were identified. This treatment is being tested in an ongoing phase II trial with a similar design, where we hope to confirm and expand on our understanding of the mechanisms underlying resistance to therapy.
INSTITUTE
University of Colorado Denver
LAST_NAME
Culp-Hill
FIRST_NAME
Rachel
ADDRESS
12801 E 17th Ave L18-9403D, Aurora, Colorado, 80045, USA
Towards a mechanistic understanding of patient response to neoadjuvant SBRT with anti-PDL1 in human HPV-unrelated locally advanced HNSCC: Phase I/Ib trial results (Part 2)
STUDY_SUMMARY
Five-year survival for HPV-unrelated head and neck squamous cell carcinomas (HNSCC) remains below 50%. We assessed the safety of administering combination hypofractionated stereotactic body radiation therapy (SBRT) with anti-PDL-1 neoadjuvantly followed by adjuvant anti-PDL-1 with standard of care therapy (n=21). The primary endpoint of the study was safety, which was met. Secondary endpoints included radiographic, pathologic, and objective response, locoregional control (LRC), progression-free survival (PFS), and overall survival (OS). Among evaluable patients at early median follow-up of 16 months (448 days), OS was 83.3%, LRC and PFS were 83.3%, and major pathological response (MPR) or complete response (CR) was 75%. Circulating CD8/Treg ratio, CD4 effector memory T cells, and TCR repertoire emerged as biologic correlates of response to therapy. Using high-dimensional multi-omics and spatial data as well as biological correlatives pre- and post-treatment, three major changes were noted in responders within the tumor microenvironment (TME) (and within the blood) post-treatment: 1) an increase in effector T cells; 2) a decrease in immunosuppressive cells; and 3) an increase in antigen presentation. Non-responders appeared to fail due to a lack of one of these three identified steps needed for priming and maintaining activation of T cells. Multiple correlates for response, along with subsets of non-responders that may benefit from additional or alternative immunotherapies, were identified. This treatment is being tested in an ongoing phase II trial with a similar design, where we hope to confirm and expand on our understanding of the mechanisms underlying resistance to therapy.
INSTITUTE
University of Colorado Denver
LAST_NAME
Culp-Hill
FIRST_NAME
Rachel
ADDRESS
12801 E 17th Ave L18-9403D, Aurora, Colorado, 80045, USA
Metabolomics dataset of optogenetic axon regenerative mouse model post optic nerve crush
STUDY_SUMMARY
This metabolite dataset was collected from bacterial channel rhodopsin expressing transgenic mouse models subjected to optic nerve crush (ONC) with subsequent stimulation with light (promoting regeneration) or non-stimulation (lacking axon regeneration). ONC retains retinal ganglion cells within the retina, while degenerating axons. In transgenic bacterial channel rhodopsin expressing cells, light stimulation promotes regeneration. Genetically matched wild-type uninjured optic nerves, or control transgenic mice, were also analyzed. Metabolites were carefully extracted from finely minced optic nerve tissue using a solvent system (initial separation using 1:1 Methanol and H2O and second extraction using 8:1:1 of Acetonitrile:Acetone:Methanol). Untargeted liquid chromatography-mass spectrometry (LC-MS/MS) profiling was performed using fractionation on a Vanquish Horizon Binary UHPLC. Subsequent analyses were performed on an inline coupled Q-Exactive Orbitrap instrument. Metabolites were identified using Compound DiscovererTM software. Statistical analysis was performed using MetaboAnalyst 5.0.
Global, distinctive and personal changes in molecular and microbial profiles induced by specific fibers in humans (Untargeted)
STUDY_SUMMARY
Dietary fibers act through the microbiome and improve cardiovascular health, metabolic disorders and cancer prevention. To understand health benefits of dietary fiber supplementation we investigated two popular purified fibers, arabinoxylan (AX) and long-chain inulin (LCI), and a mixture of five fibers. We present multi-omic signatures of metabolomics, lipidomics, proteomics, metagenomics, a cytokine panel and clinical measurements on healthy and insulin resistant participants. Each fiber is associated with fiber-dependent biochemical and microbial responses. AX consumption associates with a significant reduction in LDL and an increase in bile acids, contributing to its observed cholesterol reduction. LCI is associated with an increase in Bifidobacterium. However, at the highest LCI dose there is increased inflammation and elevation in the liver enzyme alanine aminotransferase. This study yields insights into the effects of fiber supplementation, it provides insights into mechanisms behind fiber induced cholesterol reduction, and it shows effects of individual, purified fibers on the microbiome.
Metabolomic analyses redefine the biological classification of pancreatic cancer: From clinical stage to metabolic subtype
STUDY_SUMMARY
Pancreatic ductal adenocarcinoma (PDAC) is characterized by high heterogeneity, and the postoperative prognosis of different patients often varies greatly. Therefore, the classification of pancreatic cancer patients and precise treatment becomes particularly important. In this study, 1H NMR spectroscopy was used to analyze the 76 PDAC serum samples and identify the potential metabolic subtypes. The metabolic characteristics of each metabolic subtype were screened out and the relationship between metabolic subtype and the long-term prognosis was further identified. The clinical stages of PDAC did not show the metabolic differences at the serum metabolomic level. And three metabolic subtypes, basic, choline-like and amino acid-enriched types, were defined by the HCA of the serum metabolites and the disturbed metabolic pathways. The characteristic metabolites of each PDAC subtype were identified, and the metabolite model was established to distinguish the PDAC patients in the different subtypes. Among the three metabolic subtypes, choline-like type displayed better long-term prognosis compared with the other two types of patients. Metabolic subtypes are of clinical importance and can fully express the heterogeneity in the actual life activities of pancreatic cancer. The excavation of metabolic subtypes based on this will be more accurate and in line with clinical reality, so as to guide clinical precision individualization treatment.
Involvement of Mieap in Cardiolipin metabolism (part I-revised)
STUDY_SUMMARY
Cardiolipin (CL) alterations cause mitochondrial dysfunction. Mieap is involved in mitochondrial quality control (MQC). To investigate whether Mieap functions in MQC via regulation of CL metabolism, quantitative assessment of total CL and comparison of CL species conducted with A549 (Ad-Mieap infected vs. non-infected) and LS174T cells (LS174T-cont vs. Mieap-KD). The A549 cells were harvested 24 hr after infection with Ad-Mieap and were compared with non-infected cells by mass spectrometric analysis. The LS174T-cont and Mieap-KD cells incubated under a normal condition were harvested and subjected to mass spectrometric analysis.
LC-MS analysis of metabolic changes induced by GPX4 inhibitor treatment in cultured HT1080 cells
STUDY_SUMMARY
HT1080 cells were treated with vehicle (DMSO), RSL3 (10 micromolar), ML210 (10 micromolar), or ML162 (10 micromolar) for 2 hours. Cellular metabolites were then extracted and analyzed by LC-MS.
Comparative NMR metabolomics of the responses of A2780 human ovarian cancer cells to clinically established Pt based drugs
STUDY_SUMMARY
Pt based drugs play a very important role in current cancer treatment; yet their cellular and mechanistic aspects are not fully understood. NMR metabolomics provides a powerful tool to investigate the metabolic perturbations induced by Pt drugs in cancer cells and decipher their meaning in relation to the presumed molecular mechanisms. We have carried out a systematic and comparative NMR metabolomics study to analyze the responses of A2780 human ovarian cancer cells to the main clinically established Pt drugs, i.e. cisplatin, carboplatin and oxaliplatin, with a particular attention for the oxaliplatin/cisplatin comparison in view of recently described mechanistic differences. Notably, NMR analysis revealed some moderate and consistent changes in the metabolomic profiles of A2780 cells treated with the 3 Pt drugs with respect to controls but only very small differences among them. Beyond the expected alterations at the level of the nucleic acids the observed changes highlight in all cases induction of a significant ER stress. Owing to the clinical relevance of platinum resistance the behavior of a cisplatin resistant A2780 cancer cell line upon cisplatin treatment was also evaluated.
Recent work indicates that Toll/Interleukin-1 receptor (TIR) domain-containing proteins degrade NAD. Given increasing attention to how the gut microbiome contributes to NAD metabolism and healthy growth, we examined the representation and expressed NADase activities of its TIR domains. The NADase activities of 151 bacterial TIRs were characterized in vitro. Gut bacterial strains representing the diversity of TIRs observed in the microbiome, and activities observed in vitro, were introduced into germ-free mice fed defined diets. Mass spectrometry of cecal metabolites disclosed that a product of TIR NADase activity, variant cyclic-ADPR-x, distinguished colonized from germ-free animals. Mass spectrometry and microbial RNA-Seq of gnotobiotic mice colonized with one, two and all members of this bacterial consortium, identified Bacteroides xylanisolvens as the principal in vivo source of v-cADPR-x. Guided by bioinformatic analyses of biochemically validated TIR domains, we determined that compared to age-matched healthy Bangladeshi children, those with acute malnutrition had significantly lower fecal levels of TIRs known or predicted to generate v-cADPR-x, and of this metabolite. These results indicate that v-cADPR-x may be an informative biomarker of healthy gut microbiome development.
Exposures to Metals in Pregnant Women and the Impact on Fetal Development and Birth Outcomes in Suriname
STUDY_TYPE
C18 Reversed-Phase Broad Spectrum Metabolomics
STUDY_SUMMARY
The Caribbean Consortium for Environmental and Occupational Health, a NIH-funded integrated research and research training program, started in 2015. The research component is a population-based prospective longitudinal environmental epidemiologic cohort study addressing the potential adverse impact of chemical and non-chemical environmental exposures in mother/child dyads in Suriname. The study determines associations between exposures to neurotoxicants (metals and pesticides) and essential elements and non-chemical stressors in pregnant women and the impact on birth and neurodevelopmental outcomes. The study population consists of culturally diverse pregnant women (n=1143; ages: 16-49 years) and their babies/children (n=1069). Data collection takes place twice prenatally, at birth, 12, 36, and 48 months. Through HHEAR, targeted and untargeted (metabolomics) analyses will characterize exposure to metals in a sub-cohort of pregnant women for whom exposure data are not yet available. This expanded exposure analysis will enable a more comprehensive cumulative risk assessment of adverse birth and neurodevelopmental outcomes in the overall cohort.
INSTITUTE
University of Pittsburgh
DEPARTMENT
Department of Environmental and Occupational Health
LAST_NAME
Lichtveld
FIRST_NAME
Maureen
ADDRESS
130 DeSoto Street, University of Pittsburgh, Pittsburgh, PA 15261
Metabolomics analysis of zebrafish response to CID661578 treatment
STUDY_SUMMARY
zebrafish larvae were treated with DMSO or CID661578 for 24 hours prior to global metabolomics analysis (n=6). Metabolites were extracted from pools of 10 zebrafish larvae at 5 dpf using a 80% methanol-based extraction method. Samples were dried in speed vac and stored in -80C freezer until ready for LC-MS analysis.
Feasibility of detecting AC and SCC using UPLC-HRMS based tissue metabolomics
STUDY_SUMMARY
UPLC-HRMS analysis was performed on AC and SCC patients. OPLSDA classfication was performed on tumor vs. ANT&DNT samples. Panels of discriminant features were identified.The biomarkers identified in discovery set samples for each binary classification were confirmed by using a set of validation samples, which were run separately. Additionally, paired analysis shows the abundance of discriminant metabolic features has significant altered in tumor tissues compared to corresponding DNT and ANT samples, indicating metabolic reprogramming during tumorigenesis in AC and SCC.
Functional metabolic molecule were identified as novel therapeutic targets to facilitate gemcitabine treatment against pancreatic cancer
STUDY_SUMMARY
With the development of frontier technologies in system biology, traditional omics-drove phenotypic studies are insufficient to decipher the diseases. Therefore, for a thorough understanding of the molecular mechanisms of diseases to investigate novel drug targets, traditional phenotypic studies must be broken through to the functional exploration of molecules. Meanwhile, the intuitive role of small molecule compounds (metabolites) in pathogenesis, precision diagnosis and therapy are gradually recognized compared to macromolecules such as DNA, RNA and proteins. Therefore, we pioneeringly proposed Spatial Temporal Operative Real Metabolomics (STORM) strategy that established a relationship between metabolic phenotypes and functions to accurately character abnormal metabolisms and further identify operative functional molecules as novel therapeutic targets. Here, given the difficulty of pancreatic cancer (PC) treatment and the high resistance of clinical drugs, we were committed to explore new targets and drugs of pancreatic cancer from a small molecular functional perspective via STORM strategy. Fortunately, based on targeted metabolomics, we found that gemcitabine, one of the most effective clinical anti-PC drugs, served as a dual modulator that promote the accumulation of functional metabolic molecules in purine metabolism to activate down-streamed kinases. And the quantitative consequences of related enzymes annotated the unique molecular mechanisms of purine metabolism regulations by gemcitabine. Collectively, we broadened the cognitions of gemcitabine in tumor inhibition, providing potential strategies for treating PC with small molecules modification. Even more importantly, with the integration of multiple frontier technologies, the STORM strategy has proven to be well adapted to the phenotypic era of functional molecules devoted to innovate molecule mechanism annotation and therapeutic discovery.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
Functional metabolic molecules were identified as novel therapeutic targets to facilitate gemcitabine treatment against pancreatic cancer (Cells metabolomics)
STUDY_SUMMARY
With the development of frontier technologies in system biology, traditional omics-drove phenotypic studies are insufficient to decipher the diseases. Therefore, for a thorough understanding of the molecular mechanisms of diseases to investigate novel drug targets, traditional phenotypic studies must be broken through to the functional exploration of molecules. Meanwhile, the intuitive role of small molecule compounds (metabolites) in pathogenesis, precision diagnosis and therapy are gradually recognized compared to macromolecules such as DNA, RNA and proteins. Therefore, we pioneeringly proposed Spatial Temporal Operative Real Metabolomics (STORM) strategy that established a relationship between metabolic phenotypes and functions to accurately character abnormal metabolisms and further identify operative functional molecules as novel therapeutic targets. Here, given the difficulty of pancreatic cancer (PC) treatment and the high resistance of clinical drugs, we were committed to explore new targets and drugs of pancreatic cancer from a small molecular functional perspective via STORM strategy. Fortunately, based on targeted metabolomics, we found that gemcitabine, one of the most effective clinical anti-PC drugs, served as a dual modulator that promote the accumulation of functional metabolic molecules in purine metabolism to activate down-streamed kinases. And the quantitative consequences of related enzymes annotated the unique molecular mechanisms of purine metabolism regulations by gemcitabine. Collectively, we broadened the cognitions of gemcitabine in tumor inhibition, providing potential strategies for treating PC with small molecules modification. Even more importantly, with the integration of multiple frontier technologies, the STORM strategy has proven to be well adapted to the phenotypic era of functional molecules devoted to innovate molecule mechanism annotation and therapeutic discovery.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
GCN2 regulates mitochondrial OXPHOS in HSPCs under proliferation conditions.
STUDY_SUMMARY
Our results revealed that among all 273 metabolites detected, the levels of metabolites involved in glucose-related glycolysis and gluconeogenesis were elevated in GCN2 deleted HSPCs. Moreover, GCN2 deletion specifically increased mitochondrial OXPHOS and suppressed anaerobic glycolysis in HSPCs.
Amino acids and TCA substrates in hematopoietic cells (Part1)
STUDY_SUMMARY
This study uses [13C,15N] labeled amino acids to study the amino acid consumption and their catabolism into tricarboxylic acid cycle substrates in hematopoietic stem cells, hemopoietic progenitors, and differentiated hematopoietic cells under different conditions, such as homeostasis and proliferation and with drug treatment.
Amino acids and TCA substrates in hematopoietic cells (Part2)
STUDY_SUMMARY
This study uses [13C,15N] labeled amino acids to study the amino acid consumption and their catabolism into tricarboxylic acid cycle substrates in hematopoietic stem cells, hemopoietic progenitors, and differentiated hematopoietic cells under different conditions, such as homeostasis and proliferation and with drug treatment.
Amino acids and TCA substrates in hematopoietic cells (Part3)
STUDY_SUMMARY
This study uses [13C,15N] labeled amino acids to study the amino acid consumption and their catabolism into tricarboxylic acid cycle substrates in hematopoietic stem cells, hemopoietic progenitors, and differentiated hematopoietic cells under different conditions, such as homeostasis and proliferation and with drug treatment.
Amino acids and TCA substrates in hematopoietic cells (Part4)
STUDY_SUMMARY
This study uses [13C,15N] labeled amino acids to study the amino acid consumption and their catabolism into tricarboxylic acid cycle substrates in hematopoietic stem cells, hemopoietic progenitors, and differentiated hematopoietic cells under different conditions, such as homeostasis and proliferation and with drug treatment.
Discovery and characterization of virulence associated functional metabolites in Escherichia coli based on functional metabolomics strategy(siderophores metabolomics-1)
STUDY_SUMMARY
Bacterial metabolites are substrates of virulence factors of uropathogenic Escherichia coli (UPEC), but the mechanism underlying the role of functional metabolites in bacterial virulence from the perspective of small molecular metabolism is unclear. In the present study, we used a strategy of functional metabolomics integrated with bacterial genetics in attempt to decipher the mechanism of virulence formation in Escherichia coli (E. coli) from the viewpoint of small molecule metabolism. We identified the virulence-associated metabolome via analysis of the primary metabolome of the pathogenic UTI89 strain and the non-pathogenic MG1655 strain. Then, the iron-mediated virulence associated metabolome was identified by an iron fishing strategy. Also, the mechanism of siderophores in regulating pathogenicity in different environments was explored by investigating the effect of iron on siderophore biosynthesis. Finally, by knocking out genes related to siderophore biosynthesis, siderophore transport and iron utilization, siderophores dependent iron-regulating virulence associated metabolome, including 18 functional metabolites, was identified and verified to be involved in the regulation of bacterial virulence. Based on this we found that these functional metabolites regulated the virulence of E. coli by targeting multiple metabolic pathways in an iron-siderophores dependent manner. Moreover, a quantitative proteomics approach was implemented to further elucidate the mechanism of functional metabolites and functional proteins in modulating bacterial virulence. And our findings demonstrated that functional proteins regulated the virulence of E. coli by mediating iron binding, iron-siderophore transmembrane transport, and the biosynthesis and expression of functional metabolites. Interestingly, we found that functional metabolites enhance the virulence of E. coli by specifically modulating the key metabolic pathways involved in purine metabolism, proline metabolism, arginine metabolism and pyrimidine metabolism. Taken together, our study identified for the first time 18 functional metabolites regulating the of E. coli virulence, greatly enriching our understanding of the mechanism of functional metabolites that regulate the E. coli virulence by targeting primary metabolism, which will largely contribute to the development of new strategies to target virulence-based diagnosis and therapy of infections caused by different pathogens.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Discovery and characterization of virulence associated functional metabolites in Escherichia coli based on functional metabolomics strategy(siderophores metabolomics-2)
STUDY_SUMMARY
Bacterial metabolites are substrates of virulence factors of uropathogenic Escherichia coli (UPEC), but the mechanism underlying the role of functional metabolites in bacterial virulence from the perspective of small molecular metabolism is unclear. In the present study, we used a strategy of functional metabolomics integrated with bacterial genetics in attempt to decipher the mechanism of virulence formation in Escherichia coli (E. coli) from the viewpoint of small molecule metabolism. We identified the virulence-associated metabolome via analysis of the primary metabolome of the pathogenic UTI89 strain and the non-pathogenic MG1655 strain. Then, the iron-mediated virulence associated metabolome was identified by an iron fishing strategy. Also, the mechanism of siderophores in regulating pathogenicity in different environments was explored by investigating the effect of iron on siderophore biosynthesis. Finally, by knocking out genes related to siderophore biosynthesis, siderophore transport and iron utilization, siderophores dependent iron-regulating virulence associated metabolome, including 18 functional metabolites, was identified and verified to be involved in the regulation of bacterial virulence. Based on this we found that these functional metabolites regulated the virulence of E. coli by targeting multiple metabolic pathways in an iron-siderophores dependent manner. Moreover, a quantitative proteomics approach was implemented to further elucidate the mechanism of functional metabolites and functional proteins in modulating bacterial virulence. And our findings demonstrated that functional proteins regulated the virulence of E. coli by mediating iron binding, iron-siderophore transmembrane transport, and the biosynthesis and expression of functional metabolites. Interestingly, we found that functional metabolites enhance the virulence of E. coli by specifically modulating the key metabolic pathways involved in purine metabolism, proline metabolism, arginine metabolism and pyrimidine metabolism. Taken together, our study identified for the first time 18 functional metabolites regulating the of E. coli virulence, greatly enriching our understanding of the mechanism of functional metabolites that regulate the E. coli virulence by targeting primary metabolism, which will largely contribute to the development of new strategies to target virulence-based diagnosis and therapy of infections caused by different pathogens.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Discovery and characterization of virulence associated functional metabolites in Escherichia coli based on functional metabolomics strategyļ¼pellets metabolomics-1ļ¼
STUDY_SUMMARY
Bacterial metabolites are substrates of virulence factors of uropathogenic Escherichia coli (UPEC), but the mechanism underlying the role of functional metabolites in bacterial virulence from the perspective of small molecular metabolism is unclear. In the present study, we used a strategy of functional metabolomics integrated with bacterial genetics in attempt to decipher the mechanism of virulence formation in Escherichia coli (E. coli) from the viewpoint of small molecule metabolism. We identified the virulence-associated metabolome via analysis of the primary metabolome of the pathogenic UTI89 strain and the non-pathogenic MG1655 strain. Then, the iron-mediated virulence associated metabolome was identified by an iron fishing strategy. Also, the mechanism of siderophores in regulating pathogenicity in different environments was explored by investigating the effect of iron on siderophore biosynthesis. Finally, by knocking out genes related to siderophore biosynthesis, siderophore transport and iron utilization, siderophores dependent iron-regulating virulence associated metabolome, including 18 functional metabolites, was identified and verified to be involved in the regulation of bacterial virulence. Based on this we found that these functional metabolites regulated the virulence of E. coli by targeting multiple metabolic pathways in an iron-siderophores dependent manner. Moreover, a quantitative proteomics approach was implemented to further elucidate the mechanism of functional metabolites and functional proteins in modulating bacterial virulence. And our findings demonstrated that functional proteins regulated the virulence of E. coli by mediating iron binding, iron-siderophore transmembrane transport, and the biosynthesis and expression of functional metabolites. Interestingly, we found that functional metabolites enhance the virulence of E. coli by specifically modulating the key metabolic pathways involved in purine metabolism, proline metabolism, arginine metabolism and pyrimidine metabolism. Taken together, our study identified for the first time 18 functional metabolites regulating the of E. coli virulence, greatly enriching our understanding of the mechanism of functional metabolites that regulate the E. coli virulence by targeting primary metabolism, which will largely contribute to the development of new strategies to target virulence-based diagnosis and therapy of infections caused by different pathogens.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Discovery and characterization of virulence associated functional metabolites in Escherichia coli based on functional metabolomics strategyļ¼pellets metabolomics-2ļ¼
STUDY_SUMMARY
Bacterial metabolites are substrates of virulence factors of uropathogenic Escherichia coli (UPEC), but the mechanism underlying the role of functional metabolites in bacterial virulence from the perspective of small molecular metabolism is unclear. In the present study, we used a strategy of functional metabolomics integrated with bacterial genetics in attempt to decipher the mechanism of virulence formation in Escherichia coli (E. coli) from the viewpoint of small molecule metabolism. We identified the virulence-associated metabolome via analysis of the primary metabolome of the pathogenic UTI89 strain and the non-pathogenic MG1655 strain. Then, the iron-mediated virulence associated metabolome was identified by an iron fishing strategy. Also, the mechanism of siderophores in regulating pathogenicity in different environments was explored by investigating the effect of iron on siderophore biosynthesis. Finally, by knocking out genes related to siderophore biosynthesis, siderophore transport and iron utilization, siderophores dependent iron-regulating virulence associated metabolome, including 18 functional metabolites, was identified and verified to be involved in the regulation of bacterial virulence. Based on this we found that these functional metabolites regulated the virulence of E. coli by targeting multiple metabolic pathways in an iron-siderophores dependent manner. Moreover, a quantitative proteomics approach was implemented to further elucidate the mechanism of functional metabolites and functional proteins in modulating bacterial virulence. And our findings demonstrated that functional proteins regulated the virulence of E. coli by mediating iron binding, iron-siderophore transmembrane transport, and the biosynthesis and expression of functional metabolites. Interestingly, we found that functional metabolites enhance the virulence of E. coli by specifically modulating the key metabolic pathways involved in purine metabolism, proline metabolism, arginine metabolism and pyrimidine metabolism. Taken together, our study identified for the first time 18 functional metabolites regulating the of E. coli virulence, greatly enriching our understanding of the mechanism of functional metabolites that regulate the E. coli virulence by targeting primary metabolism, which will largely contribute to the development of new strategies to target virulence-based diagnosis and therapy of infections caused by different pathogens.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Optimization of Imputation Strategies for High-Resolution Gas Chromatography-Mass Spectrometry (HR GC-MS) Metabolomics Data
STUDY_SUMMARY
Gas chromatography-coupled mass spectrometry (GC-MS) has been used in biomedical research to analyze volatile, non-polar, and polar metabolites in a wide array of sample types. Despite advances in technology, missing values are still common in metabolomics datasets and must be properly handled. We evaluated the performance of ten commonly used missing value imputa-tion methods with metabolites analyzed on an HR GC-MS instrument. By introducing missing values into the complete (i.e., data without any missing values) NIST plasma dataset we demon-strate that Random Forest (RF), Glmnet Ridge Regression (GRR), and Bayesian Principal Com-ponent Analysis (BPCA) shared the lowest Root Mean Squared Error (RMSE) in technical repli-cate data. Further examination of these three methods in data from baboon plasma and liver samples demonstrated they all maintained high accuracy. Overall, our analysis suggests that any of the three imputation methods can be applied effectively to untargeted metabolomics datasets with high accuracy. However, it is important to note that imputation will alter the correlation structure of the dataset, and bias downstream regression coefficients and p-values.
INSTITUTE
Wake Forest School of Medicine
LAST_NAME
Ampong
FIRST_NAME
Isaac
ADDRESS
Center for Precision Medicine, Department of Internal Medicine, Section on Molecular Medicine, Wake Forest University, Winston-Salem, North Carolina, United States
Metabolite variation in the blood serum of HIV patients carrier treated with effervecent glutamine
STUDY_TYPE
Liquid NMR experiments
STUDY_SUMMARY
It was demonstrated that effervescent glutamine supplementation in people living with HIV/AIDS increased the amount of CD4+ T lymphocytes. , decreased the serum levels of biomarkers of inflammation, and introduced health benefits. Herein, NMR spectroscopy was applied to evaluate if oral ingestion of glutamine (12.5 g) for 30 days can be measured through serum metabolite variations and if the reported benefits might correlate to small metabolites' changes in a chosen cohort of individuals. The HIV/AIDS group has been carefully selected and studied before and after the 30-days of supplementation with the glutamine, together with the matched non-HIV carrier patients' group. The group of HIV/AIDS individuals presented lower (p < 0.05) levels of choline, creatine, pyruvate, glutamate, lysine, and tyrosine when compared to the non-HIV carrier patients. On the other hand, serum concentrations of glucose, lipids, lactate, glutamine, phenylalanine, and threonine were higher (p < 0.05) in HIV/AIDS individuals. The variation in metabolites as a result of treatment with glutamine is consistent with the improvements observed in these patients and may lead us to suggest the introduction of the effervescent glutamine supplementation to antiretroviral therapies in people living with HIV/AIDS.
Alignment and Analysis of a Disparately Acquired Multi-Batch Metabolomics Study of Maternal Pregnancy Samples (Part 1).
STUDY_SUMMARY
This is an untargeted RPLC-MS metabolomics study of mother-infant pairs from the State of Michigan. Maternal plasma was collected in the first and third trimesters of gestation, as well as umbilical cord blood, with the aim of studying metabolic changes throughout pregnancy and influences on the infant's metabolome. The data is split into two experiment subsets, assayed with different H2O-methanol gradients and mass spectrometers 2-3 years apart, inducing major chromatographic and signal measurement shifts. Analysis steps consisted of disparate LC-MS alignment and data normalization, followed by differential timepoint comparisons and partial correlation network construction.
INSTITUTE
University of Michigan
DEPARTMENT
Computational Medicine and Bioinformatics
LAST_NAME
Habra
FIRST_NAME
Hani
ADDRESS
100 Washtenaw Ave., Palmer Commons Bldg. Ann Arbor, MI 48109-2218
Alignment and Analysis of a Disparately Acquired Multi-Batch Metabolomics Study of Maternal Pregnancy Samples. (Part 2)
STUDY_SUMMARY
This is an untargeted RPLC-MS metabolomics study of mother-infant pairs from the State of Michigan. Maternal plasma was collected in the first and third trimesters of gestation, as well as umbilical cord blood, with the aim of studying metabolic changes throughout pregnancy and influences on the infant's metabolome. The data is split into two experiment subsets, assayed with different H2O-methanol gradients and mass spectrometers 2-3 years apart, inducing major chromatographic and signal measurement shifts. Analysis steps consisted of disparate LC-MS alignment and data normalization, followed by differential timepoint comparisons and partial correlation network construction.
INSTITUTE
University of Michigan
DEPARTMENT
Computational Medicine and Bioinformatics
LAST_NAME
Habra
FIRST_NAME
Hani
ADDRESS
100 Washtenaw Ave., Palmer Commons Bldg. Ann Arbor, MI 48109-2218
Targeted Microchip Capillary Electrophoresis-Orbitrap Mass Spectrometry Metabolomics to Monitor Ovarian Cancer Progression (calibration standards)
STUDY_SUMMARY
The lack of effective screening strategies for high-grade serous carcinoma (HGSC), a subtype of ovarian cancer (OC) responsible for 80% of OC related deaths, emphasizes the need for new diagnostic markers and a better understanding of disease pathogenesis. Capillary electrophoresis (CE) coupled with high-resolution mass spectrometry (HRMS) offers high selectivity and sensitivity, thereby increasing metabolite coverage and consequently enhancing biomarker discovery. Recent advances in CE-MS include small, chip-based CE systems coupled with nanoelectrospray ionization (nanoESI) to provide rapid, high-resolution analysis of biological specimens. Here, we describe the development of a targeted microchip (Āµ) CE-HRMS method to analyze 40 target metabolites in serum samples from a triple-mutant (TKO) mouse model of HGSC, with an acquisition time of only 3 min. Extracted ion electropherograms showed sharp, highly resolved peak shapes, even for structural isomers such as leucine and isoleucine. All analytes maintained good linearity with an average R2 of 0.994, while detection limits were in the nM range. Thirty metabolites were detected in mice serum, with recoveries ranging from 78 to 120 %, indicating minimal ionization suppression and good accuracy. We applied the ĀµCE-HRMS method to sequentially-collected serum samples from TKO and TKO-control mice. Time-resolved analysis revealed characteristic temporal trends for amino acids, nucleosides, and amino acids derivatives associated with HGSC progression. Comparison of the ĀµCE-HRMS dataset with non-targeted ultra-high performance liquid chromatography (UHPLC) ā MS results revealed identical temporal trends for the 5 metabolites detected on both platforms, indicating the ĀµCE-HRMS method performed satisfactorily in terms of capturing metabolic reprogramming due to HGSC progression, while reducing the total analysis time 3-fold.
Targeted Microchip Capillary Electrophoresis-Orbitrap Mass Spectrometry Metabolomics to Monitor Ovarian Cancer Progression. (Ovarian cancer mouse data)
STUDY_SUMMARY
The lack of effective screening strategies for high-grade serous carcinoma (HGSC), a subtype of ovarian cancer (OC) responsible for 80% of OC related deaths, emphasizes the need for new diagnostic markers and a better understanding of disease pathogenesis. Capillary electrophoresis (CE) coupled with high-resolution mass spectrometry (HRMS) offers high selectivity and sensitivity, thereby increasing metabolite coverage and consequently enhancing biomarker discovery. Recent advances in CE-MS include small, chip-based CE systems coupled with nanoelectrospray ionization (nanoESI) to provide rapid, high-resolution analysis of biological specimens. Here, we describe the development of a targeted microchip (Āµ) CE-HRMS method to analyze 40 target metabolites in serum samples from a triple-mutant (TKO) mouse model of HGSC, with an acquisition time of only 3 min. Extracted ion electropherograms showed sharp, highly resolved peak shapes, even for structural isomers such as leucine and isoleucine. All analytes maintained good linearity with an average R2 of 0.994, while detection limits were in the nM range. Thirty metabolites were detected in mice serum, with recoveries ranging from 78 to 120 %, indicating minimal ionization suppression and good accuracy. We applied the ĀµCE-HRMS method to sequentially-collected serum samples from TKO and TKO-control mice. Time-resolved analysis revealed characteristic temporal trends for amino acids, nucleosides, and amino acids derivatives associated with HGSC progression. Comparison of the ĀµCE-HRMS dataset with non-targeted ultra-high performance liquid chromatography (UHPLC) ā MS results revealed identical temporal trends for the 5 metabolites detected on both platforms, indicating the ĀµCE-HRMS method performed satisfactorily in terms of capturing metabolic reprogramming due to HGSC progression, while reducing the total analysis time 3-fold.
Investigation of Gastrolobium bilobum metabolism in formation of fluoroacetate
STUDY_SUMMARY
This data contains untargeted LC-MS/MS analysis of gastrolobium bilobum plants fed isotopically labeled precursors to investigate synthesis of fluoroacetate.
Mitochondrial respiration in B lymphocytes is essential for humoral immunity by controlling flux of the TCA cycle
STUDY_SUMMARY
The function of mitochondrial respiration during B cell fate decisions and differentiation 55 remained equivocal. This study reveals that selection for mitochondrial fitness occurs during B 56 cell activation and is essential for subsequent plasma cell differentiation. By expressing a 57 mutated mitochondrial helicase in transitional B cells, we depleted mitochondrial DNA during 58 B cell maturation, resulting in reduced oxidative phosphorylation. Although no changes in 59 follicular B cell development were evident, germinal centers, class switch recombination to 60 IgG, plasma cell maturation and humoral immunity were diminished. Defective oxidative 61 phosphorylation led to aberrant flux of the tricarboxylic acid cycle and lowered the amount of 62 saturated phosphatidic acid. Consequently, mTOR activity and BLIMP1 induction were 63 curtailed whereas HIF1 _and glycolysis were amplified. Exogenous phosphatidic acid 64 increased mTOR activity in activated B cells. Hence, mitochondrial function is required and 65 selected for in activated B cells for the successful generation of functional plasma cells.
INSTITUTE
University of Erlangen-NĆ¼rnberg
DEPARTMENT
Division of Molecular Immunology.UniversitƤtsklinikum Erlangen, Nikolaus Fibinger Zentrum
Endo- and Exometabolome Crosstalk in Mesenchymal Stem Cells Undergoing Osteogenic Differentiation (Lipid Samples)
STUDY_SUMMARY
The holistic nature of NMR enabled the time-course evolution of cholesterol, mono- and polyunsaturated fatty acids (including Ļ-6 and Ļ-3 fatty acids), several phospholipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelins, and plasmalogens), and mono- and triglycerides to be followed. Lipid changes occurred almost exclusively between days 1 and 7, followed by a tendency for lipidome stabilization after day 7. On average, phospholipids and longer and more unsaturated fatty acids increased up to day 7, probably related to plasma membrane fluidity. Articulation of lipidome changes with previously reported polar endometabolome profiling and with exometabolome changes reported here in the same cells, enabled important correlations to be established during hAMSC osteogenic differentiation. Our results supported hypotheses related to the dynamics of membrane remodelling, anti-oxidative mechanisms, protein synthesis, and energy metabolism. Importantly, the observation of specific up-taken or excreted metabolites paves the way for the identification of potential osteoinductive metabolites useful for optimized osteogenic protocols.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry
LABORATORY
CICECO - Aveiro Institute of Materials
LAST_NAME
Bispo
FIRST_NAME
Daniela S.C.
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, Aveiro, 3810-193, Portugal
Endo- and Exometabolome Crosstalk in Mesenchymal Stem Cells Undergoing Osteogenic Differentiation (Media Samples)
STUDY_SUMMARY
The holistic nature of NMR enabled the time-course evolution of cholesterol, mono- and polyunsaturated fatty acids (including Ļ-6 and Ļ-3 fatty acids), several phospholipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelins, and plasmalogens), and mono- and triglycerides to be followed. Lipid changes occurred almost exclusively between days 1 and 7, followed by a tendency for lipidome stabilization after day 7. On average, phospholipids and longer and more unsaturated fatty acids increased up to day 7, probably related to plasma membrane fluidity. Articulation of lipidome changes with previously reported polar endometabolome profiling and with exometabolome changes reported here in the same cells, enabled important correlations to be established during hAMSC osteogenic differentiation. Our results supported hypotheses related to the dynamics of membrane remodelling, anti-oxidative mechanisms, protein synthesis, and energy metabolism. Importantly, the observation of specific up-taken or excreted metabolites paves the way for the identification of potential osteoinductive metabolites useful for optimized osteogenic protocols.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry
LABORATORY
CICECO - Aveiro Institute of Materials
LAST_NAME
Bispo
FIRST_NAME
Daniela S.C.
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, Aveiro, 3810-193, Portugal
The Carbohydrate Sensing Transcription Factor ChREBP Links Mitochondrial Lipidomes to Mitochondrial Dynamics and Progression of Diabetic Nephropathy
STUDY_TYPE
Biomarker
STUDY_SUMMARY
Despite recent advances, diabetic nephropathy (DN) remains a major public health concern. The precise underlying molecular mechanisms of DN remain elusive. Accumulating recent evidence suggests that mitochondrial integrity and lipid metabolism in podocytes significantly contribute to the pathogenesis of DN. However, the interplay between these two key metabolic regulators of DN is not fully understood. This study examines the role of ChREBP (carbohydrate-response element-binding protein), a master regulator of lipogenesis, on mitochondrial morphology and progression of DN. Our findings suggest that diabetic mice with podocyte-specific deletion of ChREBP are protected against mitochondrial fragmentation and progression of DN. Using liquid chromatography coupled with mass spectrometry, we identified the central role of ChREBP-induced plasmalogen phospholipids in linking mitochondrial lipidomes with mitochondrial dynamics in DN.
Mice were euthanized at 20 weeks of age, and various tissues were obtained for metabolomic analyses including whole bone, bone marrow, and isolated cortical bone. Metabolites were extracted and underwent LC-MS analysis using an Agilent LC 1290 coupled to an Agilent 6538 QTOF
Metabolomic differences detected in female C57BL/6 mice
STUDY_SUMMARY
Mice were euthanized at 20 weeks of age, and humeri were obtained for metabolomic analyses. Once obtained, marrow was flushed in some humeri resulting in 3 sample types including whole bone, bone marrow, and isolated cortical bone. Metabolites were extracted and underwent LC-MS analysis using an Agilent LC 1290 coupled to an Agilent 6538 QTOF
Untargeted MS-based metabolomics analysis of the responses to drought stress in Quercus ilex leaf seedlings, and identification of putative compounds related to tolerance
STUDY_TYPE
Untargeted MS-based metabolomics
STUDY_SUMMARY
The effect and responses to drought stress have been analyzed in Quercus ilex seedlings by using a non-targeted metabolomic approach, implementing previous studies pub-lished in which other -omics platforms, transcriptomics, and proteomics, have been em-ployed. This work is aimed at characterizing the Q. ilex leaf metabolome, determining possible mechanisms and molecular markers of drought tolerance, and identifying puta-tive bioactive compounds. Six-month-old seedling leaves, subjected to drought stress im-posed by water withholding under high temperature and irradiance conditions, were col-lected when leaf fluorescence decreased by 20 % (day 17) and 45 % (day 24) relative to ir-rigated seedlings. A total of 3934 compounds were resolved, with 616 being variable, and 342 identified, belonging to five chemical families. Out of the identified compounds 33 were variable, mostly corresponding to amino acids, carboxylic acids, benzenoids, flavo-noids and isoprenoids. Epigallocatechin, ellagic acid, pulegone, indole-3-acrylic acid and dihydrozeatin-O-glucoside were up-accumulated under drought conditions at both sam-pling times. An integrated multi-omics analysis of phenolic compounds and related en-zymes was performed, revealing that some enzymes involved in the flavonoid pathways (chalcone synthase, anthocyanidin synthase and anthocyanidin reductase) were up-accumulated at day 24 in non-irrigated seedlings. Some putative markers of drought tol-erance to drought in Q. ilex are proposed for assisting breeding programs based on the se-lection of elite genotypes.
INSTITUTE
new
DEPARTMENT
Universidad de CĆ³rdoba
LABORATORY
Agroforestry and Plant Biochemistry, Proteomics and Systems Biology
In vivo commensal control of Clostridioides difficile virulence
STUDY_TYPE
Gnotobiotic mouse infection model
STUDY_SUMMARY
Gnotobiotic mouse infection model with C. difficile strain ATCC43255 and co-colonization studies with the commensals Paraclostridium bifermentans or Clostridium sardiniense
Sphingomyelin depletion inhibits CXCR4 dynamics and CXCL12-mediated directed cell migration in human T cells
STUDY_SUMMARY
Sphingolipids, ceramides and cholesterol are integral components of cellular membranes, and they also play important roles in signal transduction by regulating the dynamics of membrane receptors through their effects on membrane fluidity. Here, we combined biochemical and functional assays with single-molecule dynamic approaches to demonstrate that the local lipid environment regulates CXCR4 organization and function and modulates chemokine-triggered directed cell migration. Prolonged treatment of T cells with neutral sphingomyelinase promoted the complete and sustained breakdown of sphingomyelins and the accumulation of the corresponding ceramides, which altered both membrane fluidity and CXCR4 nanoclustering and dynamics. Under these conditions CXCR4 retained some CXCL12-mediated signaling activity but failed to promote efficient directed cell migration. Our data underscore a critical role for the local lipid composition at the cell membrane in regulating the lateral mobility of chemokine receptors, and their ability to dynamically increase receptor density at the leading edge to promote efficient cell migration
INSTITUTE
Universidad CEU San Pablo
LAST_NAME
Gonzalez-Riano
FIRST_NAME
Carolina
ADDRESS
km 0, Universidad CEU-San Pablo UrbanizaciĆ³n MonteprĆncipe. M-501
Integrative Exposomic, Transcriptomic, Epigenomic Analyses of Human Placental Samples Links Understudied Chemicals to Preeclampsia
STUDY_SUMMARY
Background Environmental health research has recently undergone a dramatic shift, with ongoing technological advancements allowing for broader coverage of exposure and molecular biology signatures. Approaches to integrate such measures are still needed to increase understanding between systems-level exposure and biology. Objectives We address this gap by evaluating placental tissues to identify novel chemical-biological interactions associated with preeclampsia. This study tests the hypothesis that understudied chemicals are present in the human placenta and associated with preeclampsia-relevant disruptions, including overall case status (preeclamptic vs. normotensive patients) and underlying transcriptomic/epigenomic signatures. Methods A non-targeted analysis based on high-resolution mass spectrometry was used to analyze placental tissues from a cohort of 35 patients with preeclampsia (n = 18) and normotensive (n = 17) pregnancies. Molecular feature data were queried against chemicals within the U.S. Environmental Protection Agencyās DSSTox database, and prioritized for confirmation based on association with preeclampsia case status and confidence of chemical identification. All molecular features were evaluated for relationships to mRNA, microRNA, and CpG methylation (i.e., multi-omic) signature alterations involved in preeclampsia. Results A total of 183 molecular features were identified with significantly differentiated abundance in placental extracts of preeclamptic patients; these features clustered into distinct chemical groupings using unsupervised methods. Of these features, 53 were identified (mapping to 40 distinct chemicals) using chemical standards, fragmentation spectra, and chemical metadata. In general, human metabolites had the largest feature intensities and strongest associations with preeclampsia-relevant multi-omic changes. Exogenous drugs were second most abundant and had fewer associations with multi-omic changes. Other exogenous chemicals (non-drugs) were least abundant and had the fewest associations with multi-omic changes. Conclusions These global data trends suggest that human metabolites are heavily intertwined with biological processes involved in preeclampsia etiology, while exogenous chemicals may still impact select transcriptomic/epigenomic processes. This study serves as a demonstration of merging systems exposures with systems biology to better understand chemical-disease relationships.
Metabolomics analysis of mouse liver with or without SIRT5 deficiency
STUDY_SUMMARY
8 Wild-type (WT) and 8 Sirt5-/- (SIRT5 KO) mice on 129 background were maintained on a standard chow diet (Teklad Global 18% Protein Rodent Diet, ENVIGO, Cat.#2018) until they were put on euthanized. Liver metabolites were extracted (5-10 mg) were extracted using 80% methanol/water as the extraction solvent. Metabolite extract was split into two tubes (one for polar metabolite analysis and the other one for acyl-CoA analysis), and then extraction solvent was evaporated using a speed vacuum concentrator. Dry pellets were stored in ā80 Ā°C freezer until ready for LC-MS analysis. For acyl-CoA analysis, dry pellets were reconstituted into 30 Ī¼L of sample solvent (water containing 50 mM ammonium acetate) per 6 mg tissue, and 8 Ī¼L was injected into the LC-MS. For non-acyl-CoA polar metabolite analysis, pellets were reconstituted into 30 Ī¼L of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v/v) per 3 mg tissue, and 3 Ī¼L was injected into the LC-MS.
Data from plasma metabolome analysis of APP-KI and Wild type mice treated with B. breve MCC1274
STUDY_SUMMARY
Recently, we showed that administration of B. breve MCC1274 reduced amyloid-beta production, inhibited microglial activation, and suppressed inflammatory responses in the brains of APP knock-in (AppNL-G-F) mice. To elucidate the mechanism of action of this probiotic strain in an Alzheimer's disease-like model, we orally fed 3-month-old mice with B. breve MCC1274 or saline for 4 months and comprehensively investigated metabolites in plasma using CE-FTMS and LC-TOFMS.
INSTITUTE
Morinaga milk industry CO., LTD.
DEPARTMENT
Next generation science institute
LABORATORY
Frontier research section
LAST_NAME
Ohno
FIRST_NAME
Kazuya
ADDRESS
1-83 5-Chome Higashihara, Zama-city, Kanagawa-Pref. Japan
Virus-host interaction studies of P. plecoglossicida strain (NZBD9) and Epinephelus coioides.
STUDY_SUMMARY
E. coioides of the infection group were intrapleurally injected with 105 colony-forming units per fish (cfu/fish) of P. plecoglossicida (wild-type strain or clpV-RNAi strain), while control E. coioides received an equivalent volume of phosphate-buffered saline (PBS). The temperature of the water was maintained at 18 Ā± 1 Ā°C during the infection experiment. The spleens of E. coioides were collected at 6, 12, 24, 48, 72 and 96 hours post injection (hpi).
Western diet (WD) and gut microbiota interplay drives the development of non-alcoholic fatty liver disease (NAFLD) and its progression to non-alcoholic steatohepatitis (NASH). However, the metabolic mediators contributing to NASH remain to be identified. In this study, we explored the change of liver metabolites in a diet-induced mouse NASH model. Identification of these metabolites will help to uncover the pathogenesis of NASH and explore the new therapeutic treatment.
Effect of long-term exposure to graphene on skin cell metabolism
STUDY_SUMMARY
Graphene-derived materials are a family of nanomaterials with multiple potential applications in different fields such as biomedicine. It is therefore essential to understand their interaction with cellular barriers such as skin. In this work we evaluated the metabolic changes in human skin cells (HaCaT) exposed to different GRMs for 7 and 30 days. Objectives Endogenous metabolic profiles of control and graphene-treated keratinocytes have been studied using ultra-high performance liquid chromatography ā mass spectrometry (UHPLC-MS). Keratinocytes were treated with graphene oxide (GO) from two different suppliers and with few layer graphene (FLG). Samples were collected one week and one month after the start of the treatment. The general aim of the project was to evaluate potential metabolic differences between: 1) Graphene-treated keratinocytes and control keratinocytes at one week; 2) Graphene-treated keratinocytes and control keratinocytes at one month; 3) Control keratinocytes at 1 month and 1 week; 4) Graphene-treated keratinocytes at 1 month and 1 week. Experimental Procedures A successful metabolic profiling experiment relies on the ability to determine changes in an organismās biofluid or tissue complement of metabolites. Mass spectrometry coupled to ultra-high performance liquid chromatography (UHPLC-MS) is well suited to such analyses due to its high sensitivity, large coverage over different classes of metabolites, high throughput capacity, and wide dynamic range. In this study, one UHPLC-MS based platform was used to analyse endogenous analytes for inclusion in subsequent statistical analysis procedures used to study metabolic differences between the groups of samples. Results The oxidation degree and size of the GRMs is determinant in the effect on cell metabolism, as well as the exposure time. Thus, one of the materials used generated a change in the energy metabolism of the cells, significantly increasing the level of different Krebs cycle metabolites.
Untargeted serum metabolomic profiling for early detection of Schistosoma mekongi infection in mouse model
STUDY_SUMMARY
Mekong schistosomiasis is a parasitic disease caused by blood flukes in the Lao Peopleās Democratic Republic and in Cambodia. The standard method for diagnosis of schistosomiasis is detection of parasite eggs from patient samples. However, this method is not sufficient to detect asymptomatic patients, low egg numbers, or early infection. Therefore, diagnostic methods with higher sensitivity at the early stage of the disease are needed to fill this gap. The aim of this study was to identify potential biomarkers of early schistosomiasis using an untargeted metabolomics approach. Serum of uninfected and S. mekongi-infected mice was collected at 2, 4, and 8 weeks post-infection. Samples were extracted for metabolites and analyzed with a liquid chromatography-tandem mass spectrometer. Metabolites were annotated with the MS-DIAL platform and analyzed with Metaboanalyst bioinformatic tools. Multivariate analysis distinguished between metabolites from the different experimental conditions. Biomarker screening was performed using three methods: correlation coefficient analysis; feature important detection with a random forest algorithm; and receiver operating characteristic (ROC) curve analysis. Three compounds were identified as potential biomarkers at the early stage of the disease: heptadecanoyl ethanolamide; picrotin; and theophylline. The levels of these three compounds changed significantly during early-stage infection, and therefore these molecules may be promising schistosomiasis markers. These findings may help to improve early diagnosis of schistosomiasis, thus reducing the burden on patients and limiting spread of the disease in endemic areas.
INSTITUTE
Princess Srisavangavadhana College of Medicine, Chulabhorn Royal Academy
Chemotaxonomic patterns in intracellular metabolites of marine microbial plankton
STUDY_SUMMARY
Targeted and untargeted metabolomes were generated for a variety of marine microbial taxa including eukaryotic phytoplankton, cyanobacteria, archaea, and heterotrophic bacteria. Microbial metabolism generates small organic molecules that reflect both the biochemical and physiological diversity as well as the taxonomic specificity of biological processes. These small molecules serve as the conduit for taxon-specific signaling and exchange. We used liquid chromatography-mass spectrometry (LC-MS)-based metabolomics to taxonomically categorize metabolites that include small molecules in central and secondary metabolism across 42 taxa representing numerically dominant and metabolically important lineages of microbial autotrophs and heterotrophs.
Global metabolomics analysis of neutrophils isolated from tumor
STUDY_SUMMARY
Pathologically activated neutrophils (PMN), termed myeloid-derived suppressor cells (PMN-MDSCs), are major negative regulators of anti-tumor immunity. The mechanisms responsible for the pathological activation of neutrophils upon infiltration into tumors are not well defined, thus limiting the selective targeting of these cells. Tumor cells and immune cells engage in bi-directional manipulation of their respective metabolism, thereby altering cell function to facilitate tumor progression. Targeting the metabolism of responding immune cells can improve cancer treatment when combined with existing therapeutic strategies. Here, we investigated the role of metabolism in the immunoinhibitory actions of tumor PMN-MDSCs.
Glycerate Production from Intestinal Fructose Metabolism Elevated by Dietary Fat Induces Glucose Intolerance Through Ī²-cell Damage
STUDY_SUMMARY
Dietary fructose, especially in the context of a high-fat western diet, has been linked to type 2 diabetes. Although the effect of fructose on liver metabolism has been extensively studied, a significant portion of the fructose is first metabolized in the small intestine. Here we report that dietary fat enhances intestinal fructose metabolism, which releases glycerate into the blood. High systemic glycerate levels reduce pancreatic islet sizes and Ī²-cell content, thus inducing glucose intolerance. Our findings provide an additional link between dietary fructose and diabetes that is modulated by dietary fat.
INSTITUTE
Duke University
LAST_NAME
Wong
FIRST_NAME
Chi Wut
ADDRESS
CEIMAS, 101 Science Dr. Room 2141, Durham, NC, 27709, USA
CFAP418 participates in membrane-associated cellular processes through binding lipids during ciliogenesis
STUDY_SUMMARY
Ciliopathies and retinal degenerative diseases are heterogeneous groups of genetic diseases. CFAP418 is a causative gene of both diseases, and its sequence is evolutionarily conserved. Here, we employ affinity purification coupled with mass spectrometry and quantitative lipidomic, proteomic, and phosphoproteomic approaches to address the function of CFAP418 in retinas. We show CFAP418 unexpectedly binds to lipid metabolism precursor phosphatidic acid (PA) and mitochondrion-specific lipid cardiolipin but does not form a tight and static complex with proteins. Loss of Cfap418 leads to a widespread disruption of membrane lipid homeostasis and changes in protein-membrane association, which subsequently causes mitochondrial morphological and functional defects and membrane remodeling abnormalities in multiple vesicular trafficking pathways. The signaling of PA-binding protein kinase Ca is increased. Our results indicate that membrane lipid imbalance is a new pathological mechanism underlying inherited ciliopathies and retinal degenerations, which is associated with other known causative RAB28 and BBS genes.
INSTITUTE
University of Utah - Metabolomics Core
LAST_NAME
Maschek
FIRST_NAME
John
ADDRESS
Emma Eccles Jones Medical Science Building, 15 N Medical Dr East, Salt Lake City, UT, 84112, USA
Remote solid cancers rewire hepatic nitrogen metabolism via host nicotinamide-N-methyltransferase
STUDY_SUMMARY
Cancers disrupt host homeostasis in various manners but the identity of host factors underlying such disruption remains largely unknown. Here we show that nicotinamide-N-methyltransferase (NNMT) is a novel host factor that mediates metabolic dysfunction in the livers of cancer-bearing mice. Multiple solid cancers distantly increase expression of Nnmt and its product 1-methylnicotinamide (MNAM) in the liver. Multi-omics analyses reveal suppression of the urea cycle accompanied by accumulation of amino acids, and enhancement of uracil biogenesis in the livers of cancer-bearing mice. Importantly, genetic deletion of Nnmt leads to alleviation of these metabolic abnormalities, and buffers cancer-dependent weight loss and reduction of the voluntary wheel-running activity. Our data also demonstrate that MNAM is capable of affecting urea cycle metabolites in the liver. These results suggest that cancers up-regulate the hepatic NNMT pathway to rewire liver metabolism towards uracil biogenesis rather than nitrogen disposal via the urea cycle, thereby disrupting host homeostasis. Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides,etc.) were analyzed via IC/HR/MS/MS. Cationic polar metabolites (i.e., amino acids, bases, nucleosides, NAM, SAM, MNAM, SAH, me2PY, me4PY, etc) were analyzed via PFPP-LC/HR/MS/MS.
TMEM41B and VMP1 modulate cellular lipid and energy metabolism for facilitating Dengue virus infection
STUDY_TYPE
untargeted analysis
STUDY_SUMMARY
Lipid metabolism is an intricate yet crucial cellular process co-opted by multiple viruses for replication and biogenesis. Transmembrane Protein 41B (TMEM41B) and Vacuole Membrane Protein 1 (VMP1) are two recently identified ER-resident lipid scramblases that play a role in autophagosome formation and cellular lipid metabolism. Importantly, TMEM41B is also a newly validated host dependency factor required for productive infection of several medically important enveloped RNA viruses, such as flaviviruses and human coronaviruses. However, the exact underlying mechanism of TMEM414B in modulating viral infections remains an open question. Here, we uncovered that TMEM41B and VMP1 deficiencies severely impaired replication of flavivirus and human coronavirus via multiple parallel cellular mechanisms. In accordance with previous reports, we validated that both TMEM41B and VMP1 are indispensable for all four serotypes of dengue virus (DENV) and human coronavirus OC43 (HCoV-OC43) to infect human cells, but not chikungunya virus, an alphavirus. Impaired dengue virus replication in TMEM41B and VMP1 deficient cells could induce a robust activation of innate immune RNA sensing as evidenced by hyperactivation of RIG-I and MDA5. However, this phenomenon was a consequence but not the root cause of the diminished viral replication. Notably, the impact of TMEM41B deficiency on DENV replication could be reversed by complementing the cells using exogenous unsaturated fatty acids, indicating a metabolic role for TMEM41B in flavivirus infection. Furthermore, we found that derailed cellular energy metabolism could be a contributing factor to block DENV infection as TMEM41B and VMP1 deficient cells harbored higher levels of compromised mitochondria that exhibited aberrant functions in facilitating beta-oxidation. Using lipidome and metabolome profiling of TMEM41B and VMP1 deficient cells, we further revealed that each of these genetic deficiencies result in distinctive cellular metabolic dysregulations, underlining their necessity for a balanced metabolic landscape, and strengthening the metabolic role of these ER membrane proteins in facilitating virus infection. Our results highlighted that TMEM41B and VMP1 are required for homeostasis of cellular metabolism, and this metabolic role contributes to their essentiality in facilitating DENV infection.
INSTITUTE
Singapore-MIT Alliance for Research and Technology (SMART Centre)
Untargeted metabolomics profiling of mice urine after 5 Gy and 7.5 Gy total body radiation exposure at 24 hours.
STUDY_TYPE
Untargeted LCMS
STUDY_SUMMARY
Untargeted metabolomics profiling of mice urine after 5 Gy and 7.5 Gy total body radiation exposure at 24 hours. Metabolic profiling of urine samples were performed by TripleTOF 5600+ (AB Sciex) in both ionization mode
INSTITUTE
Institute of Nuclear Medicine and Allied Sciences, Defence Research and Development Organisation, INDIA
Cancers disrupt host homeostasis in various manners but the identity of host factors underlying such disruption remains largely unknown. Here we show that nicotinamide-N-methyltransferase (NNMT) is a novel host factor that mediates metabolic dysfunction in the livers of cancer-bearing mice. Multiple solid cancers distantly increase expression of Nnmt and its product 1-methylnicotinamide (MNAM) in the liver. Multi-omics analyses reveal suppression of the urea cycle accompanied by accumulation of amino acids, and enhancement of uracil biogenesis in the livers of cancer-bearing mice. Importantly, genetic deletion of Nnmt leads to alleviation of these metabolic abnormalities, and buffers cancer-dependent weight loss and reduction of the voluntary wheel-running activity. Our data also demonstrate that MNAM is capable of affecting urea cycle metabolites in the liver. These results suggest that cancers up-regulate the hepatic NNMT pathway to rewire liver metabolism towards uracil biogenesis rather than nitrogen disposal via the urea cycle, thereby disrupting host homeostasis. Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides, etc.) were analyzed via IC/HR/MS/MS. Cationic polar metabolites (i.e., amino acids, bases, nucleosides, NAM, SAM, MNAM, SAH, me2PY, me4PY, etc) were analyzed via PFPP-LC/HR/MS/MS.
Multi-omics analyses of 398 foxtail millet accessions reveal genomic regions associated with domestication, metabolite traits and anti-inflammatory effects
STUDY_SUMMARY
Foxtail millet, domesticated from the wild species green foxtail, provides a rich source of phytonutrients for humans. To evaluate how breeding changed the metabolome of foxtail millet grains, we generated and analyzed datasets encompassing genomes, transcriptomes, metabolomes and anti-inflammatory indices from 398 foxtail millet accessions. We identified hundreds of common variants that influence numerous secondary metabolites, with significant heterogeneity in the natural variation of metabolites and their underlying genetic architectures between different sub-groups of foxtail millet. The combined results from variations in genome, transcriptome and metabolome illustrated how breeding has altered foxtail millet metabolite content. Selection for alleles of genes associated with yellow grains led to altered metabolite profiles, such as carotenoids and endogenous hormones. The importance of PSY1 (phytoene synthase 1) for millet color was validated using CRISPR-Cas9. The in vitro cell inflammation assay showed that 83 metabolites have anti-inflammatory effects. This multi-omics study illustrates how the breeding history of foxtail millet has impacted metabolites. It provides some fundamental resources for understanding how grain quality could be associated with different metabolites, and highlights future perspectives on millet genetic research and metabolome-assisted improvement.
INSTITUTE
Shanxi Agricultural University
DEPARTMENT
College of Life Sciences
LABORATORY
Shanxi Key Laboratory of Minor Crop Germplasm Innovation and Molecular Breeding
Identifying putative key metabolites from fingerprinting metabolomics for the authentication of rice origin: A case study of Sengcu rice
STUDY_TYPE
MS Untargeted analysis
STUDY_SUMMARY
The expanding scale and nature of rice fraud in the global food system has caused major economic and human health concerns. Herein, an untargeted metabolomics approach based on the UHPLC-Q-Orbitrap-HRMS system was utilized for the discrimination between authentic and commercial Sengcu rice, a local specialty cultivated by terraced farming in northern Vietnam. A total of 8398 positive and 5250 negative mode compounds were introduced to multivariate statistical analyses for the construction of classification models. The first two principal components explaining 52% of the total variance in both datasets exhibited distinguished clusters of authentic against commercial Sengcu rice. Partial least squares-discriminant analysis models were optimized to obtain the optimal number of retained components, the optimal number of variables retained in each component and the best prediction distance type for model evaluation. One component containing five positive (DMG, RSA, RCA, PAL and BOSe) and six negative mode variables (PXP, RXP, TDHP, ISS, MXP and RGB) was sufficient to discriminate between authentic and commercial Sengcu rice. The classification error rate was less than 1.1310-4, as determined from repeated k-fold cross validation. These putative signature metabolites clearly separated authentic and commercial Sengcu rice in the hierarchical clustering models. In addition, the isolated metabolites also reflected the cultivation practices of terraced farming of authentic Sengcu rice. Overall, we have proposed an effective method for the identification of key metabolites from fingerprinting metabolomics, and it could serve as a fundamental approach for other in-depth food authentication studies.
INSTITUTE
Institute of Chemistry, Vietnam Academy of Science and Technology
LABORATORY
Laboratory of Environmental and Bioorganic Chemistry
Cecal metabolome of specific-pathogen-free mice fed with five distinct rodent diets with varying fiber content and source.
STUDY_SUMMARY
In an effort to discern the effects of diverse fibers on host immunity, we employed five distinct rodent diets with varying fiber content and source in specific-pathogen-free.
Serum metabolome of specific-pathogen-free mice fed with five distinct rodent diets with varying fiber content and source.
STUDY_SUMMARY
In an effort to discern the effects of diverse fibers on host immunity, we employed five distinct rodent diets with varying fiber content and source in specific-pathogen-free.
Cecal metabolome of gnotobiotic (containing a 14-member synthetic human gut microbiota), and germ-free mice fed with two distinct rodent diets with varying fiber content.
STUDY_SUMMARY
In an effort to discern the effects of diverse fibers on host immunity, we employed five distinct rodent diets with varying fiber content in gnotobiotic (containing a 14-member synthetic human gut microbiota), and germ-free mice.
Unveiling the mechanism of action of nature-inspired anti-cancer compounds using a multi-Omics approach
STUDY_SUMMARY
Novel anti-cancer compounds SIMR-3058 and SIMR-3066 were tested against MCF-7. The 2020 global cancer registry has ranked breast cancer (BCa) as the most commonly diagnosed type of cancer and the most common cause of cancer-related deaths, especially in women worldwide. Increased resistance and significant side effects continue to limit the efficacy of anti-BCa drugs, hence the need to identify new drug targets and to develop novel compounds to overcome these limitations. Nature-inspired anti-cancer compounds are becoming increasingly popular since they often provide a relatively safe and effective alternative. In this study, we employed multi-omics techniques to gain insights into the novel and potentially relevant mechanism of action of two recently identified nature- inspired anti-cancer compounds (SIMR 3066 and SIMR 3058). Discovery proteomics analysis combined with LC-MS/MS-based untargeted metabolomics analysis was performed on compound-treated vs. DMSO-treated control MCF-7 cells. Downstream protein functional analysis showed that most of the responsive proteins were functionally associated with antigen processing and neutrophil degranulation, RNA catabolism and protein folding as well as cytoplasmic vesicle lumen and mitochondrial matrix formation. Consistent with the proteomics findings, metabolomic pathway analysis suggested that SIMR compounds could alter metabolic pathways such as glycolysis, the Krebs cycle and oxidative phosphorylation. Furthermore, metabolomics-based enriched-for-action pathway analysis showed that the of two SIMR compounds associate with mercaptopurine and thioguanine and azathiprine. Lastly, joint proteomics and metabolomics analysis revealed that treatment of BCa with SIMR3066 disrupts several signaling pathways including such p53-mediated apoptosis and the circadian entertainment pathway. Overall, the multi-omics we used in this study seems potent at probing the mechanism of action of novel anti-cancer agents.
Identifying a tryptophan derivative in hydrogen peroxide-treated cell culture medium
STUDY_SUMMARY
Reactive oxygen species (ROS) are by-products of metabolism of oxygen and they play an important role in normal homeostasis and cell signaling, as well as in the initiation of diseases including cancer when their production is upregulated. Thus, it is imperative to understand the cellular and molecular basis by which ROS impact on various biological and pathological processes. Here, we identified 2-oxindole, a tryptophan derivative, was a major catabolic product in hydrogen peroxide-treated cell culture medium. We used 2-oxindole to study its role in regulating AhR signaling and tryptophan metabolic pathways. We found that 2-oxindole significantly increased the activity of AhR, leading to enhanced expression of its downstream targets including cytochrome P450 genes.
INSTITUTE
NYU Langone Health
DEPARTMENT
Department of Biochemistry and Molecular Pharmacology
Effect of external high-dose rate radiation on mouse biofluid metabolomic signatures
STUDY_SUMMARY
In the event of an improvised nuclear device (IND), a complex IR exposure will occur consisting of both low (LDR) and high-dose rates (HDR). We have previously addressed LDR exposures from internal emitters or externally deposited radionuclides on biofluid small molecule signatures, but further research on the HDR component is required. Here, we exposed 8 ā 10 week old male C57BL/6 mice to a cumulative dose of 3 Gy using a reference dose rate of 0.7 Gy/min or a HDR of 7 Gy/sec, collected urine and serum at 1 and 7 d, then compared the metabolite signatures using either untargeted (urine) or targeted (serum) approaches with liquid chromatography mass spectrometry platforms.
Effect of external high-dose rate radiation on mouse biofluid metabolomic signatures (neg)
STUDY_SUMMARY
In the event of an improvised nuclear device (IND), a complex IR exposure will occur consisting of both low (LDR) and high-dose rates (HDR). We have previously addressed LDR exposures from internal emitters or externally deposited radionuclides on biofluid small molecule signatures, but further research on the HDR component is required. Here, we exposed 8 ā 10 week old male C57BL/6 mice to a cumulative dose of 3 Gy using a reference dose rate of 0.7 Gy/min or a HDR of 7 Gy/sec, collected urine and serum at 1 and 7 d, then compared the metabolite signatures using an untargeted (urine) approach with liquid chromatography mass spectrometry platforms.
Multiomics Analyses Reveal the Central Role of Pentose Phosphate Pathway in Resident Thymic Macrophages to Cope with Efferocytosis-Associated Stress
STUDY_SUMMARY
Tissue-resident macrophages (TRMs) are heterogeneous cell populations found throughout the body. Depending on their location, they perform diverse functions maintaining tissue homeostasis and providing immune surveillance. To survive and function within, TRMs adapt metabolically to the distinct microenvironments. However, little is known about the metabolic signatures of TRMs. The thymus provides a nurturing milieu for developing thymocytes yet efficiently removes those that failed the selection, relying on the TRMs ā resident thymic macrophages (TMĻs). This study harnesses multiomics analyses to characterize TMĻs and unveils their unique metabolic features. We find that the pentose phosphate pathway (PPP) is preferentially activated in TMĻs, responding to the reduction-oxidation demands associated with the efferocytosis of dying thymocytes. The blockade of PPP in MĻs leads to decreased efferocytosis, which can be rescued by ROS scavengers. Our study reveals the key role of PPP in TMĻs and underscores the importance of metabolic adaptation in supporting MĻ efferocytosis.
Age-independent Cardiac Protection by Pharmacological Activation of Beclin-1 During Endotoxemia and Its Association with Energy Metabolic Reprograming in Myocardium ā A Targeted Metabolomics Study
STUDY_SUMMARY
Background: We previously showed that Beclin-1-dependent autophagy is cardiac protective in a rodent model of endotoxemia using young adult mice. In this report, we compared the potential therapeutic effects of pharmacological Beclin-1 activating peptide, TB-peptide, on the cardiac outcomes of young adult and aged mice during endotoxemia. We further examined alterations in myocardial metabolism induced by lipopolysaccharide (LPS) challenge with and without the TB-peptide treatment. Methods and Results: C57BL/6J mice of 10-week and 24-month-old were challenged by LPS at doses at which cardiac dysfunction occurred. Following the treatment of TB-peptide or control vehicle, heart contractility, circulating cytokines, and myocardial autophagy were evaluated. A targeted metabolomics assay was applied to analyze cardiac metabolism. TB-peptide boosted autophagic response, attenuated cytokine production, and improved cardiac performance in both young and aged mice during endotoxemia. A targeted metabolomics assay was designed to detect a pool of 361 known metabolites, of which 156 were detected in at least one of the heart tissue samples. LPS-induced impairments were found in glucose and amino acid (AA) metabolisms in mice of all ages, and TB-peptide provided ameliorative effects to rescue these alterations. However, lipid metabolites were upregulated in the young group but moderately downregulated in the aged by LPS, suggesting an age-dependent response. TB-peptide mitigated LPS-mediated trend of lipids in the young mice but provided little effect on the aged ones. Conclusion: Pharmacological activation of Beclin-1 by TB-peptide protects the heart in both young and aged population during endotoxemia, suggest a therapeutic potential for sepsis-induced cardiomyopathy. Metabolomics analysis suggests that this age-independent protection by TB-peptide is associated with reprograming of energy production via glucose and AA metabolisms.
INSTITUTE
Loyola University Chicago Stritch School of Medicine
Impact of nitisinone on the cerebrospinal fluid metabolome of a murine model of alkaptonuria
STUDY_SUMMARY
Background: Nitisinone induced hypertyrosinaemia is well documented in Alkaptonuria (AKU), and there is uncertainty over whether it may contribute to a decline in cognitive function and or mood by altering neurotransmitter metabolism. The aim of this work was to evaluate the impact of nitisinone on the cerebrospinal fluid (CSF) metabolome in a murine model of AKU, with a view to providing additional insight into metabolic changes that occur following treatment with nitisinone. Methods: 17 CSF samples were collected from BALB/c Hgd-/-mice (n=8, treated with nitisinone ā 4 mg/L and n=9, no treatment). Samples were diluted 1:1 with deionised water and analysed using a 1290 Infinity II liquid chromatography system coupled to a 6550 quadrupole time-of-flight mass spectrometry (Agilent, Cheadle, UK). Raw data were processed using a targeted feature extraction algorithm and an established in-house accurate mass retention time database. Matched entities (Ā±10 ppm theoretical accurate mass and Ā±0.3 minutes retention time window) were filtered based on their frequency and variability. Experimental groups were compared using a moderated t-test with Benjamini-Hochberg false-discovery rate adjustment. Results: Tyrosine, acetyl-tyrosine, Ī³-glutamyl-tyrosine, p-hydroxyphenylacetic acid and 3-(4-hydroxyphenyl)lactic acid were shown to increase in abundance (log2 fold change 2.6-6.9, 3/5 were significant p<0.05) in the mice that received nitisinone. Several other metabolites of interest were matched but no significant differences were observed, including the aromatic amino acids phenylalanine and tryptophan, and monoamine metabolites adrenaline, 3-methoxy-4-hydroxyphenylglycol and octopamine. Conclusions: Evaluation of the CSF metabolome of a murine model of AKU showed a significant difference in the abundance of a limited number of metabolites. None of these have been reported in CSF from a murine model of AKU previously. Moreover this study confirms that some monoamine metabolites do not appear to be altered following nitisinone therapy.
INSTITUTE
University of Liverpool Institute of Life Course & Medical Sciences
LAST_NAME
Davison
FIRST_NAME
Andrew
ADDRESS
1. Department of Clinical Biochemistry and Metabolic Medicine, Liverpool Clinical Laboratories, Liverpool University Hospitals NHS Foundation Trust, Liverpool, UK; 2. Department of Musculoskeletal and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool, UK 3. School of Exercise Science, Liverpool John Moores University, Liverpool, UK
Global, distinctive and personal changes in molecular and microbial profiles induced by specific fibers in humans (Targeted)
STUDY_SUMMARY
Dietary fibers act through the microbiome and improve cardiovascular health, metabolic disorders and cancer prevention. To understand health benefits of dietary fiber supplementation we investigated two popular purified fibers, arabinoxylan (AX) and long-chain inulin (LCI), and a mixture of five fibers. We present multi-omic signatures of metabolomics, lipidomics, proteomics, metagenomics, a cytokine panel and clinical measurements on healthy and insulin resistant participants. Each fiber is associated with fiber-dependent biochemical and microbial responses. AX consumption associates with a significant reduction in LDL and an increase in bile acids, contributing to its observed cholesterol reduction. LCI is associated with an increase in Bifidobacterium. However, at the highest LCI dose there is increased inflammation and elevation in the liver enzyme alanine aminotransferase. This study yields insights into the effects of fiber supplementation, it provides insights into mechanisms behind fiber induced cholesterol reduction, and it shows effects of individual, purified fibers on the microbiome.
Piperaquine-resistant PfCRT mutations differentially impact drug transport, hemoglobin catabolism and parasite physiology in Plasmodium falciparum asexual blood stages.
STUDY_SUMMARY
The emergence of multidrug resistance in Plasmodium falciparum parasites presents a significant obstacle to the malaria elimination agenda. Resistance to piperaquine (PPQ), an important first-line partner drug, has spread across Southeast Asia where it has contributed to widespread treatment failures. The genetic cause of resistance to PPQ is attributable to a novel set of amino acid substitutions in the P. falciparum chloroquine resistance transporter (PfCRT). In this study, we used magnetically-purified trophozoite extracts from seven different lines comprising five genetically-modified and two field isolates. Three independent extractions with triplicate technical repeats were run by mass spectrometry on positive and negative modes and spectral peak raw data analyzed to obtain the fold change difference between the samples and parental control, Dd2Dd2crt. We show that PPQ-resistant, PfCRT mutant asexual blood stage parasites accumulate higher levels of hemoglobin-derived peptides than do their PPQ-sensitive counterparts.
INSTITUTE
The Pennsylvania State University
DEPARTMENT
Department of Biochemistry and Molecular Biology
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millenium Science Complex, University Park, PA 16802
Amelioration of developmental programming of NAFLD in weanling liver using PQQ
STUDY_TYPE
Diet and PQQ treatment
STUDY_SUMMARY
Maternal obesity and consumption of a high-fat diet significantly elevate risk for pediatric non-alcoholic fatty liver disease (NAFLD), affecting 10% of children in the US. Almost half of these children are diagnosed with nonalcoholic steatohepatitis (NASH), a leading etiology for liver transplant. Animal models show that signs of liver injury and perturbed lipid metabolism asso-ciated with NAFLD begin in utero; however, safe dietary therapeutics to blunt developmental programming of NAFLD are unavailable. Using a mouse model of maternal Western-style diet (WD), we previously showed that pyrroloquinoline quinone (PQQ), a potent dietary antioxidant, protected offspring of WD-fed dams from development of NAFLD and NASH. Here, we used untargeted mass spectrometry-based lipidomics to delineate lipotoxic effects of WD on offspring liver and identify lipid targets of PQQ. PQQ exposure during pregnancy altered hepatic lipid profiles of WD-exposed offspring, upregulating peroxisome proliferator-activated receptor (PPAR) Ī± signaling and mitochondrial fatty acid oxidation to markedly attenuate triglyceride accumulation beginning in utero. Surprisingly, the abundance of very long-chain ceramides, important in promoting gut barrier and hepatic function, was significantly elevated in PQQ-treated offspring. PQQ exposure reduced the hepatic phosphatidylcho-line/phosphatidylethanolamine (PC/PE) ratio in WD-fed offspring and improved glucose toler-ance. Notably, levels of protective n ā 3 polyunsaturated fatty acids (PUFAs) were elevated in offspring exposed to PQQ, beginning in utero, and the increase in n ā 3 PUFAs persisted into adulthood. Our findings suggest that PQQ supplementation during gestation and lactation augments pathways involved in the biosynthesis of long-chain fatty acids and plays a unique role in modifying specific bioactive lipid species critical for protection against NAFLD risk in later life.
INSTITUTE
University of Oklahoma Health Sciences Center
DEPARTMENT
Biochemistry and Molecular Biology, Harold Hamm Diabetes Center
LABORATORY
Jonscher
LAST_NAME
Jonscher
FIRST_NAME
Karen
ADDRESS
975 NE 10th Street BRC-N 362A, Oklahoma City, OK, 73104, USA
EMAIL
karen-jonscher@ouhsc.edu
PHONE
3032294620
NUM_GROUPS
3
TOTAL_SUBJECTS
9
PUBLICATIONS
Jonscher, et al FASEB J 2017; Friedman, et al Hepatol Commun 2018
Individualized exercise intervention for people with multiple myeloma improves quality of life in a randomized controlled trial
STUDY_SUMMARY
Although new treatments have improved survival for multiple myeloma (MM), quality of life remains poor for people with this incurable cancer. We conducted a multi-site randomized, waitlist-controlled trial of an individualized exercise program for people at all stages of MM (n=60). Compared to the waitlist control group, participants of the 12-week intervention had significant improvement in health-related quality of life, mediated through improved MM symptoms, cardiorespiratory fitness and bone pain, with were mostly maintained at follow-up (up to 12 months). Exploratory plasma metabolomics and lipidomics was conducted to delineate molecular mechanisms and biomarkers
Metabolic effect of the loss of mitochondrial-specific aspartyl-tRNA synthetase Das2 on mouse intestinal epithelial cells
STUDY_SUMMARY
We analysed the intestinal epithelial cell (IEC) from Dars2 fl/fl ; VillinCreERT2 tg/wt mice (n=15) and and Dars2 fl/fl ; VillinCreERT2 wt/wt mice (n=9) at 8 days upon tamoxifen injection to assess the metabolic effect of Das2 loss. Isolated IECs were divided into three technical replicates (n=69) and analysed with two analytical repeats (n=138).
Lipidomic characterization of Jurkat-derived T cell line in which the chaperonin complex CCT has been partially silenced
STUDY_TYPE
Untargeted Lipidomics
STUDY_SUMMARY
When the chaperonin complex CCT is partially silenced in Jurkat-derived T cell line J77 E61, we observe changes in the production of exosomes and in their composition. Thus, to explore the bases of these alterations and find possible new mechanisms of exosome biosynthesis regulation we characterized the lipidome content in cells where the chaperonin complex CCT was partially silent (CCT) and controls (CTRL). We analyzed 5 biological replicates containing 3 Ć 106 cells using LC-MS in positive and negative polarity mode. For quality control, 5 QCs samples and 4 blanks were also included in the analysis (total 19 samples and 2 groups).
INSTITUTE
Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC)
DEPARTMENT
Proteomics and Metabolomics Unit
LABORATORY
Metabolomics Lab
LAST_NAME
Ferrarini
FIRST_NAME
Alessia
ADDRESS
Calle de Melchor FernƔndez Almagro, 3, Madrid, Madrid, 28029, Spain
An early-life microbiota metabolite protects against obesity via intestinal PPAR-gamma
STUDY_SUMMARY
The mechanisms by which the early-life microbiota protects against environmental factors that promote childhood obesity remain largely unknown. Using a mouse model in which young mice are simultaneously exposed to antibiotics and a high-fat diet, we show that Lactobacillus species, predominant members of the small intestine microbiota, regulate intestinal epithelial cells (IECs) to limit diet-induced obesity during early-life. A Lactobacillus-derived metabolite, phenyllactic acid (PLA), protected against metabolic dysfunction caused by early-life exposure to antibiotics and a high-fat diet by increasing the abundance of peroxisome proliferator activated receptor gamma (PPAR-gamma) in the small intestine IECs. Therefore, PLA is a microbiota-derived metabolite that activates protective pathways in the small intestine epithelium to regulate fat absorption and prevent obesity during early life.
Interplay of CodY and CcpA in regulating central metabolism and biofilm formation in S. aureus
STUDY_TYPE
Research
STUDY_SUMMARY
Staphylococcus aureus is a medically important pathogen that exhibit high metabolic versatility allowing it to infect various niches within a host. S. aureus utilizes two major transcriptional regulators, CodY and CcpA, to remodel metabolic and virulence gene expression in response to changing environmental conditions. Previous studies revealed that inactivation of either codY or ccpA has a pronounced impact on different aspects of staphylococcal physiology and pathogenesis. To determine the contribution and interplay of these two regulators in modulating central metabolism, virulence, and biofilm development we constructed and characterized codY ccpA double mutant in S. aureus UAMS-1. In line with previous studies, we found that CcpA and CodY control cellular metabolic status by altering carbon flow through the central and overflow metabolic pathways. Our results demonstrate that ccpA inactivation impairs biofilm formation and decreases incorporation of eDNA into the biofilm matrix while disrupting codY resulted in a robust structured biofilm tethered together with eDNA and PIA. Interestingly, inactivation of both codY and ccpA decreases biofilm biomass and neglects eDNA release in the double mutant. Compared to inactivation of codY, the codY ccpA mutant did not overexpress toxins but maintained overexpression of amino acid metabolism pathways. Furthermore, codY ccpA mutant produced higher amounts of PIA, in contrast to the wild-type strain and ccpA mutant. Overall, results of this study suggest that interplay between CodY and CcpA modulates central metabolism to optimize growth on preferred carbon sources while repressing virulence gene expression until nutrient limitation requires scavenging nutrients from the host.
INSTITUTE
University of Nebraska Medical Center
DEPARTMENT
Pathology and Microbiology
LAST_NAME
Sadykov
FIRST_NAME
Marat
ADDRESS
UNMC Department of Pathology and Microbiology 985900 Nebraska Medical Center Omaha, NE 68198-5900
Metabolomic profiling reveals muscle metabolic changes following iliac arteriovenous fistula creation in mice
STUDY_TYPE
Study of the skeletal muscle metabolome in mice with iliac arteriovenous fistula via 1H NMR
STUDY_SUMMARY
In the present study, we hypothesize that the creation of an iliac AVF would result in significant alterations to the limb muscle metabolome. Recently, our group developed a new murine model to address the pathophysiology of access-related hand dysfunction (ARHD) in mice, where AVF creation is performed in the iliac artery/vein. Because of the anatomical location of the AVF creation, this model produces clinically relevant changes in the mouse hindlimb including hemodynamic alterations, muscle weakness, and mitochondrial function impairment.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Metabolomic profiling reveals muscle metabolic changes following iliac arteriovenous fistula creation in mice (Aqueous)
STUDY_SUMMARY
In the present study, we hypothesize that the creation of an iliac AVF would result in significant alterations to the limb muscle metabolome. Recently, our group developed a new murine model to address the pathophysiology of access-related hand dysfunction (ARHD) in mice, where AVF creation is performed in the iliac artery/vein. Because of the anatomical location of the AVF creation, this model produces clinically relevant changes in the mouse hindlimb including hemodynamic alterations, muscle weakness, and mitochondrial function impairment.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Ryan
FIRST_NAME
Terence
ADDRESS
1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
EMAIL
ryant@ufl.edu
NUM_GROUPS
4
TOTAL_SUBJECTS
34
NUM_MALES
All
STUDY_COMMENTS
Metabolomic study via NMR
PUBLICATIONS
Frontiers
STUDY_TYPE
Study of the skeletal muscle metabolome in mice with iliac arteriovenous fistula via 1H NMR
Mass Spectrometry Imaging of Lipids In A Gut Epithelial Cell Model
STUDY_TYPE
Mass spectromery imaging of cells.
STUDY_SUMMARY
Scope: The Caco2/HT29-MTX co-culture system is widely used as a cell model of the intestinal epithelium. Although the gut epithelium plays an important role in the uptake of free fatty acids and the resynthesis of triglycerides the lipid distribution profile of the co-culture system is not well understood. Desorption electrospray ionization (DESI) is a mass spectrometry (MS) technique which has been widely used to study the main classes of lipid molecules on different tissue surfaces. This has been used to map lipid species and their distribution in Caco2 and HT29-MTX co-culture system. Methods and results: Caco2 and HT29-MTX cells were seeded on coverslips either singly or as cocultures in ratios of 75:25 and 50:50. Cells were cultured for 21 days before MS imaging using a DESI source in both the positive and negative ionization modes. The identity of selected lipids was confirmed in negative and positive ionisation modes using tandem MS. Although many lipids were common to both cell lines, there were distinctive patterns in the lipidomes. Thus, the lipidome of Caco2 cells was more heterogeneous and rich in cholesterol esters and triglycerides whilst HT29-MTX cells has a distinctive lipidome relating to phosphatidylethanolamines, phosphatidylinositols and odd chain lipids, including C17 fatty acids. Conclusion: DESI-MSI has shown that Caco2 and HT29-MTX cells have distinctive lipidomes which are still evident when the cells are cocultured. It has potential to both allow further validation of these widely used cell models and provide insights into how dietary components may modify lipid metabolism in future.
INSTITUTE
Manchester Institute of Biotechnology, University of Manchester
LAST_NAME
Mattar
FIRST_NAME
Hadeer
ADDRESS
Manchester Institute of Biotechnology, Princess Street, Manchester, UK, M1 7DN
Amelioration of developmental programming of NAFLD in fetal liver using PQQ
STUDY_TYPE
Diet and PQQ treatment
STUDY_SUMMARY
Maternal obesity and consumption of a high-fat diet significantly elevate risk for pediatric non-alcoholic fatty liver disease (NAFLD), affecting 10% of children in the US. Almost half of these children are diagnosed with nonalcoholic steatohepatitis (NASH), a leading etiology for liver transplant. Animal models show that signs of liver injury and perturbed lipid metabolism asso-ciated with NAFLD begin in utero; however, safe dietary therapeutics to blunt developmental programming of NAFLD are unavailable. Using a mouse model of maternal Western-style diet (WD), we previously showed that pyrroloquinoline quinone (PQQ), a potent dietary antioxidant, protected offspring of WD-fed dams from development of NAFLD and NASH. Here, we used untargeted mass spectrometry-based lipidomics to delineate lipotoxic effects of WD on offspring liver and identify lipid targets of PQQ. PQQ exposure during pregnancy altered hepatic lipid profiles of WD-exposed offspring, upregulating peroxisome proliferator-activated receptor (PPAR) Ī± signaling and mitochondrial fatty acid oxidation to markedly attenuate triglyceride accumulation beginning in utero. Surprisingly, the abundance of very long-chain ceramides, important in promoting gut barrier and hepatic function, was significantly elevated in PQQ-treated offspring. PQQ exposure reduced the hepatic phosphatidylcho-line/phosphatidylethanolamine (PC/PE) ratio in WD-fed offspring and improved glucose toler-ance. Notably, levels of protective n ā 3 polyunsaturated fatty acids (PUFAs) were elevated in offspring exposed to PQQ, beginning in utero, and the increase in n ā 3 PUFAs persisted into adulthood. Our findings suggest that PQQ supplementation during gestation and lactation augments pathways involved in the biosynthesis of long-chain fatty acids and plays a unique role in modifying specific bioactive lipid species critical for protection against NAFLD risk in later life.
INSTITUTE
University of Oklahoma Health Sciences Center
DEPARTMENT
Biochemistry and Molecular Biology, Harold Hamm Diabetes Center
LABORATORY
Jonscher
LAST_NAME
Jonscher
FIRST_NAME
Karen
ADDRESS
975 NE 10th Street BRC-N 362A, Oklahoma City, OK, 73104, USA
EMAIL
karen-jonscher@ouhsc.edu
PHONE
3032294620
NUM_GROUPS
3
TOTAL_SUBJECTS
9
PUBLICATIONS
Jonscher, et al FASEB J 2017; Friedman, et al Hepatol Commun 2018
Amelioration of developmental programming of NAFLD in adult liver using PQQ
STUDY_TYPE
Pre-natal and Post-natal Diet and PQQ treatment
STUDY_SUMMARY
Maternal obesity and consumption of a high-fat diet significantly elevate risk for pediatric non-alcoholic fatty liver disease (NAFLD), affecting 10% of children in the US. Almost half of these children are diagnosed with nonalcoholic steatohepatitis (NASH), a leading etiology for liver transplant. Animal models show that signs of liver injury and perturbed lipid metabolism asso-ciated with NAFLD begin in utero; however, safe dietary therapeutics to blunt developmental programming of NAFLD are unavailable. Using a mouse model of maternal Western-style diet (WD), we previously showed that pyrroloquinoline quinone (PQQ), a potent dietary antioxidant, protected offspring of WD-fed dams from development of NAFLD and NASH. Here, we used untargeted mass spectrometry-based lipidomics to delineate lipotoxic effects of WD on offspring liver and identify lipid targets of PQQ. PQQ exposure during pregnancy altered hepatic lipid profiles of WD-exposed offspring, upregulating peroxisome proliferator-activated receptor (PPAR) Ī± signaling and mitochondrial fatty acid oxidation to markedly attenuate triglyceride accumulation beginning in utero. Surprisingly, the abundance of very long-chain ceramides, important in promoting gut barrier and hepatic function, was significantly elevated in PQQ-treated offspring. PQQ exposure reduced the hepatic phosphatidylcho-line/phosphatidylethanolamine (PC/PE) ratio in WD-fed offspring and improved glucose toler-ance. Notably, levels of protective n ā 3 polyunsaturated fatty acids (PUFAs) were elevated in offspring exposed to PQQ, beginning in utero, and the increase in n ā 3 PUFAs persisted into adulthood. Our findings suggest that PQQ supplementation during gestation and lactation augments pathways involved in the biosynthesis of long-chain fatty acids and plays a unique role in modifying specific bioactive lipid species critical for protection against NAFLD risk in later life.
INSTITUTE
University of Oklahoma Health Sciences Center
DEPARTMENT
Biochemistry and Molecular Biology, Harold Hamm Diabetes Center
LABORATORY
Jonscher
LAST_NAME
Jonscher
FIRST_NAME
Karen
ADDRESS
975 NE 10th Street BRC-N 362A, Oklahoma City, OK, 73104, USA
EMAIL
karen-jonscher@ouhsc.edu
PHONE
3032294620
NUM_GROUPS
4
TOTAL_SUBJECTS
24
NUM_MALES
24
PUBLICATIONS
Jonscher, et al FASEB J 2017; Friedman, et al Hepatol Commun 2018
The effects of obesity microbiota produced metabolites on colorectal carcinogenesis in murine models
STUDY_SUMMARY
Obesity is a risk factor for colorectal cancer (CRC). We aim to study the effects and mechanisms of gut microbiota of obese subjects in contributing to CRC progression. Conventional AOM-treated and ApcMin/+ mice receiving feces from obese individuals showed significantly increased colon tumor formation compared with those receiving feces from control subjects. AOM-treated mice receiving feces from obese (OB-M) exhibited microbiota dysbiosis with enriched potential pathobionts Erysipelotrichaceae bacterium GAM147, Turicibacter sp. H121, Mucinivorans hirudinis, and depleted symbionts Bacteroides vulgatus, Faecalibaculum rodentium, Bifidobacterium spp. and Lactobacillus delbrueckii. The OB-M group also showed altered gut metabolites including elevated phenylacetic acid, and depleted genipin. Moreover, OB-M group showed impaired intestinal barrier function and significant upregulation of pro-inflammatory cytokines and activation of oncogenic Wnt signaling pathway. In conclusion, gut microbiota from obese individuals promotes colorectal carcinogenesis. Microbiota modulation in obese individuals may provide new insight into obesity-driven CRC prevention and therapy.
INSTITUTE
The Chinese University of Hong Kong
LAST_NAME
Kang
FIRST_NAME
Xing
ADDRESS
Rm806, Li Ka Shing Medical Science Building, PWH, Shatin, Hong Kong
Metabolic profiling at COVID-19 onset shows disease severity and sex-specific dysregulation
STUDY_SUMMARY
In this study CE-MS and GC-MS based metabolomics was used to analyze COVID-19 disease. In total, 144 individuals classified as healthy, asymptomatic/mils, moderate and severe according to the highest COVID-19 severity status, were analyzed.
Untargeted lipidomics studies in the course of dermatitis onset and progression
STUDY_SUMMARY
We applied untargeted lipidomic analysis to the atopic dermatitis-like dermatitis model (Spade mice) to capture the comprehensive lipidome profile in the course of dermatitis onset and progression. Spade mice harbor a single amino acid mutation in Jak1 that causes hyperactivation, leading to Th2 dermatitis. Progressive dermatitis develops as desquamation and redness of the ears at approximately 8 weeks of age. At 10 weeks of age, serum IgE and IgG1 levels are increased, and Th2 cytokines, such as IL-4, IL-5, and IL-13, produced by CD4+ cells are upregulated, followed by elevated serum histamine levels at 12 weeks of age. Skin lesions manifest as epidermal hyperplasia at 8 weeks of age, while there are few morphological changes at 4 weeks of age. TEWL, the readout for barrier function, is significantly elevated in Spade mice at 4 weeks of age, suggesting that barrier defects had occurred before disease onset. WT and Spade skin tissues in P0, 4, 8, and 10 weeks of age were applied to untargeted lipidomics. Over 700 skin lipids including glycerophospholipids, ceramides, neutral lipids, and fatty acids were successfully annotated, and many of them were found to be significantly changed after dermatitis onset as determined by pruritus and erythema. Among them, the levels of Cer[NdS] containing very long-chain (C22 or more) fatty acids were significantly downregulated before AD onset.
INSTITUTE
Graduate School of Pharmaceutical Sciences, Keio University
LAST_NAME
Iino
FIRST_NAME
Yudai
ADDRESS
1-5-30, Shibakoen, Minato-ku, Tokyo, 105-8512, Japan
Metabolic signature of idiopathic inflammatory myopathy
STUDY_SUMMARY
Objective of this study was to find Metabolic signature of idiopathic inflammatory myopathy (IIM). We used the serum samples of healthy control, IIM, ankylosing spondylitis. Metabolites were quantified using biocrates p180 kit.
INSTITUTE
Seoul National University
LAST_NAME
Kang
FIRST_NAME
Jihyun
ADDRESS
502ho Intergrative Biomedical Education Research Building, 101 Deahakro, Jongro-gu, Seoul, 03080, Korea, South Korea
Metabolic signature of C-protein induced myositis mouse model
STUDY_SUMMARY
Objective of this study was to find Metabolic signature of C-protein induced myositis mouse model. We used the tissue samples of C-protein induced myositis mouse model and control using biocrates p180 kit.
INSTITUTE
Seoul National University
LAST_NAME
Kang
FIRST_NAME
Jihyun
ADDRESS
502ho Intergrative Biomedical Education Research Building, 101 Deahakro, Jongro-gu, Seoul, 03080, Korea, South Korea
Untargeted metabolomics of Pinus pinaster needles under heat and drought stress
STUDY_TYPE
Untargeted MS-based metabolomics
STUDY_SUMMARY
Current projections for global climate change predict an increase in the intensity and frequency of heat waves and droughts. The improvement in our understanding of the mechanisms of how trees precisely can predict environmental threats and cope with these stresses benefits our natural selection or genetic improvement to the maintenance of forest sustainability. In this work, we investigate the metabolic changes in heat and drought combined stress in Pinus pinaster plantlets. Maritime pine is a coniferous tree with native populations distributed across the European Atlantic and Mediterranean basins and the north of Africa ranging from cool moist to warm dry climates. This species shows high plasticity and a contrasting adaptive capacity and resilience. This plasticity in the response to stress exposure may be associated with a differential ability to modulate their secondary metabolism. For this reason, the current study aims to investigate the gradual and synergetic metabolomic response using liquid chromatography coupled to mass spectrometry (LC-MS) based on untargeted metabolomic profiling of four stress levels. These metabolic profiles were supported by physiological and biochemical determinations. Our results showed that the metabolic profiles induced by low-stress exposition represent an adaptive conditioning mode with metabolome changes that help seedlings to cope with upcoming stress. The metabolism pathways involved in this response were mainly included in amino acid metabolism and carbohydrate metabolism leading to an enhanced accumulation of phenolics, flavonoids, and terpenoids. However, when the plantlets were exposed to higher-stress exposition, the secondary metabolites that starred the response are more complex and decorated, such as alkaloids, lignans, and glycosyloxyflavones. Those changes could help to maintain homeostasis and control the response magnitude on establishing and facilitating the plantletsā survival. Overall, our findings provide new insights into the responsive mechanisms of the maritime pine under heat and drought stress in terms of metabolic profiles.
The metabolomics effect of Foxa2 knockdown on FH-deficient cells
STUDY_SUMMARY
To assess whether there is a functional convergence between the two proteins, we performed metabolomics on Fh1fl/fl, Fh1-/-CL1 and Fh1-/-CL19 cells treated with either siNT or siFoxa2. 5 x 104 Fh1fl/fl, 1 x 105 Fh1-/-CL1 and 1 x 105 Fh1-/-CL19 cells were plated the onto 6-well plate and reverse transfected with siRNA. Before extraction, cells were counted using a separate counting plate. There are a total of 30 samples prepared from three cell lines and 6 biological conditions.
Analysis of metabolite relative abundance in blood plasma from 316 individuals with trisomy 21 (T21, Down syndrome) and 103 euploid controls. This dataset is part of the Human Trisome Project run by the Linda Crnic Institute for Down Syndrome at the University of Colorado Anschutz Medical Campus. http://www.trisome.org/
INSTITUTE
University of Colorado Denver
LABORATORY
PIs - Joaquin Espinosa and Angelo D'Alessandro
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Multi-omics profiling of 5 Arabidopsis accessions in response to combined water and nitrogen deficiencies.
STUDY_TYPE
Multi-omics analysis
STUDY_SUMMARY
In this study, we build an in-depth multi-omics profile of 5 Arabidopsis accessions, in order to find the inner relations of N deficit, mild drought, and combined stress response at different scales. We used our own phenotyping platform (Phenoscope) to handle hundreds of Arabidopsis plants across 20 modalities (5 Genotypes x 2 Treatments of Water availability x 2 Treatments of Nitrogen availablity), phenotype those plants for growth-related traits and ultimately harvest the samples used in transcriptomics, metabolomics and physiological profiling. Then we have analysed those data to reveal in particular the specificities of the response to stress in each accession, as well as the W x N interactions leading to patterns that are specific to the combined stress response. The metabolomics data is particularly useful to highlight for instance the very specific profile of the Cvi-0 accession overall, or the larger extent of the metabolite responsiveness to N-deficiency with respect to mild drought. The share of the stress response that is specific to the combined stress condition is also quite variable between the 5 accessions.
Metabolic profiling of induced acute pancreatitis and pancreatic cancer progression in a mutant Kras mouse model
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Untargeted Nuclear Magnetic Resonance (NMR) metabolomics of polar extracts from the pancreata of a caerulin-induced mouse model of pancreatitis (Pt) and of a transgenic mouse model of pancreatic cancer (PCa) were used to find metabolic markers of Pt and to characterize the metabolic changes accompanying PCa progression. Using multivariate analysis a 10-metabolite metabolic signature specific to Pt tissue was found to distinguish the benign condition from both normal tissue and precancerous tissue (low grade pancreatic intraepithelial neoplasia, PanIN, lesions). The mice pancreata showed significant changes in the progression from normal tissue, through low-grade and high-grade PanIN lesions to pancreatic ductal adenocarcinoma (PDA). These included increased lactate production, amino acid changes consistent with enhanced anaplerosis, decreased concentrations of intermediates in membrane biosynthesis (phosphocholine and phosphoethanolamine) and decreased glycosylated uridine phosphates, reflecting activation of the hexosamine biosynthesis pathway and protein glycosylation.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Single Cell Spatial Analysis Reveals the Topology of Immunomodulatory Purinergic Signaling in Glioblastoma
STUDY_SUMMARY
Abstract from manuscript "Glioblastoma develops an immunosuppressive microenvironment that fosters tumorigenesis and resistance to current therapeutic strategies. Here we use multiplexed tissue imaging and single-cell RNA-sequencing to characterize the composition, spatial organization, and clinical significance of extracellular purinergic signaling in glioblastoma. We show that glioblastoma exhibit strong expression of CD39 and CD73 ectoenzymes, correlating with increased adenosine levels. Microglia are the predominant source of CD39, while CD73 is principally expressed by tumor cells, particularly in tumors with amplification of EGFR and astrocyte-like differentiation. Spatially-resolved single-cell analyses demonstrate strong spatial correlation between tumor CD73 and microglial CD39, and that their spatial proximity is associated with poor clinical outcomes. Together, this data reveals that tumor CD73 expression correlates with tumor genotype, lineage differentiation, and functional states, and that core purine regulatory enzymes expressed by neoplastic and tumor-associated myeloid cells interact to promote a distinctive adenosine-rich signaling niche and immunosuppressive microenvironment potentially amenable to therapeutic targeting. "
Endothelial Sirtuin1 Suppresses Whole-body Insulin Sensitivity by Modulating the Secretome
STUDY_TYPE
End point
STUDY_SUMMARY
Sirtuin1 (Sirt1) in skeletal muscle (SK) and fat protects against metabolic damage by stimulating insulin sensitivity. Here we report that mice with selective deletion of endothelial Sirt1 (E-Sirt1-KO) paradoxically exhibit heightened whole-body insulin sensitivity. Akt phosphorylation, glucose uptake, and glycolysis are boosted in SK and brown adipose tissue (BAT) of E-Sirt1-KO mice. E-Sirt1-KO mice have higher energy expenditure and are partially protected from high-fat diet-induced insulin resistance. Enhanced insulin sensitivity and peripheral tissue Akt phosphorylation in E-Sirt1-KO mice is transferrable to wild-type mice via the systemic circulation after surgical parabiosis. Silencing of Sirt1 in endothelial cells upregulates transcription of the F-actin-binding protein thymosin beta-4 (TĪ²4), whose secretion activates Akt in skeletal myotubes. Sirt1 downregulation stimulates endothelial TĪ²4 transcription through inhibition of autophagy and upregulation of nuclear factor-kappa B signaling. Thus, unlike Sirt1 in skeletal muscle and fat, endothelial Sirt1 curtails whole-body insulin sensitivity by inhibiting expression of secreted TĪ²4
Effect of Hira loss in the metabolic landscape of Fh1-deficient cells
STUDY_SUMMARY
Tumour initiation and progression requires the metabolic rewiring of cancer cells. Fumarate hydratase (FH), a mitochondrial enzyme that catalyses the reversible hydration of fumarate to malate in the TCA cycle, has been identified as a bona fide tumour suppressor . FH loss predisposes to Hereditary Leiomyomatosis and Renal Cell Carcinoma (HLRCC), a cancer syndrome characterized by the presence of benign tumours of the skin and uterus, and a highly aggressive form of renal cancer. Its loss leads to aberrant accumulation of fumarate, an oncometabolite that drives malignant transformation . Even though the link between FH loss, fumarate accumulation and HLRCC is well-known, the associated tumorigenic mechanism is it is still not fully understood. Indeed, although HLRCC tumours metastasize even when small, Fh1-deficient mice develop premalignant cysts in the kidneys, rather than overt carcinomas. Interestingly, these cysts are positive for the key tumour suppressor p21. Since p21 expression is a central trigger of cellular senescence, it is postulated that this process could be an obstacle for tumorigenesis in Fh1-deficient cells. Consistent with this hypothesis, HLRCC patients harbour the epigenetic suppression of p16, another key player of senescence. Here, we have confirmed that additional oncogenic events independent from a senescence bypass are required to allow full-blown transformation in FH deficient cells. Moreover, a genome wide CRISPR/Cas9 screen identified HIRA as a target that, when ablated, increases proliferation and invasion in Fh1-deficient cells. Moreover, Fh1 and Hira-deficient cells lead to the development of tumours and invasive features in the kidney in vivo. Strikingly, Hira depletion in Fh1 deficient cells controls the activation of a MYC and E2F-dependent transcriptional and metabolic program, which is known to play different oncogenic roles during tumour initiation and progression. Of note, the activation of these programs is independent of H3.3 deposition into the chromatin, known to be controlled by HIRA. Overall, we have identified a novel oncogenic event occurring in FH deficient tumours, which will be instrumental for understanding mechanisms of tumorigenesis in HLRCC and the development of targeted treatments.
INSTITUTE
CECAD Research Center, University Hospital Cologne
LAST_NAME
Yang
FIRST_NAME
Ming
ADDRESS
Joseph-Stelzmann-StraĆe 26, CECAD Research Center, Kƶln, Koeln, 50931, Germany
Lipolysis-derived Lipids Determine Autophagy Initiation during Fasting
STUDY_SUMMARY
For survival, autophagy is a crucial intracellular self-degradation process to provide energy sources, helping adapt to nutrient deprivation. Although nutrient availability is a key determinant of autophagy initiation, it remains elusive underlying mechanism(s) of perceiving nutritional scarcity by which cells timely turn on autophagy as the last self-destructive process for energy supply. Here, we showed that PKA-dependent lipolysis can block the initiation of futile autophagy during short-term nutritional deprivation by repressing AMPK. Using Raman microscopy imaging and metabolomics, we found that autophagy occurred by reduction in available free fatty acids (FFAs) for energy sources. By modulating genes involved in lipolysis and fatty acid oxidation, we found that the use of lipolysis-derived FFAs precedes autophagy initiation. The dysregulated autophagy suppression during short-term fasting decreased motility and lifespan extension of worms. Taken together, these data suggest that PKA is a pivotal factor to orchestrate sophisticated catabolic pathways, preferring the use of PKA-mediated lipolytic products to repress futile autophagic degradation during short-term fasting through AMPK inhibition.
INSTITUTE
Seoul National University
LAST_NAME
Ji
FIRST_NAME
Yul
ADDRESS
San 56-1, Sillim-Dong, Kwanak-Gu, Seoul, Seoul, 08826, Korea, South
Metabolomic analysis to assess response to immunotherapy for malignant brain tumors: Part 1
STUDY_SUMMARY
An effective immune response in patients with cancer treated with immunotherapy includes dendritic cell (DC) activation and migration followed by stimulation of CD8 and CD4 T cells. This then leads to the activation, proliferation and further activation of other immune cell populations including NK cells or immunosuppressive populations such as Tregs and myeloid derived suppressor cells (MDSCs). These studies were carried out utilizing murine brain tumor models treated with an RNA DC vaccine platform. We hypothesized that metabolomic analyses of urines would be sensitive to the action of this diverse set of immune cells. The objective of this study was to evaluate the feasibility of using metabolomics to follow immune responses after immunotherapy. We chose NMR as our analytical technique of choice, as it has many favorable qualities that make it ideal for analyses of urine.
Metabolomic analysis to assess response to immunotherapy for malignant brain tumors: Part 2
STUDY_SUMMARY
An effective immune response in patients with cancer treated with immunotherapy includes dendritic cell (DC) activation and migration followed by stimulation of CD8 and CD4 T cells. This then leads to the activation, proliferation and further activation of other immune cell populations including NK cells or immunosuppressive populations such as Tregs and myeloid derived suppressor cells (MDSCs). These studies were carried out utilizing murine brain tumor models treated with an RNA DC vaccine platform. We hypothesized that metabolomic analyses of urines would be sensitive to the action of this diverse set of immune cells. The objective of this study was to evaluate the feasibility of using metabolomics to follow immune responses after immunotherapy. We chose NMR as our analytical technique of choice, as it has many favorable qualities that make it ideal for analyses of urine.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 042
LAST_NAME
Khattri
FIRST_NAME
Ram
ADDRESS
1200 Newell Dr., ARB 240, Gainesville, FL, 32611, USA
Metabolomic analysis to assess response to immunotherapy for malignant brain tumors: Part 3
STUDY_SUMMARY
An effective immune response in patients with cancer treated with immunotherapy includes dendritic cell (DC) activation and migration followed by stimulation of CD8 and CD4 T cells. This then leads to the activation, proliferation and further activation of other immune cell populations including NK cells or immunosuppressive populations such as Tregs and myeloid derived suppressor cells (MDSCs). These studies were carried out utilizing murine brain tumor models treated with an RNA DC vaccine platform. We hypothesized that metabolomic analyses of urines would be sensitive to the action of this diverse set of immune cells. The objective of this study was to evaluate the feasibility of using metabolomics to follow immune responses after immunotherapy. We chose NMR as our analytical technique of choice, as it has many favorable qualities that make it ideal for analyses of urine.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 042
LAST_NAME
Khattri
FIRST_NAME
Ram
ADDRESS
1200 Newell Dr., ARB 240, Gainesville, FL, 32611, USA
Revealing the Social Biomarkers of Residual Feed Intake by Using 16s rRNA and LC-MS/MS in Duroc Pig
STUDY_SUMMARY
Feed efficiency (FE) is a typical social affected trait. However, the mechanisms involved are not fully elucidated. According to the rank of residual feed intake (RFI)ās the social genetic effect (SGE), ten high and low pigs were selected, named LRI and HRI groups. The sampling of jejunal chyme after slaughter. 16S rRNA and LC-MS/MS were conducted to investigate the relationship between the gut microbiome or metabolites and the SGE of RFI. The results showed significant differences between HRI and LRI groups. Compared with the HRI group, Escherichia, Eubacterium, and Gemmiger were enriched in the LRI group (P < 0.01), whereas the abundance of Fusobacterium, Eubacterium, and Desulfovibrio in the HRI group were significantly higher than that in the LRI group (P < 0.01). In the metabolome, we found that Glycine, L-lysine, and L-tryptophan were positively correlated with RFIās SGE. KEGG pathway analysis revealed that most differential metabolites were involved in amino acid metabolism. The Pearson correlation analysis of the candidate social biomarkers was carried out. Amino acid metabolites were discovered to have significant correlations with Escherichia and Fusobacterium. Therefore, Escherichia and Fusobacterium may influence the SGE of RFI through amino acid metabolism, thereby affecting feed efficiency.
Effects of Medwakh Smoking on Oxidative Stress and Inflammation Among Youth in UAE using Liquid chromatography-mass spectrometry
STUDY_SUMMARY
Oxidative stress and inflammation are closely related, and inter-dependent pathophysiological processes. Oxidative stress plays a key role in the pathophysiology of several chronic inflammatory conditions that causes cellular alterations such as lipid damage, increase of cell permeability, apoptosis/cell death, alteration in growth factors, and signaling pathways.An imbalance between the production of reactive oxygen species (ROS) and their neutralization by the antioxidant system results in the onset of oxidative stress which subsequently leads to the release of chronic inflammatory mediators such as pro-inflammatory cytokines, which causes damages to DNA, proteins, and lipids.LC-MSMS is widely used for non-targeted and non-invasive metabolomics of biological samples including saliva in a variety of conditions such as inflammatory conditions, cancers, diabetes, etc. However, to the best of our knowledge, there are currently no metabolomics-based studies identifying salivary biomarkers associated with inflammation and oxidative stress utilizing HPLC-ESI-QTOF-MS. Therefore, in this study, we aim to discover the biochemical and physiological changes induced by medwak smoking and explore the metabolomics profile of medwak smokers with a focus on the markers of oxidative stress and inflammation in comparison to nonsmokers.
Human fecal metabolome profiles under 3 different dietary terms
STUDY_SUMMARY
The gut microbiota produce numerous metabolites that affect host physiology, although the effects of the daily diet on human intestinal metabolome profiles and robustness are not well understood. Here we investigated the robustness of the human intestinal environment through gut microbiome and metabolome profiles in response to daily dietary fluctuations. We analyzed 176 fecal samples from 25 healthy Japanese individuals under three dietary conditions, including heterogeneous and homogeneous diets. Human intestinal metabolome and microbiome profiles were unique to each individual and were robust under daily dietary fluctuation in most cases. Our findings provide insight into the use of intestinal environment information for clinical studies.
INSTITUTE
Keio University
DEPARTMENT
Institute for Advanced Biosciences
LAST_NAME
Fukuda
FIRST_NAME
Shinji
ADDRESS
Kakuganji 246-2, Mizukami, Tsuruoka City Yamagata,Japan
Metabolic changes in Alzheimer patient-derived induced neurons and the effects of PKM2 modulation and hypoxia on their metabolic landscape
STUDY_SUMMARY
We have obtained fibroblast cultures from old adult human non-demented control donors and Alzheimer patients (AD). The fibroblasts were reprogrammed into directly induced neurons (iNs) to serve as an adult-like and age-equivalent model for aging and neurodegeneration. Metabolomic landscape and glucose flux in control versus AD were assessed. Additionally, their response to PKM2 modulation (shikonin 10 ĀµM or PKM2 overexpression) and hypoxia (CoDo treatment) were assessed.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
The effects of PKM2 modulation and hypoxia on the metabolic landscape of Alzheimer patient-derived induced neurons
STUDY_SUMMARY
We have obtained fibroblast cultures from old adult human non-demented control donors and Alzheimer patients (AD). The fibroblasts were reprogrammed into directly induced neurons (iNs) to serve as an adult-like and age-equivalent model for aging and neurodegeneration. Their response to PKM2 modulation (shikonin 10 ĀµM or PKM2 overexpression) and hypoxia (CoDo treatment) were assessed.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Time course GCMS based untargeted metabolomics in the presence of glucose and glycerol
STUDY_SUMMARY
Our preliminary analysis identifies that glycerol enhanced DHA accumulation in native isolate Aurantiochytrium limacinum. To evaluate temporal changes in the presence of glycerol, time-course metabolite profiling was done in the presence of glycerol and glucose at three different time-points i.e., 0, 48 and 96 h using GC-MS. We identified nearly 40 metabolites, among which metabolites belonging to pentose phosphate pathway, TCA cycle and amino acid metabolism were significantly differentially regulated which suggests its role in glycerol induced DHA accumulation.
INSTITUTE
International Centre for Genetic Engineering and Biotechnology
LAST_NAME
Jutur
FIRST_NAME
Pannaga Pavan
ADDRESS
Omics of Algae Lab, 2nd Floor, New Building, ICGEB, Aruna Asaf Ali Marg, New Delhi
Non-targeted metabolomics screen comparing metabolite profiles of serum from PDAC-bearing mice that received metronidazole using high-resolution, high-performance LC-MS/MS analysis.
STUDY_SUMMARY
The composition of the gut microbiome controls innate and adaptive immunity and has emerged as a key regulator of tumor growth and the success of immune checkpoint blockade (ICB) therapy. However, the underlying mechanisms remain unclear. Pancreatic ductal adenocarcinoma (PDAC) tends to be refractory to therapy, including ICB. We found that the gut microbe-derived metabolite trimethylamine N-oxide (TMAO) enhances anti-tumor immunity to PDAC. Delivery of TMAO given intraperitoneally or via dietary choline supplement to PDAC-bearing mice reduces tumor growth and is associated with an immunostimulatory tumor-associated macrophage (TAM) phenotype and activated effector T cell response in the tumor microenvironment. Mechanistically, TMAO signals through potentiating type-I interferon (IFN) pathway and confers anti-tumor effects in a type-I IFN dependent manner. Notably, delivering TMAOprimed macrophages alone produced similar anti-tumor effects. Combining TMAO with ICB (anti-PD1 and/or anti-Tim3) significantly reduced tumor burden and improved survival beyond TMAO or ICB alone. Finally, the levels of trimethylamine (TMA)- producing bacteria and of CutC gene expression correlate with improved survivorship and response to anti-PD1 in cancer patients. Together, our study identifies the gut microbial metabolite TMAO as an important driver of anti-tumor immunity and lays the groundwork for new therapeutic strategies.
Non-targeted metabolomics screen comparing metabolite profiles of serum from PDAC-bearing mice that received 1% choline supplementation or control diet using high-resolution, high-performance LC-MS/MS analysis.
STUDY_SUMMARY
The composition of the gut microbiome controls innate and adaptive immunity and has emerged as a key regulator of tumor growth and the success of immune checkpoint blockade (ICB) therapy. However, the underlying mechanisms remain unclear. Pancreatic ductal adenocarcinoma (PDAC) tends to be refractory to therapy, including ICB. We found that the gut microbe-derived metabolite trimethylamine N-oxide (TMAO) enhances anti-tumor immunity to PDAC. Delivery of TMAO given intraperitoneally or via dietary choline supplement to PDAC-bearing mice reduces tumor growth and is associated with an immunostimulatory tumor-associated macrophage (TAM) phenotype and activated effector T cell response in the tumor microenvironment. Mechanistically, TMAO signals through potentiating type-I interferon (IFN) pathway and confers anti-tumor effects in a type-I IFN dependent manner. Notably, delivering TMAOprimed macrophages alone produced similar anti-tumor effects. Combining TMAO with ICB (anti-PD1 and/or anti-Tim3) significantly reduced tumor burden and improved survival beyond TMAO or ICB alone. Finally, the levels of trimethylamine (TMA)- producing bacteria and of CutC gene expression correlate with improved survivorship and response to anti-PD1 in cancer patients. Together, our study identifies the gut microbial metabolite TMAO as an important driver of anti-tumor immunity and lays the groundwork for new therapeutic strategies.
Effect of Hira loss in the metabolic landscape of Fh1-deficient cells Part 2
STUDY_SUMMARY
Tumour initiation and progression requires the metabolic rewiring of cancer cells. Fumarate hydratase (FH), a mitochondrial enzyme that catalyses the reversible hydration of fumarate to malate in the TCA cycle, has been identified as a bona fide tumour suppressor . FH loss predisposes to Hereditary Leiomyomatosis and Renal Cell Carcinoma (HLRCC), a cancer syndrome characterized by the presence of benign tumours of the skin and uterus, and a highly aggressive form of renal cancer. Its loss leads to aberrant accumulation of fumarate, an oncometabolite that drives malignant transformation . Even though the link between FH loss, fumarate accumulation and HLRCC is well-known, the associated tumorigenic mechanism is it is still not fully understood. Indeed, although HLRCC tumours metastasize even when small, Fh1-deficient mice develop premalignant cysts in the kidneys, rather than overt carcinomas. Interestingly, these cysts are positive for the key tumour suppressor p21. Since p21 expression is a central trigger of cellular senescence, it is postulated that this process could be an obstacle for tumorigenesis in Fh1-deficient cells. Consistent with this hypothesis, HLRCC patients harbour the epigenetic suppression of p16, another key player of senescence. Here, we have confirmed that additional oncogenic events independent from a senescence bypass are required to allow full-blown transformation in FH deficient cells. Moreover, a genome wide CRISPR/Cas9 screen identified HIRA as a target that, when ablated, increases proliferation and invasion in Fh1-deficient cells. Moreover, Fh1 and Hira-deficient cells lead to the development of tumours and invasive features in the kidney in vivo. Strikingly, Hira depletion in Fh1 deficient cells controls the activation of a MYC and E2F-dependent transcriptional and metabolic program, which is known to play different oncogenic roles during tumour initiation and progression. Of note, the activation of these programs is independent of H3.3 deposition into the chromatin, known to be controlled by HIRA. Overall, we have identified a novel oncogenic event occurring in FH deficient tumours, which will be instrumental for understanding mechanisms of tumorigenesis in HLRCC and the development of targeted treatments. Part 2 of this study emoployed a second FH-null clone to complement Part 1 of the study.
Spatially resolved characterization of tissue metabolic compartments in fasted and high-fat diet livers
STUDY_SUMMARY
Cells adapt their metabolism to physiological stimuli, and metabolic heterogeneity exists between cell types, within tissues, and subcellular compartments. The liver plays an essential role in maintaining whole-body metabolic homeostasis and is structurally defined by metabolic zones. These zones are well-understood on the transcriptomic level, but have not been comprehensively characterized on the metabolomic level. Mass spectrometry imaging (MSI) can be used to map hundreds of metabolites directly from a tissue section, offering an important advance to investigate metabolic heterogeneity in tissues compared to extraction-based metabolomics methods that analyze tissue metabolite profiles in bulk. We established a workflow for the preparation of tissue specimens for matrix-assisted laser desorption/ionization (MALDI) MSI that can be implemented to achieve broad coverage of central carbon, nucleotide, and lipid metabolism pathways. Herein, we used this approach to visualize the effect of nutrient stress and excess on liver metabolism. Our data revealed a highly organized metabolic tissue compartmentalization in livers, which becomes disrupted under high fat diet. Fasting caused changes in the abundance of several metabolites, including increased levels of fatty acids and TCA intermediates while fatty livers had higher levels of purine and pentose phosphate-related metabolites, which generate reducing equivalents to counteract oxidative stress. This spatially conserved approach allowed the visualization of liver metabolic compartmentalization at 30 Āµm pixel resolution and can be applied more broadly to yield new insights into metabolic heterogeneity in vivo.
Cataboslim of branched-chain amino acids (BCAAs) in renal cells HK2 and 786-O
STUDY_SUMMARY
The objective of this experiment is to compare the catabolism of branched-chain amino acids (BCAAs) in human renal epithelial cell line HK2 versus ccRCC cell lines 786-O, 786-M1A and 786-M2A using 13C6-labelled leucine and isoleucine stable isotope tracers. To this end, we incubated the above cell lines with 13C6-leucine and 13C6-isoleucine in Plasmax media for 27h. Data were generated from 5 independent cultures. This is Part I of the study and the experimental number is MS42.
INSTITUTE
CECAD Research Center, University Hospital Cologne
LAST_NAME
Yang
FIRST_NAME
Ming
ADDRESS
Joseph-Stelzmann-StraĆe 26, CECAD Research Center, Kƶln, Koeln, 50931, Germany
Glutaminolysis contribution to the carbon backbone of aspartate through ATP Citrate Lyase (ACLY) in ccRCC
STUDY_SUMMARY
The objective of this experiment is to test the contribution of the carbons from glutamine to generation of aspartate via ATP citrate lyase (ACLY) in human epithelial renal cells HK2 and ccRCC cell lines 786-O and 786-M1A. To test this hypothesis, we incubated all cells with 13C5-glutamine in Plasmax media with or without a pharmacological inhibitor of ACLY. This is Part 8 of the study and the experimental number is MS58.
Glutaminolysis contribution to the carbon backbone of aspartate and glutamate in ccRCC
STUDY_SUMMARY
The objective of this experiment is to test the contribution of the carbons derived from glutamine to the generation of aspartate and glutamate in human epithelial renal cells HK2 and ccRCC cell lines 786-O and 786-M1A. To test this hypothesis, we incubated all cells with 13C5-glutamine in Plasmax media with or without a pharmacological inhibitor of glutaminase CB-839. This is Part 7 of a study and the experimental number is MS57.
Metabolic profiling of mouse tissues and tissue interstitial fluids
STUDY_SUMMARY
Tissue and tissue interstitial fluids was collected from mice of a hybrid C57BL/6J;129/SvJ background of about 12 weeks of age (8 in total) and used to profile the metabolic content. This is Part 6 of a study and the experimental number is MS56.
Intracellular metabolic profile of renal cells cultured in Plasmax
STUDY_SUMMARY
The objective of this experiment is to analyse the metabolic profiles of human renal epithelial cells HK2 and ccRCC cell lines 786-O, 786-M1A and 786-M2A in Plasmax media. The experiment was conducted on three different days using cells with different passage numbers. This is Part 5 of a study and the experimental number is MS55.
Time sensitive contribution of the BCAA catabolism to the TCA cycle carbons in HK2, 786-O, OS-RC-2 and RFX-631
STUDY_SUMMARY
The objective of this experiment is to test the contribution of the branched chain amino acids catabolism to the carbons used in the TCA cycle. To test this hypothesis, we incubated human renal epithelial cells (HK2) and ccRCC cell lines (786-O, 786-M1A, OS-RC-2, OS-LM1, RFX-631) with 13C6-leucine and 13C6-isoleucine in Plasmax media for 10 mins, 1 hour and 3 hours. Data were generated from 5 independent cultures. This is Part 4 of the study and the experiment number is MS52.
Exometabolomics of HK2, 786-O cells cultured in Plasmax media
STUDY_SUMMARY
The objective of this study is to analyse the exometabolomics of human epithelial renal cell line HK2 and clear cell renal cell carcinoma (ccRCC) cell lines 786-O, 786-M1A, 786-M2A, OS-RC-2, OS-LM1 and RFX-631 that are cultured with the Plasmax media. This is part 3 of the study, and the experimental number is MS51.
Assessing the contribution of branched-chain amino acid (BCAA)-derived nitrogen to amino acid biosynthesis in renal cell lines HK2 and 786-O.
STUDY_SUMMARY
The objective of this experiment is to test the contribution of branched-chain amino acids (BCAA)-derived nitrogen to the generation of de novo amino acids in renal cells through transamination reactions. To test this hypothesis, we incubated human renal epithelial cell line HK2 and ccRCC cell lines 786-O, 786-M1A and 786-M2A with 15N-leucine and 15N-isoleucine in Plasmax media for 27h. Data were generated from 5 independent cultures. This is Part 2 of the study and the experimental number is MS48.
Estrogen receptor a deficiency in cardiac myocytes reprograms heart-derived extracellular vesicle proteome and induces obesity in female mice (Part 2)
STUDY_SUMMARY
Dysregulation of ERĪ± has been linked with increased metabolic and cardiovascular disease risk. Uncovering the impact of ERĪ± deficiency in specific tissues has implications for understanding the role of ERĪ± in normal physiology and disease, the increased disease risk in postmenopausal women, and the design of tissue-specific ERĪ±-based therapies for a range of pathologies including cardiac disease and cancer. Cardiac myocyte-specific ERļ” knockout mice (ERĪ±HKO) were generated to assess the role of ERĪ± in the heart. Female ERĪ±HKO mice displayed a modest cardiac phenotype, but unexpectedly, the most striking phenotype was obesity in female ERĪ±HKO but not male ERļ”HKO mice. In female ERĪ±HKO mice we identified cardiac dysfunction, mild glucose and insulin intolerance, and reduced ERĪ± gene expression in skeletal muscle and white adipose tissue (WAT). Gene expression, protein, lipidomic and metabolomic analyses showed evidence of contractile and/or metabolic dysregulation in heart, skeletal muscle and WAT. We also show that extracellular vesicles (EVs) collected from the perfusate of isolated hearts from female ERĪ±HKO mice have a distinct proteome, and these EVs have the capacity to reprogram the proteome of a skeletal muscle cell including proteins linked with ERĪ±, fatty acid regulation, lipid metabolism and mitochondrial function. This study uncovers a cardiac-initiated and sex-specific cardiometabolic phenotype that is regulated by ERĪ±.
INSTITUTE
Baker Heart and Diabetes Institute
DEPARTMENT
Discovery and Preclinical Science
LABORATORY
Cardiac Hypertrophy
LAST_NAME
Tham
FIRST_NAME
Yow Keat
ADDRESS
75 Commercial Rd, Melbourne, Victoria, 3004, Australia
Estrogen receptor Ī± deficiency in cardiac myocytes reprograms heart-derived extracellular vesicle proteome and induces obesity in female mice (Part 1)
STUDY_SUMMARY
Dysregulation of ERĪ± has been linked with increased metabolic and cardiovascular disease risk. Uncovering the impact of ERĪ± deficiency in specific tissues has implications for understanding the role of ERĪ± in normal physiology and disease, the increased disease risk in postmenopausal women, and the design of tissue-specific ERĪ±-based therapies for a range of pathologies including cardiac disease and cancer. Cardiac myocyte-specific ER knockout mice (ERĪ±HKO) were generated to assess the role of ERĪ± in the heart. Female ERĪ±HKO mice displayed a modest cardiac phenotype, but unexpectedly, the most striking phenotype was obesity in female ERĪ±HKO but not male ERĪ±HKO mice. In female ERĪ±HKO mice we identified cardiac dysfunction, mild glucose and insulin intolerance, and reduced ERĪ± gene expression in skeletal muscle and white adipose tissue (WAT). Gene expression, protein, lipidomic and metabolomic analyses showed evidence of contractile and/or metabolic dysregulation in heart, skeletal muscle and WAT. We also show that extracellular vesicles (EVs) collected from the perfusate of isolated hearts from female ERĪ±HKO mice have a distinct proteome, and these EVs have the capacity to reprogram the proteome of a skeletal muscle cell including proteins linked with ERĪ±, fatty acid regulation, lipid metabolism and mitochondrial function. This study uncovers a cardiac-initiated and sex-specific cardiometabolic phenotype that is regulated by ERĪ±.
INSTITUTE
Baker Heart and Diabetes Institute
LAST_NAME
Tham
FIRST_NAME
Yow Keat
ADDRESS
75 Commercial Rd, Melbourne, Victoria, 3004, Australia
Metabolomics of bone marrow-derived dendritic cells conditioned with H. polygyrus bakery non-polar metabolites
STUDY_SUMMARY
Bone marrow-derived dendritic cells were incubated with 50 micro g/mL of H. polygyrus bakery non-polar metabolites in RPMI 1640 containing 10% FBS, 1% of 100X penicillin/streptomycin, 1% of 100 mM sodium pyruvate, and 20 ng/mL GM-CSF at 37Ā°C, 5% CO2 for 4 or 20 h. Supernatants and cell-free media controls were collected and incubated with ice-cold HPLC-grade methanol for 30 min on ice, centrifuged at 10,000 x g for 10 min at 4Ā°C and stored at -80Ā°C until analysis. The profiling of nonpolar metabolites was performed by LC-MS/MS analysis of the deproteinated conditioned media by injecting 3 mL of sample onto a Dionex UHPLC system equipped with an Agilent Eclipse C18 (2.1 x 15 mm, 1.8 mm) column incubated at 45oC. Metabolites were resolved with a 30 min linear running 0-80 % using the buffers system 0.05 % formic acid and 0.05 % formic acid in acetonitrile at a flowrate of 300 mL/min. The column effluent was introduced by electrospray ionization onto a ThermoScientific Velos LTQ Orbitrap Analyzer using a spray voltage of 3.6 kV, a source heater temperature of 350oC, and a sheath gas flow of 40 L/min. Survey scans were performed using the Orbitrap mass spectrometer and the 10 most intense ions were selected for fragmentation using a 30-40 V stepped collision induced dissociation energy. Fragmentation products were analyzed in the linear ion trap mass spectrometer. Fragmentation was used to perform XCMS online database (https://xcmsonline.scripps.edu) search to identify possible metabolites.
Metabolomics Analysis of HOG-EV and HOG-R132H Cells with and without BAY 2402234 Treatment
STUDY_TYPE
Metabolomics Analysis
STUDY_SUMMARY
HOG cells were plated in 6-well plates (0.5 Ć 10^6 cells per well). 24 hours later, HOG-EV or HOG-R132H cells were treated for 24 hours with 10 nM BAY 2402234 or DMSO. Cells were then harvested for LC-MS analysis.
INSTITUTE
UT Southwestern Medical Center
DEPARTMENT
Children's Research Institute
LABORATORY
McBrayer Laboratory
LAST_NAME
McBrayer
FIRST_NAME
Samuel
ADDRESS
6000 Harry Hines Boulevard, NL10.110K, Dallas, TX 75235, USA
The goal of this work was to analyze metabolic changes in yeast with the mct1 gene knock-out or CTP1 overexpression conditions using liquid chromatography-mass spectrometry (LC-MS).
INSTITUTE
University of Utah
DEPARTMENT
Biochemistry
LABORATORY
Rutter
LAST_NAME
Berg
FIRST_NAME
Jordan
ADDRESS
15 N Medical Drive East RM 5520, Salt Lake City, UT 84112-5650 USA
Batch variation of large scale LC-MS metabolomics analysis of human plasma samples
STUDY_SUMMARY
Metabolomics holds the promise to measure and quantify small molecules comprehensively in biological systems, and LC-MS (liquid chromatography coupled mass spectrometry) has become the leading technology in the field. Significant challenges still exist in the computational processing of data from LC-MS metabolomic experiments into metabolite features, including provenance and reproducibility of the current software tools. Current dataset, named HZV029, comprises of 268 data files, from two QC (human pooled plasma) samples that were analyzed repeatedly over 17 batches, on a Thermo Scientific Orbitrap ID-X Tribrid mass spectrometer, coupled with dual liquid chromatography via a Transcend LX-2 System. It provides a unique opportunity to be used as a benchmark dataset to evaluate reproducibility via available choices of different processing tools without knowing the ground truth about these QC samples.
A metabolic map of the DNA damage response identifies PRDX1 in nuclear ROS scavenging and aspartate synthesis
STUDY_SUMMARY
Targetted metabolomics in U2OS PRDX1 WT and PRDX1-/- While cellular metabolism impacts the DNA damage response, a systematic understanding of the metabolic requirements that are crucial for DNA damage repair has yet to be achieved. Here, we investigate the metabolic enzymes and processes that are essential when cells are exposed to DNA damage. By integrating functional genomics with chromatin proteomics and metabolomics, we provide a detailed description of the interplay between cellular metabolism and the DNA damage response. Subsequent analysis identified Peroxiredoxin 1, PRDX1, as fundamental for DNA damage repair. During the DNA damage response, PRDX1 translocates to the nucleus where it is required to reduce DNA damage-induced nuclear reactive oxygen species levels. Moreover, PRDX1 controls aspartate availability, which is required for the DNA damage repair-induced upregulation of de novo nucleotide synthesis. Loss of PRDX1 leads to an impairment in the clearance of Ī³Ī2ĪĪ§ nuclear foci, accumulation of replicative stress and cell proliferation defects, thus revealing a crucial role for PRDX1 as a DNA damage surveillance factor.
INSTITUTE
CRG
DEPARTMENT
GRSC
LABORATORY
Sdelci_lab
LAST_NAME
Kourtis
FIRST_NAME
Savvas
ADDRESS
Carrer del Dr. Aiguader, 88, 08003 Barcelona, Barcelona, barcelona, 08003, Spain
EMAIL
savvas.kourtis@crg.eu
STUDY_TYPE
Targetted metabolomics in U2OS PRDX1 WT and PRDX1-/-
Application of Artificial Intelligence to Plasma Metabolomics Profiles to Predict Response to Neoadjuvant Chemotherapy in Triple-Negative Breast Cancer
STUDY_SUMMARY
Summary: There is a need for biomarkers predictive of response to neoadjuvant chemotherapy (NACT) in triple-negative breast cancer (TNBC). We previously obtained evidence that a polyamine signature in the blood is associated with TNBC development and progression. In this study, we evaluated whether plasma polyamines and other metabolites may identify TNBC patients who are unlikely to respond to NACT. Pre-treatment plasma levels of acetylated polyamines were elevated in TNBC patients that had moderate to extensive tumor burden (RCB-II/III) following NACT compared to those that achieved a complete pathological response (pCR/RCB-0) or had minimal residual disease (RCB-I). We further applied artificial intelligence to comprehensive metabolic profiles to identify additional metabolites associated with treatment response. A deep learning model (DLM) consisting of two polyamines as well as nine additional metabolites was developed for improved prediction of RCB-II/III. The DLM has potential clinical value for identifying TNBC patients who are unlikely to respond to NACT and who may benefit from other treatment modalities.
The impact of IgE in gut and serum metabolomes in a murine experimental model of allergic enteritis
STUDY_TYPE
Case-control study
STUDY_SUMMARY
The pathological mechanism of the gastrointestinal forms of food allergies is less understood in comparison to other clinical phenotypes, such as asthma, and anaphylaxis, partly due to difficulty in the access to intestinal tissues and because of a highly complex interplay between microbiota and intestinal mucosa. Importantly, a high level of IgE is a poor prognostic factor in gastrointestinal allergies. This study aimed to investigate how IgE influences the development of intestinal inflammation and the metabolome in allergic enteritis (AE), using IgE knock-in (IgEki) mice expressing high levels of IgE. Ovalbumin-sensitized and egg-white diet fed (OVA/EW) BALB/c WT mice developed moderate AE, whereas OVA/EW IgEki mice induced more aggravated intestinal inflammation with enhanced eosinophil accumulation.
INSTITUTE
Institute of Applied Molecular Medicine
LAST_NAME
Zubeldia-Varela
FIRST_NAME
Elisa
ADDRESS
Avda. Monteprincipe s/n 28668 Boadilla del Monte, Madrid, EspaƱa
EMAIL
elisa.zubeldiavarela@ceu.es
PHONE
Tlf: 91 372 47 00 ext. 14675
NUM_GROUPS
4 groups: BALB/c wild type (WT) and IgE knock-in mice (C.Ighg1tm1.1Pyu) in the BALB/c background, both sensitised and non-sensitised to ovalbumin.
Metabolomic analysis of brain cortex from neuronal specific Depdc5 knockout in fed and fasting state
STUDY_TYPE
Depdc5 KO fed vs. fasted comparison
STUDY_SUMMARY
Caloric restriction and acute fasting are known to reduce seizures but through unclear mechanisms. In this study, we demonstrate that mTORC1 signaling is reduced after acute fasting of mice. In neurons, mTORC1 is most sensitive to withdrawal of leucine, arginine, and glutamine, which is dependent on DEPDC5. We performed metabolomic analysis of brain cortex from neuronal specific Depdc5 knockout in fed and fasting state. The Depdc5 neuronal specific knockout mice are resistant to sensing significant fluctuations in brain amino acid levels after fasting. These results establish that acute fasting reduces seizure susceptibility in a DEPDC5-dependent manner.
INSTITUTE
Northwestern University Feinberg School of Medicine
DEPARTMENT
Medicine
LABORATORY
Chandel Lab
LAST_NAME
Chandel
FIRST_NAME
Navdeep
ADDRESS
303 E Superior St, Chicago, Illinois, 60611, USA
EMAIL
nav@northwestern.edu
PHONE
3125032549
NUM_GROUPS
4
TOTAL_SUBJECTS
40
NUM_MALES
20
NUM_FEMALES
20
PUBLICATIONS
DEPDC5-dependent mTORC1 signaling mechanisms are critical for the anti-seizure effects of acute fasting
LC-HRMS based plasma metabolomics analysis for biomarker discovery of neuroblastoma: 3-O-methyldopa is a new biomarker of poor prognosis of metastatic disease
STUDY_TYPE
Biomarker Discovery
STUDY_SUMMARY
In this paper we show for the first time a metabolomic-based biomarker discovery using HRMS applied to plasma of NB patients and its validation on a second independent cohort of patients using a different analytical method.
Insights into plant lipid metabolism using stable isotopes and high resolution mass spectrometry
STUDY_TYPE
Stable isotope enriched lipidomics
STUDY_SUMMARY
Data analysis and mass spectrometry tools have advanced significantly in the last decade. This ongoing revolution has elevated the status of analytical chemistry within the big-data omics era. High resolution mass spectrometers (HRMS) can now distinguish different metabolites with mass to charge ratios (i.e. m/z) that differ by 0.01 Da or less. This unprecedented level of resolution not only enables identification of previously unknown compounds but also presents an opportunity to establish active metabolic pathways through quantification of isotope enrichment. Studies with stable isotope tracers continue to contribute to our knowledge of biological pathways in human, plant and bacterial species, however most current studies have been based on targeted analyses. The capacity of HRMS to resolve near-overlapping isotopologues and identify compounds with high mass precision presents a strategy to assess āactiveā pathways de novo from data generated in an untargeted way, that is blind to the metabolic network and therefore unbiased. Currently, identifying metabolic features, enriched with stable isotopes, at an āomicsā level remains an experimental bottleneck, limiting our capacity to understand biological network operation at the metabolic level. We developed data analysis tools that: i) use labeling information and exact mass to determine the elemental composition of each isotopically enriched ion, ii) apply correlation-based approaches to cluster metabolite peaks with similar patterns of isotopic labels and, iii) leverage this information to build directed metabolic networks de novo. Using Camelina sativa, an emerging oilseed model, we demonstrate the power of stable isotope labeling in combination with imaging and HRMS to reconstruct lipid metabolic networks in developing seeds and are currently addressing questions about lipid and central metabolism. Tools developed in this study will have a broader application to assess context specific operation of metabolic pathways.
Use of HRMS and Dual Isotope Labels to Resolve Difficult-to Measure Fluxes
STUDY_TYPE
Stable isotope enriched Metabolomics
STUDY_SUMMARY
Data analysis and mass spectrometry tools have advanced significantly in the last decade. This ongoing revolution has elevated the status of analytical chemistry within the big-data omics era. High resolution mass spectrometers (HRMS) can now distinguish different metabolites with mass to charge ratios (i.e. m/z) that differ by 0.01 Da or less. This unprecedented level of resolution not only enables identification of previously unknown compounds but also presents an opportunity to establish active metabolic pathways through quantification of isotope enrichment. Studies with stable isotope tracers continue to contribute to our knowledge of biological pathways in human, plant and bacterial species, however most current studies have been based on targeted analyses. The capacity of HRMS to resolve near-overlapping isotopologues and identify compounds with high mass precision presents a strategy to assess āactiveā pathways de novo from data generated in an untargeted way, that is blind to the metabolic network and therefore unbiased. Currently, identifying metabolic features, enriched with stable isotopes, at an āomicsā level remains an experimental bottleneck, limiting our capacity to understand biological network operation at the metabolic level. We developed data analysis tools that: i) use labeling information and exact mass to determine the elemental composition of each isotopically enriched ion, ii) apply correlation-based approaches to cluster metabolite peaks with similar patterns of isotopic labels and, iii) leverage this information to build directed metabolic networks de novo. Using Camelina sativa, an emerging oilseed model, we demonstrate the power of stable isotope labeling in combination with imaging and HRMS to reconstruct lipid metabolic networks in developing seeds and are currently addressing questions about lipid and central metabolism. Tools developed in this study will have a broader application to assess context specific operation of metabolic pathways.
ACSS2 Regulates HIF-2Ī± Degradation through the E3-Ubiquitin Ligase MUL1 in Clear Cell Renal Cell Carcinoma
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Clear cell renal cell carcinoma (ccRCC) is an aggressive kidney cancer driven by VHL loss and aberrant HIF-2Ī± signaling. Acetate metabolism may contribute to this axis by ACSS2-dependent acetylation of HIF-2Ī± and may provide opportunities to intervention. Here we tested the effects of pharmacological and genetic manipulation of ACSS2 on HIF-2Ī±, ccRCC cells, and tumors. ACSS2 inhibition led to HIF-2Ī± degradation and suppressed ccRCC growth in vitro, in vivo, and in primary cell cultures of ccRCC patient tumors. This treatment resulted in reduced glucose and cholesterol metabolism, mitochondrial biogenesis and altered cristae deformation, that are consistent with loss of HIF-2Ī±. Mechanistically, HIF-2Ī± protein levels are regulated through proteolytic degradation and we found, in parallel to VHL, HIF-2Ī± stability was dependent on ACSS2 activity to prevent direct interaction with the E3 ligase MUL1. These findings highlight ACSS2 as a critical upstream regulator of pathogenically stabilized HIF-2Ī±, and provides a mechanism that may be exploited to overcome resistance to HIF-2Ī± inhibitor therapies.
Hypoxia promotes osteogenesis via regulating the acetyl-CoA-mediated mito-nuclear communication.
STUDY_SUMMARY
Bone-mesenchymal stem cells (MSCs) reside in a hypoxic niche that maintains their differentiation potential. Although the role of hypoxia (low oxygen concentration) in the regulation of stem cell function has been previously reported, with normoxia (high oxygen concentration) leading to impaired osteogenesis, the molecular events triggering changes in stem cell fate decisions in response to high oxygen remain elusive. Here, we study the impact of normoxia on the mito-nuclear communication with regards to stem cell differentiation. We show that normoxia-cultured MSCs undergo profound transcriptional alterations which cause irreversible osteogenesis defects. Mechanistically, high oxygen promotes chromatin compaction and histone hypo-acetylation, particularly on promoters and enhancers of osteogenic genes. Although normoxia induces metabolic rewiring resulting in high acetyl-CoA levels, histone hypo-acetylation occurs due to trapping of acetyl-CoA inside mitochondria, owing to lower CiC activity. Strikingly, restoring the cytosolic acetyl-CoA pool remodels the chromatin landscape and rescues the osteogenic defects. Collectively, our results demonstrate that the metabolism-chromatin-osteogenesis axis is heavily perturbed in response to high oxygen and identify CiC as a novel, oxygen-sensitive regulator of the MSC function.
Lipidomics analysis of Friedreich's ataxia (FRDA) (part II)
STUDY_TYPE
Untargeted and targeted (PRM) analysis
STUDY_SUMMARY
Friedreichās Ataxia (FRDA) is an autosomal neurodegenerative disease caused by the deficiency of protein frataxin. Frataxin functions in the assembly of iron-sulfur clusters that are important for iron homeostasis and metabolic functions. To identify metabolic features that can be used for potential biomarkers in FRDA plasma, we performed a targeted multi-omics (metabolomics, lipidomics, and proteomics) analysis using discovery-validation cohort design. Muti-omics analysis revealed that FRDA patients had dysregulated sphingolipid metabolism, phospholipid metabolism, citric acid cycle, amino acid metabolism, and apolipoprotein metabolism. Sphingolipid dysfunctions were revealed by decreased very long chain ceramides but unchanged long chain ceramides in FRDA plasma, which resulted in the increased ratio of long chain ceramides to very long chain ceramides. Decreased very long chain ceramides distinguished FRDA patients from healthy controls and showed good predictive capacities with AUC values from 0.75 to 0.85. Furthermore, by performing lipidomic and stable isotope tracing experiment in induced pluripotent stem cell differentiated cardiomyocytes (iPSC-CMs, we demonstrated that frataxin deficiency affected ceramide synthase (CerS2), and preferentially enriched long chain ceramides and depleted very long chain ceramides. Moreover, ceramide metabolism was differentially regulated in a tissue-specific manner. Finally, machine learning model increased the prediction of FRDA using the combination of three metabolites (AUC > 0.9). In conclusion, decreased very long chain ceramides are potential biomarkers and therapeutic target in FRDA patients.
Metabolomics analysis of Friedreich's ataxia (FRDA) (part I)
STUDY_TYPE
Untargeted and targeted (PRM) analysis
STUDY_SUMMARY
Friedreichās Ataxia (FRDA) is an autosomal neurodegenerative disease caused by the deficiency of protein frataxin. Frataxin functions in the assembly of iron-sulfur clusters that are important for iron homeostasis and metabolic functions. To identify metabolic features that can be used for potential biomarkers in FRDA plasma, we performed a targeted multi-omics (metabolomics, lipidomics, and proteomics) analysis using discovery-validation cohort design. Muti-omics analysis revealed that FRDA patients had dysregulated sphingolipid metabolism, phospholipid metabolism, citric acid cycle, amino acid metabolism, and apolipoprotein metabolism. Sphingolipid dysfunctions were revealed by decreased very long chain ceramides but unchanged long chain ceramides in FRDA plasma, which resulted in the increased ratio of long chain ceramides to very long chain ceramides. Decreased very long chain ceramides distinguished FRDA patients from healthy controls and showed good predictive capacities with AUC values from 0.75 to 0.85. Furthermore, by performing lipidomic and stable isotope tracing experiment in induced pluripotent stem cell differentiated cardiomyocytes (iPSC-CMs, we demonstrated that frataxin deficiency affected ceramide synthase (CerS2), and preferentially enriched long chain ceramides and depleted very long chain ceramides. Moreover, ceramide metabolism was differentially regulated in a tissue-specific manner. Finally, machine learning model increased the prediction of FRDA using the combination of three metabolites (AUC > 0.9). In conclusion, decreased very long chain ceramides are potential biomarkers and therapeutic target in FRDA patients.
Deciphering the metabolomic differences between two fast-growing cyanobacteria, S.elongatus PCC 11801 and 11802 via metabolite profiling
STUDY_SUMMARY
The study aims to identify the metabolic differences between two promising fast-growing, non-model cyanobacterial strains, S. elongatus PCC 11801 and PCC 11802. To this end, experiments were carried out to measure metabolite levels in the two cyanobacterial strains grown in shake flasks at a similar light intensity of approx. 300-350 Āµmole photons.m-2. s-1. The samples for metabolomics analysis were collected during the exponential growth phase at an optical cell density of 0.5-0.6. Isotopic ratio method was utilized to compare the metabolite levels and delineate the differences in their metabolic pathways.
INSTITUTE
Indian Institute of Technology Bombay
DEPARTMENT
Chemical Engineering
LAST_NAME
Wangikar
FIRST_NAME
Pramod
ADDRESS
Biosystems and Bioengineering Lab, Department of Chemical Engineering, IIT Bombay, Powai, Mumbai, Maharashtra, India -400076
Longitudinal fecal metabolomic profiles from mothers and their infants in the EDIA study
STUDY_SUMMARY
In a cohort consisting of 32 mother-infant dyads, we profiled the fecal metabolome at birth and at 3 and 6 months of infant age. Metagenomes from the same samples were also generated.
The Microbiota and Health Study (clinicaltrials.gov: NCT02361164) was a longitudinal, community-based cohort study in Nandipara, a peri-urban community of Dhaka, Bangladesh conducted from April 2013 to October 2016. 267 newborns born to healthy mothers were followed from birth to two years of age. Fecal samples were collected at birth, during subsequent scheduled visits, and when possible during illness episodes. Active surveillance of diarrheal and respiratory infections was conducted by a community-based team of nurses supervised by a physician. Fecal samples of 222 participants were analyzed by metabolomic profiling.
Quantitative multi-Omics analysis of paclitaxel-loaded Poly(lactide-co-glycolide) nanoparticles for identification of potential biomarkers for head and neck cancer
STUDY_SUMMARY
The narrow therapeutic index and significant potential for toxicity of chemotherapeutic drugs are two of the factors that restrict their use. Because of the usage of nanoparticles (NPs) as carriers for chemotherapeutic agents, the therapeutic efficacy of these treatments has been significantly boosted. This was accomplished by increasing the bioavailability of the pharmaceuticals and changing the bio-distribution profile of the drugs. Untargeted metabolomics has recently risen to the forefront as a potentially useful method for better comprehending the growth of tumours and the treatment outcomes of many kinds of cancer cells. In the current study, we used LCMS/MS-based untargeted metabolomics to identify differences in the metabolic profile of head and neck squamous cell carcinomas FaDu that were treated with the anticancer drug paclitaxel (PTX) delivered as free drug versus paclitaxel-loaded poly(lactide-co-glycolide) nanoparticles (PXT-PLGA-NPs). The experimental design consisted of four groups: those treated with DMSO (serving as a control), those treated with drug-free PXT, those treated with PXT-PLGA-NPs, and those treated with PLGA-NPs that lacked PTX. MetaboScape (V4, Bruker Daltonics) was used as the platform for the data analysis, and the results were compared to the Bruker Human Metabolome Data Base (HMDB) spectrum library 2.0. We found a total of 162 metabolites with a high level of confidence ascribed to them. The principal component analysis of the metabolites showed that PTX-free drugs grouped along with PXT-PLGA-NPs, but the control and PLGA-NPs without PXT clustered apart from drug-treated cells but together with each other. In further group pairwise comparisons, it was shown that 37 metabolites were substantially dysregulated (p 0.05) between the PTX-free medication and the PXT-PLGA-NPs. Out of these, it is important to call attention to the metabolites that became more abundant as a result of treatment with PXT-PLGA-NPs. These include 5-Thymidyclic acid with a 7.8-fold change (FC) and 3,4,5-Trimethoxycinnamic acid, both of which have been linked in the past to effective anticancer drug treatment (Quinn et al. 2015; Anantharaju et al. 2017). The findings suggest a more successful anti-drug therapy that makes use of NP, and also indicate a number of metabolites that have the potential to serve as indicators for determining how well an antidrug treatment is working. Our previous findings are consistent with these findings.
Sex-dependent effects of FGF21 on hepatic metabolism
STUDY_SUMMARY
To investigate sex-dependent effects of fibroblast growth factor-21 (FGF21), diet-induced obese male and female wildtype mice were administered two acute doses of FGF21 or saline vehicle. Mice were euthanized, livers collected, and submitted for untargeted metabolomics analysis.
Ramadan diurnal intermittent fasting is associated with significant plasma metabolomics changes in overweight and obese subjects: A prospective cohort study
STUDY_SUMMARY
During the holy month of Ramadan, adult healthy Muslims are mandated to abstain from dawn to sunset, with free eating night hours that may extend up to 12 hours. The current work was designed to investigate the metabolomics changes incurred upon the observance of Ramadan diurnal intermittent fasting (RDIF). Twenty-five metabolically healthy participants with overweight and obesity (7 females and 18 males, with a mean age of 39.48Ā±10.0 years) were recruited for the study and were followed before and at the end of RDIF month. Dietary, anthropometric, biochemical, and physical activity assessments were performed before and at the end of the fasting month. The metabolomic assay was performed using liquid chromatography-mass spectrometry for the two-time points. Metabolomics assay revealed a significant reduction in a few metabolites. The analysis revealed that 27 metabolites differed significantly (P<0.05) between pre-and post-RDIF. Among the differentially abundant metabolites, 23 showed a decrease with fasting, these included several amino acids such as aspartame, tryptophan, phenylalanine, histidine, and other metabolites including valeric acid, and cortisol. On the other hand, only four metabolites showed increased levels with RDIF including traumatic acid, 2-pyrrolidinone, PC(18:1(9Z)/18:1(9Z)), and L-sorbose. The MetaboAnalystĀ® platform reported that the top enriched metabolic pathways included: (1) histidine metabolism; (2) folate biosynthesis (3) phenylalanine, tyrosine, and tryptophan biosynthesis; (3) aminoacyl-tRNA biosynthesis; (3) caffeine metabolism (4) vitamin B6 metabolism; and several other pathways relating to lipid metabolisms such as arachidonic acid metabolism, glycerophospholipid metabolism, and linoleic acid metabolism. In conclusion, RDIF entails significant changes in various metabolic pathways that reflect different dietary and lifestyle behaviors practiced during the fasting month.
Untargeted metabolomics on plasma from children with asthma, comparing exacerbation-prone to non-exacerbation-prone
STUDY_TYPE
Untargeted MS
STUDY_SUMMARY
Plasma was collected via venipuncture from 215 children, age 6-17 years, with physician-diagnosed asthma. Exacerbation-prone asthma was defined as having had three or more exacerbations treated with systemic corticosteroids in the previous calendar year, with each exacerbation separated by at least two weeks.
Lipidomics analysis on Arabidopsis autophagy mutants
STUDY_SUMMARY
Autophagy is an essential cellular process in eukaryotes that degrades and recycles macromolecules and organelles. Defects in autophagy is known to affect metabolism, including the lipidome. Genetic approaches have identified a series of AuTophaGy-related (ATG) genes in Arabidopsis. In this study we used WT (ecotype Col-0) and two Arabidopsis autophagy-defective mutants, atg7 and atg9 to perform a multi-omics study on the effect of nitrogen starvation treatment, which induces autophagy. Specifically, we have quantified ~100 lipids from leaf and root tissues of WT, atg7 and atg9 mutant plants, under either autophagy-inducing conditions (-N) or normal nitrogen conditions (+N). The lipid species we quantified include: DGDG, MGDG, LPC, LPE, PE, LPG, PC, PA, PG, PI, and PS. Our study sheds lights on the understanding of the relationships between autophagy and metabolism, especially lipid metabolism.
Metabolomic profiles of T. spiralis-infected mouse serum at 0, 2, 4, 8 weeks
STUDY_SUMMARY
Trichinellosis is the zoonosis affected people worldwide, caused by parasitic nematode in Genus Trichinella. After ingesting raw meat containing infective larvae of Trichinella spp., patients may show signs of myalgia, headaches, facial and periorbital edema. In severe cases, patients develop myocarditis, heart failure, and possibly death. The standard method for diagnosis of Trichinella infection is immunological techniques, which lack of sensitivity and timeliness. Metabolomics has been extensively used to identify compounds with diagnostic potential in many diseases, however, there is no study regarding biomarker discovery in trichinellosis yet. Therefore, this study aims to identify potential biomarkers of trichinellosis using metabolomics. Mice were infected with larvae stage of T. spiralis and their serum were collected before, 2 weeks, 4 weeks, and 8 weeks after infection. Metabolites in serum were extracted and identified using mass spectrometer in untargeted manner. Metabolomic data was annotated with XCMS online platform and analyzed with Metaboanalyst version 5.0. A total of 4,688 and 5,533 metabolite features were identified from positive and negative mode, respectively. The 1,139 features were significantly changed metabolites and further used for pathway analysis and biomarker selection. Glycerophospholipid metabolism was the major pathway affected by Trichinella infection and these lipid species were the main lipid class identified. The Receiver operating characteristic (ROC) revealed 247 molecules with diagnostic power of trichinellosis. Phosphatidylserine was the major lipid class from ROC analysis, for example, PS(12:0/15:0), PS(18:0/19:0)[U]. Our study suggested glycerophospholipid and phosphatidylserine species as the potential markers of trichinellosis. Findings of this study are the initial step for biomarker discovery in trichinellosis, which would be a benefit for improvement of disease diagnosis in the future.
INSTITUTE
Princess Srisavangavadhana College of Medicine, Chulabhorn Royal Academy
Maternal obesity alters offspring liver and skeletal muscle metabolism in early post-puberty despite maintaining a normal post-weaning dietary lifestyle
STUDY_SUMMARY
Maternal obesity (MO) during pregnancy is linked to increased and premature risk of age-related metabolic diseases in the offspring. However, the underlying molecular mechanisms still remain not fully understood. Using a well-established baboon model of MO, we analyzed tissue biopsies and plasma samples obtained from post-pubertal offspring (3-6.5y at sample collection) of MO mothers (n=19) and from control animals born to mothers fed a standard diet (CON, n=13). All offspring ate normal chow diet after weaning. With an untargeted gas chromatography-mass spectrometry metabolomics profiling, we quantified a total of 351 liver, 316 skeletal muscle and 423 plasma metabolites. We found 58 metabolites significantly altered in liver and 46 in skeletal muscle of MO offspring, including 8 metabolites shared between both tissues. Male and female-specific metabolites in opposite direction of change were found in liver and skeletal muscle of MO offspring. Several tissue-specific and 4 shared metabolic pathways were identified from these dysregulated metabolites. Interestingly, none of the tissue-specific metabolic alterations reflected in plasma. Our results identify tissue metabolites and pathways in post-pubertal MO offspring in a sex-specific manner.
INSTITUTE
Wake Forest School of Medicine
LAST_NAME
Ampong
FIRST_NAME
Isaac
ADDRESS
Center for Precision Medicine, Department of Internal Medicine, Section on Molecular Medicine, Wake Forest University, Winston-Salem, North Carolina, United States
Functional metabolic molecules were identified as novel therapeutic targets to facilitate gemcitabine treatment against pancreatic cancer (Cells metabolomics with ATP)
STUDY_SUMMARY
With the development of frontier technologies in system biology, traditional omics-drove phenotypic studies are insufficient to decipher the diseases. Therefore, for a thorough understanding of the molecular mechanisms of diseases to investigate novel drug targets, traditional phenotypic studies must be broken through to the functional exploration of molecules. Meanwhile, the intuitive role of small molecule compounds (metabolites) in pathogenesis, precision diagnosis and therapy are gradually recognized compared to macromolecules such as DNA, RNA and proteins. Therefore, we pioneeringly proposed Spatial Temporal Operative Real Metabolomics (STORM) strategy that established a relationship between metabolic phenotypes and functions to accurately character abnormal metabolisms and further identify operative functional molecules as novel therapeutic targets. Here, given the difficulty of pancreatic cancer (PC) treatment and the high resistance of clinical drugs, we were committed to explore new targets and drugs of pancreatic cancer from a small molecular functional perspective via STORM strategy. Fortunately, based on targeted metabolomics, we found that gemcitabine, one of the most effective clinical anti-PC drugs, served as a dual modulator that promote the accumulation of functional metabolic molecules in purine metabolism to activate down-streamed kinases. And the quantitative consequences of related enzymes annotated the unique molecular mechanisms of purine metabolism regulations by gemcitabine. Collectively, we broadened the cognitions of gemcitabine in tumor inhibition, providing potential strategies for treating PC with small molecules modification. Even more importantly, with the integration of multiple frontier technologies, the STORM strategy has proven to be well adapted to the phenotypic era of functional molecules devoted to innovate molecule mechanism annotation and therapeutic discovery.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Lu
FIRST_NAME
Haitao
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Metabolomics and metagenomics of pediatric obesity (Serum)
STUDY_SUMMARY
Pediatric obesity has grown as important global health problem in the world. The pediatric obesity affects all the organs and it is closely linked to risks of metabolic diseases such as diabetes, cardiovascular disease, and mental disease. However, the mechanism of both the pediatric obesity and treatment of the obesity remains unclear. Therefore, we investigated metabolomic pathways related to the pediatric obesity and the treatment through metabolomics and metagenomics approaches.
INSTITUTE
Seoul National University College of Medicine and Hospital
LAST_NAME
Lee
FIRST_NAME
Yujin
ADDRESS
101, Daehak-ro, Jongno-gu, Seoul, Republic of Korea
Lipidomic profiling reveals age-dependent changes in complex plasma membrane lipids that regulate neural stem cell aging
STUDY_SUMMARY
The aging brain exhibits a decline in the regenerative populations of neural stem cells (NSCs), which may underlie age-associated defects in sensory and cognitive functions1-4 . While mechanisms that restore old NSC function have started to be identified5-9 , the role of lipids ā especially complex lipids ā in NSC aging remains largely unclear. Using lipidomic profiling by mass spectrometry, we identify age-related lipidomic signatures in young and old quiescent NSCs in vitro and in vivo. These analyses reveal drastic changes in several complex membrane lipid classes, including phospholipids and sphingolipids in old NSCs. Moreover, polyunsaturated fatty acids (PUFAs) strikingly increase across complex lipid classes in quiescent NSCs during aging. Lipidomic profiling of isolated plasma membrane vesicles shows that agerelated differences in complex lipid levels and side chain composition are largely occurring in plasma membrane lipids. Experimentally, we find that aging is accompanied by modifications in membrane biophysical properties, with a decrease in plasma membrane order in old quiescent NSCs in vitro and in vivo. To determine the functional role of plasma membrane lipids in aging NSCs, we perform genetic and supplementations studies. Knockout of Mboat2, which encodes a phospholipid acyltransferase, exacerbates age-related lipidomic changes in old quiescent NSCs and impedes their ability to activate. As Mboat2 expression declines with age, Mboat2 deficiency may drive NSC decline during aging. Interestingly, supplementation of plasma membrane lipids derived from young NSCs boosts the ability of old quiescent NSCs to activate. Our work could lead to lipid-based strategies for restoring the regenerative potential of NSCs in old individuals, which has important implications to counter brain decline during aging.
Lipidomic profiling reveals age-dependent changes in complex plasma membrane lipids that regulate neural stem cell aging
STUDY_SUMMARY
The aging brain exhibits a decline in the regenerative populations of neural stem cells (NSCs), which may underlie age-associated defects in sensory and cognitive functions1-4 . While mechanisms that restore old NSC function have started to be identified5-9 , the role of lipids ā especially complex lipids ā in NSC aging remains largely unclear. Using lipidomic profiling by mass spectrometry, we identify age-related lipidomic signatures in young and old quiescent NSCs in vitro and in vivo. These analyses reveal drastic changes in several complex membrane lipid classes, including phospholipids and sphingolipids in old NSCs. Moreover, polyunsaturated fatty acids (PUFAs) strikingly increase across complex lipid classes in quiescent NSCs during aging. Lipidomic profiling of isolated plasma membrane vesicles shows that agerelated differences in complex lipid levels and side chain composition are largely occurring in plasma membrane lipids. Experimentally, we find that aging is accompanied by modifications in membrane biophysical properties, with a decrease in plasma membrane order in old quiescent NSCs in vitro and in vivo. To determine the functional role of plasma membrane lipids in aging NSCs, we perform genetic and supplementations studies. Knockout of Mboat2, which encodes a phospholipid acyltransferase, exacerbates age-related lipidomic changes in old quiescent NSCs and impedes their ability to activate. As Mboat2 expression declines with age, Mboat2 deficiency may drive NSC decline during aging. Interestingly, supplementation of plasma membrane lipids derived from young NSCs boosts the ability of old quiescent NSCs to activate. Our work could lead to lipid-based strategies for restoring the regenerative potential of NSCs in old individuals, which has important implications to counter brain decline during aging.
Lipidomic profiling reveals age-dependent changes in complex plasma membrane lipids that regulate neural stem cell aging
STUDY_SUMMARY
The aging brain exhibits a decline in the regenerative populations of neural stem cells (NSCs), which may underlie age-associated defects in sensory and cognitive functions1-4 . While mechanisms that restore old NSC function have started to be identified5-9 , the role of lipids ā especially complex lipids ā in NSC aging remains largely unclear. Using lipidomic profiling by mass spectrometry, we identify age-related lipidomic signatures in young and old quiescent NSCs in vitro and in vivo. These analyses reveal drastic changes in several complex membrane lipid classes, including phospholipids and sphingolipids in old NSCs. Moreover, polyunsaturated fatty acids (PUFAs) strikingly increase across complex lipid classes in quiescent NSCs during aging. Lipidomic profiling of isolated plasma membrane vesicles shows that agerelated differences in complex lipid levels and side chain composition are largely occurring in plasma membrane lipids. Experimentally, we find that aging is accompanied by modifications in membrane biophysical properties, with a decrease in plasma membrane order in old quiescent NSCs in vitro and in vivo. To determine the functional role of plasma membrane lipids in aging NSCs, we perform genetic and supplementations studies. Knockout of Mboat2, which encodes a phospholipid acyltransferase, exacerbates age-related lipidomic changes in old quiescent NSCs and impedes their ability to activate. As Mboat2 expression declines with age, Mboat2 deficiency may drive NSC decline during aging. Interestingly, supplementation of plasma membrane lipids derived from young NSCs boosts the ability of old quiescent NSCs to activate. Our work could lead to lipid-based strategies for restoring the regenerative potential of NSCs in old individuals, which has important implications to counter brain decline during aging.
Lipidomic profiling reveals age-dependent changes in complex plasma membrane lipids that regulate neural stem cell aging
STUDY_SUMMARY
The aging brain exhibits a decline in the regenerative populations of neural stem cells (NSCs), which may underlie age-associated defects in sensory and cognitive functions1-4 . While mechanisms that restore old NSC function have started to be identified5-9 , the role of lipids ā especially complex lipids ā in NSC aging remains largely unclear. Using lipidomic profiling by mass spectrometry, we identify age-related lipidomic signatures in young and old quiescent NSCs in vitro and in vivo. These analyses reveal drastic changes in several complex membrane lipid classes, including phospholipids and sphingolipids in old NSCs. Moreover, polyunsaturated fatty acids (PUFAs) strikingly increase across complex lipid classes in quiescent NSCs during aging. Lipidomic profiling of isolated plasma membrane vesicles shows that agerelated differences in complex lipid levels and side chain composition are largely occurring in plasma membrane lipids. Experimentally, we find that aging is accompanied by modifications in membrane biophysical properties, with a decrease in plasma membrane order in old quiescent NSCs in vitro and in vivo. To determine the functional role of plasma membrane lipids in aging NSCs, we perform genetic and supplementations studies. Knockout of Mboat2, which encodes a phospholipid acyltransferase, exacerbates age-related lipidomic changes in old quiescent NSCs and impedes their ability to activate. As Mboat2 expression declines with age, Mboat2 deficiency may drive NSC decline during aging. Interestingly, supplementation of plasma membrane lipids derived from young NSCs boosts the ability of old quiescent NSCs to activate. Our work could lead to lipid-based strategies for restoring the regenerative potential of NSCs in old individuals, which has important implications to counter brain decline during aging.
Investigating metabolic pathways of pediatric obesity (urine)
STUDY_SUMMARY
The pediatric obesity influences all the organs and is closely linked to an increased risk of diverse diseases such as diabetes, cardiovascular, stroke, social problems and depression. Therefore, there is needed to diverse study for effective methods for the prevention and treatment of pediatric obesity. Diverse evidences suggest that gut microbiome and its metabolites affect metabolic disease such as obesity, diabetes, and heart disease. Previous studies in human and fecal transplantation experiments in animal models identified connections between the metabolic diseases and gut microbiota [4]. Moreover, metabolites can be fulfilled as diagnostic, prognostic, and therapeutic targets for diseases. Thus, approaches using metagenomics and metabolomics have the potential to provide new insights into metabolomic pathways of the diseases and help personalized and efficient treatments. In this study, we aimed to investigate metabolomic pathways of pediatric obesity analyzing both metabolome and microbiome profiles. In addition, we proceeded obese intervention with obese children to examine underlying metabolomic pathways related to effect of the intervention.
INSTITUTE
Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital
LAST_NAME
Lee
FIRST_NAME
Yujin
ADDRESS
101 Daehak-ro, Jongno-gu, Seoul 110-799, Korea, Seoul, Seoul, 03080, Korea, South
Metabolomics and metagenomics of pediatric obesity (Feces)
STUDY_SUMMARY
Pediatric obesity has grown as important global health problem in the world. The pediatric obesity affects all the organs and it is closely linked to risks of metabolic diseases such as diabetes, cardiovascular disease, and mental disease. However, the mechanism of both the pediatric obesity and treatment of the obesity remains unclear. Therefore, we investigated metabolomic pathways related to the pediatric obesity and the treatment through metabolomics and metagenomics approaches.
INSTITUTE
Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital
LAST_NAME
Lee
FIRST_NAME
Yujin
ADDRESS
101 Daehak-ro, Jongno-gu, Seoul 110-799, Korea, Seoul, Seoul, 03080, Korea, South
Intermittent fasting induces rapid hepatocyte proliferation to maintain the hepatostat
STUDY_SUMMARY
Nutrient availability fluctuates in most natural populations, forcing organisms to undergo periods of fasting and re-feeding. It is unknown how dietary change influences liver homeostasis. Here, we show that a switch from ad libitum feeding to intermittent fasting (IF) promotes rapid hepatocyte proliferation. Mechanistically, IF- induced hepatocyte proliferation is driven by the combined action of intestinally produced, systemic endocrine FGF15 and localized WNT signaling. IF proliferation re-establishes a constant liver-to-body-mass ratio during periods of fasting and re-feeding, a process termed the hepatostat. This study provides the first example of dietary influence on adult hepatocyte proliferation, and challenges the widely held view that liver tissue is mostly quiescent unless chemically or mechanically injured.
Transcriptomic-Metabolomic Profiling in Mouse Lung Tissues Reveals Sex- and Strain-Based Differences
STUDY_SUMMARY
There has been a heighted interest in understanding the influences of sex and strain on variety of diseases states. We assessed the metabolomics profiles of male and female C57B6/J as well as male and female C3H/HeN. We pretreated each of these mice groups with 25mg/kg of Aurothioglucose (ATG) prior to being exposed to either 100% fitered air, or >95% oxygen for 96 hours. These studies begin to address the underlying altered metabolomic signaling that is influenced by sex and/or strain.
INSTITUTE
Univeristy of Oklahoma Health Sciences Center
DEPARTMENT
Section of Neonatal-Perinatal Medicine
LABORATORY
Clinical Biomarkers Laboratory
LAST_NAME
Tipple
FIRST_NAME
Trent
ADDRESS
1200 North Everett Drive, ETNP 7504 Oklahoma City, Oklahoma 73104-5407
EMAIL
Trent-Tipple@ouhsc.edu
PHONE
405-271-5215
TOTAL_SUBJECTS
48
STUDY_COMMENTS
Clinical Biomarker Laboratory pooled plasma samples were used as standards
Multi-omic analysis reveals bacteria may have a role in dental erosion
STUDY_TYPE
Research Study
STUDY_SUMMARY
NMR was performed on 11 saliva samples; 5 from participants classified as having dental erosion and 6 from healthy control participants with no dental erosion to assess the differences in metabolome between the two groups. NMR analysis alone revealed no significant differences between the dental erosion and healthy controls. However, bacterial mRNA sequencing of the oral microbiome from the same saliva samples was performed and the bacterial gene expression profiles was correlated to metabolite concentrations in the groups. The dental erosion group had strong correlations between metabolites associated with protein degradation and amino acid fermentation (formate, butyrate, propionate, 5-aminopentanoate, acetate, glycine, phenylalanine, dimethyl sulfone) and increased activity of species including 4 Prevotella species, Actinomyces graevenitzii, Tannerella species, and 2 Selenomas species, to name a few. Whereas in the healthy control group, the only positive correlations between metabolite concentrations and bacterial activity was for urea and 5-aminopentanoate; urea was positively correlated with Aggregatibacter actinomycetecomytans, Lysinibacillus fusiformis, and Veillonella tobetsuensis, and 5-aminopentanoate was positively correlated with 3 different Leptotrichia species, Streptococcus parasanguinis, and 2 Prevotella species.
INSTITUTE
King's College London
LAST_NAME
Cleaver
FIRST_NAME
Leanne
ADDRESS
Floor 17, Tower Wing, Guy's Hospital, King's College London, Great Maze Pond
KÄ«lauea lava fuels phytoplankton bloom in the North Pacific Ocean - study of particulate metabolites
STUDY_TYPE
Study of particulate metabolites in phytoplankton blooms.
STUDY_SUMMARY
From June to August 2018, the eruption of the KÄ«lauea volcano on the island of Hawaiāi injected millions of cubic meters of molten lava into the nutrient-poor waters of the North Pacific Subtropical Gyre. The lava-impacted seawater was characterized by high concentrations of metals and nutrients that stimulated phytoplankton growth, resulting in an extensive plume of chlorophyll a that was detectable by satellite. Samples for particulate metabolites were collected from different stations surrounding the lava flowing into the ocean to see how marine microorganisms respond to exogenous inputs of nutrients and metals.
INSTITUTE
University of Washington
DEPARTMENT
School of Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Lionheart
FIRST_NAME
Regina
ADDRESS
1400 NE Campus Parkway, Seattle, Washington, 98195, USA
Evaluation of the effects of chitosan films as a replacement for conventional sulphur dioxide treatment of white wines
STUDY_TYPE
1H-NMR metabolomics
STUDY_SUMMARY
In this study, 1H-NMR metabolomics was used to evaluate the effects of using chitosan-genipin (Ch-Ge) films as replacement of sulfur dioxide (SO2) in white wines preservation, to circumvent adverse health consequences caused by SO2 intake, on the final compositional profile of white wines. To do so, differently sized Ch-Ge films (25 and 100 cm2) were tested, as well as SO2-tretment and untreated wines. The obtained data added important knowledge on the potential use of Ch-Ge films, particularly those of higher surface areas, as replacements for the use of SO2 in wine conservation, based on the changes noted in metabolite composition and their putative explanations in terms of wine chemical and biochemical characteristics.
INSTITUTE
University of Aveiro
DEPARTMENT
CICECO ā Aveiro Institute of Materials, Department of Chemistry
LABORATORY
Metabolomics group
LAST_NAME
Rodrigues
FIRST_NAME
Joao E.
ADDRESS
Departamento de QuĆmica, Universidade de Aveiro, Campus de Santiago
Autophagy-related protein PIK3C3 maintains healthy brown and white adipose tissues to prevent metabolic diseases
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Adequate mass and function of adipose tissues (ATs) play an essential role in preventing metabolic perturbations. Pathological reduction of ATs in lipodystrophy leads to an array of metabolic diseases. Understanding the underlying mechanisms may benefit the development of effective therapies. Several cellular processes, including autophagy, function collectively to maintain AT homeostasis. Here, we investigated the impact of adipocyte-specific deletion of the autophagy-related lipid kinase PIK3C3 on AT homeostasis and systemic metabolism in mice. We report that PIK3C3 functions in all ATs and that its absence disturbs adipocyte autophagy and hinders adipocyte differentiation, survival, and function with differential effects on brown and white ATs. These abnormalities caused loss of white ATs, whitening followed by loss of brown ATs, and impaired browning of white ATs. Consequently, mice exhibited compromised thermogenic capacity and developed dyslipidemia, hepatic steatosis, insulin resistance and type 2 diabetes. While these effects of PIK3C3 contrast previous findings with the autophagy-related protein ATG7 in adipocytes, mice with a combined deficiency in both factors revealed a dominant role of the PIK3C3-deficient phenotype. We also found that dietary lipid excess exacerbates AT pathologies caused by PIK3C3 deficiency. Surprisingly, glucose tolerance was spared in adipocyte-specific PIK3C3-deficient mice, a phenotype that was more evident during dietary lipid excess. These findings reveal a crucial yet complex role for PIK3C3 in ATs and suggest the potential of targeting this factor for therapeutic intervention in metabolic diseases.
Targeted plasma metabolomics in pediatric NAFLD patients
STUDY_SUMMARY
Several adult omics studies have been conducted to understand the pathophysiology of nonalcoholic fatty liver disease (NAFLD). However, the histological features of children are different from those of adults, and the onset and progression of pediatric NAFLD are not fully understood. In this study, we aimed to evaluate the metabolome profile and metabolic pathway changes associated with pediatric NAFLD to elucidate its pathophysiology. We analyzed the metabolic profiles of healthy control, lean NAFLD, overweight control and overweight NAFLD groups of children and adolescent participants (n = 165) by assessing plasma samples, and identified 18 NAFLD-specific metabolic features and metabolic changes in lipid, glutathione-related amino acid, and branched-chain amino acid metabolism by comparing control and NAFLD group in pediatric population with overweight. Metabolome changes in the plasma of pediatric patients with NAFLD are associated with the pathophysiology of the disease and can be utilized as a less-invasive approach to diagnose the disease.
INSTITUTE
Seoul National University College of Medicine and Hospital
DEPARTMENT
Department of Clinical Pharmacology and Therapeutics
LAST_NAME
Chae
FIRST_NAME
Woori
ADDRESS
101 Daehak-ro, Jongno-gu, Seoul 03080, Republic of Korea
Stage 41 tadpoles were injected with 4 nmol of the glycolysis inhibitor, 2-deoxyglucose (2DG), or a tracer control to evaluate consequences of inhibition on metabolites 24 hours after treatment began. Similarly, tadpoles were incubated in DMSO control or 1 of 2 G6PD inhibitors (Dehydroepiandrosterone and g6pdi), to similarly assess the consequences of inhibiting the pentose phosphate pathway. We find that inhibition of glucose metabolism with 2DG results in a decrease in downstream glycolytic intermediates, confirming a reduction in activity of this pathway. G6PD inhibition was not as clear as changes were less consistent across treatments and downstream metabolites did not behave in a coordinated way, though impacts on other metabolic processes by these inhibitors may be fruitful for exploring how they perturb metabolism in the tadpoles.
INSTITUTE
University of Washington
LAST_NAME
Patel
FIRST_NAME
Jeet
ADDRESS
1705 NE Pacific St., HSB J-Wing, J405, Seattle, Washington, 98195, USA
To identify changes in metabolites that correlate with progression of tail regeneration, we collected a timecourse of tissues containing 250 um of tissue anterior to the wound site as well as all regenerating tissue at 0, 3, and 24 hours post amputation. We also collected the posterior 500 um of the developing tail to represent the metabolic profile of uninjured tissues. Tissues from 25 individuals were collected and frozen in more than 5-8 minutes per replicate before processing as in the methods. 104 metabolites were identified in these samples and relative peak intensities were compared to identify changes in abundance corresponding to regeneration. 42 differentially abundant metabolites were found using MetaboAnalyst, the majority of which were increased 24 hours post amputation. Further investigation of these 24 hours post amputation enriched metabolites revealed that these metabolites were largely associated with increased growth and nucleotide metabolism. This finding is in line with the growth of new tissue seen at this timepoint and also suggests that generation of nucleotides are a major factor in sustaining this growth.
INSTITUTE
University of Washington
LAST_NAME
Patel
FIRST_NAME
Jeet
ADDRESS
1705 NE Pacific St., HSB J-Wing, J405, Seattle, Washington, 98195, USA
Metabolic changes in seeds of malting barley produced under drought or elevated temperature
STUDY_TYPE
Barley seed phenotyping and GC-MS based metabolomic analysis
STUDY_SUMMARY
Plants of a āHana-typeā landrace (B1) were taller, flowered earlier and produced heavier, larger and more vigorous seeds that resisted ageing longer compared to a semi-dwarf breeding line (B2). Drought significantly reduced seed yield in both genotypes, and elevated temperature reduced seed size. Genotype B2 showed partial thermodormancy that was alleviated by drought and elevated temperature, in line with lower abundance of the TF ABI5, a key regulator of seed dormancy and vigour. Metabolite profiling revealed clear differences between the embryos of B1 and B2. Drought, but not elevated temperature, affected the metabolism of amino acids, organic acids, osmolytes and nitrogen assimilation, in the seeds of both genotypes.
INSTITUTE
INRAE
LAST_NAME
CLEMENT
FIRST_NAME
Gilles
ADDRESS
Route de ST-Cyr, Versailles, Ile de France, 78026, France
Multi-Omics analysis revealed a significant alteration of critical metabolic pathways due to sorafenib-resistance in Hep3B cell lines
STUDY_TYPE
MS- comparative metabolomic analysis
STUDY_SUMMARY
Hepatocellular carcinoma (HCC) is the second prominent cause of cancer-associated death worldwide. Usually, HCC is diagnosed in advanced stages, where sorafenib, a multiple target ty-rosine kinase inhibitor, is used as the first line of treatment. Unfortunately, resistance to sorafenib is usually encountered within six months of the treatment. Therefore, there is a critical need to identify the underlying reasons for drug resistance. In the present study, we investigated the proteomic and metabolomics alterations accompanying to sorafenib resistance in hepatocellular carcinoma Hep3B cells by employing ultra-high-performance liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS). The Bruker Human Metabolome Database (HMDB) library was used to identify the differentially abundant metabolites through MetaboScape 4.0 software (Bruker). For protein annotation and identification, the Uniprot proteome for Homo sapiens (Human) da-tabase was utilised through MaxQuant. The results revealed that 27 metabolites and 18 proteins were significantly dysregulated due to sorafenib resistance in Hep3B cells compared to the parental phenotype. D-alanine, L-proline, o-tyrosine, succinic acid and phosphatidylcholine (PC, 16:0/16:0) were among the significantly altered metabolites. Ubiquitin carboxyl-terminal hydrolase isozyme L1, mitochondrial superoxide dismutase, UDP-glucose-6-dehydrogenase, sorbitol dehydrogenase and calpain small subunit 1 were among the significantly altered proteins. The findings revealed that resistant Hep3B cells demonstrated significant alterations in amino acid and nucleotide met-abolic pathways, energy production pathways and other pathways related to cancer aggressive-ness, migration, proliferation, and drug-resistance. Joint pathway enrichment analysis unveiled unique pathways, including the antifolate resistance pathway and other important pathways that maintain cancer cells' survival, growth, and proliferation. Collectively, the results identified po-tential biomarkers for sorafenib-resistant HCC and gave insights into their role in chemotherapeutic drug resistance, cancer initiation, progression, and aggressiveness, which may contribute to better prognosis and chemotherapeutic outcomes.
1-deoxysphingolipid synthesis compromises anchorage-independent growth and plasma membrane endocytosis in cancer cells
STUDY_SUMMARY
Serine palmitoyltransferase (SPT) predominantly incorporates serine and fatty acyl-CoAs into diverse sphingolipids that serve as structural components of membranes and signaling molecules within or amongst cells. However, SPT also uses alanine as a substrate in the contexts of low serine availability, alanine accumulation, or diseasecausing mutations in hereditary sensory neuropathy type I (HSAN1), resulting in the synthesis and accumulation of 1-deoxysphingolipids. These species promote cytotoxicity in neurons and impact diverse cellular phenotypes, including suppression of anchorage-independent cancer cell growth. While altered serine and alanine can promote 1-deoxysphingolipid synthesis, they impact numerous other metabolic pathways important for cancer cells. Here we combined isotope tracing, quantitative metabolomics, and functional studies to better understand the mechanistic drivers of 1- deoxysphingolipid toxicity in cancer cells. Both alanine treatment and SPTLC1 C133W expression induce 1-deoxy(dihydro)ceramide synthesis and accumulation but fail to broadly impact intermediary metabolism, abundances of other lipids, or growth of adherent cells. However, spheroid culture and soft agar colony formation were compromised when endogenous 1-deoxysphingolipid synthesis was induced via SPTLC1 C133W expression. Consistent with these impacts on anchorageindependent cell growth, we observed that 1-deoxysphingolipid synthesis reduced plasma membrane endocytosis. These results highlight a potential role for SPT promiscuity in linking altered amino acid metabolism to plasma membrane endocytosis.
INSTITUTE
Salk Institute for Biological Studies
LABORATORY
Molecular and Cell Biology Laboratory (Christian Metallo)
LAST_NAME
Cordes
FIRST_NAME
Thekla
ADDRESS
10010 N Torrey Pines Rd, La Jolla, CA 92037, United States
Insights from hippocampal neurogenesis and brain tumor development in a mouse model of experimental colitis induced by dextran sodium sulfate
STUDY_SUMMARY
We here reported investigations on a model of chemically induced experimental colitis by oral administration of sodium dextran sulfate (DSS) in C57BL/6 mice. We investigated, in vivo, the crosstalk between the intestine and the brain, evaluating the consequences of intestinal inflamma-tion on neuroinflammation and hippocampal adult neurogenesis. By using different DSS admin-istration strategies, we were able to induce acute or chronic colitis simulating clinical character-istics observed in IBD patients
INSTITUTE
Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile
Machine Learning Reveals Lipidome Dynamics in a Mouse Model of Ovarian Cancer
STUDY_SUMMARY
Ovarian cancer (OC) is one of the deadliest cancers affecting the female reproductive system. It presents little or no symptoms at the early stages, and typically unspecific symptoms at later stages. Of the OC subtypes, high-grade serous carcinoma (HGSC) is responsible for most OC deaths. However, very little is known about the metabolic course of this disease. In this longitudinal study, we investigated the temporal course of lipidome changes in a Dicer-Pten Double-Knockout (DKO) HGSC mouse model using machine and statistical learning approaches. Early progression of HGSC was marked by increased levels of phosphatidylcholines and phosphatidylethanolamines. In contrast, later stages were marked by more diverse lipids alterations, including fatty acids and their derivatives, triglycerides, ceramides, hexosylceramides, sphingomyelins, lysophosphatidylcholines, and phosphatidylinositols. These alterations provided evidence of perturbations in cell membrane stability, proliferation, and survival and candidates for early-stage and prognostic markers in humans.
Skin-to-blood pH shift triggers metabolome and proteome global remodelling in Staphylococcus epidermidis
STUDY_TYPE
NMR Metabolomics combine with proteomics to study pH adaptation of Staphylococcus epidermidis 19N
STUDY_SUMMARY
Staphylococcus epidermidis (SE) is one of the most common bacteria of the human skin microbiota. Despite its role as a commensal, SE has emerged as an opportunistic pathogen, associated with 80% of medical devices related infections. Moreover, these bacteria are extremely difficult to treat due to their ability to form biofilms and accumulate resistance to almost all classes of antimicrobials developed so far. Thus new preventive and therapeutic strategies are urgently needed. In spite of its clinical importance, the molecular mechanisms associated with SE colonisation and disease are still poorly understood. A deeper understanding of the metabolic and cellular processes associated with response to environmental factors characteristic of SE ecological niches in health and disease might provide new clues on colonisation and disease processes. Here we studied the impact of pH conditions, mimicking the skin pH (5.5) and blood pH (7.4), in a S. epidermidis commensal strain, belonging to the B clonal lineage, by means of next-generation proteomics and 1H NMR-based metabolomics. Moreover, we evaluated the metabolic changes occurring when a sudden pH change arise, simulating the skin barrier break produced by a catheter. We found that exposure of S. epidermidis to skin pH induced oxidative phosphorylation and biosynthesis of peptidoglycan, lipoteichoic acids and betaine. In contrast, at blood pH, the incorporation of monosaccharides and its oxidation by glycolysis and fermentation was promoted. Additionally, several proteins related to virulence and immune evasion, namely extracellular proteases and membrane iron transporters were more abundant at blood pH. In the situation of an abrupt skin-to-blood pH shift we observed the decrease in the osmolyte betaine and changes in the levels of several metabolites and proteins involved in redox cell homeostasis. Our results suggest that at the skin pH S. epidermidis cells are metabolically more active and adhesion is promoted, while at blood pH, metabolism is tuned down and cells have a more virulent profile. pH increase during commensal-to-pathogen conversion appears to be a critical environmental signal to the remodelling of the S. epidermidis metabolism towards a more pathogenic state. Targeting S. epidermidis proteins induced by a low alkaline pH and local acidification of medical devices microenvironment might be new strategies to treat and prevent S. epidermidis infections.
INSTITUTE
ITQB NOVA
LABORATORY
Proteomics of Non-Model Organisms
LAST_NAME
GonƧalves
FIRST_NAME
LuĆs
ADDRESS
Avenida Republica, Oeiras, Not USCanada, 2780-157 Oeiras, Portugal
In vitro studies associated oxidative phosphorylation (OXPHOS) with anti-inflammatory macrophages, while pro-inflammatory macrophages rely on glycolysis. However, the metabolic needs of macrophages in tissues (TMFs) to fulfil their homeostatic activities are incompletely understood. Here, we identified OXPHOS as highly discriminating process among TMFs from different tissues in homeostasis by analysis of RNAseq data, in both human and mouse. Impairing OXPHOS in TMFs via Tfam deletion differentially affected TMF populations. Tfam deletion resulted in reduction of alveolar macrophages (AMs) due to impaired lipid-handling capacity, leading to increased cholesterol content and cellular stress, causing cell cycle arrest in vivo. In obesity, Tfam depletion selectively ablated pro-inflammatory lipid-handling white adipose tissue macrophages (WAT-MFs), preventing insulin resistance and hepatosteatosis. Thus, OXPHOS, rather than glycolysis, distinguishes TMF populations and is critical for the maintenance of TMFs with a high lipid-handling activity, including pro-inflammatory WAT-MFs. This could provide a selective therapeutic targeting tool.
INSTITUTE
Spanish National Center for Cardiovascular Research (CNIC)
DEPARTMENT
Novel mechanisms of atherosclerosis
LABORATORY
Immunobiology
LAST_NAME
Mastrangelo
FIRST_NAME
Annalaura
ADDRESS
Calle de Melchor FernƔndez Almagro, 3, Centro Nacional de Investigaciones Cardiovasculares
Metabolite patterns between isogenic normal hiPSCs and Trisomy hiPSC
STUDY_SUMMARY
We wanted to compare metabolites patterns between isogenic normal hiPSCs and Trisomy hiPSCs. All hiPSCs cell lines were generated from same mosaic Down symdrome patient.
Detection of Methyl jasmonate (MeJA) in Plant root VOCs
STUDY_SUMMARY
Methyl jasmonate (MeJA) is a well-known plant hormone known for plant defense and plant-plant signaling. However, most of the studies are focussed on its aboveground presence and functions. Here we report that MeJA is also released by plant roots in a volatile form. More importantly, it is shown in Arabidopsis growing in natural conditions in soil.
The āForensOMICSā approach to forensic post-mortem interval estimation: combining metabolomics, lipidomics and proteomics for the analysis human skeletal remains
STUDY_SUMMARY
The combined use of multiple omics methods to answer complex system biology questions is growing in biological and medical sciences, as the importance of studying interrelated biological processes in their entirety is increasingly recognized. We applied a combination of metabolomics, lipidomics and proteomics to human bone to investigate the potential of this multi-omics approach to estimate the time elapsed since death (i.e., the post-mortem interval, PMI). This āForensOMICSā approach has the potential to improve accuracy and precision of PMI estimation of skeletonized human remains, thereby helping forensic investigators to establish the timeline of events surrounding death. Anterior midshaft tibial bone was collected from four female body donors in a fresh stage of decomposition before placement of the bodies to decompose outdoors at the human taphonomy facility managed by the Forensic Anthropological Center at Texas State (FACTS). Bone samples were again collected at selected PMIs (219, 790, 834 and 872 days). Liquid chromatography mass spectrometry (LC-MS) was used to obtain untargeted metabolomic, lipidomic and proteomic profiles from the pre- and post-placement bone samples. Multivariate analysis was used to investigate the three omics blocks by means of Data Integration Analysis for Biomarker discovery using Latent variable approaches for Omics studies (DIABLO), to identify the reduced number of markers that could effectively describe post-mortem changes and classify the individuals based on their PMI. The resulting model showed that pre-placement bone metabolome, lipidome and proteome profiles were clearly distinguishable from post-placement profiles. Metabolites associated with the pre-placement samples, suggested an extinction of the energetic metabolism and a switch towards another source of fuelling (e.g., structural proteins). We were able to identify certain biomolecules from the three groups that show excellent potential for estimation of the PMI, predominantly the biomolecules from the metabolomics block. Our findings suggest that, by targeting a combination of compounds with different post-mortem stability, in future studies we could be able to estimate both short PMIs, by using metabolites and lipids, and longer PMIs, by including more stable proteins.
Genetically defined human GBM organoids reveal principles of GBM development and actionable targets
STUDY_SUMMARY
Recent advances in glioblastoma (GBM) studies provide a comprehensive catalog of its genetic aberrations and cellular heterogeneity. However, a solid understanding of genotype-based analysis of cancer pathway dependency and actionable target identification is required to transform GBM treatment into a personalized era. Here, we generated a spectrum of mutant iPSCs harboring frequent GBM mutations with CRISPR/Cas9 and profiled the organoids (LEGO: Laboratory Engineered Glioblastoma Organoid) derived from these iPSCs temporally on transcriptome, methylome, metabolome, lipidome, proteome, and phospho-proteome levels. We found that LEGOs form brain tumors in vivo and recapitulate critical features of human GBM. The multi-omics analysis discovered essential milestones driven by genetic heterogeneity during GBM progressions, such as lineage alteration, methylome rewriting, and metabolome/lipidome reprogramming, in concordance with altered pathway activity and drug response. This study provides a tool and research path to realizing genome-based personalized GBM therapy using novel advanced models.
Multiplatform mass spectrometry-based analysis of Leishmania donovani infected macrophages at different time points after infection
STUDY_TYPE
Mutiplatform mass spectrometry-based metabolomics, time course experiment
STUDY_SUMMARY
The project aims to measure targeted and non-targeted metabolite data of intracellular extracts of uninfected and Leishmania-donovani infected macrophages at 0, 12, 36 and 72 hours post infection using a multiplatform mass spectrometry approach combining CE-TOF/MS (polar metabolites), LC-QTOF/MS (non-polar metabolites) and LC-QqQ/MS (polar metabolites) to characterize the dynamics of metabolic alterations ocurring in the human macrophage upon L. donovani infection.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
Chemistry and Biochemistry
LABORATORY
Centre for Metabolomics and Bioanalysis (CEMBIO)
LAST_NAME
FernĆ”ndez GarcĆa
FIRST_NAME
Miguel
ADDRESS
Centro de MetabolĆ³mica y BioanĆ”lisis (CEMBIO), Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, UrbanizaciĆ³n MonteprĆncipe, 28660 Boadilla del Monte. EspaƱa
Metabolic impact of anticancer drugs Pd2Spermine and Cisplatin on the polar metabolome of brain from cell-derived xenograft mouse model of Triple-Negative Breast Cancer (part 1)
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Palladium (Pd(II))-complexes are alternatives due to similar metal coordination and promising cytotoxic properties. Metabolomics can measure the metabolic response of drug-exposed tissues, unveiling insight into drug mechanisms and new markers of drug efficacy/toxicity. The present 1H NMR metabolomics study aims to characterize the in vivo response of the impact of a Pd(II)-complex with polyamine spermine (Pd2Spm), compared to cDDP, on polar metabolism of brain from cell-derived xenograft mouse model of Triple-Negative Breast Cancer.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Integrated metabolic and inflammatory signatures associated with severity, fatality, and recovery of COVID-19
STUDY_TYPE
Research
STUDY_SUMMARY
Severe manifestations of coronavirus disease 2019 (COVID-19) and mortality have been associated with physiological alterations that provide insights into the pathogenesis of the disease. Moreover, factors that drive recovery from COVID-19 can be explored to identify correlates of protection. The cellular metabolism represents a potential target to improve survival upon severe disease, but the associations between the metabolism and the inflammatory response during COVID-19 are not well defined. We analyzed blood laboratorial parameters, cytokines, and metabolomes of 150 individuals with mild to severe disease, of which 33 progressed to a fatal outcome. A subset of 20 individuals was followed-up after hospital discharge and recovery of acute disease. We used hierarchical community networks to integrate metabolomics profiles with cytokines and markers of inflammation, coagulation, and tissue damage. Infection by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) promotes significant alterations in the plasma metabolome, whose activity varies according to disease severity and correlates with oxygen saturation. Differential metabolism underlying death was marked by amino acids and related metabolites, such as glutamate, tryptophan and oxoproline; and lipids, including progesterone, phosphocholine and lysophosphatidylcholines (lysoPCs). Individuals that recovered from severe disease displayed persistent alterations enriched for metabolism of purines, phosphatidylinositol phosphate and glycolysis. Recovery of mild disease was associated with vitamin E metabolism. Data integration shows that the metabolic response is a hub connecting other biological features during disease and recovery. Infection by SARS-CoV-2 induces concerted activity of metabolic and inflammatory responses that depend on disease severity and collectively predict clinical outcomes of COVID-19.
INSTITUTE
Institute of Tropical Pathology and Public Health - Federal University of GoiƔs
LAST_NAME
Gardinassi
FIRST_NAME
Luiz Gustavo
ADDRESS
R. 235 s/n - Institute of Tropical Pathology and Public Health - Federal University of GoiƔs
Quantification of Dissolved Metabolites in Environmental Samples through Cation-Exchange Solid Phase Extraction (CX-SPE) paired with Liquid Chromatography-Mass Spectrometry
STUDY_TYPE
Method Development for Dissolved Metabolomics in Seawater
STUDY_SUMMARY
Small, biologically produced, organic molecules called metabolites play key roles in microbial systems where they directly mediate exchanges of nutrients, energy, and information. However, the study of dissolved polar metabolites in seawater and other environmental matrices has been hampered by analytical challenges including high inorganic ion concentrations, low analyte concentrations, and high chemical diversity. Here we show that a cation-exchange solid phase extraction (CX-SPE) sample preparation approach separates positively charged and zwitterionic metabolites from seawater and freshwater samples, allowing their analysis by liquid chromatography-mass spectrometry (LC-MS). We successfully extracted 69 known compounds from an in-house compound collection and evaluated the performance of the method by establishing extraction efficiencies and limits of detection (pM to low nM range) for these compounds. CX-SPE extracted a range of compounds including amino acids and compatible solutes, resulted in very low matrix effects, and performed robustly across large variations in salinity and dissolved organic matter (DOM) concentration. We compared CX-SPE to an established solid phase extraction procedure (PPL-SPE) and demonstrate that these two methods extract fundamentally different fractions of the dissolved metabolite pool with CX-SPE extracting compounds that are on average smaller and more polar. We use CX-SPE to analyze four environmental samples from distinct aquatic biomes, producing some of the first CX-SPE dissolved metabolomes. Quantified compounds ranged in concentration from 0.0093 nM to 49 nM and were composed primarily of amino acids (0.15 ā 16 nM) and compatible solutes such as TMAO (0.89 ā 49 nM) and glycine betaine (2.8 ā 5.2 nM).
INSTITUTE
University of Washington
DEPARTMENT
Oceanography
LABORATORY
Ingalls Lab
LAST_NAME
Sacks
FIRST_NAME
Joshua
ADDRESS
Ocean Sciences Building, 1492 NE Boat St. Seattle, WA 98105
Metabolic impact of anticancer drugs Pd2Spermine and Cisplatin on the lipophilic metabolome of brain from cell-derived xenograft mouse model of Triple-Negative Breast Cancer (part 2)
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Palladium (Pd(II))-complexes are alternatives due to similar metal coordination and promising cytotoxic properties. Metabolomics can measure the metabolic response of drug-exposed tissues, unveiling insight into drug mechanisms and new markers of drug efficacy/toxicity. The present 1H NMR metabolomics study aims to characterize the in vivo response of the impact of a Pd(II)-complex with polyamine spermine (Pd2Spm), compared to cDDP, on nonpolar metabolism of brain from cell-derived xenograft mouse model of Triple-Negative Breast Cancer.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Metabolic impact of anticancer drugs Pd2Spermine and Cisplatin on the polar metabolome of liver from cell-derived xenograft mouse model of Triple-Negative Breast Cancer (part 1)
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Palladium (Pd(II))-complexes are alternatives due to similar metal coordination and promising cytotoxic properties. Metabolomics can measure the metabolic response of drug-exposed tissues, unveiling insight into drug mechanisms and new markers of drug efficacy/toxicity. The present 1H NMR metabolomics study aims to characterize the in vivo response of the impact of a Pd(II)-complex with polyamine spermine (Pd2Spm), compared to cDDP, on polar metabolism of liver from cell-derived xenograft mouse model of Triple-Negative Breast Cancer.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Metabolic impact of anticancer drugs Pd2Spermine and Cisplatin on the lipophilic metabolome of liver from cell-derived xenograft mouse model of Triple-Negative Breast Cancer (part 2)
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Palladium (Pd(II))-complexes are alternatives due to similar metal coordination and promising cytotoxic properties. Metabolomics can measure the metabolic response of drug-exposed tissues, unveiling insight into drug mechanisms and new markers of drug efficacy/toxicity. The present 1H NMR metabolomics study aims to characterize the in vivo response of the impact of a Pd(II)-complex with polyamine spermine (Pd2Spm), compared to cDDP, on lipophilic metabolism of liver from cell-derived xenograft mouse model of Triple-Negative Breast Cancer.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Comprehensive biotransformation analysis of phenylalanine-tyrosine metabolism reveals alternative routes of metabolite clearance in nitisinone-treated alkaptonuria (Serum metabolomic analysis)
STUDY_SUMMARY
Background: Metabolomic analyses in alkaptonuria (AKU) have recently revealed alternative pathways in phenylalanine-tyrosine (phe-tyr) metabolism from biotransformation of homo-gentisic acid (HGA), the active molecule in this disease. The aim of this research was to study the phe-tyr metabolic pathway and whether the metabolites upstream of HGA, increased in nitisinone-treated patients, also undergo phase 1 and 2 biotransformation reactions. Methods: Metabolomic analyses were performed on serum and urine from patients partaking in the SONIA 2 phase 3 international randomised-controlled trial of nitisinone in AKU (EudraCT no. 2013-001633-41). Serum and urine samples were taken from the same patients at baseline (pre-nitisinone) then at 24 and 48 months on nitisinone treatment (patients N = 47 serum; 53 urine) or no treatment (patients N = 45 serum; 50 urine). Targeted feature extraction was per-formed to specifically mine data for the entire complement of theoretically predicted phase 1 and 2 biotransformation products derived from phenylalanine, tyrosine, 4-hydroxyphenylpyruvic acid and 4-hydroxyphenyllactic acid, in addition to phenylalanine-derived metabolites with known increases in phenylketonuria. Results: In total, we ob-served 13 phase 1 and 2 biotransformation products from phenylalanine through to HGA. Each of these products were observed in urine and two were detected in serum. The derivatives of the metabolites upstream of HGA were markedly increased in urine of nitisinone-treated patients (fold change 1.2-16.2) and increases in 12 of these compounds were directly proportional to the degree of nitisinone-induced hypertyrosinaemia (correlation coefficient with serum tyrosine = 0.2-0.7). Increases in the urinary phenylalanine metabolites were also observed across consecutive visits in the treated group. Conclusions: Nitisinone treatment results in marked increases in a wider network of phe-tyr metabolites than shown before. This network comprises alternative biotransformation products from the major metabolites of this pathway, produced by reactions including hydration (phase 1) and bioconjugation (phase 2) of acetyl, methyl, acetylcysteine, glucuronide, glycine and sulfate groups. We propose that these alternative routes of phe-tyr metabolism, predominantly in urine, minimise tyrosinaemia as well as phenylalanaemia.
INSTITUTE
University of Liverpool Institute of Life Course & Medical Sciences
DEPARTMENT
Department of Musculoskeletal & Ageing Science
LAST_NAME
Brendan
FIRST_NAME
Norman
ADDRESS
William Henry Duncan Building, 6 West Derby Street, Liverpool, UK. L7 8TX
Comprehensive biotransformation analysis of phenylalanine-tyrosine metabolism reveals alternative routes of metabolite clearance in nitisinone-treated alkaptonuria (Urine metabolomic analysis)
STUDY_SUMMARY
Background: Metabolomic analyses in alkaptonuria (AKU) have recently revealed alternative pathways in phenylalanine-tyrosine (phe-tyr) metabolism from biotransformation of homo-gentisic acid (HGA), the active molecule in this disease. The aim of this research was to study the phe-tyr metabolic pathway and whether the metabolites upstream of HGA, increased in nitisinone-treated patients, also undergo phase 1 and 2 biotransformation reactions. Methods: Metabolomic analyses were performed on serum and urine from patients partaking in the SONIA 2 phase 3 international randomised-controlled trial of nitisinone in AKU (EudraCT no. 2013-001633-41). Serum and urine samples were taken from the same patients at baseline (pre-nitisinone) then at 24 and 48 months on nitisinone treatment (patients N = 47 serum; 53 urine) or no treatment (patients N = 45 serum; 50 urine). Targeted feature extraction was per-formed to specifically mine data for the entire complement of theoretically predicted phase 1 and 2 biotransformation products derived from phenylalanine, tyrosine, 4-hydroxyphenylpyruvic acid and 4-hydroxyphenyllactic acid, in addition to phenylalanine-derived metabolites with known increases in phenylketonuria. Results: In total, we ob-served 13 phase 1 and 2 biotransformation products from phenylalanine through to HGA. Each of these products were observed in urine and two were detected in serum. The derivatives of the metabolites upstream of HGA were markedly increased in urine of nitisinone-treated patients (fold change 1.2-16.2) and increases in 12 of these compounds were directly proportional to the degree of nitisinone-induced hypertyrosinaemia (correlation coefficient with serum tyrosine = 0.2-0.7). Increases in the urinary phenylalanine metabolites were also observed across consecutive visits in the treated group. Conclusions: Nitisinone treatment results in marked increases in a wider network of phe-tyr metabolites than shown before. This network comprises alternative biotransformation products from the major metabolites of this pathway, produced by reactions including hydration (phase 1) and bioconjugation (phase 2) of acetyl, methyl, acetylcysteine, glucuronide, glycine and sulfate groups. We propose that these alternative routes of phe-tyr metabolism, predominantly in urine, minimise tyrosinaemia as well as phenylalanaemia.
INSTITUTE
University of Liverpool Institute of Life Course & Medical Sciences
DEPARTMENT
Department of Musculoskeletal & Ageing Science
LAST_NAME
Brendan
FIRST_NAME
Norman
ADDRESS
William Henry Duncan Building, 6 West Derby Street, Liverpool, UK. L7 8TX
NAD(P) deficiency plays an important role in the restraint-stress-induced depression in the rat model
STUDY_SUMMARY
The metabolic dysfunction or irreversible metabolic changes from stress may cause body vulnerability, potentially leading to the onset of psychiatric and non-psychiatric illnesses. Nevertheless, little is known about the biochemical events that cause depression due to stress. Our study employed open field test, plasma adrenocorticotropic hormone (ACTH) and corticosterone determination, serum biochemical analysis, quantitative PCR, immunoblotting, enzyme activity assay, and NMR-based metabolomics to analyze and identify the biochemical variations of body fluids (serum and urine) and tissues (brain, kidney, liver, lung, and spleen) in an acute restraint stress-induced rat model of depression. Our data suggested that the post-stress effects on biochemical alterations involved different biochemical pathways, including regulating the NAD(P) pool, glucose homeostasis, biosynthesis and degradation of heme, and uric acid production and metabolism. The urinary excretion of nicotinate and nicotinamide N-oxide increased significantly. Thus, we conclude that the depletion of NAD(P) precursors may occur in response to restraint stress. Our results show a close association between NAD(P) deficiency and post-stress metabolic dysfunction, which would provide a ground for developing recovery-promoting micronutrients in treating depression.
INSTITUTE
Anhui Science and Technology University
LAST_NAME
Li
FIRST_NAME
Jinquan
ADDRESS
No. 9, Donghua Road, Fengyang, Anhui Province, 233100, China
Characterizing the intrauterine environment via untargeted metabolomics profiling of maternal blood collected during pregnancy (Healthy Start Cohort)-Part 1
STUDY_TYPE
Prospective cohort study
STUDY_SUMMARY
Healthy Start is a prospective, pre-birth cohort study that recruited pregnant participants from outpatient prenatal clinics at the University of Colorado Hospital between 2009 and 2014. Eligible participants were 16 years or older with singleton pregnancies, no history of stillbirth or extremely preterm birth (<25 weeks of gestation) and no serious medical conditions, and had not yet completed 24 weeks of gestation at the time of enrollment. Mothers completed two study visits during pregnancy (median gestational ages 17 and 27 weeks). Mother-child pairs were assessed at birth for neonatal outcomes, and are currently still being followed through 8-10 years postpartum (NB: outcomes for this proposal are at birth). For the present project, we will use data from 1,297 mother-child pairs with adequate maternal plasma at 17 and 27 gestational weeks for untargeted metabolomics profiling. Please contact Wei Perng at wei.perng@cuanschutz.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Healthy Start is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
University of Colorado Anschutz Medical Campus
DEPARTMENT
Epidemiology; Lifecourse Epidemiology of Adiposity and Diabetes (LEAD) Center
LAST_NAME
Perng
FIRST_NAME
Wei
ADDRESS
1890 North Revere Court, Campus Box F426, Rm 1014, Aurora, CO 80010
Characterizing the intrauterine environment via untargeted metabolomics profiling of maternal blood collected during pregnancy (Healthy Start Cohort)-Part 2
STUDY_SUMMARY
Healthy Start is a prospective, pre-birth cohort study that recruited pregnant participants from outpatient prenatal clinics at the University of Colorado Hospital between 2009 and 2014. Eligible participants were 16 years or older with singleton pregnancies, no history of stillbirth or extremely preterm birth (<25 weeks of gestation) and no serious medical conditions, and had not yet completed 24 weeks of gestation at the time of enrollment. Mothers completed two study visits during pregnancy (median gestational ages 17 and 27 weeks). Mother-child pairs were assessed at birth for neonatal outcomes, and are currently still being followed through 8-10 years postpartum (NB: outcomes for this proposal are at birth). For the present project, we will use data from 1,297 mother-child pairs with adequate maternal plasma at 17 and 27 gestational weeks for untargeted metabolomics profiling. Please contact Wei Perng at wei.perng@cuanschutz.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Healthy Start is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
University of Colorado Anschutz Medical Campus
DEPARTMENT
Epidemiology; Lifecourse Epidemiology of Adiposity and Diabetes (LEAD) Center
LAST_NAME
Perng
FIRST_NAME
Wei
ADDRESS
1890 North Revere Court, Campus Box F426, Rm 1014, Aurora, CO 80010
Serum metabolomics profiling identifies new predictive biomarkers for disease severity in COVID-19 patients
STUDY_SUMMARY
Over the last three years, numerous groups have reported on different predictive models of disease severity in COVID-19 patients. However, almost all such models, which relied on serum biomarkers, clinical data or a combination of both, were subsequently deemed as cumbersome, inadequate and/or subject to bias. Moreover, although serum metabolomics profiling has shown significant differences among patients with different degrees of disease severity, the use of serum metabolomics profiling to identify prognostic biomarkers has, so far, been neglected. Herein, we sought to develop highly predictive models of disease severity by integrating routine laboratory findings and serum metabolomics profiling which identified several metabolites including K_4_aminophenol, acetaminophen and cytosine as potential biomarkers of disease severity in COVID-19 patients. Two models were subsequently developed and internally validated on the basis of ROC-AUC values. The predictive accuracy of the first model was 0.998 (95% CI: 0.992 to 1.000) with an optimal cut-off risk score of 4 biomarkers from among 8 linearly-related biomarkers (D-dimer, ferritin, neutrophil counts, Hp, sTfR, K_4_aminophenol, acetaminophen and cytosine). The predictive accuracy of the second model was 0.996 (95% CI: 0.989 to 1.000) with an optimal cut-off risk score of 3 biomarkers from among 6 biomarkers (D-dimer, ferritin, neutrophil counts, Hp, sTfR and cytosine). The two models are of high predictive power, need a small number of variables that can be acquired at minimal cost and effort, and can be applied independent of non-empirical clinical data. In conclusion, the metabolomics profiling data and the modeling work stemming from it, as presented here, could further explain the cause of COVID-19 disease prognosis and patient management.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Soares
FIRST_NAME
Nelson
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Integrated metabolomics and lipidomics study of patients with atopic dermatitis in response to dupilumab
STUDY_SUMMARY
Background: Atopic dermatitis (AD) is one of the most common chronic inflammatory skin diseases. Dupilumab, a monoclonal antibody that targets the interleukin (IL)-4 and IL-13 receptors, has been widely used in AD because of its efficacy. However, metabolic changes occurring in patients with AD in response to dupilumab remains unknown. In this study, we integrated metabolomics and lipidomics analyses with clinical data to explore potential metabolic alterations associated with dupilumab therapeutic efficacy. In addition, we investigate whether the development of treatment side effects was linked to the dysregulation of metabolic pathways. Methods: A total of 33 patients with AD were included in the current study, with serum samples collected before and after treatment with dupilumab. Comprehensive metabolomic and lipidomic analyses have previously been developed to identify serum metabolites (including lipids) that vary among treatment groups. An orthogonal partial least squares discriminant analysis model was established to screen for differential metabolites and metabolites with variable importance in projection > 1 and p < 0.05 were considered potential metabolic biomarkers. MetaboAnalyst 5.0 was used to identify related metabolic pathways. Patients were further classified into two groups, well responders (n = 19) and poor responders (n = 14), to identify differential metabolites between the two groups. Results: The results revealed significant changes in serum metabolites before and after 16 weeks of dupilumab treatment. Variations in the metabolic profile were more significant in the well-responder group than in the poor-responder group. Pathway enrichment analysis revealed that differential metabolites derived from the well-responder group were mainly involved in glycerophospholipid metabolism, valine, leucine and isoleucine biosynthesis, the citrate cycle, arachidonic acid metabolism, pyrimidine metabolism, and sphingolipid metabolism. Conclusion: Serum metabolic profiles of patients with AD varied significantly after treatment with dupilumab. Differential metabolites and their related metabolic pathways may provide clues for understanding the effects of dupilumab on patient metabolism.
INSTITUTE
Peking Union Medical College Hospital, Chinese Academy of Medical Sciences
Fitm2 is required for ER homeostasis and normal function of murine liver
STUDY_SUMMARY
The ER-resident protein fat-inducing transcript 2 (FIT2) catalyzes acyl-CoA cleavage in vitro and is required for endoplasmic reticulum (ER) homeostasis and normal lipid storage in cells. The gene encoding FIT2 is essential for the viability of mice and worms. Whether FIT2 acts as an acyl-CoA diphosphatase in vivo and how this activity affects liver, where the protein was discovered, are unknown. Here, we report that hepatocyte-specific Fitm2 knockout (FIT2-LKO) mice fed a chow diet exhibited elevated acyl-CoA levels, ER stress, and signs of liver injury. These mice also had more triglycerides in their livers than control littermates due, in part, to impaired secretion of triglyceride-rich lipoproteins and reduced capacity for fatty acid oxidation. Challenging FIT2-LKO mice with a high-fat diet worsened hepatic ER stress and liver injury, but unexpectedly reversed the steatosis phenotype, similar to what is observed in FIT2-deficient cells loaded with fatty acids. Our findings support the model that FIT2 acts as an acyl-CoA diphosphatase in vivo and is crucial for normal hepatocyte function and ER homeostasis in murine liver.
INSTITUTE
Harvard School of Public Health
LAST_NAME
Bond
FIRST_NAME
Laura
ADDRESS
665 Huntington Ave., Building 2, 3rd Floor, Room 311 | Boston, MA 02115
White-nose syndrome disrupts the splenic lipidome of little brown bats (Myotis lucifugus) at early disease stages
STUDY_SUMMARY
The fungal disease of bats, white-nose syndrome (WNS), is caused by the pathogen Pseudogymnoascus destructans (Pd). WNS-positive little brown bats (Myotis lucifugus) can exhibit an immune response during infection that include increases in cytokine and pro-inflammatory mediator gene levels. While bioactive lipid mediators (oxylipins) formed by enzymatic oxidation of polyunsaturated fatty acids (PUFAs) can contribute to this type of immune response, their role in WNS pathophysiology have not been investigated. Nonenzymatic conversion of PUFAs can also occur due to reactive oxygen species (ROS), however, these enantiomeric isomers will lack the same signaling properties. In this study, we performed a series of targeted lipidomic approaches on laboratory Pd-inoculated bats to assess changes in their splenic lipidome, including the formation of lipid mediators at early stages of WNS. Hepatic lipids previously identified were also resolved to a higher structural detail. We compared WNS-susceptible M. lucifugus to a WNS-resistant species, the big brown bat (Eptesicus fuscus). Altered splenic lipid levels were only observed in M. lucifugus, with lower total levels of glycerophospholipids (GPs) and free fatty acids (FFAs) in the Pd-inoculated group compared to the sham-inoculated group. Lower concentrations of splenic GPs were observed in lipid compounds containing 18:2 or saturated acyl chains. Differences in splenic FFAs included both omega-3 (including docosahexaenoic acid [DHA]) and omega-6 compounds. Increased levels of an enantiomeric monohydroxy DHA (4-hydroxydocosahexaenoic [HDoHE], 10-HDoHE, and 13-HDoHE) mixture, suggesting nonenzymatic formation, along with 6-keto-PGF1a were found. Changes in previously identified hepatic lipids were confined to omega-3 constituents. Together, these results suggest that increased oxidative stress, but not an inflammatory response, is occurring in bats at early stages of WNS that proceeds fat depletion.
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Metabolomics profiling of full extracts of bacterial culture supernatants.
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
Separate LC-MS runs of AhR-active and inactive bacterial culture supernatants were used to identify differentially expressed compounds in the active bacterial strains
Metabolomics profiling of the secondary fractions 4 of bacterial culture supernatants.
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
Separate LC-MS runs of AhR-active and inactive HPLC fractions of bacterial culture supernatants were used to identify differentially expressed compounds in the active bacterial strains.
Targeting malaria parasites with novel derivatives of azithromycin
STUDY_SUMMARY
The spread of artemisinin resistant Plasmodium falciparum parasites is of global concern and highlights the need to identify new antimalarials for future treatments. Azithromycin, a macrolide antibiotic used clinically against malaria, kills parasites via two mechanisms: ādelayed deathā by inhibiting the bacterium-like ribosomes of the apicoplast, and āquick-killingā that kills rapidly across the entire blood stage development. Here, 22 azithromycin analogues were explored for delayed death and quick-killing activities against P. falciparum (the most virulent human malaria) and P. knowlesi (a monkey parasite that frequently infects humans). Seventeen analogues showed improved quick-killing against both Plasmodium species, with up to 38 to 20-fold higher potency over azithromycin after less than 48 or 28 hours of treatment for P. falciparum and P. knowlesi, respectively. Lead analogues had limited activity against the related parasite Toxoplasma gondii and were >5-fold more selective against malaria than human cells. Quick-killing analogues maintained activity throughout the blood stage lifecycle including ring stages of P. falciparum parasites (<12 hrs treatment). Isopentenyl pyrophosphate supplemented parasites that lacked an apicoplast were equally sensitive to quick-killing analogues, confirming that the quick killing activity of these drugs was not directed at the apicoplast. Metabolomic profiling of parasites subjected to the lead analogue revealed a similar profile to chloroquine treatment, suggesting that the food-vacuole is a likely target of this drugs activity. The azithromycin analogues characterised in this study expanded the structural diversity over previously reported quick-killing compounds and provide new starting points to develop azithromycin analogues with quick-killing antimalarial activity.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
The objective of this study was to assess the impact of Salmonella bacteriophage treatment on microbiome in the ceca and serum of the broilers during the rearing period
INSTITUTE
La Agenzia nazionale per le nuove tecnologie, l'energia e lo sviluppo econĆ³mico sostenibile
Cecal Microbiota in Phage-treated Salmonella-infected broilers
STUDY_SUMMARY
Analysis of the gastrointestinal microbiota of Salmonella-infected broilers has been carried out after the application of encapsulated bacteriophages in the feed, during the production cycle under farm conditions
Serum metabolome in Phage-treated Salmonella-infected broilers
STUDY_SUMMARY
Analysis of the serum metabolome of Salmonella-infected broilers has been carried out after the application of encapsulated bacteriophages in the feed, during the production cycle under farm conditions
Dietary inclusion of nitrite-containing frankfurter exacerbates colorectal cancer pathology, increases oxidative stress, alters metabolism and causes gut dybiosis in APCmin mice
STUDY_SUMMARY
Colorectal cancer (CRC) is the second most prevelant malignancy in Europe and diet is an important modifiable risk factor. Processed meat consumption, including meats with preservative salts such as sodium nitrite, have been implicated in CRC pathogenesis. This study investigated how the CRC pathology and metabolic status of adenomatous polyposis coli (APC) multiple intestinal neoplasia (min) mice was perturbed following 8 weeks of pork meat consumption.Dietary inclusions (15%) of either nitrite-free pork, nitrite-free sausage or nitrite-containing sausage (frankfurter) were compared against a parallel control group (100% chow). Comprehensive studies investigated: gastrointestinal tract histology (tumours, aberant crypt foci (ACF) and mucin deplin foci (MDF), lipid peroxidation (urine and serum), faecal microbiota and serum metabolomics (599 metabolites).
INSTITUTE
Institute for Global Food Security
LAST_NAME
Pan
FIRST_NAME
Xiaobei
ADDRESS
19 Chlorine Gardens, Belfast, Antrim, BT9 5DL, United Kingdom
Differential requirements for mitochondrial electron transport chain components in the adult murine liver - Lactate/Pyruvate Tolerance Test
STUDY_SUMMARY
Hepatic Cox10 knockout mice were injected with a lactate/pyruvate (10:1) solution with 40% of the mixture labeled with [U-13C]. After 30 minutes, plasma and liver tissue was harvested. Metabolites were extracted, dried, and derivatized to form methoxime-TBDMS adducts.
INSTITUTE
The University of Texas Southwestern Medical Center at Dallas
Differential requirements for mitochondrial electron transport chain components in the adult murine liver - in vivo glucose tracing
STUDY_SUMMARY
Wild-type and knockout mice (Ndufa9 and Cox10) were implanted with a jugular vein catheter and infused with [U-13C] glucose for 3 hours. Plasma and liver tissue was collected and analyzed via GCMS.
INSTITUTE
The University of Texas Southwestern Medical Center at Dallas
Differential requirements for mitochondrial electron transport chain components in the adult murine liver - Untargeted Metabolomics (qTOF)
STUDY_SUMMARY
Wild-type and knockout mice (Ndufa9 and Cox10) livers were harvested on liquid nitrogen. Samples were crushed on liquid nitrogen, lysed in 80% ACN, and immediately injected onto the QE.
INSTITUTE
The University of Texas Southwestern Medical Center at Dallas
Mass spectroscopyābased proteomics and metabolomics analysis of tripleāpositive breast cancer cells treated with tamoxifen
STUDY_SUMMARY
HER2-enriched breast cancer with high levels of hormone receptor expression, known as "triple positive" breast cancer, may represent a new entity with a relatively favourable prognosis against which the combination of chemotherapy, HER-2 inhibition, and endocrine treatment may be considered overtreatment. We explored the effect of the anticancer drugs tamoxifen and trastuzumab, both separately and in combination, on the integrated proteomic and metabolic profile of "triple positive" breast cancer cells (BT-474). Method We employed ultra-high-performance liquid chromatography-quadrupole time of flight mass spectrometry using a Bruker timsTOF to investigate changes in BT-474 cell line treated with either tamoxifen, trastuzumab or a combination. Differentially abundant metabolites were identified using the Bruker Human Metabolome Database metabolite library and proteins using the Uniprot proteome for Homo sapiens using MetaboScape and MaxQuant, respectively, for identification and quantitation. Results A total of 77 proteins and 85 metabolites were found to significantly differ in abundance in BT-474 treated cells with tamoxifen 5 Ī¼M/and or trastuzumab 2.5 Ī¼M. Findings suggest that by targeting important cellular signalling pathways which regulate cell growth, apoptosis, proliferation, and chemoresistance, these medicines have a considerable anti-growth effect in BT-474 cells. Pathways enriched for dysregulation include RNA splicing, neutrophil degranulation and activation, cellular redox homeostasis, mitochondrial transmembrane transport, ferroptosis and necroptosis, ABC transporters and central carbon metabolism. Conclusion Our findings in protein and metabolite level research revealed that anti-cancer drug therapy had a significant impact on the key signalling pathways and molecular processes in triple positive BT-474 cell lines.
INSTITUTE
University of Sharjah
DEPARTMENT
Sharjah Institute for Medical Research
LABORATORY
Biomarker Discovery Group
LAST_NAME
Soares
FIRST_NAME
Nelson
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
Mass spectroscopyābased proteomics and metabolomics analysis of tripleāpositive breast cancer cells treated with trastuzumab
STUDY_SUMMARY
HER2-enriched breast cancer with high levels of hormone receptor expression, known as "triple positive" breast cancer, may represent a new entity with a relatively favourable prognosis against which the combination of chemotherapy, HER-2 inhibition, and endocrine treatment may be considered overtreatment. We explored the effect of the anticancer drugs tamoxifen and trastuzumab, both separately and in combination, on the integrated proteomic and metabolic profile of "triple positive" breast cancer cells (BT-474). Method We employed ultra-high-performance liquid chromatography-quadrupole time of flight mass spectrometry using a Bruker timsTOF to investigate changes in BT-474 cell line treated with either tamoxifen, trastuzumab or a combination. Differentially abundant metabolites were identified using the Bruker Human Metabolome Database metabolite library and proteins using the Uniprot proteome for Homo sapiens using MetaboScape and MaxQuant, respectively, for identification and quantitation. Results A total of 77 proteins and 85 metabolites were found to significantly differ in abundance in BT-474 treated cells with tamoxifen 5 Ī¼M/and or trastuzumab 2.5 Ī¼M. Findings suggest that by targeting important cellular signalling pathways which regulate cell growth, apoptosis, proliferation, and chemoresistance, these medicines have a considerable anti-growth effect in BT-474 cells. Pathways enriched for dysregulation include RNA splicing, neutrophil degranulation and activation, cellular redox homeostasis, mitochondrial transmembrane transport, ferroptosis and necroptosis, ABC transporters and central carbon metabolism. Conclusion Our findings in protein and metabolite level research revealed that anti-cancer drug therapy had a significant impact on the key signalling pathways and molecular processes in triple positive BT-474 cell lines.
INSTITUTE
University of Sharjah
DEPARTMENT
Sharjah Institute for Medical Research
LABORATORY
Biomarker Discovery Group
LAST_NAME
Soares
FIRST_NAME
Nelson
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
Mass spectroscopyābased proteomics and metabolomics analysis of tripleāpositive breast cancer cells treated with tamoxifen and trastuzumab
STUDY_SUMMARY
HER2-enriched breast cancer with high levels of hormone receptor expression, known as "triple positive" breast cancer, may represent a new entity with a relatively favourable prognosis against which the combination of chemotherapy, HER-2 inhibition, and endocrine treatment may be considered overtreatment. We explored the effect of the anticancer drugs tamoxifen and trastuzumab, both separately and in combination, on the integrated proteomic and metabolic profile of "triple positive" breast cancer cells (BT-474). Method We employed ultra-high-performance liquid chromatography-quadrupole time of flight mass spectrometry using a Bruker timsTOF to investigate changes in BT-474 cell line treated with either tamoxifen, trastuzumab or a combination. Differentially abundant metabolites were identified using the Bruker Human Metabolome Database metabolite library and proteins using the Uniprot proteome for Homo sapiens using MetaboScape and MaxQuant, respectively, for identification and quantitation. Results A total of 77 proteins and 85 metabolites were found to significantly differ in abundance in BT-474 treated cells with tamoxifen 5 Ī¼M/and or trastuzumab 2.5 Ī¼M. Findings suggest that by targeting important cellular signalling pathways which regulate cell growth, apoptosis, proliferation, and chemoresistance, these medicines have a considerable anti-growth effect in BT-474 cells. Pathways enriched for dysregulation include RNA splicing, neutrophil degranulation and activation, cellular redox homeostasis, mitochondrial transmembrane transport, ferroptosis and necroptosis, ABC transporters and central carbon metabolism. Conclusion Our findings in protein and metabolite level research revealed that anti-cancer drug therapy had a significant impact on the key signalling pathways and molecular processes in triple positive BT-474 cell lines.
INSTITUTE
University of Sharjah
DEPARTMENT
Sharjah Institute for Medical Research
LABORATORY
Biomarker Discovery Group
LAST_NAME
Soares
FIRST_NAME
Nelson
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
Untargeted Fecal Metabolomic Analyses Across an Industrialization Gradient Reveal Shared Metabolites and Impact of Industrialization on Fecal Microbiome-Metabolome Interactions
STUDY_SUMMARY
The metabolome is a central determinant of human phenotypes and includes the plethora of small molecules produced by host and microbiome, or taken up from exogenous sources. However, studies of the metabolome have so far focused predominantly on urban, industrialized populations. Through an untargeted metabolomic analysis of 90 fecal samples from human individuals from Africa and the Americasāthe birthplace and the last continental expansion of our species, respectivelyāwe characterized a shared human fecal metabolome. The majority of detected metabolite features were ubiquitous across populations, despite any geographic, dietary, or behavioral differences. Such shared metabolite features included hyocholic acid and cholesterol. However, any characterization of the shared human fecal metabolome is insufficient without exploring the influence of industrialization. Here, we show chemical differences along an industrialization gradient, where the degree of industrialization correlates with metabolomic changes. We identified differential metabolite features like amino acid-conjugated bile acids and urobilin as major metabolic correlates of these behavioral shifts. Additionally, co-analyses with over 5,000 publicly available human fecal samples and co-occurrence probability analyses with the gut microbiome highlight connections between the human fecal metabolome and gut microbiome. Our results indicate that industrialization significantly influences the human fecal metabolome, but diverse human lifestyles and behavior still maintain a shared human fecal metabolome. This study represents the first characterization of the shared human fecal metabolome through untargeted analyses of populations along an industrialization gradient.
13C NMR metabolomics: integrating J-resolved STOCSY and INADEQUATE
STUDY_SUMMARY
Robust annotation of metabolites remains a challenging task in metabolomics. This study introduces an approach that uses 13C homonuclear J-resolved experiment (JRES), statistical total correlation spectroscopy (STOCSY), and 2D incredible natural abundance double-quantum experiment (INADEQUATE) complementarily, to obtain robust molecular structure information based on 13C NMR with less experiment time. This approach was tested using the endometabolome from a model marine phytoplankton strain, varying the settings of incubation temperature, nutrient condition, and the presence of co-culturing bacteria.
Metabolomics study comparing SCAP KO and WT B cells
STUDY_TYPE
Purified mouse B cells, stimulated ex vivo
STUDY_SUMMARY
Splenic B cells isolated from SCAP+/+ CD19-Cre (WT) and SCAPfl/fl CD19-Cre (KO) mice were stimulated with LPS or anti-CD40 for 24 and 48 hours. Cells were then analyzed by metabolomics. Metabolomics reveals global metabolic changes in SCAP deficient B cells.
SETD1A regulates transcriptional pause release of heme biosynthesis genes in leukemia
STUDY_SUMMARY
Histone methyltransferase SETD1A is critical for acute myeloid leukemia (AML) cell survival, but the molecular mechanism driving SETD1A gene regulation remains elusive. To delineate the role of SETD1A, we utilize a protein degrader technology to induce rapid SETD1A degradation in AML cell lines. SETD1A degradation results in immediate downregulation of transcripts associated with DNA repair and heme biosynthesis pathways. CRISPR-based functional analyses and metabolomics reveal an essential role of SETD1A to maintain mitochondrial respiration in AML cells. These SETD1A targets are enriched in head-to-head (H2H) genes. SETD1A degradation disrupts a non-enzymatic SETD1A domain-dependent cyclin K function, increases the Ser5P RNA polymerase II (RNAP2) at TSS, and induces the promoter-proximal pausing of RNAP2 in a strand-specific manner. This study reveals a non-enzymatic role for SETD1A in transcriptional pause release and provides insight into the mechanism of RNAP2 pausing and its function in cancer.
INSTITUTE
Chiba University
LAST_NAME
Hoshii
FIRST_NAME
Takayuki
ADDRESS
1-8-1 Inohana Chuo-ku, Chiba, Chiba, 2608670, Japan
Exploration of age-dependent changes of phospholipid profiles in C. elegans depleted of tif-IA and ncl-1
STUDY_SUMMARY
Analysis of phosphatidylcholines, phosphatidylethanolamines, phosphatidylinositols, phosphatidylserines and phosphatidylglycerols in young (d2), middle age (d6) and old age (d12) C. elegans as well as animals with tif-IA or ncl-1 knockdown.
DJ1 KO was generated in BJsips iPSC and differentiated into midbrain organoids with the respective iPSC controls. The midbrain organoids were collected at day 40, 100 and 200 after differentiation.
Metabolome and transcriptome analysis of oral mucosa of HIV+ patients reveal a role for polyamine metabolic pathway in T cell dysfunction
STUDY_SUMMARY
Metabolic changes of immune cells contribute to both physiological and pathophysiological outcomes of immune reactions. How viruses alter the metabolic states of mucosal T cells and the precise mechanisms underlying the persisting immune dysfunction during chronic viral infections are key questions that have not been fully addressed. Here, by comparing transcriptome and salivary metabolome profiles of the uninfected individuals and people living with HIV (PLWH) on treatment, we found a role of polyamine metabolism in immune perturbations of the oral mucosa of HIV+ patients. Flow cytometry analysis confirmed the higher expression of ornithine decarboxylase (ODC-1) and eukaryotic translation initiation factor 5A (EIF5A), the polyamine metabolism intermediates in CD4+ T cells in PLWH. Mechanistic studies using an in vitro human tonsil organoid infection model revealed that HIV infection of activated T cells also resulted in increased polyamine synthesis, which was dependent on the activities of caspase-1, IL-1Ī², and ODC-1. HIV-1 also led to elevated dysfunctional regulatory T cells (TregDys) /Thelper 17 (Th17) cell ratios as well as heightened expression of ODC-1, EIF5A, and hypusinated EIF5A. Blockade of caspase-1, ODC-1, and EIF5A hypusination and not HIF-1āŗ or NLRP3 reversed the frequency of TregDys showing the direct impact of polyamine pathway in Treg dysfunction during HIV-1 infection. The addition of exogenous polyamines increased TregDys percentages independent of HIV-1 infection in vitro. Finally, oral mucosal TregDys/Th17 ratios and CD4 hyperactivation positively correlated with the increases in salivary putrescine levels, which were found to be elevated in the saliva of PLWH. Thus, by revealing the role of aberrantly increased polyamine synthesis during HIV infection, our study unveils a new mechanism by which chronic viral infections could drive distinct T cell effector programs and Treg dysfunction.
INSTITUTE
Case Western Reserve University
DEPARTMENT
Biological Sciences
LABORATORY
Pushpa Pandiyan
LAST_NAME
Pandiyan
FIRST_NAME
Pushpa
ADDRESS
Department of Biological Sciences, School of Dental Medicine, Case Western Reserve University, Cleveland, Ohio, 44106
Early-stage responses to Plasmodiophora brassicae at the metabolome levels in clubroot resistant and susceptible oilseed Brassica napus
STUDY_TYPE
Timecourse experiment
STUDY_SUMMARY
A total of 36 samples comprised two types of genotypes [CR (5 individuals pooled in each biological replicate) and CS (5 individuals pooled in each biological replicate)], two treatments (inoculated and uninoculated) and three biological replicates generated from three independent experiments and collected at 1-, 4-, and 7āDPI were used to extract primary and secondary metabolites and analyse the differences among the treatments.
Comprehensive characterization of putative genetic influences on plasma metabolome in a pediatric cohort
STUDY_SUMMARY
Background: The human exposome is composed of diverse metabolites and small chemical compounds originated from endogenous and exogenous sources, respectively. Genetic and environmental factors influence metabolite levels while the extent of genetic contributions across metabolic pathways is not yet known. Untargeted profiling of human metabolome using high-resolution mass spectrometry (HRMS) combined with genome-wide genotyping allows comprehensive identification of genetically influenced metabolites. As such previous studies of adults discovered and replicated genotype-metabotype associations. However, these associations have not been characterized in children. Results: We conducted the largest genome by metabolome-wide association study to date of children (N=441) using 619,688 common genetic variants and 14,342 features measured by HRMS. Narrow-sense heritability (h2) estimates of plasma metabolite concentrations using genomic relatedness matrix restricted maximum likelihood (GREML) method showed a bimodal distribution with high h2 (>0.8) for 15.9% of features and low h2 (<0.2) for most of features (62.0%). The features with high h2 were enriched for amino acid and nucleic acid metabolism while carbohydrate and lipid concentrations showed low h2. For each feature, a metabolite quantitative trait locus (mQTL) analysis was performed to identify genetic variants that were potentially associated with plasma levels. Fifty-four associations among 29 features and 43 genetic variants were identified at a genome-wide significance threshold p < 3.5x10-12 (= 5 x 10-8/14,342 features). Previously reported associations such as UGT1A1 and bilirubin; PYROXD2 and methyl lysine; ACADS and butyrylcarnitine were successfully replicated in our pediatric cohort. We found potential candidates for novel associations including CSMD1 and a monostearyl alcohol triglyceride; CALN1 and a triglyceride; RBFOX1 and dimethylarginine. A gene-level enrichment analysis using MAGMA revealed highly interconnected modules for ADP biosynthesis, sterol synthesis, and long-chain fatty acid transport in the gene-feature network. Conclusion: Comprehensive profiling of plasma metabolome across age groups combined with genome-wide genotyping revealed a wide range of genetic influence on diverse chemical species and metabolic pathways. The developmental trajectory of a biological system is shaped by gene-environment interaction especially in early life. Therefore, continuous efforts on generating metabolomics data in diverse human tissue types across age groups are required to understand gene-environment interaction toward healthy aging trajectories.
Plasma metabolomic profiling of individuals with autism spectrum disorder and their family members.
STUDY_SUMMARY
Autism spectrum disorder (ASD) is a common neurodevelopmental condition affecting 2.3% of 8-year-old children and is attributable to polygenic risks in most cases. Gene discovery studies catalogued >1000 genes with de novo, rare and common genetic variants that are likely associated with ASD; however, the candidate genes are rarely translated to diagnostic and treatment biomarkers. As such no pharmacological treatment option is available for targeting core symptoms. Neural circuits involved in verbal/nonverbal communications and social interaction are likely changed, which may be caused by an excitatory-inhibitory (E-I) imbalance in individuals with ASD. To date, clinical trials targeting excitatory glutamatergic or inhibitory GABAergic receptors showed mixed results. These early clinical trials highlight the unmet need of biomarkers for target populations and outcome indicators. We investigated whether plasma biomarkers would be associated with genetic risk factors and core symptoms of ASD. Plasma samples were collected for metabolomics profiling from the Autism Genetics Resource Exchange (AGRE). Detailed phenotype information is available at NIMH Data Archive (Collection ID: 4214) and can be accessed using NDAR GUID for the individuals.
Phospholipase D3 impact on the endolysosomal lipidome
STUDY_SUMMARY
Neurons rely on the endo-lysosomal network for the maintenance of lipid turnover, removal of dysfunctional organelles and the recycling of proteins. These mechanisms appear to go awry in late-onset Alzheimerās disease (LOAD). Interestingly, GWA-studies identified risk genes for LOAD linked to endocytic transport regulation (BIN1-CD2AP-PICALM-RIN3-SORL1) and lysosomes (PLD3). Phospholipase D3, also known as PLD3, is a single-pass type II membrane protein that is majorly localized to lysosomes, making it one of the few (or only) risk factors that potentially links lysosomal dysfunction directly to LOAD initiation and progression. CRISPR/Cas9 gene editing was used to generate PLD3 knockout SH-SY5Y cells that were subsequently stably rescued with wild-type PLD3 and coding-variants (M6R & V232M). All cell lines were evaluated for morphological and functional alterations of the endolysosomal compartment, including lipid profiling of endolysosomes magnetically isolated from the different cell lines, as previously described (DOI: 10.1016/j.xpro.2020.100122). A prior isolation step has the unique advantage that it provides spatial resolution to the identified dysregulated networks or compositions. We observe a marked accumulation of storage lipids in endolysosomal isolates; chiefly attributed to cholesterol ester (CE) accretion. A significantly lowered monoacylglycerol level and increased phosphatidylinositol level point to an affected transport/sorting (vesicle/tubule formation).
Pathogenic Auxilin mutations affect lipids that are critical to Synaptojanin function
STUDY_SUMMARY
Mass spectrometry was performed on heads of 15DO flies homogenized in 100 Āµl D-PBS (Dulbeccoās phosphate-buffered saline without Mg2+ and Ca2+) by Lipotype. Fifteen fly heads were pooled from three independent crosses for each analysis, and mass spectrometry was performed on n=7 for wild-type, nā„4 analyses for dAuxWT/WT, dAuxRG/RG, dAuxWT/F956x and dAuxRG/F956x and n=3 for dAuxWT/F956x and dAuxRG/F956x overexpressing Synj.
INSTITUTE
VIB-KU Leuven
LAST_NAME
Jacquemyn
FIRST_NAME
Julie
ADDRESS
Medical Sciences Building 7-25, Edmonton, Alberta, T6H 0L2, Canada
Characterizing the intrauterine environment via untargeted metabolomics profiling of maternal blood collected during pregnancy (Project Viva)
STUDY_TYPE
Prospective cohort study
STUDY_SUMMARY
Project Viva: Pregnant women were enrolled in Project Viva between 1999 and 2002 at their first prenatal visit at one of 8 obstetric clinics of Atrius Harvard Vanguard Medical Associates, a multispecialty group practice in eastern Massachusetts. Eligible mothers were fluent in English, had singleton gestations, were <22 weeks gestation, and had no plans to move away from the study area. Research staff performed in-person study visits with participating mothers in the first (median gestational age 9.9 weeks) and second (median gestational age 28.1 weeks) trimesters of pregnancy, and with mothers and children during the first few days after delivery, during infancy (median age 6.3 months), in early childhood (median age 3.3 years), mid-childhood (median age 7.7 years), and adolescence (median age13 years). In this analysis, we will use data from 188 mother-child pairs in Project Viva with available information on prenatal PFAS concentrations, available umbilical cord serum samples at delivery, and outcomes of interest. Please contact MollyAn Killingbeck at mollyan.killingbeck@point32health.org for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Project Viva is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
The GNHS was an onging community-based prospective cohort study.In GNHS,a total of 4,048 Chinese participants aged 40-75 years living in the urban area of Guangzhou, China, for at least 5 years were recruited between 2008-2013. Participants were followed up every three years. During the period 2008-2019, blood samples were collected at up to three time points. In 2014-2019 follow-up visits, stool samples were collected at up to two time points.The information of diet, sociodemographic factors, lifestyle factors, and medications, etc. was collected at the baseline and follow-up visits. The aim of this study was to investigate the relationships among human nutrition, enviromental factors, gut microbiome and human diseases.
INSTITUTE
Westlake University
LAST_NAME
Ju-Sheng
FIRST_NAME
Zheng
ADDRESS
Westlake University,18 Shilongshan Rd, Cloud Town, Hangzhou, China
Interplay Between Cruciferous Vegetables and the Gut Microbiome: A Multi-Omic Approach
STUDY_TYPE
Ex Vivo Fecal Incubation
STUDY_SUMMARY
Cruciferous vegetable consumption has been associated with a decreased risk of multiple types of cancers, thus presenting a cost-effective, non-pharmacological approach to cancer prevention through dietary intervention. Broccoli sprouts and Brussels sprouts are among the leading cruciferous vegetables under study and contain some similar and some distinct phytochemicals which can activate different, but complementary, mechanisms to promote health. While the cancer-preventative effects of cruciferous vegetables are typically attributed to glucosinolates and their metabolic products, isothiocyanates and indoles, other components of cruciferous vegetables could play a synergistic role in conferring cancer-protective and health promoting effects. Additionally, metabolism of phytochemicals from cruciferous vegetables by the gut microbiome could further lead to the production, inactivation, or clearance of bioactive dietary components. The gut microbiome is essential to the production of bioactive compounds from various food sources. For example, with soy isoflavones and pomegranate urolithins, the presence or absence of specific microbial taxa directly dictates which metabolites are produced (resulting in a metabotype). A similar paradigm could be extended to cruciferous vegetables in which the gut microbiome may play an important role in driving inter-individual metabolism of glucosinolates and isothiocyanates. We recently reported (Bouranis et. al, 2021, Nutrients) that the gut microbiome composition can influence production of glucosinolate-derived nitriles from cruciferous vegetables, showing that the presence or absence of specific microbes can influence the abundance of a single metabolite. Thus, we sought to take an untargeted approach to investigate other phytochemicals from cruciferous vegetables which the gut microbiome could play a role in generating. To investigate plant- and microbe-derived metabolites of cruciferous vegetable digestion and capture information about the microbiome, we utilized an ex vivo fecal incubation system. Broccoli sprouts and Brussels sprouts were in vitro digested using an oral, gastric, and intestinal phase. For fecal bacterial cultivation a 20% fecal slurry (w/v) was made from fecal material from 10 healthy volunteers (6 female, and 4 male, age 17-51, Lee Biosolutions) and sterile PBS (0.1 M pH 7). 500 ĀµL of fecal slurry was mixed with 10 mL of Brain Heart Infusion Broth (BHI) with hemin and vitamin K, per the manufacturerās recommendation, and either 500 Āµl of filter sterilized in vitro digested broccoli sprouts (Broc), 500 ĀµL of filter sterilized in vitro digested Brussels sprouts (Brus), 500 ĀµL of Broc and 500 ĀµL of Brus were added (Combo) or a negative control in vitro digestion (NC). NC contained reverse osmosis water, equivalent in volume to the water content of broccoli sprouts and underwent the same in vitro digestion procedure as described above with the same enzymes, chemicals and equipment. Broc and Brus digests were scaled to be equivalent in concentration to a human consuming Ā½ cup of broccoli or Brussels sprouts, or in the case of the combination, Ā½ cup of broccoli sprouts and Ā½ cup of Brussels sprouts. This combination was included as Broc and Brus contain many similar but also some distinct phytochemicals and thus by combining the vegetables we increased the dose and broadened the range of phytochemicals from cruciferous vegetables which can be achieved in the kitchen as a mixed vegetable dish. Fecal cultures were incubated at 37Ā°C for 24 h in anaerobic conditions.
INSTITUTE
Oregon State University
DEPARTMENT
Linus Pauling Institute
LABORATORY
Emily Ho
LAST_NAME
Bouranis
FIRST_NAME
John
ADDRESS
371 Linus Pauling Science Center, 2900 SW Campus Way, Corvallis, OR, 97331, USA
Myriocin rescue of serine-associated hepatic lipid diversity
STUDY_SUMMARY
We analyzed hepatic polar and lipid metabolites in mice (C57BL/6J) in response to two dietary interventions. The first was: 1) low fat (LFD), 2) serine/glycine-free LFD (-SG LFD), high fat (HFD), and serine/glycine-free HFD (-SG HFD). And the second was: 1) low fat (LFD), high fat (HFD), serine/glycine-free HFD (-SG HFD), and serine/glycine-free HFD (-SG HFD) with myriocin (0.3 mg/kg every other day) treatment. The goal of the study was to determine the impact of dietary serine/glycine restriction and myriocin treatment on the hepatic lipidome.
INSTITUTE
Salk Institute for Biological Studies
DEPARTMENT
Molecular and Cell Biology Laboratory
LABORATORY
Metallo Lab
LAST_NAME
Handzlik
FIRST_NAME
Michal
ADDRESS
10010 N Torrey Pines Rd, La Jolla, California, 92037, USA
Metabolomic analysis of colorectal cancer cells using mass spectrometry
STUDY_SUMMARY
Colorectal cancer (CRC) is one of the most prevalent tumors, with a high mortality rate. Nearly half of CRC patients develop metastasis, which accounts for as many as 90% of CRC-related deaths. In the metastasis process, cancer cells exhibit altered dependency on specific metabolic pathways and some of the metabolites discovered might be useful as potential diagnostic biomarkers. To identify metabolic pathway dependencies in CRC metastasis, mass spectrometry-based untargeted metabolomic analysis was performed in two pairs of CRC cell lines with different metastatic abilities. Each pair of cell lines was comprised of primary and metastatic colorectal cancer cell lines (SW480 vs. SW620; HT-29 vs. COLO 205). Relative levels of intracellular metabolites distinguished high-metastatic CRC cells from low-metastatic CRC cells.
INSTITUTE
Nanjing Medical University
LAST_NAME
Zhang
FIRST_NAME
Wenjun
ADDRESS
101Longmian Avenue, Jiangning District, Nanjing 211166, P.R. China
The gut microbiome has sexually dimorphic effects on bone tissue energy metabolism and multiscale bone quality in C57BL/6J mice
STUDY_SUMMARY
Male and female germ-free and conventional were euthanized at 20 weeks of age, and various tissues were obtained to assess differences in bone properties associated with microbiome status and sex. For metabolomic analyses, the humeri was obtained, bone marrow was flushed using PBS, and subchondral bone metabolites were extracted using a methanol:acetone precipitation approach. Next, samples were injected, ionized, and data was acquired using LC-MS. Data was processed and converted using MSConvert and XCMS. Following this step, metabolic phenotypes of mice that differ by microbiome status and sex were investigated using MetaboAnalyst. Differences in metabolism were related to lipid, energy, and amino acid metabolism. Specifically, females, both conventional and germ-free, had dysregulated lipid metabolism, whereas, males, both conventional and germ-free had dysregulated amino acid metabolism. Other differences detected between male and female mice that differ by germ-free status corresponded to differences in bone strength, bone marrow adiposity, and osteocyte density.
The impact of myriocin and dietary serine restriction on paw skin sphingolipid diversity
STUDY_SUMMARY
We analyzed paw skin polar and lipid metabolites in mice (C57BL/6J) in response to two dietary interventions. The first was: 1) low fat (LFD), 2) serine/glycine-free LFD (-SG LFD), high fat (HFD), and serine/glycine-free HFD (-SG HFD). And the second was: 1) low fat (LFD), high fat (HFD), serine/glycine-free HFD (-SG HFD), and serine/glycine-free HFD (-SG HFD) with myriocin (0.3 mg/kg every other day) treatment. The goal of the study was to determine the impact of dietary serine/glycine restriction and myriocin treatment on the hepatic lipidome.
INSTITUTE
Salk Institute for Biological Studies
DEPARTMENT
Molecular and Cell Biology Laboratory
LABORATORY
Metallo Lab
LAST_NAME
Handzlik
FIRST_NAME
Michal
ADDRESS
10010 N Torrey Pines Rd, La Jolla, California, 92037, USA
Stress-Induced Mucosal Layer Disruption Drives Gut Dysbiosis and Depressive-like Behaviors
STUDY_SUMMARY
Depression is a common mental health condition with a large social and economic impact. While depression etiology is multifactorial, chronic stress is a well-accepted contributor to disease onset. In addition, depression is associated with altered gut microbial signatures that can be replicated in animal models. While targeted restoration of the microbiome has been shown to reduce depressive-like behaviors in mice, the complexity and diversity of the human microbiome has complicated therapeutic intervention in patients. To circumvent these limitations, there is a critical need for identifying pathways responsible for microbiome dysbiosis. Here, for the first time, we identify the changes in host physiology that induce microbiome dysbiosis. Specifically, we show that a component of mucosal layer, the transmembrane protein mucin 13, can regulate microbiome composition. Using a model of chronic stress to induce behavioral and microbial changes in mice, we show a significant reduction in mucin 13 expression across the intestines that occurs independently of the microbiome. Furthermore, deleting Muc13 leads to gut dysbiosis, and baseline behavioral changes normally observed after stress exposure. Together, these results validate the hypothesis that mucosal layer disruption is an initiating event in stress-induced dysbiosis and offer mucin 13 as a potential new therapeutic target for microbiome dysbiosis in stress-induced depression. For the first time, our data provide an upstream and conserved target for treating microbiome dysbiosis, a result with sweeping implications for diseases presenting with microbial alterations.
INSTITUTE
University of Virginia
LAST_NAME
Rivet-Noor
FIRST_NAME
Courtney
ADDRESS
409 Lane Road, Charlottsville, Virginia, 22903, USA
Lipidomics study of the cells defective in peroxisome division
STUDY_SUMMARY
Proliferation of peroxisomes is accompanied by the growth and division of pre-existing peroxisomes. Pex11Ī² induces the elongation of peroxisome membrane and then dynamin-like GTPase, DLP1, elicits the peroxisomal division. Nucleoside diphosphate kinase 3 (NME3) generates GTP for the DLP1 activity. Deficiencies of either of the factors induce abnormal morphology of peroxisomes. In this study, we assessed the phospholipid compositions in cells lacking each of the different division factors. In the fibroblasts from the patients deficient in DLP1, NME3, or Pex11Ī², docosahexaenoic acid (DHA, C22:6)-containing phospholipids were found to be decreased. Conversely, the levels of several fatty acids such as arachidonic acid (AA, C20:4) and oleic acid (C18:1) were elevated. Mouse embryonic fibroblasts from Drp1- and Pex11Ī²-knockout mice also showed a decrease in the levels of phospholipids containing DHA and AA.
Impact of Visceral Leishmaniasis on Local Organ Metabolism in Hamsters
STUDY_SUMMARY
Leishmania is an intracellular parasite with different species pathogenic to humans and causing the disease leishmaniasis. Leishmania donovani causes visceral leishmaniasis (VL) that manifests as hepatosplenomegaly, fever, pancytopenia and hypergammaglobulinemia. If left without treatment, VL can cause death, especially in immunocompromised people. Current treatments have often significant adverse effects, and resistance has been reported in some countries. Determining the metabolites perturbed during VL can lead us to find new treatments targeting disease pathogenesis. We therefore compared metabolic perturbation between L. donovani-infected and uninfected hamsters across organs (spleen, liver, and gut). Metabolites were extracted, analyzed by liquid chromatography-mass spectrometry, and processed with MZmine and molecular networking to annotate metabolites. We found few metabolites commonly impacted by infection across all three sites, including glycerophospholipids, ceramides, acylcarnitines, peptides, purines and amino acids. In accordance with VL symptoms and parasite tropism, we found a greater overlap of perturbed metabolites between spleen and liver compared to spleen and gut, or liver and gut. Targeting pathways related to these metabolite families would be the next focus that can lead us to find more effective treatments for VL.
Comparative metabolite profiling of glycolytic and sulfoglycolytic E. coli
STUDY_SUMMARY
Glycolytic E. coli (E. coli grown on glucose as a sole carbon source) and sulfoglycolytic E. coli (E. coli grown on the sulfosugar sulfoquinovose as a sole carbon source) were grown to mid-log phase in M9 minimal medium. Samples were harvested at mid-log phase and analysed using GC-EI-QqQ-MS.
INSTITUTE
University of Melbourne
LAST_NAME
Williams
FIRST_NAME
Spencer
ADDRESS
05, 532, David Penington Building, Parkville, 3010, VIC, Australia
Proteins and RNA are able to phase separate from the aqueous cellular environment to form sub-cellular compartments called condensates. This process results in a protein-RNA mixture that is chemically distinct from the surrounding aqueous phase. Here we use mass spectrometry to characterize the metabolomes of condensates. To test this, we prepared mixtures of phase-separated proteins and cellular metabolites and identified metabolites enriched in the condensate phase. These proteins included SARS-CoV-2 nucleocapsid, as well as low complexity domains of MED1 and HNRNPA1.
Identify putative volatile biomarkers of Valley fever using a murine lung infection model
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
Coccidioides immitis and Coccidioides posadasii are soil-dwelling fungi of arid regions in North and South America that are responsible for Valley fever (coccidioidomycosis). Forty percent of patients with Valley fever exhibit symptoms ranging from mild, self-limiting respiratory infections, to severe, life-threatening pneumonia that requires treatment. Misdiagnosis as bacterial pneumonia commonly occurs in symptomatic Valley fever cases, resulting in inappropriate treatment with antibiotics, increased medical costs, and delay in diagnosis. In this study, we explored the feasibility of developing breath-based diagnostics for Valley fever using a murine lung infection model. To investigate potential volatile biomarkers of Valley fever that arise from host-pathogen interactions, we infected C57BL/6J mice with C. immitis RS and C. posadasii Silveira via intranasal inoculation. We collected bronchoalveolar lavage fluid (BALF) for cytokine profiling and for untargeted volatile metabolomics via solid phase microextraction (SPME) and two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GCĆGC-TOFMS). We identified 36 volatile organic compounds (VOCs) that were significantly correlated to cytokine abundances and clustered mice by disease severity. These 36 VOCs were also able to separate mice with a moderate to high disease severity by infection strain. The data presented here show that Coccidioides and/or the host produce volatile metabolites that may yield biomarkers for a Valley fever breath test that can detect Coccidioidal infection and provide clinically relevant information on disease severity.
Characterizing the intrauterine environment via untargeted metabolomics profiling of maternal blood collected during pregnancy (Atlanta ECHO Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The Atlanta ECHO Cohort is comprised of African American mother-child dyads who reside in the greater metropolitan Atlanta area. Enrollment begins during pregnancy from prenatal care clinics affiliated with two metro hospital systems. During pregnancy, data and biospecimens are obtained during an initial study visit (between 8-14 weeks gestation) and during a second study visit (between 24-30 weeks gestation). In the perinatal period, we collect the residual newborn blood spot (obtained from the neonate during the delivery hospitalization for state metabolic screening). Details of pregnancy complications, birth, and neonatal outcomes are ascertained via medical record abstraction. Children are then followed annually via in-person clinical assessments and maternal questionnaire data. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The Atlanta ECHO Cohort is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
Emory University
DEPARTMENT
School of Medicine, Department of Gynecology & Obstetrics; School of Public Health, Department of Environmental Health
LAST_NAME
Dunlop
FIRST_NAME
Anne
ADDRESS
101 Woodruff Circle, Rm 4303 Woodruff Memorial Building, Atlanta, GA 30322
Proteins and RNA are able to phase separate from the aqueous cellular environment to form sub-cellular compartments called condensates. This process results in a protein-RNA mixture that is chemically distinct from the surrounding aqueous phase. Here we use mass spectrometry to characterize the metabolomes of condensates. To test this, we prepared mixtures of phase-separated proteins and cellular metabolites and identified metabolites enriched in the condensate phase. These proteins included SARS-CoV-2 nucleocapsid, as well as low complexity domains of MED1 and HNRNPA1.
Proteins and RNA are able to phase separate from the aqueous cellular environment to form sub-cellular compartments called condensates. This process results in a protein-RNA mixture that is chemically distinct from the surrounding aqueous phase. Here we use mass spectrometry to characterize the metabolomes of condensates. To test this, we prepared mixtures of phase-separated proteins and cellular metabolites and identified metabolites enriched in the condensate phase. These proteins included SARS-CoV-2 nucleocapsid, as well as low complexity domains of MED1 and HNRNPA1.
Combination of TP-252 and Naproxen elicit tumor protective Eicosanoid changes.
STUDY_SUMMARY
Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. Patients with the genetic disorder Familial Adenomatous Polyposis (FAP) develop hundreds to thousands of polyps that unless removed by prophylactic colectomy will progress to CRC at an early age. Non-steroidal anti-inflammatory drugs (NSAIDs) and -3 marine polyunsaturated fatty acids (PUFA), such as eicosapentaenoic acid (EPA), have been evaluated for their chemopreventive potential in delaying the onset of CRC in high-risk patients. In this study, we determined whether the NSAID, naproxen, alone or in combination with a chemically-stable form of EPA (TP-252), affects tumor formation in the ApcPirc rat model. When compared to control diet, animals fed naproxen or HD TP-252 had 66%, and 82% fewer tumors respectively. However, when fed a combination of naproxen and HD TP-252, animals exhibited a 95% reduction in tumor formation and a 98% reduction in tumor volume, respectively. To elucidate potential mechanisms of tumor protection, a comprehensive, targeted lipidomic analysis was performed on colonic mucosa to determine changes in eicosanoid metabolism. Animals receiving TP-252 alone or in combination with naproxen had significantly reduced mucosal levels of pro-inflammatory -6 eicosanoids (PGE2, 5-HETE, and 14,15-DiHETrE), along with a simultaneous increase in anti-inflammatory EPA-derived -3 eicosanoids. Our colonic mucosal lipidomic analysis also uncovered several potential pharmacodynamic (PD) lipid biomarkers, including resolvin E2, 9-HEPE, 12-HEPE and 18-HEPE, that were increased in both the tissue and plasma of rats receiving TP-252 and were significantly correlated with tumor protection. Further studies with this drug combination should be focused on dose optimization and the role of EPA-derived lipid mediators in CRC initiation and progression.
Stool short chain fatty acid (SCFA) levels in peanut allergy
STUDY_SUMMARY
Prior evidence supports differential levels of short chain fatty acids in the stool of human beings with allergy and murine models of allergy. Here we performed a targeted study of selected short chain fatty acid levels in stool samples collected from children with allergy risk factors. Sample processing included homogenization of stool samples, inclusion of internal standards, and derivitization for liquid chromatography tandem mass spectrometry.
INSTITUTE
Icahn School of Medicine at Mount Sinai
LAST_NAME
Bunyavanich
FIRST_NAME
Supinda
ADDRESS
1 Gustave L. Levy Pl, New York, NY 10029
EMAIL
supinda.bunyavanich@mssm.edu
PHONE
Stool metabolite levels in individuals with peanut allergy were measured.
Isolated murine skeletal muscles utilize pyruvate over glucose for oxidation-Part 1
STUDY_TYPE
Study of the different substrate by isolated skeletal muscle at room temperature via C-13 isotopomer analysis
STUDY_SUMMARY
Preclinical studies of muscle contractile function often employ ex vivo preparations of the soleus and/or extensor digitorum longus (EDL) muscles which are relatively easy to prepare and represent slow and fast fiber properties, respectively. Therefore, the current study sought to examine the utility of this preparation for understanding the metabolic fuel utilization in isolated resting mouse muscles at room temperature. 13C-labeling in both muscle types was performed using three fuels: glucose, pyruvate, and acetate, followed by NMR-based metabolomics analyses. Incubating 13C-labeled substrates in the isolated skeletal muscles makes it possible to examine TCA cycle flux and substrate selection by these muscles.
INSTITUTE
University of Florida
DEPARTMENT
Applied Physiology and Kinesiology
LABORATORY
Rm 42 and Rm 43
LAST_NAME
Khattri
FIRST_NAME
Ram
ADDRESS
1864 Stadium RD, Gainesville, FL, 32611, USA
EMAIL
rbk11@ufl.edu
PHONE
3307856045
NUM_GROUPS
4
TOTAL_SUBJECTS
18
STUDY_COMMENTS
Southeastern Center for Integrated Metabolomics (SECIM) (ERB), NIH AR U54 AR052646 (Physiological Assessment Core, ERB), and Wellstone Muscular Dystrophy Cooperative Research Center Grant (NIAMS: U54AR052646/P50 AR052646). The AMRIS Facility is supported by the National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida.
Effect of hypoxia and reoxygenation on the juvenile hooded seal brain lipidome
STUDY_SUMMARY
Brain samples from juvenile hooded seals (Cystophora cristata) were subjected to hypoxia and reoxygenation in vitro and lipid composition was compared.
The autophagy-flux-promoting protein TFG (Trk-fused gene) is up-regulated during B cell differentiation into plasma cells and supports survival of CH12 B cells. We hypothesized that quantitative proteomics analysis of CH12tfgKO B cells with intact or blocked autophagy-lysosome flux (via NH4Cl) will identify mechanisms of TFG-dependent autophagy, plasma cell biology and B cell survival. Analysis of CH12WT B cells in the presence of NH4Cl will identify proteins whose presence is continuously regulated by lysosomes independent of TFG. We determined hundreds of proteins to be controlled by TFG and/or NH4Cl. Notably, NH4Cl treatment alone increased the abundance of a cluster of cytosolic and mitochondrial translational proteins while it also reduced a number of proteins. Within the B cell relevant protein pool, BCL10 was reduced, while JCHAIN was increased in CH12tfgKO B cells. Furthermore, TFG regulated the abundance of transcription factors, such as JUNB, metabolic enzymes, such as the short-chain fatty acid activating enzyme ACOT9 or the glycolytic enzyme ALDOC. Gene ontology enrichment analysis revealed that TFG-regulated proteins localized to mitochondria and membrane-bounded organelles. Due to these findings we performed shotgun lipidomics of glycerophospholipids, uncovering that a particular phosphatidylethanolamine (PE) species, PE 32:0, which lipidates LC3 most efficiently, was less abundant while phosphatidylglycerol (PG) was more abundant in CH12tfgKO B cells. In line with the role of PG as precursor for Cardiolipin (CL), the CL content was higher in CH12tfgKO B cells and addition of PG liposomes to B cells increased the amount of CL. We propose a role for TFG in B cell activation and plasma cell biology via regulation of proteins involved in germinal center and plasma cell development, such as BCL10 or JCHAIN, as well as in lipid homeostasis, mitochondria and metabolism.
INSTITUTE
University of Cologne
DEPARTMENT
Faculty of Medicine and University Hospital of Cologne, Cluster of Excellence Cellular Stress Responses in Aging-associated Diseases (CECAD)
UCP2-dependent redox-sensing in POMC neurons regulates feeding
STUDY_SUMMARY
Paradoxically, glucose, the primary driver of satiety, activates a small population of anorexigenic POMC neurons. Here we show that lactate levels in the circulation and in the cerebrospinal fluid are elevated in fed state and addition of lactate to glucose activates the majority of POMC neurons while increasing cytosolic NADH generation, mitochondrial respiration and extracellular pyruvate levels. Inhibition of lactate dehydrogenases diminishes mitochondrial respiration, NADH production, and POMC neuronal activity. However, inhibition of the mitochondrial pyruvate carrier has no effect. POMC-specific downregulation of Ucp2 (Ucp2PomcKO), a molecule regulated by fatty acid metabolism and shown to play a role as transporter in the malate-aspartate shuttle, abolishes lactate- and glucose-sensing of POMC neurons. Ucp2PomcKO mice have impaired glucose metabolism and are prone to obesity on a high fat diet. Altogether, our data show that lactate through redox signaling and blocking mitochondrial glucose utilization activates POMC neurons to regulate feeding and glucose metabolism.
INSTITUTE
Columbia University
LAST_NAME
Diano
FIRST_NAME
Sabrina
ADDRESS
1150 St. Nicholas Avenue Russ Berrie Medical Science Pavilion Rm 405 New York, NY, 10032
[U-13C]glucose tracing in naĆÆve vs. activated CD8+ T cells
STUDY_SUMMARY
NaĆÆve vs. 24 hr plate-bound anti-CD3 and soluble anti-CD28 activated CD8+ T cells were pulsed with [U-13C]glucose. Intracellular glucose-derived glutamate levels were quantified using MS.
[U-13C]glucose tracing in NT, AOA or EGCG treated activated CD8+ T cells
STUDY_SUMMARY
CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28 for 24 hours without (NT), or with AOA (250uM) or EGCG (500uM) treatment. CD8+ T cells were pulsed with [U-13C]glucose. Intracellular glucose-derived glutamate levels were quantified using MS.
[U-13C]glucose tracing in activated WT, GOT1 or GLUD1 knockout CD8+ T cells
STUDY_SUMMARY
WT, GOT1 knockout or GLUD1 knockout CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28 for 24 hours. CD8+ T cells were pulsed with [U-13C]glucose. Intracellular glucose-derived glutamate, serine and Ī±-ketoglutarate levels were quantified using MS.
[U-13C]glutamine tracing in activated WT, GOT1 or GLUD1 knockout CD8+ T cells
STUDY_SUMMARY
WT, GOT1 knockout or GLUD1 knockout CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28 for 24 hours. CD8+ T cells were pulsed with [U-13C]glutamine for 4-6 hours. Intracellular glutamine-derived glutamate and Ī±-ketoglutarate levels were quantified using MS.
Single cell lipidome analysis of phosphatidylcholines and spingomyelins from HepG2 and C2C12 cells
STUDY_TYPE
Quantitative single cell lipidomics
STUDY_SUMMARY
We have developed a high-throughput method for the detection and quantification of a wide range of phosphatidylcholine (PC) and sphingomyelin (SM) species from single cells that combines fluorescence-assisted cell sorting (FACS) with automated chip-based nanoelectrospray ionization (nanoESI) and shotgun lipidomics. Using this method we can detect and perform relative quantitation on more than >50 different PC and SM species from immortalised human cells, and can easily distinguish between cells of different lineages (e.g. hepatocarcinoma HePG2 vs C2C12 myoblasts) and cells treated with exogenous fatty acids.
INSTITUTE
Victor Chang Cardiac Research Institute
LABORATORY
Cellular Bioenergetics Laboratory
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW, 2010, Australia
Single cell lipidome analysis of phosphatidylcholines and spingomyelins from prostate cells
STUDY_TYPE
Quantitative single cell lipidomics
STUDY_SUMMARY
We have developed a high-throughput method for the detection and quantification of a wide range of phosphatidylcholine (PC) and sphingomyelin (SM) species from single cells that combines fluorescence-assisted cell sorting (FACS) with automated chip-based nanoelectrospray ionization (nanoESI) and shotgun lipidomics. Using this method we can detect and perform relative quantitation on more than >50 different PC and SM species from immortalised human cells, and can easily distinguish between tumorigenic and non-tumorigenic prostate cell lines.
INSTITUTE
Victor Chang Cardiac Research Institute
LABORATORY
Cellular Bioenergetics Laboratory
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW, 2010, Australia
Sperm Environmental Epigenetics and Development Study (SEEDS)
STUDY_SUMMARY
Infertility is one of the most common reproductive health disorders affecting 16% of couples in the U.S. Most concerning are the new meta-analysis data showing that sperm counts among men in developed countries have declined over 50% in the past four decades. With no sign of reversing this downward trajectory, we may not only be facing a fertility crisis, but low sperm count also has wider public health implications, including increased risks in morbidity and mortality. Given this dramatic decrease in sperm quality over a short period, genetic influences are likely not attributable, but rather, environmental factors encountered over the life-course. The objective of this pilot project is to determine the feasibility of generating metabolomic data from human seminal plasma collected as part of the ongoing SEEDS cohort.
The impact of acute Colony Stimulating Factor 1 treatment on serum and liver metabolites in fed and fasted mice LIVER
STUDY_TYPE
Drug treatment
STUDY_SUMMARY
The aim of the study was to investigate the impact of expanding tissue macrophage populations on systemic metabolism. Groups of male mice were treated with 4 x daily injections of Colony Stimulating Factor (1), followed by normal feeding or a 24 h fast from day 6 to day 7. Serum and liver were collected for GC-MS metabolomic analysis on a Shimadzu TQ8050.
The impact of acute Colony Stimulating Factor 1 treatment on serum and liver metabolites in fed and fasted mice ā SERUM
STUDY_TYPE
Drug treatment
STUDY_SUMMARY
The aim of the study was to investigate the impact of expanding tissue macrophage populations on systemic metabolism. Groups of male mice were treated with 4 x daily injections of Colony Stimulating Factor (1), followed by normal feeding or a 24 h fast from day 6 to day 7. Serum and liver were collected for GC-MS metabolomic analysis on a Shimadzu TQ8050.
High-resolution metabolomics analysis of NLRP3 inflammasome activated macrophages
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Activating macrophage NLRP3 inflammasome can promote excessive inflammation, with severe cell and tissue damage and organ dysfunction. Here, we show that pharmacological or genetic inhibition of pyruvate dehydrogenase kinase (PDHK) significantly attenuates NLRP3 inflammasome activation in murine and human macrophages and septic mice by lowering caspase-1 cleavage and IL-1beta secretion. Inhibiting PDHK reverses NLRP3 inflammasome-induced metabolic reprogramming, enhances autophagy, promotes mitochondrial fusion over fission, preserves cristae ultrastructure, and attenuates mitochondrial ROS production. The suppressive effect of PDHK inhibition on the NLRP3 inflammasome is independent of its canonical role as a pyruvate dehydrogenase regulator. We suggest that PDHK inhibition improves mitochondrial fitness by reversing NLRP3 inflammasome activation in acutely inflamed macrophages.
Extracellular pyruvate secretion by activated CD8+ T cells revealed by [U-13C]glucose tracing
STUDY_SUMMARY
WT OT-I CD8+ T cells were activated in [U-13C]glucose for 24 hours. Blank [U-13C]glucose media or media post cell culture were collected for mass spectrometry analysis. Percent contribution of carbon flux from [U-13C]glucose to pyruvate were analyzed.
Extracellular metabolome of activated CD8+ T cells
STUDY_SUMMARY
WT OT-I CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28. Blank media or media post 10hr, 30hr or 48hr cell culture were collected for mass spectrometry analysis.
Metabolomics analysis of WT vs. GOT1 knockout CD8+ T cells
STUDY_SUMMARY
WT or GOT1 knockout OT-I CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28 for 24 hours. Cells were collected for mass spectrometry analysis.
Metabolomics analysis of WT or GOT1 knockout CD8+ T cells cultured in serine-replete or serine-free media
STUDY_SUMMARY
WT or GOT1 knockout CD8+ T cells were activated with plate-bound anti-CD3 or soluble anti-CD28, in serine-replete or serine-free media for 24 hours. Intracellular metabolome were assessed by MS.
Hepatic Phosphatidylcholine Catabolism Driven by PNPLA7 and PNPLA8 Supplies Endogenous Choline to Replenish the Methionine Cycle with Methyl Groups(Pnpla8-knockout)
STUDY_SUMMARY
Choline supplies methyl groups for regeneration of methionine and the methyl donor S-adenosylmethionine in the liver. Here we demonstrate that the catabolism of membrane phosphatidylcholine (PC) into water-soluble glycerophosphocholine (GPC) by the phospholipase/lysophospholipase PNPLA8-PNPLA7 axis enables endogenous choline stored in hepatic PC to be utilized in methyl metabolism. PNPLA7-deficient mice show marked decreases in hepatic GPC, choline, and several metabolites related to the methionine cycle, accompanied by various signs of methionine insufficiency including growth retardation, hypoglycemia, hypolipidemia, increased energy consumption, reduced adiposity, increased FGF21, and an altered histone/DNA methylation landscape. Moreover, PNPLA8-deficient mice recapitulate most of these phenotypes. In contrast to wild-type mice fed a methionine/choline-deficient diet, both knockout strains display a decreased hepatic triglyceride likely via reductions of lipogenesis and GPC-derived glycerol flux. Collectively, our findings highlight the biological importance of phospholipid catabolism driven by PNPLA8/PNPLA7 in methyl group flux and triglyceride synthesis in the liver.
INSTITUTE
Tokyo Metropolitan Institute of Medical Science
LAST_NAME
Hirabayashi
FIRST_NAME
Tetsuya
ADDRESS
2-6-1 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan
Hepatic Phosphatidylcholine Catabolism Driven by PNPLA7 and PNPLA8 Supplies Endogenous Choline to Replenish the Methionine Cycle with Methyl Groups (Pnpla7-knockout)
STUDY_SUMMARY
Choline supplies methyl groups for regeneration of methionine and the methyl donor S-adenosylmethionine in the liver. Here we demonstrate that the catabolism of membrane phosphatidylcholine (PC) into water-soluble glycerophosphocholine (GPC) by the phospholipase/lysophospholipase PNPLA8-PNPLA7 axis enables endogenous choline stored in hepatic PC to be utilized in methyl metabolism. PNPLA7-deficient mice show marked decreases in hepatic GPC, choline, and several metabolites related to the methionine cycle, accompanied by various signs of methionine insufficiency including growth retardation, hypoglycemia, hypolipidemia, increased energy consumption, reduced adiposity, increased FGF21, and an altered histone/DNA methylation landscape. Moreover, PNPLA8-deficient mice recapitulate most of these phenotypes. In contrast to wild-type mice fed a methionine/choline-deficient diet, both knockout strains display a decreased hepatic triglyceride likely via reductions of lipogenesis and GPC-derived glycerol flux. Collectively, our findings highlight the biological importance of phospholipid catabolism driven by PNPLA8/PNPLA7 in methyl group flux and triglyceride synthesis in the liver.
INSTITUTE
Tokyo Metropolitan Institute of Medical Science
LAST_NAME
Hirabayashi
FIRST_NAME
Tetsuya
ADDRESS
2-6-1 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan
Targeted metabolomics analysis of WT and GSDMDKO macrophages
STUDY_TYPE
MS analysis
STUDY_SUMMARY
Activating macrophage NLRP3 inflammasome can promote excessive inflammation, with severe cell and tissue damage and organ dysfunction. Here, we show that pharmacological or genetic inhibition of pyruvate dehydrogenase kinase (PDHK) significantly attenuates NLRP3 inflammasome activation in murine and human macrophages and septic mice by lowering caspase-1 cleavage and IL-1beta secretion. Inhibiting PDHK reverses NLRP3 inflammasome-induced metabolic reprogramming, enhances autophagy, promotes mitochondrial fusion over fission, preserves cristae ultrastructure, and attenuates mitochondrial ROS production. The suppressive effect of PDHK inhibition on the NLRP3 inflammasome is independent of its canonical role as a pyruvate dehydrogenase regulator. We suggest that PDHK inhibition improves mitochondrial fitness by reversing NLRP3 inflammasome activation in acutely inflamed macrophages.
Glucose flux analysis of NLRP3 inflammasome activated macrophages
STUDY_TYPE
Basic research
STUDY_SUMMARY
Activating macrophage NLRP3 inflammasome can promote excessive inflammation, with severe cell and tissue damage and organ dysfunction. Here, we show that pharmacological or genetic inhibition of pyruvate dehydrogenase kinase (PDHK) significantly attenuates NLRP3 inflammasome activation in murine and human macrophages and septic mice by lowering caspase-1 cleavage and IL-1beta secretion. Inhibiting PDHK reverses NLRP3 inflammasome-induced metabolic reprogramming, enhances autophagy, promotes mitochondrial fusion over fission, preserves cristae ultrastructure, and attenuates mitochondrial ROS production. The suppressive effect of PDHK inhibition on the NLRP3 inflammasome is independent of its canonical role as a pyruvate dehydrogenase regulator. We suggest that PDHK inhibition improves mitochondrial fitness by reversing NLRP3 inflammasome activation in acutely inflamed macrophages.
Metabolomic and Cultivation Insights into the Tolerance of the Spacecraft-Associated Acinetobacter Towards Kleenol 30, a Cleanroom Floor Detergent
STUDY_TYPE
research
STUDY_SUMMARY
To ensure cleanliness, NASA spacecraft are assembled in cleanroom facilities, where floors are routinely cleansed with Kleenol 30 (K30), an alkaline detergent. Through metabolomic and cultivation approaches, we show that cultures of spacecraft-associated Acinetobacter tolerate up to 1% v/v K30 and are fully inhibited at ā„2%; in comparison, NASA cleanrooms are cleansed with 0.8% K30. For A. johnsonii 2P08AA (isolated from a cleanroom floor), cultivations with 0.1% v/v K30 yield (1) limited changes in the intracellular metabolome and (2) increases in extracellular sugar acids, monosaccharides, organic acids, and fatty acids. For A. radioresistens 50v1 (isolated from a spacecraft surface), cultivations yield (1) differential changes in intracellular amino acids, compatible solutes, nucleotide-related metabolites, dicarboxylic acids, and saturated fatty acids and (2) substantial yet differential impacts to extracellular sugar acids, monosaccharides, and organic acids. These combined results suggest that (1) K30 manifests strain-dependent impacts on the intracellular metabolomes and (2) K30 influences extracellular trace element acquisition in both strains. Hence, this work lends support towards the hypothesis that repeated cleansing during spacecraft assembly serve as selective pressures that promote tolerances towards the cleaning conditions.
The goal of this project was to identify bacterial transporters responsible for uptake of environmentally relevant marine metabolites. We used the model marine heterotrophic bacterium Ruegeria pomeroyi DSS-3, for which an arrayed library of single gene knockout mutants has been generated by selecting isolated from a barcoded transposon mutant library (BasSeq). Knockout mutants of putative transporters were grown on minimal medium with a single substrate as sole carbon source. Mutant defect was assessed by comparing the substrate drawdown of isolated mutants to drawdown by a pooled mutant library (BarSeq), a proxy for wildtype fitness.
Deep multi-omic profiling reveals extensive mitochondrial remodeling driven by glycemia in early diabetic kidney disease
STUDY_SUMMARY
Changes in mitochondrial energy metabolism are thought to be central to the development of diabetic kidney disease (DKD); however, whether this response is explicitly driven by systemic glucose concentrations remains unknown. Here, we show that titrating blood glucose concentrations in vivo directly impacts mitochondrial morphology and bioenergetics and remodels the mitochondrial proteome in the kidney in early DKD. Mitoproteomic analysis revealed profound metabolic disturbances induced by severe hyperglycemia, including upregulation of enzymes involved in the TCA cycle and fatty acid metabolism, enhanced ketogenesis as well as extensive dysregulation of the mitochondrial SLC25 carrier family. The metabolite and lipid landscape were perturbed by severe hyperglycemia; untargeted metabolomics and lipidomics confirmed the enrichment of TCA cycle metabolites, an increase in triglyceride concentrations, and extensive and specific cardiolipin remodeling. Lowering blood glucose to moderate hyperglycemia stabilized all three omic landscapes, partially prevented changes in mitochondrial morphology and bioenergetics, and improved kidney injury. This study provides insights into altered substrate utilization and energy generation in the kidney early in diabetes, during moderate and severe hyperglycemia and has implications for therapeutic strategies aiming at the reinvigoration of mitochondrial function and signaling in diabetes.
Metabolomics of B-cell Acute Lymphoblastic Leukemia in Response to Adipocyte Conditioned Media
STUDY_TYPE
Untargeted MS
STUDY_SUMMARY
Adipocyte conditioned media (ACM), stromal cell conditioned media (SCM) and unconditioned media (UCM) were added to B-cell Acute Lymphoblastic Leukemia cells (REH and RCH-AcV) either with or without methotrexate (MTX). The metabolomic profiles of the cells was determined by mass spectrometry.
Lipidomics study of curcumin and epigallocatechin gallate on IgE-mediated degranulation of RBL-2H3 cells
STUDY_SUMMARY
This study provides novel insights into the potential anti-allergic mechanism of food-derived curcumin and EGCG, and then facilitates the discovery of drug target and the development of diagnostic methods for allergic diseases.
INSTITUTE
College of Marine Food and Biological Engineering, Jimei University
LAST_NAME
Yang
FIRST_NAME
Zhiqiang
ADDRESS
No. 185 Yinjiang Road, Jimei District, Xiamen City, Fujian Province, China
Metabolomics in small-spotted catshark reproduction- Seminal plasma Semi-polar
STUDY_SUMMARY
this study aimed to elucidate the influence of the environment (wild vs. aquarium) on the metabolic signatures in the seminal plasma (wild-captured vs. aquarium-housed).
Metabolomics in small-spotted catshark reproduction- Seminal plasma no-polar
STUDY_SUMMARY
this study aimed to elucidate the influence of the environment (wild vs. aquarium) on the metabolic signatures in the seminal plasma (wild-captured vs. aquarium-housed).
Metabolomics in small-spotted catshark reproduction- Blood plasma Semi-polar
STUDY_SUMMARY
this study aimed to elucidate the influence of the environment (wild vs. aquarium) on the metabolic signatures in the blood plasma (wild-captured vs. aquarium-housed).
Metabolomics in small-spotted catshark reproduction- Blood plasma No-polar
STUDY_SUMMARY
this study aimed to elucidate the influence of the environment (wild vs. aquarium) on the metabolic signatures in the blood plasma (wild-captured vs. aquarium-housed).
Alterations in SHH signal transduction introduce a state of hypometabolism in sporadic Parkinson's disease
STUDY_SUMMARY
Induced pluripotent stem cells (iPSC) derived from sporadic Parkinson's disease patients and healthy control subjects were used for disease modeling. iPSC were differentiated towards midbrain dopaminergic neurons. For metabolic analysis, midbrain neuronal precursor cells were cultivated in growth medium supplemented with either 1.25 mM [U-13C]-glutamine or 21.25 mM [U-13C]-glucose. Metabolites were extracted and analyzed using GC-MS. The MetaboliteDetector software was used to analyze chromatograms, calculate mass isotopomer distributions (MIDs) and perform relative comparison of metabolite levels.
The metabolomic resetting effect of DMXAA in cisplatin-induced injured mouse kidney
STUDY_SUMMARY
A single injection of cisplatin was used to induce AKI in mice. To assess the effect of DMXAA, the mice were injected intraperitoneally with DMXAA or vehicle one hour before cisplatin injection, repeated every 24 hours. The mice were euthanized 72 hours after cisplatin injection, and renal tissues were collected for MS analysis.
INSTITUTE
Children's Hospital of Nanjing Medical University
LAST_NAME
Lu
FIRST_NAME
Lingling
ADDRESS
Guangzhou Road 72, Nanjing, Jiangsu, 210000, China
Evaluation of Two Simultaneous Metabolomic and Proteomic Extraction Protocols Assessed by Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry
STUDY_SUMMARY
Untargeted multi-omics analysis of plasma is an emerging tool for the identification of novel biomarkers for evaluating disease prognosis and for a better understanding of molecular mechanisms underlying human disease. The successful application of metabolomic and proteomic approaches relies on reproducibly quantifying a wide range of metabolites and proteins. Herein, we report the results of untargeted metabolomic and proteomic analyses from blood plasma samples following analyte extraction by two frequently used solvent systems: chloro-form/methanol and methanol-only. Whole blood samples were collected from participants (n=6) at University Hospital Sharjah (UHS) hospital, then plasma was separated and extracted by two methods i. methanol precipitation and, ii. 4:3 methanol:chloroform extraction. The coverage and reproducibility of the two methods were assessed by ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). The study revealed that metabolite extraction by methanol-only showed greater reproducibility for both metabolomic and proteomic quantifications than did methanol/chloroform, while yielding similar peptide coverage. However, coverage of extracted metabolites was higher with the methanol/chloroform precipitation.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
The NMR-based metabolomics analysis of 95 C. albicans isolates using the culture supernatants collected after 14h of incubation. The processed data are used as binns or as annotated and quantified metabolites data for study of the isolates variability within and between genomic clades.
Untargeted metabolomics analysis of DSS-treated murine intestinal microbiota cultivated in continuous bioreactors
STUDY_SUMMARY
We cultivated murine faecal microbiota in six replicate continuous bioreactors. Three bioreactors were supplemented with 1% DSS and compared to the untreated control bioreactors.
Short-chain fatty acid (SCFA) analysis of DSS-treated murine intestinal microbiota using continuous cultivation in bioreactors
STUDY_SUMMARY
We cultivated murine faecal microbiota in six replicate continuous bioreactors. Three bioreactors were supplemented with 1% DSS and compared to the untreated control bioreactors.
A Mammalian Conserved Circular RNA CircLARP2 Regulates Hepatocellular Carcinoma Metastasis and Lipid Metabolism (Part 2)
STUDY_SUMMARY
Circular RNAs (circRNAs) have emerged as crucial regulators in physiology and human diseases. However, evolutionarily conserved circRNAs with potent functions in cancers are rarely reported. Here, we identified a mammalian conserved circRNA circLARP2 that played critical roles in hepatocellular carcinoma (HCC). With clinical specimens, we found that patients with high circLARP2 levels in HCC had advanced prognostic stage and poor overall survival. CircLARP2 facilitated HCC metastasis and lipid accumulation in HCC cell lines. CircLARP2 was one of the rare ones that were identified in HCC metastasis and conserved in mammals, which enabled further studies with animal models. CircLARP2-deficient mice exhibited reduced metastasis and less lipid accumulation in an induced HCC model. We provided lines of evidence at molecular, cellular, and whole organismal levels, to support that circLARP2 functioned as a protein sponge of AUF1. CircLARP2 sequestered AUF1 from binding to LKB1 mRNA, which led to decreased LKB1 mRNA stability and lower LKB1 protein levels. LKB1 as a kinase promoted the phosphorylation of AMPK and then the phosphorylation of ACC, the rate limiting enzyme of fatty acid synthesis. Knockdown of Lkb1 with AAV8-shLkb1 in mice HCC model also proved that Lkb1 was a key element in the regulation. Through this AUF1-LKB1-AMPK-ACC pathway, circLARP2 promoted HCC metastasis and lipid accumulation.
p53 K316P mutation results in increased liver triglyceride levels and increased rates of de novo lipogenesis.
STUDY_SUMMARY
Our lab generated the p53 K316P mouse, which mimicks a common amino acid change found in bats. The K316P mutation, found in the nuclear localization signal of p53, results in increased cytoplasmic localization of p53. We found that K316P mutant mice develop several metabolic phenotypes, including increased body fat percentage, and increased liver lipid levels. In order to determine the mechanism through which K316P mutation increases liver lipid levels, we performed metabolomic analysis of mouse livers from WT and K316P mutant mice. Mouse livers were isolated from four wild type (WT) and four p53 K316P (M) mice for lipidomic analysis. Samples were isolated and flash frozen in liquid nitrogen. Lipids were then extracted from each liver sample and analyzed using mass spectrometry.
System-level analysis of flux regulation of yeast show that glycolytic flux is controlled by allosteric regulation and enzyme phosphorylation
STUDY_SUMMARY
Energy metabolism is central for cellular function and has therefore evolved to be tightly regulated such that energy production can be balanced to energy demand. Energy is being produced in the central carbon metabolism (CCM) and even though there has been extensive studies on how fluxes through the different pathways in this part of metabolism are regulated. There is little understanding of how fluxes are affected by posttranslational modifications and by allosteric regulators. Here we integrated multi-omics data (intracellular metabolome, extracellular metabolome, proteome, phosphoproteome, and fluxome) under 9 different chemostat conditions for building a mathematical model that could map functional regulatory events (FREs) in the Saccharomyces cerevisiae. Using hierarchical analysis combined with the mathematical model, we observed pathway and metabolism-specific flux regulation mechanisms in the CCM. We also found that the glycolytic flux increased with specific growth rate, and this increase was accompanied by a decrease of both metabolites derived FREs and protein phosphorylation level.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
LAST_NAME
Chen
FIRST_NAME
Min
ADDRESS
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology
Lipidomics of Tango2 Deficient and Wildtype Zebrafish Muscle Tissue
STUDY_TYPE
Untargeted Lipidomics
STUDY_SUMMARY
Rhabdomyolysis is a clinical emergency characterized by severe muscle damage, resulting in the release of intracellular muscle components which leads to myoglobinuria and in severe cases, acute kidney failure. Rhabdomyolysis is caused by genetic factors that are linked to increased disease susceptibility in response to extrinsic triggers. Recessive mutations in TANGO2 result in episodic rhabdomyolysis, metabolic crises, encephalopathy, and cardiac arrhythmia. The underlying mechanism contributing to disease onset in response to specific triggers remains unclear. To address these challenges, we created a zebrafish model of Tango2 deficiency. Here we demonstrate that the loss of Tango2 in zebrafish results in growth defects, early lethality, and increased susceptibility of muscle defects similar to TANGO2 patients. Detailed analyses of skeletal muscle revealed defects in the sarcoplasmic reticulum and mitochondria at the onset of disease development. The sarcoplasmic reticulum (SR) constitutes the primary lipid biosynthesis site and regulates calcium handling in skeletal muscle to control excitation-contraction coupling. Tango2 deficient SR exhibits increased sensitivity to calcium release that was partly restored by inhibition of Ryr1-mediated Ca2+ release in skeletal muscle. Using lipidomics, we identified alterations in the glycerolipid state of tango2 mutants which is critical for membrane stability and energy balance. Therefore, these studies provide insight into key disease processes in Tango2 deficiency and have increased our understanding of the impacts of specific defects on predisposition to environmental triggers in TANGO2-related disorders.
INSTITUTE
University of North Carolina at Chapel Hill
DEPARTMENT
Chemistry
LABORATORY
MS Core Laboratory
LAST_NAME
Wallace
FIRST_NAME
Emily
ADDRESS
131 South Rd
EMAIL
emdiane@email.unc.edu
PHONE
7042453664
NUM_GROUPS
2
TOTAL_SUBJECTS
5
NUM_MALES
N/A
NUM_FEMALES
N/A
STUDY_COMMENTS
Zebrafish were all 4 weeks old when tissue was harvested, sex is determined at 4 weeks old.
Metabolomics of Adipocyte-Conditioned Media Compared to Stromal Cell- and Un-conditioned Media
STUDY_TYPE
Untargeted MS
STUDY_SUMMARY
The metabolomics profiles of adipocyte conditioned media (ACM), stromal cell conditioned media (SCM) and unconditioned media (UCM) were analyzed by untargeted mass spectrometry.
Alcohol dehydrogenase 1B is crucial for adipocyte homeostasis
STUDY_SUMMARY
Background. Alcohol dehydrogenase (ADH1B), encoded by the ADH1B gene, is a cytosolic enzyme mainly known for its role in ethanol catabolism in the liver. A few studies have paved the way to show an equally important role of ADH1B in adipocytes. This study aimed to better identify the cellular mechanisms and signaling pathways involving ADH1B in adipose tissue and to determine if ADH1B variants might contribute to adipose tissue dysfunction. Results. We showed that CRISPR-Cas9-mediated ADH1B knockout (KO) in human adipose stem cells (ASC) abolished adipocyte differentiation and decreased insulin response. This was accompanied by oxidative stress, altered mitochondrial functions, and cellular senescence. Lipidomic analysis revealed that ADH1B deficiency results in a major remodeling of lipid composition in ASC. An ADH1B homozygous loss-of-function variant was also identified in a patient presenting with a lipodystrophic and insulin resistant syndrome associated with major liver dysfunction, leading to early death. Discussion. This translational study underlines the crucial role of ADH1B in adipose tissue. It unveils cellular mechanisms accounting for its key role in adipogenesis, and adipocyte homeostasis. This study also identifies ADH1B as a candidate gene in monogenic forms of lipodystrophic and insulin resistant syndromes.
Human primary astrocytes finger- and footprinting metabolomics indicate biochemical alterations under ayahuasca treatment
STUDY_SUMMARY
Ayahuasca (Aya) is a psychotropic Amazonian beverage formulated from the combination of the Banisteriopsis caapi vine and the Psychotria viridis leaves in a water decoction. Aya is legally used in Latin American countries and used in Brazil for religious, cultural, and therapeutic purposes. Its properties constitute a bio-psycho-social-spiritual model involving effects from Ī²-carboline-derived alkaloids, present in the vine, and N,N-dimethyltryptamine (DMT), a tryptamine-derived alkaloid present in the leaves, which act together in the central nervous system (CNS). Few technical-scientific studies have been conducted to understand the effects of this brew in the metabolism. Therefore, this work aims to investigate an in vitro primary astrocyte lineage model by untargeted metabolomics evaluations of two cellular subfractions: secretome and intracellular content after Aya treatment, where DMT and other Ī²-carbolines were previously quantified. Metabolomics analysis was performed by UHPLC-MS/MS, followed by MS-Dial data processing and statistical analysis to identify metabolites and biochemical alterations related to Aya treatment. Aya doses were applied to the cell cultures considering DMT concentrations of 1, 10 and 20 ĀµM, which are in agreement with non-toxic and toxic DMT threshold assays in primary human astrocyte cells viability
Four-dimensional trapped ion mobility spectrometry lipidomics for high throughput clinical profiling of human blood samples
STUDY_TYPE
Clinical Lipidomics
STUDY_SUMMARY
Implementation of operational workflows using untargeted high-resolution ion mobility mass spectrometry in clinical lipidomics require reproducible, high-throughput lipid extraction, high-quality lipid annotation, absolute quantification, and cross-validation. We present a high-throughput 4D-PASEF MS platform suitable for clinical plasma lipidomic profiling encompassing automated extraction, accurate lipid annotation, and reproducible quantification. Newly generated 4D-PASEF lipid descriptors (m/z, RT, CCS, MS2) of 200 lipid standards and of 493 lipid signals curated from reference plasma, along with qualification of reproducible features in replicate lipid extracts and dilution analyses enabled highly confident annotation, reaching 100% confidence, of 370 lipids from NIST SRM 1950 plasma and 364 lipids from NIST SRM 1951 serum. 359 plasma lipids were reproducibly quantified using absolute quantification, cross-validated by inter-instrument studies, and supported by inter-laboratory data. The high-throughput 4D-PASEF lipidomics platform was demonstrated by reproducible identification of intra-individual multidien lipidome phenotype in plasma, serum, blood, venous, and finger-prick dried blood spots.
Deep multi-omic profiling reveals extensive mitochondrial remodeling driven by glycemia in early diabetic kidney disease (Mitochondria)
STUDY_SUMMARY
Changes in mitochondrial energy metabolism are thought to be central to the development of diabetic kidney disease (DKD); however, whether this response is explicitly driven by systemic glucose concentrations remains unknown. Here, we show that titrating blood glucose concentrations in vivo directly impacts mitochondrial morphology and bioenergetics and remodels the mitochondrial proteome in the kidney in early DKD. Mitoproteomic analysis revealed profound metabolic disturbances induced by severe hyperglycemia, including upregulation of enzymes involved in the TCA cycle and fatty acid metabolism, enhanced ketogenesis as well as extensive dysregulation of the mitochondrial SLC25 transporter family. The metabolite and lipid landscape were perturbed by severe hyperglycemia; untargeted metabolomics and lipidomics confirmed the enrichment of TCA cycle metabolites, an increase in triglyceride concentrations, and extensive and specific cardiolipin remodeling. Lowering blood glucose to moderate hyperglycemia stabilized all three omic landscapes, partially prevented changes in mitochondrial morphology and bioenergetics, and improved kidney injury. This study provides insights into altered substrate utilization and energy generation in the kidney early in diabetes, during moderate and severe hyperglycemia and has implications for therapeutic strategies aiming at the reinvigoration of mitochondrial function and signaling in diabetes.
Profiling Metabolites and Lipoproteins in COMETA, an Italian Cohort of COVID-19 patients-part 2
STUDY_SUMMARY
Here, we report a detailed and comprehensive characterization of the metabolomic and lipoproteomic fingerprint of plasma samples of a high number of hospitalized COVID-19 patients, with different disease severities, infected with different viral variants and with different vaccination statues. Our data deeply extend a first metabolomic/lipoproteomic characterization of the disease published at the beginning of 2022(Ghini V. et al., Plos Path 2022, 18(4):e1010443) performed on a much small number of subjects of the same cohort, infected before a significant spread of the Delta variant and before the administration of the vaccines.
INSTITUTE
University of Florence
LAST_NAME
Ghini
FIRST_NAME
Veronica
ADDRESS
via Luigi Sacconi, 6, Sesto Fiorentino, Firenze, 50019, Italy
Stool global metabolite levels in peanut allergy (Part 2)
STUDY_SUMMARY
Prior evidence supports differential levels of short chain fatty acids and other metabolites in the stool of humans and murine models of food allergy. Here we measure global metabolite levels in stool samples collected from children with allergy risk factors. Sample processing included polar metabolite extraction, scaling, and analysis with a global polar liquid chromatography tandem mass spectrometry platform.
Silicon ameliorates clubroot responses in canola (Brassica napus): A āmulti-omicsā-based investigation into possible mechanisms
STUDY_TYPE
Timecourse experiment
STUDY_SUMMARY
Clubroot disease, caused by Plasmodiophora brassicae Woronin results in severe yield losses in Brassica crops, including canola. Silicon (Si) mitigates several stresses and enhances plant resistance to phytopathogens. We investigated the effects of Si on clubroot disease symptoms in canola at two concentrations of Si (Si1.0 and Si0.5). In addition, the effects of Si on P. brassicae-induced gene expression, endogenous levels of phytohormones and metabolites were also studied. Si application reduced clubroot symptoms and improved plant growth under greenhouse conditions. Pathogen-induced transcript-level changes were affected by Si treatment to P. brassicae with genes related to antioxidant activity, phytohormone biosynthesis and signalling, nitrogen metabolism and secondary metabolism exhibiting differential expression. Endogenous levels of several phytohormones (e.g., auxin, cytokinin, salicylic acid and abscisic acid), amino acids and secondary metabolites (e.g., glucosinolates) were affected by Si. This is the first report that Si ameliorates clubroot symptoms and its possible mode of action.
INSTITUTE
Trent University
DEPARTMENT
Biology
LABORATORY
Emery Lab
LAST_NAME
Kisiala
FIRST_NAME
Anna
ADDRESS
1600 West Bank Drive, Trent University, Peterborough, ON, K9L 0G2, Canada
Spatial, temporal, and inter-subject variation of the metabolome along the human upper intestinal tract (MS HILIC negative data)
STUDY_SUMMARY
Most utilization of human diets occurs in the small intestine, which remains largely unstudied. Here, we used a novel non-invasive, ingestible sampling device to probe the spatiotemporal variation of upper intestinal luminal contents during routine daily digestion in 15 healthy subjects. We analyzed 274 intestinal samples and 60 corresponding stool homogenates by combining five metabolomics assays and 16S rRNA sequencing. We identified 1,909 metabolites, including sulfonolipids and novel bile acids. Stool and intestinal metabolomes differed dramatically. Food metabolites displayed known differences and trends in dietary biomarkers, unexpected increases in dicarboxylic acids along the intestinal tract, and a positive association between luminal keto acids and fruit intake. Diet-derived and microbially linked metabolites accounted for the largest inter-subject differences. Interestingly, subjects exhibited large variation in levels of bioactive fatty acid esters of hydroxy fatty acids (FAHFAs) and sulfonolipids. Two subjects who had taken antibiotics within 6 months prior to sampling showed markedly different patterns in these and other microbially related metabolites; from this variation, we identified Blautia species as most likely to be involved in FAHFA metabolism. Thus, in vivo sampling of the human small intestine under physiologic conditions can reveal links between diet, host and microbial metabolism.
Multi-omic Analysis of ClpP Activation in Triple-Negative Breast Cancer Cells
STUDY_SUMMARY
The ClpP activators ONC201 and related small molecules (TR compounds, Madera Therapeutics), have demonstrated significant anti-cancer potential in an array of in vitro cell models and in vivo studies, including clinical trials for refractory solid tumors. Though progress has been made in identifying specific phenotypic outcomes following ClpP activation, the exact mechanism by which ClpP activation leads to broad anti-cancer activity has yet to be fully elucidated. In this study, we utilized a multi-omics approach to identify the ClpP-dependent proteomic, transcriptomic, and metabolomic changes resulting from ONC201 or the TR compound TR-57 in triple-negative breast cancer cells (TNBC). Applying mass spectrometry- based methods of proteomics and metabolomics, we identified ~8000 proteins and 588 metabolites, respectively. From proteomics data, approximately 3400 (ONC201) and 3000 (TR-57) proteins increased and ~4600 (ONC201) and ~4800 (TR-57) proteins decreased in this study. Additionally, gene ontological analysis revealed strong similarities between proteins up- or downregulated by ONC201 or TR-57 treatment. Notably, this included the downregulation of many mitochondrial processes and proteins, including mitochondrial translation and mitochondrial matrix proteins. We also performed a large-scale transcriptomic analysis of WT SUM159 cells, identifying ~7700 transcripts (~3600 and 3800 increasing, ~4000 and 3900 decreasing in ONC201 and TR-57 treated cells, respectively). Less than 21% of these genes were affected by these compounds in ClpP null cells. Gene ontological analysis of these data demonstrated additional similarity of response to ONC201 and TR-57. Many of the same gene ontology processes and cellular components were identified, including a decrease in transcripts related to the mitochondrial inner membrane and matrix, the cell cycle, and the nucleus, as well as increases in other nuclear transcripts and transcripts related to metal-ion binding. Comparative analysis demonstrated a highly analogous response in all -omics datasets. Analysis of metabolites also revealed significant similarities between ONC201 and TR-57 with increases in Ī±-ketoglutarate and 2-hydroxyglutaric acid and decreased levels of ureidosuccinic acid, L-ascorbic acid, L-serine, and cytidine observed following ClpP activation in TNBC cells. Further analysis identified multiple pathways that were specifically impacted by ClpP activation, including ATF4 activation, heme biosynthesis, and the citrulline/urea cycle. In summary the results of our studies demonstrate that ONC201 and TR-57 induce highly similar and broad effects against multiple mitochondrial processes required for cell proliferation.
INSTITUTE
University of North Carolina at Chapel Hill
LAST_NAME
Graves
FIRST_NAME
Lee
ADDRESS
4111 Genetic Medicine Building, 120 Mason Farm Rd, Chapel Hill, NC 27514
Spatial, temporal, and inter-subject variation of the metabolome along the human upper intestinal tract (MS RP positive data)
STUDY_SUMMARY
Most utilization of human diets occurs in the small intestine, which remains largely unstudied. Here, we used a novel non-invasive, ingestible sampling device to probe the spatiotemporal variation of upper intestinal luminal contents during routine daily digestion in 15 healthy subjects. We analyzed 274 intestinal samples and 60 corresponding stool homogenates by combining five metabolomics assays and 16S rRNA sequencing. We identified 1,909 metabolites, including sulfonolipids and novel bile acids. Stool and intestinal metabolomes differed dramatically. Food metabolites displayed known differences and trends in dietary biomarkers, unexpected increases in dicarboxylic acids along the intestinal tract, and a positive association between luminal keto acids and fruit intake. Diet-derived and microbially linked metabolites accounted for the largest inter-subject differences. Interestingly, subjects exhibited large variation in levels of bioactive fatty acid esters of hydroxy fatty acids (FAHFAs) and sulfonolipids. Two subjects who had taken antibiotics within 6 months prior to sampling showed markedly different patterns in these and other microbially related metabolites; from this variation, we identified Blautia species as most likely to be involved in FAHFA metabolism. Thus, in vivo sampling of the human small intestine under physiologic conditions can reveal links between diet, host and microbial metabolism.
Spatial, temporal, and inter-subject variation of the metabolome along the human upper intestinal tract (MS RP negative data)
STUDY_SUMMARY
Most utilization of human diets occurs in the small intestine, which remains largely unstudied. Here, we used a novel non-invasive, ingestible sampling device to probe the spatiotemporal variation of upper intestinal luminal contents during routine daily digestion in 15 healthy subjects. We analyzed 274 intestinal samples and 60 corresponding stool homogenates by combining five metabolomics assays and 16S rRNA sequencing. We identified 1,909 metabolites, including sulfonolipids and novel bile acids. Stool and intestinal metabolomes differed dramatically. Food metabolites displayed known differences and trends in dietary biomarkers, unexpected increases in dicarboxylic acids along the intestinal tract, and a positive association between luminal keto acids and fruit intake. Diet-derived and microbially linked metabolites accounted for the largest inter-subject differences. Interestingly, subjects exhibited large variation in levels of bioactive fatty acid esters of hydroxy fatty acids (FAHFAs) and sulfonolipids. Two subjects who had taken antibiotics within 6 months prior to sampling showed markedly different patterns in these and other microbially related metabolites; from this variation, we identified Blautia species as most likely to be involved in FAHFA metabolism. Thus, in vivo sampling of the human small intestine under physiologic conditions can reveal links between diet, host and microbial metabolism.
Spleen-derived macrophage from WT or Eif2ak2-/- (gene encoding PKR protein kinase) mice are treated with a synthetic RNA mimetic (polyinosinic:polycytidylic acid) to activate the kinase and metabolites were collected for analysis. The data identified 325 putative metabolites in the cell extracts, with a large number of significant differences between the Eif2ak2- /- and WT sample groups. Metabolite levels are predominantly suppressed in the WT compared to the Eif2ak2-/- cells, with depletion of specific metabolites in amino acid, carbohydrate, lipid and nucleotide pathways, while several amino acid metabolites were significantly elevated in the WT cells compared to the Eif2ak2-/-. The changes appear to delineate a pseudo-starvation response in the WT cells. Phosphate energy metabolism is altered with decreased creatine and phosphocreatine and a compensatory increase in phosphorylated guanidinoacetate in the WT compared to the Eif2ak2- /- cells. There appears to be a constraint in glycolysis in the WT cells, most clearly in the pentose phosphate pathway.
INSTITUTE
Hudson
DEPARTMENT
CIIID
LABORATORY
Molecular Immunology
LAST_NAME
Sadler
FIRST_NAME
Anthony
ADDRESS
27-31 Wright st, Clayton, VIC 3168
EMAIL
Anthony.sadler@hudson.org.au
PHONE
+61 4 85722722
NUM_GROUPS
2
TOTAL_SUBJECTS
NA
NUM_MALES
NA
NUM_FEMALES
NA
STUDY_COMMENTS
KO vs WT
PUBLICATIONS
Suppression of the nucleic acid precursor ribose 5-phosphate by RNA-mediated antiviral immunity
Mass spectrometry dataset of LC-MS Lipidomics Analysis of Xenopus Laevis Optic Nerve
STUDY_SUMMARY
CNS injuries of the anuran amphibian, Xenopus laevis, are uniquely befitted for studying the molecular compositions of neuronal regeneration of retinal ganglion cells (RGC) due to a functional recovery of optic axons disparate to adult mammalian analogues. RGCs and their optic nerve axons undergo irreversible neurodegeneration in glaucoma and associated optic neuropathies, resulting in blindness in mammals. Conversely, Xenopus demonstrates RGC lifetime-spanning regenerative capabilities after optic nerve crush, inciting opportunities to compare de novo regeneration and develop efficient pharmaceutical approaches for vision restoration. Studies revealing lipidome alterations during optic nerve regeneration are sparse and could serve as a solid foundation for these underlying molecular changes. We profile the lipid changes in a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that were either left untreated (naĆÆve) or had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). Matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 7,12,18, and 27 days post optic nerve crush. Following euthanasia, the optic nerves were collected for lipidomic analysis. A modified Bligh and Dyer method [PMID: 13671378] was used for lipid extraction, followed by untargeted mass spectrometry lipid profiling with a Q-Exactive Orbitrap Liquid Chromatography-Mass Spectrometer (LC MS-MS) coupled with Vanquish Horizon Binary UHPLC LC-MS system.
Multiple sclerosis (MS) is a chronic autoimmune disease that affects the myelination of the neurons present in the central nervous system (CNS). The exact etiology of MS development is unclear, but various environmental and genetic factors might play a role in initiating the disease. Current treatments for MS enhance the quality of life and reduce the symptoms. One of these treatments is dimethyl fumarate (DMF), commercially known as Tecfidera. Experimental autoimmune encephalomyelitis (EAE) is a mouse model that is used to study the pathophysiology of MS disease as well as the effects of possible therapeutic agents. In this study, we investigated the effects of SIMR1707 which is a novel compound designed at Sharjah Institute for Medical Research (SIMR). . Single and multiple doses of SIMR1707 demonstrated high safety in mice studies. Treatment of EAE mice with SIMR1707 was able to reduce the EAE clinical scores and maintain their body weight similar to the MS FDA-approved (DMF, Tecfidera), when they were used preventively, prophylactically, or therapeutically. The histological and immunohistochemistry evaluations showed reduced clinical features such as signs of inflammation, demyelination, and infiltration of CD3-positive T cells into the brains of the EAE mice, as compared to vehicle-treated, or untreated EAE mice. Moreover, multi-OMICS experiments including Transcriptomics, Proteomics and Metabolomics were performed to gain insights into the relevant mechanism of action of the SIMR1707 in EAE and thus its therapeutic efficacy to treat MS. Same tissue samples extracted from the cerebellum part of the brain of normal, EAE vehicle-treated, and therapeutic SIMR1707 treated mice, were subjected for the whole RNA-sequencing for transcriptomics, Nano MS for proteomics analysis and LC-MS metabolomics analysis. The multi-OMICs integrative analysis showed that the treatment with SIMR1707 downregulated key biomarkers functionally associated with top pathways including calcium signaling, PI3K/AKT, and mTOR signaling pathways, which may play important roles in EAE and MS pathophysiology. Additionally, the metabolomics-based enriched-for-action pathway analysis showed that the top significantly activated metabolites (FC > 2, p < 0.05) are cholic acid, propionic acid, sphinganine, and nutriacholic acid. Consisting with the functional enrichment pathway analysis, two potent markers, Snta1 and Fscn1, involved in the actin-binding and cytoskeleton are commonly shared between transcriptomics and proteomics and showed mRNA-protein expression correlation in SIMR1707 treated compared to vehicle EAE mice. Importantly, these two markers are involved in the MT2/AKT/GSK3 pathway and may potentially play role in MS and EAE disease
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
Proteomics and metabolomics of multiple sclerosis (Part 2)
STUDY_SUMMARY
Multiple sclerosis (MS) is a chronic autoimmune disease that affects the myelination of the neurons present in the central nervous system (CNS). The exact etiology of MS development is unclear, but various environmental and genetic factors might play a role in initiating the disease. Current treatments for MS enhance the quality of life and reduce the symptoms. One of these treatments is dimethyl fumarate (DMF), commercially known as Tecfidera. Experimental autoimmune encephalomyelitis (EAE) is a mouse model that is used to study the pathophysiology of MS disease as well as the effects of possible therapeutic agents. In this study, we investigated the effects of SIMR1707 which is a novel compound designed at Sharjah Institute for Medical Research (SIMR). . Single and multiple doses of SIMR1707 demonstrated high safety in mice studies. Treatment of EAE mice with SIMR1707 was able to reduce the EAE clinical scores and maintain their body weight similar to the MS FDA-approved (DMF, Tecfidera), when they were used preventively, prophylactically, or therapeutically. The histological and immunohistochemistry evaluations showed reduced clinical features such as signs of inflammation, demyelination, and infiltration of CD3-positive T cells into the brains of the EAE mice, as compared to vehicle-treated, or untreated EAE mice. Moreover, multi-OMICS experiments including Transcriptomics, Proteomics and Metabolomics were performed to gain insights into the relevant mechanism of action of the SIMR1707 in EAE and thus its therapeutic efficacy to treat MS. Same tissue samples extracted from the cerebellum part of the brain of normal, EAE vehicle-treated, and therapeutic SIMR1707 treated mice, were subjected for the whole RNA-sequencing for transcriptomics, Nano MS for proteomics analysis and LC-MS metabolomics analysis. The multi-OMICs integrative analysis showed that the treatment with SIMR1707 downregulated key biomarkers functionally associated with top pathways including calcium signaling, PI3K/AKT, and mTOR signaling pathways, which may play important roles in EAE and MS pathophysiology. Additionally, the metabolomics-based enriched-for-action pathway analysis showed that the top significantly activated metabolites (FC > 2, p < 0.05) are cholic acid, propionic acid, sphinganine, and nutriacholic acid. Consisting with the functional enrichment pathway analysis, two potent markers, Snta1 and Fscn1, involved in the actin-binding and cytoskeleton are commonly shared between transcriptomics and proteomics and showed mRNA-protein expression correlation in SIMR1707 treated compared to vehicle EAE mice. Importantly, these two markers are involved in the MT2/AKT/GSK3 pathway and may potentially play role in MS and EAE disease
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
Serum metabolomics and lipidomics profiling in iliac arteriovenous fistula creation in clinical patients: part 1
STUDY_TYPE
Study of the serum metabolome in patients with end stage kidney disease before and 6-weeks after iliac arteriovenous fistula creation via 1H NMR
STUDY_SUMMARY
This project is focused on a longitudinal analysis of the small molecules metabolomes in end stage renal disease patients undergoing iliac arteriovenous fistula creation using solution state NMR spectroscopy. It was hypothesized that after 6-weeks of AVF creation, these patients would present with altered serum metabolite features.
Serum metabolomics and lipidomics profiling in iliac arteriovenous fistula creation in clinical patients: part 2
STUDY_TYPE
Study of the serum metabolome in patients with end stage kidney disease before and 6-weeks after iliac arteriovenous fistula creation via 1H NMR
STUDY_SUMMARY
This project is focused on a longitudinal analysis of the small molecules metabolomes in end stage renal disease patients undergoing iliac arteriovenous fistula creation using solution state NMR spectroscopy. It was hypothesized that after 6-weeks of AVF creation, these patients would present with altered serum metabolite features.
Metabolomic changes in growth of E. colik at four timespoints in MOPS rich and minimum medium
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
Metabolomic changes were profiled using untargeted metabolomics in E. coli following its growth in two medium types. E. coli strain was grown in MOPS rich and minimum medium and samples were taken at four timepoints: the beginning of lag phase, end of lag phase, mid-log phase, and beginning of the stationary phase.
Colorectal cancer isobaric labeling for metabolite quantification
STUDY_SUMMARY
A major challenge in reducing the death rate of colorectal cancer is to screen patients using low-invasive testing. Blood test shows a high compliance rate with reduced invasiveness. In this work, a multiplex isobaric tag labeling strategy coupled with mass spectrometry is adopted to relatively quantify primary and secondary amine-containing metabolites in serum for the discovery of metabolite biomarkers of colorectal cancer. Serum samples from patients at different risk statuses and colorectal cancer growth statuses are studied. Metabolite identification is based on accurate mass matching and/or retention time of labeled metabolite standards. We quantify 40 metabolites across all the serum samples, including 18 metabolites validated with standards. We find significantly decreased levels of threonine and asparagine in the patients with growing adenomas or high-risk adenomas (p < 0.05). Glutamine levels decrease in patients with adenomas of unknown growth status or high-risk adenomas. In contrast, arginine levels are elevated in patients with low-risk adenoma. Receiver operating characteristic analysis shows high sensitivity and specificity of these metabolites for detecting growing adenomas. Based on these results, we conclude that potential metabolite biomarkers identified here contribute to distinguishing colorectal patients with growing adenomas from normal individuals and patients with unknown growth status of adenomas.
The intimate association between the endoplasmic reticulum (ER) and mitochondrial membranes at ER-mitochondria contact sites (ERMCS) serves as a platform for several critical cellular processes, in particular lipid synthesis. Enzymes involved in lipid biosynthesis are enriched at contacts and membrane lipid composition at contacts is distinct relative to surrounding membranes. How contacts are remodeled and the subsequent biological consequences of altered contacts such as perturbed lipid metabolism remains poorly understood. Here we investigate if the ER-tethered ubiquitin-X domain adaptor 8 (UBXD8) regulates the lipidome of cells. LC-MS/MS lipidomics found significant changes in distinct lipid species in UBXD8 knockout cells, in particular in saturated or mono-unsaturated lipid species. Perturbation of contacts and inherent lipid synthesis is emerging as a hallmark in a variety of human disorders such as neurodegeneration. Our results suggest that contacts are exquisitely sensitive to alterations to membrane lipid composition and saturation in a manner that is dependent on UBXD8.
The intimate association between the endoplasmic reticulum (ER) and mitochondrial membranes at ER-mitochondria contact sites (ERMCS) serves as a platform for several critical cellular processes, in particular lipid synthesis. Enzymes involved in lipid biosynthesis are enriched at contacts and membrane lipid composition at contacts is distinct relative to surrounding membranes. How contacts are remodeled and the subsequent biological consequences of altered contacts such as perturbed lipid metabolism remains poorly understood. Here we investigate if the ER-tethered ubiquitin-X domain adaptor 8 (UBXD8) regulates the lipids found in mitochondria-associated membranes (MAM). LC-MS/MS lipidomics found significant changes in distinct lipid species in the MAM fraction of UBXD8 knockout cells. Our results suggest that lipids in MAM are regulated by UBXD8.
Integrated gut microbiome and lipidomic analyses in animal models of Wilson disease reveal a role of intestine ATP7B in copper-related metabolic dysregulation (Part 1)
STUDY_SUMMARY
Although the main pathogenic mechanism of Wilson disease (WD) is related to copper accumulation in the liver and brain, there is limited knowledge about the role of ATP7B copper transporter in extra-hepatic organs, including the intestine, and how it could affect metabolic manifestations of the disease. The aims of the present study were to profile and correlate the gut microbiota and lipidome in mouse models of WD, and to study the metabolic effects of intestine-specific ATP7B deficiency in a newly generated mouse model. Animal models of WD presented reduced gut microbiota diversity compared to mice with normal copper metabolism. Comparative prediction analysis of the functional metagenome showed the involvement of several pathways including amino acid, carbohydrate, and lipid metabolisms. Lipidomic profiles showed dysregulated tri- and diglyceride, phospholipid, and sphingolipid metabolism. When challenged with a high-fat diet, Atp7bĪIEC mice confirmed profound deregulation of fatty acid desaturation and sphingolipid metabolism pathways as well as altered APOB48 distribution in intestinal epithelial cells. Gut microbiome and lipidomic analyses reveal integrated metabolic changes underlying the systemic manifestations of WD. Intestine-specific ATP7B deficit affects both intestine and systemic response to high-fat challenge. WD is as systemic disease and organ-specific ATP7B variants can explain the varied phenotypic presentations.
INSTITUTE
University of California, Davis
DEPARTMENT
Internal Medicine
LABORATORY
Medici's Lab
LAST_NAME
Sarode
FIRST_NAME
Gaurav Vilas
ADDRESS
451 E. Health Sciences Dr. Genome and Biomedical Sciences Facility Room 6404A Davis, CA 95616
Integrated gut microbiome and lipidomic analyses in animal models of Wilson disease reveal a role of intestine ATP7B in copper-related metabolic dysregulation
STUDY_SUMMARY
Although the main pathogenic mechanism of Wilson disease (WD) is related to copper accumulation in the liver and brain, there is limited knowledge about the role of ATP7B copper transporter in extra-hepatic organs, including the intestine, and how it could affect metabolic manifestations of the disease. The aims of the present study were to profile and correlate the gut microbiota and lipidome in mouse models of WD, and to study the metabolic effects of intestine-specific ATP7B deficiency in a newly generated mouse model. Animal models of WD presented reduced gut microbiota diversity compared to mice with normal copper metabolism. Comparative prediction analysis of the functional metagenome showed the involvement of several pathways including amino acid, carbohydrate, and lipid metabolisms. Lipidomic profiles showed dysregulated tri- and diglyceride, phospholipid, and sphingolipid metabolism. When challenged with a high-fat diet, Atp7bĪIEC mice confirmed profound deregulation of fatty acid desaturation and sphingolipid metabolism pathways as well as altered APOB48 distribution in intestinal epithelial cells. Gut microbiome and lipidomic analyses reveal integrated metabolic changes underlying the systemic manifestations of WD. Intestine-specific ATP7B deficit affects both intestine and systemic response to high-fat challenge. WD is as systemic disease and organ-specific ATP7B variants can explain the varied phenotypic presentations.
INSTITUTE
University of California, Davis
DEPARTMENT
Internal Medicine
LABORATORY
Medici's Lab
LAST_NAME
Sarode
FIRST_NAME
Gaurav Vilas
ADDRESS
451 E. Health Sciences Dr. Genome and Biomedical Sciences Facility Room 6404A Davis, CA 95616
Integrated gut microbiome and lipidomic analyses in animal models of Wilson disease reveal a role of intestine ATP7B in copper-related metabolic dysregulation
STUDY_SUMMARY
Although the main pathogenic mechanism of Wilson disease (WD) is related to copper accumulation in the liver and brain, there is limited knowledge about the role of ATP7B copper transporter in extra-hepatic organs, including the intestine, and how it could affect metabolic manifestations of the disease. The aims of the present study were to profile and correlate the gut microbiota and lipidome in mouse models of WD, and to study the metabolic effects of intestine-specific ATP7B deficiency in a newly generated mouse model. Animal models of WD presented reduced gut microbiota diversity compared to mice with normal copper metabolism. Comparative prediction analysis of the functional metagenome showed the involvement of several pathways including amino acid, carbohydrate, and lipid metabolisms. Lipidomic profiles showed dysregulated tri- and diglyceride, phospholipid, and sphingolipid metabolism. When challenged with a high-fat diet, Atp7bĪIEC mice confirmed profound deregulation of fatty acid desaturation and sphingolipid metabolism pathways as well as altered APOB48 distribution in intestinal epithelial cells. Gut microbiome and lipidomic analyses reveal integrated metabolic changes underlying the systemic manifestations of WD. Intestine-specific ATP7B deficit affects both intestine and systemic response to high-fat challenge. WD is as systemic disease and organ-specific ATP7B variants can explain the varied phenotypic presentations.
INSTITUTE
University of California, Davis
DEPARTMENT
Internal Medicine
LABORATORY
Medici's Lab
LAST_NAME
Sarode
FIRST_NAME
Gaurav Vilas
ADDRESS
451 E. Health Sciences Dr. Genome and Biomedical Sciences Facility Room 6404A Davis, CA 95616
Isolated murine skeletal muscles utilize pyruvate over glucose for oxidation:Part 2
STUDY_TYPE
Study of the different substrate by isolated skeletal muscle at room temperature via C-13 isotopomer analysis
STUDY_SUMMARY
Preclinical studies of muscle contractile function often employ ex vivo preparations of the soleus and/or extensor digitorum longus (EDL) muscles which are relatively easy to prepare and represent slow and fast fiber properties, respectively. Therefore, the current study sought to examine the utility of this preparation for understanding the metabolic fuel utilization in isolated resting mouse muscles at room temperature. 13C-labeling in both muscle types was performed using three fuels: glucose, pyruvate, and acetate, followed by NMR-based metabolomics analyses. Incubating 13C-labeled substrates in the isolated skeletal muscles makes it possible to examine TCA cycle flux and substrate selection by these muscles.
Dansylated Human Plasma at Known Light-Heavy Mixing Ratios
STUDY_TYPE
MS isotopic labeling analysis
STUDY_SUMMARY
Pooled human plasma was split and tagged with either light or heavy dansyl chloride and then recombined at known light-to-heavy ratios for LC-MS analysis and subsequent testing of isotopic labeling software.
INSTITUTE
University of North Carolina Greensboro
LAST_NAME
Zhang
FIRST_NAME
Qibin
ADDRESS
600 Laureate Way, Suite 2203, Kannapolis, NC 28081
Mass Spectrometry-based Proteomic and Metabolomic profiling of serum samples for discovery and validation of Tuberculosis diagnostic biomarker signature
STUDY_SUMMARY
Tuberculosis (TB) is a transmissible disease listed as one of the 10 leading causes of death worldwide (10 million infected in 2019). A swift and precise diagnosis is essential to forestall its transmission, for which is crucial the discovery of effective diagnostic biomarkers. In this study, we aimed to discover molecular biomarkers for the early diagnosis of tuberculosis. Two independent cohorts comprising 29 and 34 subjects were assayed by proteomics, and 49 were included for metabolomic analysis. All subjects were arranged into 3 experimental groups ā healthy controls (Controls), Latent TB infection (LTBI) and TB patients. LC-MS/MS blood serum protein and metabolite levels were submitted to univariate, multivariate and ROC analysis. From the 149 proteins quantified in the discovery set, 25 were found to be differentially abundant between Controls and TB patients. The AUC, specificity and sensitivity, determined by ROC statistical analysis of the model composed by four of these proteins considering both proteomic sets, were 0.96; 93% and 91%, respectively. The five metabolites (9-methyluric acid, indole-3-lactic acid, trans-3-indoleacrylic acid, hexanoylglycine and N-acetyl-L-leucine) that better discriminate the control and TB patient groups (VIP > 1.75) from a total of 92 metabolites quantified in both ionization modes, were submitted to ROC analysis. An AUC=1 was determined with all samples being correctly assigned to the respective experimental group. An integrated ROC analysis enrolling 1 protein and 4 metabolites was also performed for the common control and TB patients in the proteomic and metabolomic groups. This combined signature has correctly assigned the 12 controls and 12 patients used only for prediction (AUC=1, specificity=100% and sensitivity=100%). This multi-omics approach has revealed a biomarker signature for tuberculosis diagnosis that could be potentially used for developing a point-of-care diagnosis clinical test.
Insights from a Multi-Omics Integration (MOI) Study in Oil Palm (Elaeis guineensis Jacq.) Response to Abiotic Stresses: Part OneāSalinity
STUDY_TYPE
Multi-Omics Integration (MOI) Study
STUDY_SUMMARY
Oil palm (Elaeis guineensis Jacq.) is the number one source of consumed vegetable oil nowadays. It is cultivated in areas of tropical rainforest, where it meets its natural condition of high rainfall throughout the year. The palm oil industry faces criticism due to a series of practices that was considered not environmentally sustainable, and it finds itself under pressure to adopt new and innovative procedures to reverse this negative public perception. Cultivating this oilseed crop outside the rainforest zone is only possible using artificial irrigation. Close to 30% of the worldās irrigated agricultural lands also face problems due to salinity stress. Consequently, the research community must consider drought and salinity together when studying to empower breeding programs in order to develop superior genotypes adapted to those potential new areas for oil palm cultivation. Multi-Omics Integration (MOI) offers a new window of opportunity for the non-trivial challenge of unraveling the mechanisms behind multigenic traits, such as drought and salinity tolerance. The current study carried out a comprehensive, large-scale, single-omics analysis (SOA), and MOI study on the leaves of young oil palm plants submitted to very high salinity stress. Taken together, a total of 1239 proteins were positively regulated, and 1660 were negatively regulated in transcriptomics and proteomics analyses. Meanwhile, the metabolomics analysis revealed 37 metabolites that were upreg- ulated and 92 that were downregulated. After performing SOA, 436 differentially expressed (DE) full-length transcripts, 74 DE proteins, and 19 DE metabolites underwent MOI analysis, revealing sev- eral pathways affected by this stress, with at least one DE molecule in all three omics platforms used. The Cysteine and methionine metabolism (map00270) and Glycolysis/Gluconeogenesis (map00010) pathways were the most affected ones, each one with 20 DE molecules.
INSTITUTE
The Brazilian Agricultural Research Corporation (Embrapa)
DEPARTMENT
Embrapa Agroenergy
LABORATORY
Genetics and Plant Biotechnology
LAST_NAME
Souza Jr
FIRST_NAME
Manoel Teixeira
ADDRESS
Parque Estacao Biologica, Final Avenida W3 Norte - Asa Norte, Brasilia, Distrito Federal, 70770901, Brazil
Insights from a Multi-Omics Integration (MOI) Study in Oil Palm (Elaeis guineensis Jacq.) Response to Abiotic Stresses: Part TwoāDrought
STUDY_TYPE
Multi-Omics Integration (MOI) Study
STUDY_SUMMARY
Drought and salinity are two of the most severe abiotic stresses affecting agriculture Worldwide and bear some similarities in the response of plants to them. The first is also known as osmotic stress and shows similarities mainly with the osmotic effect, the first phase of salinity stress. Multi-Omics Integration (MOI) offers a new opportunity for the non-trivial challenge of unraveling the mechanisms behind multigenic traits, such as drought and salinity resistance. The current study carried out a comprehensive, large-scale, single-omics analysis (SOA) and MOI studies on the leaves of young oil palm plants submitted to water deprivation. After performing SOA, 1,955 DE enzymes from transcriptomics analysis, 131 DE enzymes from proteomics analysis, and 269 DE metabolites underwent MOI analysis, revealing several pathways affected by this stress, with at least one DE molecule in all three omics platforms used. Besides, the similarities and dissimilarities in the molecular response of those plants to those two abiotic stresses underwent mapping. Cysteine and methionine metabolism (map00270) was the most affected pathway in all scenarios evaluated. The correlation analysis revealed that 91.55% of those enzymes expressed under both stresses had similar qualitative profiles, corroborating the already known fact that plant responses to drought and salinity show several similarities. At last, the results shed light on some candidate genes for engineering crop species resilient to both abiotic stresses.
INSTITUTE
The Brazilian Agricultural Research Corporation (Embrapa)
DEPARTMENT
Embrapa Agroenergy
LABORATORY
Genetics and Plant Biotechnology
LAST_NAME
Souza Jr
FIRST_NAME
Manoel Teixeira
ADDRESS
Parque Estacao Biologica, Final Avenida W3 Norte - Asa Norte, Brasilia, Distrito Federal, 70770901, Brazil
MS profiling of the Long Term Evolution Experiment
STUDY_SUMMARY
The metabolome of a cell is the integration point of an organism's environment, genetics, and gene expression pattern. The metabolic phenotype can be under selection and is known to contribute to adaption. However, the metabolome's inherent networked and convoluted nature makes relating mutations, metabolic changes, and effects on fitness challenging. To overcome this challenge, we use the Long Term Evolution Experiment (LTEE) as a model to understand how mutations can transduce themselves through a cellular network, eventually affecting metabolism and perhaps fitness. We used mass-spectropscopy to broadly survey the metabolomes of both ancestors and all 12 evolved lines and combined this with genomic and expression data to suggest how mutations that alter specific reaction pathways, such as the biosynthesis of nicotinamide adenine dinucleotide, might increase fitness in the system. Our work brings the field closer to a complete genotype-phenotype map for the LTEE and a better understanding of how mutations might affect fitness through the metabolome. We used mass-spectroscopy to profile metabolic changes in the Long Term Evolution Experiment and link these change to upstream changes in gene expression and mutations.
Metabolic impacts of metformin to seasonal influenza vaccination: a pilot study of drug interaction with the immune response
STUDY_SUMMARY
We report here a double-blinded pilot study of seasonal influenza vaccination, where half of the participants received daily metformin administration. Global metabolomics was measured in the plasma samples at six timepoints. Metformin signatures were successfully identified in the metabolomics data. Statistically significant metabolite features were found both for the vaccination effect and for the drug-vaccine interactions.
Elucidating dynamic anaerobe metabolism with HRMAS 13C NMR and genome-scale modeling
STUDY_SUMMARY
Anaerobic microbial metabolism drives critical functions within global ecosystems, host-microbiota interactions, and industrial applications, yet remains ill-defined. Here we advance a versatile approach to elaborate cellular metabolism in obligate anaerobes using the pathogen Clostridioides difficile, an amino acid and carbohydrate-fermenting Clostridia. High-Resolution Magic Angle Spinning (HRMAS) Nuclear Magnetic Resonance (NMR) spectroscopy of C. difficile, grown with fermentable 13C substrates, informed dynamic flux balance analysis (dFBA) of the pathogenās genome-scale metabolism. Analyses identified dynamic recruitment of oxidative and supporting reductive pathways, with integration of high-flux amino acid and glycolytic metabolism at alanineās biosynthesis to support efficient energy generation, nitrogen handling, and biomass generation. Model predictions informed an approach leveraging the sensitivity of 13C NMR spectroscopy to simultaneously track cellular carbon and nitrogen flow from [U-13C]glucose and [15N]leucine, confirming the formation of [13C,15N]alanine. Findings identify metabolic strategies used by C. difficile to support its rapid colonization and expansion in gut ecosystems.
INSTITUTE
Brigham and Women's Hospital
DEPARTMENT
Pathology
LABORATORY
Bry Lab, Massachusetts Host-Microbiome Center; Cheng Lab, Massachusetts General Hospital
LAST_NAME
Pavao
FIRST_NAME
Aidan
ADDRESS
221 Longwood Ave, EBRC-411, Boston, MA, 02115, USA
Metabolomics analysis of heart from CHCHD10S59L/+ KI mice
STUDY_SUMMARY
Mutations in the coiled-coil-helix-coiled-coil-helix domain containing 10 (CHCHD10) gene have been associated with a large clinical spectrum including myopathy, cardiomyopathy and amyotrophic lateral sclerosis (ALS). Herein, we analyzed the metabolic changes induced by the p.S59L CHCHD10 mutation to identify new therapeutic opportunities. Using metabolomic, lipidomic and proteomic analysis we observed a strong alteration of metabolism in plasma and heart of Chchd10S59L/+ mice compared to their wild type littermates at pre-symptomatic and symptomatic stages. In plasma, levels of phospholipids were decreased while those of carnitine derivatives and most of amino acids were increased. The cardiac tissue from Chchd10S59L/+ mice showed a decreased Oxidative Phosphorylation (OXPHOS) and Ī²-oxidation proteins levels as well as tricarboxylic acid cycle (TCA) intermediates and carnitine pathway metabolism. In parallel, lipidomics analysis reveals a drastic change in the lipidome, including triglyceride, cardiolipin and phospholipids. Consistent with this energetic deficiency in cardiac tissue, we show that L-acetylcarnitine supplementation improves the mitochondrial network length in IPS-derived cardiomyocytes from a patient carrying the CHCHD10S59L/+ mutation. These data indicate that a bioenergetic intermediate such as L-acetylcarnitine may restore mitochondrial function in CHCHD10-related disease, due to the reduction in energy deficit that could be compensated by carnitine metabolic pathways.
Metabolomics analysis of plasma from CHCHD10S59L/+ KI mice
STUDY_SUMMARY
Mutations in the coiled-coil-helix-coiled-coil-helix domain containing 10 (CHCHD10) gene have been associated with a large clinical spectrum including myopathy, cardiomyopathy and amyotrophic lateral sclerosis (ALS). Herein, we analyzed the metabolic changes induced by the p.S59L CHCHD10 mutation to identify new therapeutic opportunities. Using metabolomic, lipidomic and proteomic analysis we observed a strong alteration of metabolism in plasma and heart of Chchd10S59L/+ mice compared to their wild type littermates at pre-symptomatic and symptomatic stages. In plasma, levels of phospholipids were decreased while those of carnitine derivatives and most of amino acids were increased. The cardiac tissue from Chchd10S59L/+ mice showed a decreased Oxidative Phosphorylation (OXPHOS) and Ī²-oxidation proteins levels as well as tricarboxylic acid cycle (TCA) intermediates and carnitine pathway metabolism. In parallel, lipidomics analysis reveals a drastic change in the lipidome, including triglyceride, cardiolipin and phospholipids. Consistent with this energetic deficiency in cardiac tissue, we show that L-acetylcarnitine supplementation improves the mitochondrial network length in IPS-derived cardiomyocytes from a patient carrying the CHCHD10S59L/+ mutation. These data indicate that a bioenergetic intermediate such as L-acetylcarnitine may restore mitochondrial function in CHCHD10-related disease, due to the reduction in energy deficit that could be compensated by carnitine metabolic pathways.
Discovery of phytochelatins in human urine: Evidence for function in selenium disposition and protection against cadmium
STUDY_SUMMARY
The goal of this project was to detect phytochelatins, plant-derived peptides which function as metal chelators, in human urine. Untargeted metabolomics of 143 urine samples from healthy adults was performed. Phytochelatin 2, Ī³E-C-Ī³E-C-G, was detected, and the rest of the urine metabolome was searched for phytochelatins and predicted phytochelatin metabolites which correlated with phytochelatin 2 concentrations. Phytochelatin 2 and associated metabolites were found to correlate with urinary metals, and further experiments were performed provide insight into function of dietary phytochelatins.
Metabolomic analysis in wildtype and Isg15 knockout mice after sham or transverse aortic constriction surgery
STUDY_SUMMARY
Left ventricular tissues were isolated from wildtype and Isg15 knockout mice 8 weeks after sham or transverse aortic constriction and the metabolomic differences were compared.
INSTITUTE
St. Michael's Hospital
LAST_NAME
Advani
FIRST_NAME
Andrew
ADDRESS
6-151, 61 Queen Street East, Toronto, Ontario. M5C 2t2
Ozone alters glycosphingolipid metabolism and exacerbates characteristics of asthma in mice
STUDY_SUMMARY
Asthma is a common chronic respiratory disease exacerbated by multiple environmental factors, including exposure to air pollutants such as ozone. Acute ozone exposure has previously been implicated in airway inflammation, airway hyperreactivity, and other characteristics of asthma. Altered sphingolipid metabolism following ozone exposure may contribute to the molecular mechanisms underlying these previously reported effects. This study aimed to identify changes in metabolomic profiles and characteristics of asthma in allergen-sensitized mice following ozone exposure to provide insights regarding mechanisms of ozone-induced exacerbations in asthma. Adult male and female BALB/c mice were sensitized intranasally to house dust mite allergen (HDM) on days 1, 3, and 5 followed by HDM challenge on days 12-14. Mice were subsequently exposed to ozone following each HDM challenge for 6 hr/day. Bronchoalveolar lavage, plasma, whole lung lobes, and microdissected lung airways were collected from 8 female and 8 male mice for metabolomics analysis. 6 female and 6 male mice underwent methacholine challenge using a forced oscillation technique to assess pulmonary function. HDM-sensitized male mice exposed to ozone displayed synergistically increased airway hyperreactivity as well as increased airway inflammation and eosinophilia relative to control mice. Effects in male mice were significantly more severe than the effects observed in females. Both HDM-sensitized male and female mice exposed to ozone displayed significant decreases in multiple classes of sphingolipids in microdissected airways. However, glycosphingolipids were significantly increased in females and to a lesser extent in males. These results potentially implicate glycosphingolipids in protecting against severe outcomes of ozone exposure that coincide with exacerbation of allergic asthma.
Characterizing the intrauterine environment via untargeted metabolomics profiling of maternal blood collected during pregnancy (PETALS Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
"PETALS is a prospective cohort of multi-racial/ethnic pregnant women recruited in early pregnancy at Kaiser Permanente Northern California which aimed to examine environmental factors in association with pregnancy, perinatal, and childhood outcomes. Please contact the cohort PI Assiamira Ferrara (Assiamira.Ferrara@kp.org) and Co-I Yeyi Zhu (yeyi.zhu@kp.org) for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) OIF program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. PETALS is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539"
Impacts of interactions between environmental chemical exposures and diet on gut microbiota and microbiota-derived metabolites in mothers and children (MAAP Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
In 2014, the Microbes, Allergy, Asthma and Pets (MAAP) study (P01 AI089473) was launched. All women ages 18-49 years scheduled for prenatal visits at Henry Ford Health, expected to deliver during the recruitment period at a Henry Ford Health hospital, and whose residence was in a predefined area of Detroit and metropolitan Detroit suburbs were potentially eligible. There were no medical insurance requirements. Between 2014 and 2016, 141 women were enrolled; their infant was followed through age 18 months (with continuing follow-up through early/mid childhood underway through the NIH ECHO program under the CREW birth cohort consortium (5UG3OD023282, Gern PI). Blood and stool samples were obtained from the mother prenatally and stool specimens were obtained from the infant at 5 time points (1-week, 1-, 3-, 6- and 18-months) with 16S sequencing already completed. Blood was also obtained from the infant at pre-defined intervals. Please contact Andrea Cassidy-Bushrow at acassid1@hfhs.org for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) OIF program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. MAAP is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Impacts of interactions between environmental chemical exposures and diet on gut microbiota and microbiota-derived metabolites in mothers and children (MARCH Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
MARCH is a prospective population based pregnancy cohort that recruited pregnant participants from 2017-2023 from 11 sites within Michigan. Women over 18 years of age were recruited at first prenatal visit and had 3 data collection points focused around each trimester of pregnancy. Blood, urine, and questionnaire data were collected at each prenatal visit. At birth placenta was collected and after birth, infants were assessed at 3 months, 9 months, and yearly. For more information on MARCH please contact Jean Kerver; kerverje@msu.edu This research was supported by the Environmental influences on Child Health Outcomes (ECHO) OIF program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. MARCH is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Alterations in CSF Urea Occur in Late Manifest Stage Huntington Disease
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Huntington Disease (HD) is a neurodegenerative disorder caused by expanded cytosine-adenine-guanine (CAG) repeats in the Huntingtin gene, resulting in the production of mutant huntingtin proteins (mHTT). Previous research has identified urea as a key metabolite elevated in HD animal models and post-mortem tissues of HD patients. The exact timing of these elevations in urea and the molecular mechanism(s) responsible for these disturbances remain unknown. To better understand the pathophysiologic mechanisms responsible for elevations in urea in HD, we completed a global metabolomic profile of cerebrospinal fluid (CSF) from individuals who were at several stages of disease: pre-manifest (PRE), manifest (MAN), and late-manifest (LATE) HD participants compared to controls. We found approximately 500 metabolites were significantly altered in pre-manifest participants compared to controls, although no significant difference in CSF urea or urea metabolites. Interestingly, CSF urea was only significantly elevated in LATE participants compared to controls. There were no changes in the urea metabolites, citrulline, ornithine and arginine throughout disease; however, we did observe changes in acetate, creatinine, 4-acetamidobutanoate and 4-aminobutyraldehyde which are indirect modifiers of urea. Overall, our study confirms that elevations in urea do occur in HD, albeit later in disease and that these changes may reflect more central impairments to cellular energy metabolism yet to be explored.
Characterizing the intrauterine environment via untargeted metabolomics profiling of maternal blood collected during pregnancy (ARCH Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
"The ARCH Cohort is a pregnancy cohort of approximately 1,000 women recruited at the first prenatal visit largely in Lansing, MI between 2008 and 2016. Blood was collected when possible at the onset of prenatal care and at the time of the glucose tolerance test (late 2nd, early 3rd trimester). Please contact Jean Kerver at kerverje@msu.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) OIF program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. ARCH is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539"
Zebrafish Optic Nerve Regeneration Metabolomics - 3 Days Post Crush
STUDY_SUMMARY
Zebrafish (Danio Rerio) have the capacity for successful adult optic nerve regeneration. In contrast, mammals lack this intrinsic ability and undergo irreversible neurodegeneration seen in glaucoma and other optic neuropathies. Optic nerve regeneration is often studied using optic nerve crush, a mechanical neurodegenerative model. Untargeted metabolomic studies within successful regenerative models are deficient. Evaluation of tissue metabolomic changes in active zebrafish optic nerve regeneration can elucidate prioritized metabolite pathways that can be targeted in mammalian systems for therapeutic development. Female and male (6 month to 1 year old) right Zebrafish (Tg(gap43:GFP)) optic nerves were crushed and collected three days after. Contralateral, uninjured optic nerves were collected as controls. The tissue was dissected from euthanized fish and frozen on dry ice. Samples were pooled for each category (female crush, female control, male crush, male control) and pooled at n = 31 to obtain sufficient metabolite concentrations for analysis. Optic nerve regeneration was verified by microscope visualization of GFP fluorescence. Metabolites were extracted using a Precellys Homogenizer and a serial extraction method: (1) 1:1 Methanol/Water and (2) 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. Metabolites were identified and quantified using Compound Discoverer 3.3 and isotopic internal metabolites standards.
The interaction of ceramide-1-phosphate with group IVA cytosolic phospholipase A2 modulates neutrophil polarization during inflammatory responses
STUDY_SUMMARY
Bone marrow-derived mouse neutrophils from wildtype, cPLA2alpha-knockin (interaction site on cPLA2alpha for its substrate C1P was ablated), and cPLA2alpha-knockout (cPLA2alpha gene mutated) mice were exposed to 4 hours of trans-endothelial migration and resulting eicosanoids were analyzed (eg, PGE2, PGD2, 5-HETE, and 5-oxo-ETE).
Untargeted metabolomics of miR-142 WT vs KO CML cells
STUDY_SUMMARY
MiR-142 is dynamically expressed and plays a regulatory role in hematopoiesis. Based on the simple observation that miR-142 levels are significantly lower in CD34+CD38- cells from blast crisis (BC) chronic myeloid leukemia (CML). CML patients compared with chronic phase (CP) CML patients (p=0.002), we hypothesized that miR-142 deficit plays a role in BC transformation. To test this hypothesis, we generated a miR-142 KO BCR-ABL (i.e., miR-142ā/āBCR-ABL) mouse by crossing a miR-142ā/ā mouse with a miR-142+/+BCR-ABL mouse. While the miR-142+/+BCR-ABL mice developed and died of CP CML, the miR-142ā/āBCR-ABL mice developed a BC-like phenotype in the absence of any other acquired gene mutations and died significantly sooner than miR-142+/+BCR-ABL CP controls (p=0.001). Leukemic stem cell (LSC)-enriched Lineage-Sca-1+c-Kit+ cells (LSKs) from diseased miR-142ā/āBCR-ABL mice transplanted into congenic recipients, recapitulated the BC features thereby suggesting stable transformation of CP-LSCs into BC-LSCs in the miR-142 KO CML mouse. Single cell (sc) RNA-seq profiling showed that miR-142 deficit changed the cellular landscape of the miR-142ā/āBCR-ABL LSKs compared with miR-142+/+BCR-ABL LSKs with expansion of myeloid-primed and loss of lymphoid-primed factions. Bulk RNA-seq analyses along with unbiased metabolomic profiling and functional metabolic assays demonstrated enhanced fatty acid Ī²-oxidation (FAO) and oxidative phosphorylation (OxPhos) in miR-142ā/āBCR-ABL LSKs vs miR-142+/+BCR-ABL LSKs. MiR-142 deficit enhanced FAO in miR-142ā/āBCR-ABL LSKs by increasing the expression of CPT1A and CPT1B, that controls the cytosol-to-mitochondrial acyl-carnitine transport, a critical step in FAO. MiR-142 deficit also enhanced OxPhos in miR-142ā/āBCR-ABL LSKs by increasing mitochondrial fusion and activity. As the homeostasis and activity of LSCs depend on higher levels of these oxidative metabolism processes, we then postulate that miR-142 deficit is a potentially druggable target for BC-LSCs. To this end, we developed a novel CpG-miR-142 mimic oligonucleotide (ODN; i.e., CpG-M-miR-142) that corrected the miR-142 deficit and alone or in combination with a tyrosine kinase inhibitor (TKI) significantly reduced LSC burden and prolonged survival of miR-142ā/āBCR-ABL mice. The results from murine models were validated in BC CD34+CD38- primary blasts and patient-derived xenografts (PDXs). In conclusion, an acquired miR-142 deficit sufficed in transforming CP-LSCs into BC-LSCs, via enhancement of bioenergetic oxidative metabolism in absence of any additional gene mutations, and likely represent a novel therapeutic target in BC CML.
Bioactive molecule(s) of gut bacteria of Crocodile (Crocodylus palustris) as potential pharmaceuticals
STUDY_SUMMARY
Crocodiles thrive in unsanitary conditions, feed on rotten meat, are exposed to heavy metals and are among the very few species to endure the catastrophic Cretaceous-Tertiary extinction event, and yet they can live up to 100 years. We hypothesized that crocodiles have developed mechanisms to achieve such longevity while surviving under stressful conditions. We speculate that their microbial gut flora may produce substances contributing to their āhardinessā and ālongevityā. Previously we characterized selected microbial gut bacteria colonizing the gastrointestinal tract of Crocodylus porosus (CP) using 16S rDNA sequencing. Next, bacterial conditioned media containing gut microbial metabolites were prepared. Bioassay-guided testing of selected bacterial conditioned media using LC-TIMS-QTOF MS, revealed the identity of gut microbial metabolites. Among two bacterial conditioned media, i.e., CP27 and 36, the analyses resulted in 141 highly confidently (MS/MS) identified metabolites in both samples. The pairwise comparison of the two samples indicated that 109 metabolites change significantly between them (p <0.05). Among abundant metabolites more prevalent in CP36 there were 2-Methyl-4-nitroimidazole, N-Acetyl-L-tyrosine, Acetaminophen, Trans-Ferulic acid, N, N-Dimethylformamide, Pyrocatechol, Cyclohexanone, 3, 4-Dihydrozphenylglycol, Diphenhydramine, Melatonin, Gamma āterpinene. Whereas in CP27 samples the most abundant metabolites were Carbamazepin, deoxyninosine, Cysteamine, Benzylnicotinate, 3-phenoxypropionic acid, Indole-3-carbinol, Benzaldehyde, Benzocaine, 2-Aminobenzoic acid, 3-Methylindole. Functional enrichment analysis of all identified metabolites with metabolite sets based on drug pathways showed that they were enriched for drug action of top ten pathways associating with enalapril metabolism pathway, diphenhydramine H1-Antihistamine action, enalarpil action pathway, benzocaine action pathway, mepivacaine action pathway, oxybuprocaine action pathways, nifedipine action pathway, propranolol action pathway, acetaminophen metabolism pathway, carbamazepine metabolism pathway. These findings suggest that analyses of crocodile gut bacteria may reveal potential drug leads, intensive future research is needed to realize these expectations.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Metabolic analysis of primary HPV18-genome containing human foreskin keratinocytes compared to untransfected donor-matched controls.
STUDY_SUMMARY
Six primary HPV18-genome containing human foreskin keratinocyte cell populations and six donor-matched primary untransfected human foreskin keratinocyte cell populations were grown on lethally-irradiated 3T3-J2 fibroblasts. Before harvesting the keratinocytes, the 3T3-J2 fibroblasts were washed off. Cells and spent media were harvested and frozen at -80Ā°C until processing.
INSTITUTE
University of Birmingham, UK
DEPARTMENT
Institute of Cancer and Genomic Sciences
LABORATORY
Joanna Parish
LAST_NAME
Parish
FIRST_NAME
Joanna
ADDRESS
IBR Wolfson Drive Medical School, University of Birmingham, Edgbaston
Identify putative volatile biomarkers of Valley fever using a murine lung infection model (Human studies)
STUDY_SUMMARY
Coccidioides immitis and Coccidioides posadasii are soil-dwelling fungi of arid regions in North and South America that are responsible for Valley fever (coccidioidomycosis). Forty percent of patients with Valley fever exhibit symptoms ranging from mild, self-limiting respiratory infections, to severe, life-threatening pneumonia that requires treatment. Misdiagnosis as bacterial pneumonia commonly occurs in symptomatic Valley fever cases, resulting in inappropriate treatment with antibiotics, increased medical costs, and delay in diagnosis. In this study, we explored the feasibility of developing breath-based diagnostics for Valley fever using lung specimens from persons with community-acquired pneumonia (CAP). To investigate potential volatile biomarkers of Valley fever that arise from host-pathogen interactions, we collected bronchoalveolar lavage fluid (BALF) and sputum from patients treated at Mayo Clinic in Scottsdale, Arizona for untargeted volatile metabolomics via solid phase microextraction (SPME) and two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GCĆGC-TOFMS). We identified 244 total volatile organic compounds (VOCs). Using Random Forest, we identified a subset of these VOCs that were also able to separate Coccidioides positive samples from bacteria positive samples. The data presented here show that Coccidioides and/or the host produce volatile metabolites that may yield biomarkers for a Valley fever breath test that can detect Coccidioidal infection.
APOE modulates microglial immunometabolism in response to age, amyloid pathology, and inflammatory challenge (Part 1 of 3)
STUDY_SUMMARY
The E4 allele of Apolipoprotein E (APOE) is associated with both metabolic dysfunction and a heightened pro-inflammatory response ā two findings that may be intrinsically linked through the concept of immunometabolism. Here, we combined bulk, single-cell, and spatial transcriptomics with cell-specific and spatially resolved metabolic analyses to systematically address the role of APOE across age, neuroinflammation, and AD pathology. RNAseq highlighted immunometabolic changes across the APOE4 glial transcriptome, specifically in subsets of metabolically distinct microglia enriched in the E4 brain during aging or following an inflammatory challenge. E4 microglia display increased Hif1Ī± expression, a disrupted TCA cycle, and are inherently pro-glycolytic, while spatial transcriptomics and MALDI mass spectrometry imaging highlight an E4-specific response to amyloid that is characterized by widespread alterations in lipid metabolism. Taken together, our findings emphasize a central role for APOE in regulating microglial immunometabolism.
INSTITUTE
University of Kentucky, Department of Physiology
LAST_NAME
Devanney
FIRST_NAME
Nicholas
ADDRESS
Physiology, 760 Press Ave, Healthy Kentucky Research Bldg, Rm152, Lexington, Kentucky, 40508, USA
APOE modulates microglial immunometabolism in response to age, amyloid pathology, and inflammatory challenge (Part 2 of 3)
STUDY_SUMMARY
The E4 allele of Apolipoprotein E (APOE) is associated with both metabolic dysfunction and a heightened pro-inflammatory response ā two findings that may be intrinsically linked through the concept of immunometabolism. Here, we combined bulk, single-cell, and spatial transcriptomics with cell-specific and spatially resolved metabolic analyses to systematically address the role of APOE across age, neuroinflammation, and AD pathology. RNAseq highlighted immunometabolic changes across the APOE4 glial transcriptome, specifically in subsets of metabolically distinct microglia enriched in the E4 brain during aging or following an inflammatory challenge. E4 microglia display increased Hif1Ī± expression, a disrupted TCA cycle, and are inherently pro-glycolytic, while spatial transcriptomics and MALDI mass spectrometry imaging highlight an E4-specific response to amyloid that is characterized by widespread alterations in lipid metabolism. Taken together, our findings emphasize a central role for APOE in regulating microglial immunometabolism.
INSTITUTE
University of Kentucky
DEPARTMENT
Physiology
LABORATORY
Lance Johnson; Josh Morganti
LAST_NAME
Devanney
FIRST_NAME
Nicholas
ADDRESS
Physiology, 760 Press Ave, Healthy Kentucky Research Bldg, Rm152, Lexington, Kentucky, 40508, USA
Lipidomic analysis of human brain from frontotemporal dementia cases of with GRN and C9orf72 mutations
STUDY_SUMMARY
Lipidomic analysis carried out on postmortem human brain tissue from cases with FTD carrying inherited mutations in the GRN gene, or repeat expansions in the C9orf72 gene, and age-matched control cases. Tissue was sampled from the heavily affected superior frontal grey and white matter, and less heavily affected superior parietal grey and white matter.
INSTITUTE
The University of Sydney
LAST_NAME
Don
FIRST_NAME
Anthony
ADDRESS
Office 3217, D17 Charles Perkins Centre, Camperdown, NSW, 2006, Australia
APOE modulates microglial immunometabolism in response to age, amyloid pathology, and inflammatory challenge (Part 3 of 3)
STUDY_SUMMARY
The E4 allele of Apolipoprotein E (APOE) is associated with both metabolic dysfunction and a heightened pro-inflammatory response ā two findings that may be intrinsically linked through the concept of immunometabolism. Here, we combined bulk, single-cell, and spatial transcriptomics with cell-specific and spatially resolved metabolic analyses to systematically address the role of APOE across age, neuroinflammation, and AD pathology. RNAseq highlighted immunometabolic changes across the APOE4 glial transcriptome, specifically in subsets of metabolically distinct microglia enriched in the E4 brain during aging or following an inflammatory challenge. E4 microglia display increased Hif1Ī± expression, a disrupted TCA cycle, and are inherently pro-glycolytic, while spatial transcriptomics and MALDI mass spectrometry imaging highlight an E4-specific response to amyloid that is characterized by widespread alterations in lipid metabolism. Taken together, our findings emphasize a central role for APOE in regulating microglial immunometabolism.
INSTITUTE
University of Kentucky
DEPARTMENT
Physiology
LABORATORY
Lance Johnson; Josh Morganti
LAST_NAME
Devanney
FIRST_NAME
Nicholas
ADDRESS
Physiology, 760 Press Ave, Healthy Kentucky Research Bldg, Rm152, Lexington, Kentucky, 40508, USA
Bloody Mary (BM21)- Serial mixtures of vegetable juice /water and human plasma.
STUDY_SUMMARY
Metabolomics holds the promise to measure and quantify small molecules comprehensively in biological systems, and LC-MS (liquid chromatography coupled mass spectrometry) has become the leading technology in the field. Significant challenges still exist in the computational processing of data from LC-MS metabolomic experiments into metabolite features, including provenance and reproducibility of the current software tools. We present here, an experiment designed as serial mixtures of vegetable juice/Water and human plasma at varying ratios, nicknamed āBloody Mary 21(BM21) to test semi-quantification at āomics scale. A subset of features are expected to have their peak areas correlated with the mixing ratio. This dataset provides an opportunity to be used as benchmark to assess the performance in quantification of processing softwares. Overall, the BM21 experiment included a serial mixture of human plasma (Qstd) and vegetable juice (or water), at the ratio of 1024:1, 256:1, 64:1, 16:1, 4:1, 1:1, 1:4, 1:16, 1:64, 1:256 and 1:1024. Along with the 11 serial mixture samples, 100% vegetable juice and 100% plasma were also included. All samples were analyzed in triplicates.
Organism-Wide Analysis of Sepsis Reveals Mechanisms of Systemic Inflammation
STUDY_TYPE
lipidomics analysis
STUDY_SUMMARY
Phospholipase A2 group V (PLA2G5) is a secretory and Ca2+-dependent lipolytic enzyme and is inducible during several pathologic conditions. However, it has been unknown how PLA2G5 plays a role in sepsis. To study the role of PLA2G5 in sepsis, we performed lipidomics analysis of plasma and tissues from LPS-injected mice with or without PLA2G5 blockade. Here, we showed that PLA2G5 is involved in the production of fatty acids such as oleic acid and linoleic acid, lysophospholipids such as lysophosphatidic acid, lysophosphatidylcholine, lysophatidylethanolamine, and lysophosphatidylserine species, and metabolites derived from polyunsaturated fatty acids such as arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, and linoleic acids during sepsis. Thus, PLA2G5 regulates selective lipid pathways during sepsis.
1H NMR metabolomics applied to assess the metabolic response of Ruditapes philippinarum clams to sea warming and 17-Ī±-ethinylestradiol exposure
STUDY_TYPE
1H NMR metabolomics to study the effects of warming conditions and exposure to 17-Ī±-ethinylestradiol (EE2) on the polar metabolome of Ruditapes philippinarum clams
STUDY_SUMMARY
In this study, a comprehensive untargeted 1H NMR metabolomics strategy was applied to measure the metabolic impact of sea warming, in tandem with exposure to EE2, on Ruditapes philippinarum clams. The clams were exposed to five different EE2 concentrations: 0 (control group), 5, 25, 125 and 625 ng/L; either at 17 Ā°C as control temperature or at 21 Ā°C (representing a 4 Ā°C increase, which corresponds to the worst-case warming scenario). The obtained data added important knowledge, unveiling individual metabolic effects of temperature rise and synergetic effects upon EE2 exposure, and paving the way for the definition of new metabolic markers for the monitoring of environmental stressors.
INSTITUTE
University of Aveiro
DEPARTMENT
CICECO ā Aveiro Institute of Materials, Department of Chemistry
LABORATORY
Metabolomics Group- CICECO
LAST_NAME
Rodrigues
FIRST_NAME
Joao A.
ADDRESS
University of Aveiro, Campus Universitario de Santiago, 3810-193 Aveiro, Portugal
EMAIL
joao.e.a.rodrigues@gmail.com
PHONE
00351963481841
NUM_GROUPS
10
TOTAL_SUBJECTS
103
STUDY_COMMENTS
This work was developed within the CICECO-Aveiro Institute of Materials project (UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020) financed by national funds through the FCT/MEC (PIDDAC). We are also grateful to the Portuguese National NMR Network (PTNMR), supported by FCT funds as the NMR spectrometer used is part of PTNMR and partially supported by Infrastructure Project No. 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL, and the FCT through PIDDAC). This work was also financially supported by the project BISPECIAl: BIvalveS under Polluted Environment and ClImate chAnge (POCI-01-0145-FEDER- 028425) funded by FEDER, through COMPETE2020 - Programa Operacional Competitividade e InternacionalizaĆ§Ć£o (POCI), and by national funds (OE), through FCT/MCTES. MĆ³nica G. Silva benefited from Research Grant (MSc) (BI/CESAM/0043_2019/POCI-01-0145-FEDER-028425) under the project BISPECIAl: BIvalveS under Polluted Environment and ClImate changeĀ PTDC/CTA-AMB/28425/2017Ā (POCI-01-0145-FEDER-028425).
Here, we reveal for the first time the misregulated metabolic pathways in TSC kidneys and their relevance to TSC-associated cytogenesis. To this end, we have analyzed the metabolic profile of the whole kidney as well as sorted proximal tubule cell (PTCs) extracts. The metabolomics data show that Tsc1 deletion in nephron progenitor cells changes the arginine biosynthesis pathway as well as a substantial number of metabolic pathways.
Here, we reveal for the first time the misregulated metabolic pathways in TSC kidneys and their relevance to TSC-associated cytogenesis. To this end, we have analyzed the metabolic profile of the whole kidney as well as sorted proximal tubule cell (PTCs) extracts. The metabolomics data show that Tsc1 deletion in nephron progenitor cells changes the arginine biosynthesis pathway as well as a substantial number of metabolic pathways.
Comparison of metabolite profiles from matched whole blood microsamplers, whole blood dried blood spots, and plasma.
STUDY_SUMMARY
Venous blood was collected from 54 adult female participants from the PRISM cohort. Whole blood and venous blood was aliquoted. Untargeted metabolomics was performed on whole blood collected on Mitra microsamplers (VAMS, 10 uL), whole blood dried blood spots (DBS, 5-mm punch), and 10 uL of plasma
The prawn Palaemon serratus exhibits a large distribution (occurring along the Northeastern Atlantic coast to the Mediterranean), and has thus been found suitable as model organism valuable for various ecotoxicological studies. However, little is still known about the potential input of its metabolome and particularly concerning a potential molecular sexual dimorphism observable in the different tissues of this organism. In an ecotoxicological point of view, inter-sex and inter-organ differences of the metabolomes may introduce analytical bias and impact the robustness of the analysis and its interpretation. To explore such possibilities, we obtained qualitative metabolomic data from the analysis of different organs of mature male and female Palaemon serratus. We used ultra-high-performance liquid chromatography-electrospray ionization-high resolution tandem mass spectrometry (UHPLC-ESI-HRMS on positive mode) to characterize the 75%-extracted metabolome of both gills, hepatopancreas, nervous gland, muscle and gonads.
Untargeted metabolomics of HUVECs subjected to hypoxia-reoxygenation
STUDY_SUMMARY
Acute hemorrhage commonly leads to coagulopathy and organ dysfunction or failure. Recent evidence suggests that damage to the endothelial glycocalyx contributes to these adverse outcomes. The physiological events mediating acute glycocalyx shedding are undefined, however. Here, we show that succinate accumulation within endothelial cells drives glycocalyx degradation through a membrane reorganization-mediated mechanism. We investigated this mechanism in a cultured endothelial cell hypoxia-reoxygenation model, in a rat model of hemorrhage, and in trauma patient plasma samples. We found that succinate metabolism by succinate dehydrogenase mediates glycocalyx damage through lipid oxidation and phospholipase A2-mediated membrane reorganization (increasing lysophospholipids), promoting the interaction of MMP24 and MMP25 with glycocalyx constituents. In trauma patients, we found that succinate levels were associated with glycocalyx damage and the development of coagulopathy, and that interaction of MMP24 and syndecan-1 were elevated compared to healthy controls. This establishes a novel metabolic cascade mediating the endotheliopathy of traumatic hemorrhage.
INSTITUTE
Tulane University School of Medicine
DEPARTMENT
Surgery
LABORATORY
Tulane Trauma and Critical Care Research Lab
LAST_NAME
Jackson-Weaver
FIRST_NAME
Olan
ADDRESS
1430 Tulane Ave, Department of Surgery, School of Medicine
Map of microbially induced metabolic changes across diverse body sites in mice - Mouse Data
STUDY_SUMMARY
Tissue samples from contents along the intestinal tract and systemic sites in mice that did not have any bacteria (germ free) or colonized with a simplified microbiota or more complex microbiota.
Deciphering the metabolomic differences between two fast-growing cyanobacteria, S.elongatus PCC 11801 and 11802 via 13C-metabolic flux analysis
STUDY_SUMMARY
The study aims to identify the metabolic differences between two promising fast-growing, non-model cyanobacterial strains, S. elongatus PCC 11801 and PCC 11802. To this end, dynamic 13C-labeling experiments were carried out in the two cyanobacterial strains grown in shake flasks at a similar light intensity of approx. 300-350 Āµmole photons.m-2. s-1. The samples for metabolomics analysis were collected during the exponential growth phase at an optical cell density of 0.5-0.6. The detailed protocol for experiment can be found in the protocol file.
INSTITUTE
Indian Institute of Technology Bombay
DEPARTMENT
Chemical Engineering
LAST_NAME
Wangikar
FIRST_NAME
Pramod
ADDRESS
Biosystems and Bioengineering Lab, Department of Chemical Engineering, IIT Bombay, Powai, Mumbai, Maharashtra, India -400076
Nano-hijacked myeloid cells potentiate antitumor immunity and radiotherapy for glioblastoma
STUDY_TYPE
IR versus IR + LNP
STUDY_SUMMARY
Abstract: Radiation therapy is a key component of the standard of care for glioblastoma (GBM). Although this treatment is known to trigger pro-inflammatory immune responses, it also results in several immune resistance mechanisms such as the upregulation of CD47 by tumors leading to avoidance of phagocytosis and the overexpression of PD-L1 in tumor-associated myeloid cells (TAMCs). Leveraging these RT-elicited processes, we generated a bispecific-lipid nanoparticle (B-LNP) that engaged TAMCs to glioma cells via anti-CD47/PD-L1 dual-ligation. We show that B-LNP blocked these two vital immune checkpoint molecules and promoted the phagocytic activity of TAMCs. In order to boost subsequent T cell recruitment and antitumor activity after tumor engulfment, the B-LNP was encapsulated with diABZI, a non-nucleotidyl agonist for stimulator of interferon genes (STING). In vivo treatment with the diABZI-loaded B-LNP induced a transcriptomic and metabolic switch in TAMCs, transforming them into potent antitumor effector cells, which induced T cell infiltration and activation of in the brain tumors. In preclinical murine glioma models, B-LNP therapy significantly potentiated the antitumor effects of radiotherapy, promoted brain tumor regression, and induced immunological memory against gliomas. The nano37 therapy was efficacious through both intra-tumoral and systemic delivery routes. In summary, our study shows a unique nanotechnology-based approach that hijacks multiple immune checkpoints to boost potent and long-lasting antitumor immunity against GBM.
INSTITUTE
Northwestern University, Feinberg School of Medicine
First approach on the use of samples from fecal occult blood screening kits for metabolomic analysis. Application in colorectal cancer population.
STUDY_SUMMARY
Cancer is the most common disease around the world and colorectal cancer is the second most common cancer. The early diagnosis of colorectal cancer is difficult and relies on invasive diag-nostic tools such as colonoscopy and tissue biopsy. Other non-invasive techniques such as fecal occult blood screening test (FOBT) are less sensitive and accurate. The advantage of FOBT together with high throughput technology such as metabolomics could provide the advantages of non-invasive tool and the effectiveness of detecting novel colorectal cancer markers. In this way, this work focuses on the novelty of using FOBT as samples to perform metabolomics analysis and its application on colorectal cancer population.
INSTITUTE
CIC bioGUNE - Centro de InvestigaciĆ³n Cooperativa en Biociencias
LAST_NAME
Alboniga
FIRST_NAME
Oihane
ADDRESS
Bizkaia Technology Park, Bld.800, Derio, Bizkaia, 48160, Spain
Mesenchymal stromal cell (MSC) Metabolite MS study
STUDY_SUMMARY
Metabolomics and lipidomics workflows were used to analyze Mesenchymal stromal cell (MSC) metabolites. Metabolite abundances were used to model MSC potency results in IDO and T-cell proliferation assays.
Linking bacterial metabolites to disease-associated microbes to uncover mechanisms of host-microbial interactions in intestinal inflammation. Human plasma profiling
STUDY_SUMMARY
Understanding the role of the gut microbiome in inflammatory and autoimmune diseases requires the identification of microbial molecular effectors and their link to host pathophysiology. Here, we present a framework to identify and characterize novel microbial metabolites in patient samples and to directly link their production to disease-associated microbes. We applied this approach to investigate the spectrum of disease severity and treatment response in ulcerative colitis (UC) using longitudinal metabolite and strain profiles combined with paired plasma profiles.
Linking bacterial metabolites to disease-associated microbes to uncover mechanisms of host-microbial interactions in intestinal inflammation. Human stool profiling
STUDY_SUMMARY
Understanding the role of the gut microbiome in inflammatory and autoimmune diseases requires the identification of microbial molecular effectors and their link to host pathophysiology. Here, we present a framework to identify and characterize novel microbial metabolites in patient samples and to directly link their production to disease-associated microbes. We applied this approach to investigate the spectrum of disease severity and treatment response in ulcerative colitis (UC) using longitudinal metabolite and strain profiles combined with paired plasma profiles.
Linking bacterial metabolites to disease-associated microbes to uncover mechanisms of host-microbial interactions in intestinal inflammation. Veillonella parvula cell and media profiling
STUDY_SUMMARY
Understanding the role of the gut microbiome in inflammatory and autoimmune diseases requires the identification of microbial molecular effectors and their link to host pathophysiology. Here, we present a framework to identify and characterize novel microbial metabolites in patient samples and to directly link their production to disease-associated microbes. We applied this approach to investigate the spectrum of disease severity and treatment response in ulcerative colitis (UC) using longitudinal metabolite and strain profiles combined with paired plasma profiles.
Linking bacterial metabolites to disease-associated microbes to uncover mechanisms of host-microbial interactions in intestinal inflammation. Veillonella parvula media profiling of IBD drug metabolites
STUDY_SUMMARY
Understanding the role of the gut microbiome in inflammatory and autoimmune diseases requires the identification of microbial molecular effectors and their link to host pathophysiology. Here, we present a framework to identify and characterize novel microbial metabolites in patient samples and to directly link their production to disease-associated microbes. We applied this approach to investigate the spectrum of disease severity and treatment response in ulcerative colitis (UC) using longitudinal metabolite and strain profiles combined with paired plasma profiles.
Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice.
STUDY_SUMMARY
In this study, we identify disordered cardiac retinol metabolism in type 2 diabetic male mice and patients characterized by retinol overload, all-trans retinoic acid deficiency. By supplementing type 2 diabetic male mice with retinol or all-trans retinoic acid, we demonstrate that both cardiac retinol overload and all-trans retinoic acid deficiency promote diabetic cardiomyopathy. Mechanistically, by constructing cardiomyocyte-specific conditional retinol dehydrogenase 10-knockout male mice and overexpressing retinol dehydrogenase 10 in male type 2 diabetic mice via adeno-associated virus, we verify that the reduction in cardiac retinol dehydrogenase 10 is the initiating factor for cardiac retinol metabolism disorder and results in diabetic cardiomyopathy. Therefore, we suggest that the reduction of cardiac retinol dehydrogenase 10 and its mediated disorder of cardiac retinol metabolism is a new mechanism underlying diabetic cardiomyopathy.
INSTITUTE
Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University
LAST_NAME
Yandi
FIRST_NAME
Wu
ADDRESS
74, Zhongshan second street
EMAIL
wuyd3@mail2.sysu.edu.cn
PHONE
15622158754
PUBLICATIONS
Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice.
Psychobiotic Lactobacillus plantarum JYLP-326 relieves anxiety, depression, and insomnia symptoms in test anxious college students via modulating the gut microbiota and its metabolism
STUDY_SUMMARY
Test anxiety frequently occurs in college students and harms their physical and psychological health, but suitable interventions or therapeutical strategies are still missing. The present study aims to evaluate the potential effects of Lactobacillus plantarum JYLP-326 on test anxious college students. Sixty anxious students were enrolled and randomly allocated to the placebo group and the probiotic group, which were instructed to take placebo and JYLP-326 products twice per day for three weeks, respectively. Thirty unanxious students with no treatments were assigned to a regular control group. The anxiety, depression, and insomnia questionnaires were used to measure studentsā mental states at the baseline and the end of this study. 16S rRNA sequencing and untargeted metabolomics were performed to analyze the changes in the gut microbiota and fecal metabolism. The questionnaire results suggested that JYLP-326 administration could relieve the symptoms of anxiety, depression, and insomnia in test anxious students. The gut microbiomes of the placebo group showed a significantly greater Ī± diversity index than the control group (p < 0.05). An increased abundance of Bacteroides and Roseburia was observed in the placebo group, and the relative abundance of Prevotella and Bifidobacterium decreased. Whereas JYLP-326 administration could partly restore the disturbed gut microbiota. Additionally, test anxiety was correlation with disordered fecal metabolomics such as a higher Ethyl sulfate and a lower Cyclohexylamine, which could be reversed after taking JYLP-326. Furthermore, the changed microbiota and fecal metabolites were significantly associated with anxiety-related symptoms. These results indicated that the intervention of L. plantarum JYLP-326 could be an effective strategy to alleviate anxiety, depression and insomnia in test anxious college students.
INSTITUTE
Nanchang university
LAST_NAME
Ruizhe
FIRST_NAME
Zhu
ADDRESS
National Engineering Research Center for Bioengineering Drugs and the Technologies, Institute of Translational Medicine, Nanchang University, Nanchang, China
High body temperature increases gut microbiota-dependent host resistance to influenza A virus and SARS-CoV-2 infection (Mouse)
STUDY_SUMMARY
While a common symptom of influenza and coronavirus disease 2019 (COVID-19) is fever, its physiological role on host resistance to viral infection remains less clear. Here, we demonstrate that exposure of mice to the high ambient temperature of 36 Ā°C increase host resistance to viral pathogens including influenza virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). High heat-exposed mice increase basal body temperature over 38 Ā°C to enable more bile acids production in a gut microbiota-dependent manner. The gut microbiota-derived deoxycholic acid (DCA) and its plasma membrane-bound receptor Takeda G-protein-coupled receptor 5 (TGR5) signaling increase host resistance to influenza virus infection by suppressing virus replication and neutrophil-dependent tissue damage. Furthermore, the DCA and its nuclear farnesoid X receptor (FXR) agonist protect Syrian hamster from lethal SARS-CoV-2 infection. Moreover, we demonstrate that certain bile acids are reduced in the plasma of COVID-19 patients who developed moderate I/II disease compared with minor illness group. These findings uncover an unexpected mechanism by which virus-induced high fever increases host resistance to influenza virus and SARS-CoV-2 in a gut microbiota-dependent manner.
INSTITUTE
Keio University
LAST_NAME
Fukuda
FIRST_NAME
Shinji
ADDRESS
Kakuganji 246-2, Mizukami, Tsuruoka City Yamagata,Japan
Severe COVID-19 is characterized by an increase in the number and changes in the function of innate immune cells including neutrophils. However, it is not known how the metabolome of immune cells changes in COVID-19 patients or how metabolic changes may contribute to immune dysfunction. To address these questions, we analyzed the metabolome of neutrophils from patients with severe or mild COVID-19, or healthy controls. We identified widespread dysregulation of neutrophil metabolism with disease progression including in amino acid, redox, and central carbon metabolism.
INSTITUTE
UT Southwestern Medical Center
LAST_NAME
Li
FIRST_NAME
Yafeng
ADDRESS
5323 Harry Hines Blvd, Children's Research Institute, Dallas, TX, 75309, USA
The effect of prions on cellular metabolism: The metabolic impact of the [RNQ+] prion and the native role of Rnq1p
STUDY_SUMMARY
Within the field of amyloid and prion disease there is a need for a more comprehensive understanding of the fundamentals of disease biology. In order to facilitate the progression treatment and underpin comprehension of toxicity, fundamental understanding of the disruption to normal cellular biochemistry and trafficking is needed. Here, by removing the complex biochemistry of the brain, we have utilised known prion forming strains of Saccharomyces cerevisiae carrying different conformational variants of the Rnq1p to obtain Liquid Chromatography-Mass Spectrometry (LC-MS) metabolic profiles and identify key perturbations of prion presence. These studies reveal that prion containing [RNQ+] cells display a significant reduction in amino acid biosynthesis and distinct perturbations in sphingolipid metabolism, with significant downregulation in metabolites within these pathways. Moreover, that native Rnq1p downregulates ubiquinone biosynthesis pathways within cells, suggesting that Rnq1p may play a lipid/mevalonate-based cytoprotective role as a regulator of ubiquinone production. These findings contribute to the understanding of how prion proteins interact in vivo in both their prion and non-prion confirmations and indicate potential targets for the mitigation of these effects. . We demonstrate specific sphingolipid centred metabolic disruptions due to prion presence and give insight into a potential cytoprotective role of the native Rnq1 protein. This provides evidence of metabolic similarities between yeast and mammalian cells as a consequence of prion presence and establishes the application of metabolomics as a tool to investigate prion/amyloid-based phenomena.
High body temperature increases gut microbiota-dependent host resistance to influenza A virus and SARS-CoV-2 infection (Hamster)
STUDY_SUMMARY
While a common symptom of influenza and coronavirus disease 2019 (COVID-19) is fever, its physiological role on host resistance to viral infection remains less clear. Here, we demonstrate that exposure of mice to the high ambient temperature of 36 Ā°C increase host resistance to viral pathogens including influenza virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). High heat-exposed mice increase basal body temperature over 38 Ā°C to enable more bile acids production in a gut microbiota-dependent manner. The gut microbiota-derived deoxycholic acid (DCA) and its plasma membrane-bound receptor Takeda G-protein-coupled receptor 5 (TGR5) signaling increase host resistance to influenza virus infection by suppressing virus replication and neutrophil-dependent tissue damage. Furthermore, the DCA and its nuclear farnesoid X receptor (FXR) agonist protect Syrian hamster from lethal SARS-CoV-2 infection. Moreover, we demonstrate that certain bile acids are reduced in the plasma of COVID-19 patients who developed moderate I/II disease compared with minor illness group. These findings uncover an unexpected mechanism by which virus-induced high fever increases host resistance to influenza virus and SARS-CoV-2 in a gut microbiota-dependent manner.
INSTITUTE
Keio University
LAST_NAME
Fukuda
FIRST_NAME
Shinji
ADDRESS
Kakuganji 246-2, Mizukami, Tsuruoka City Yamagata,Japan
Impact of in-utero exposures to per- and polyfluoroalkyl substances on the human fetal liver metabolome
STUDY_SUMMARY
Background Per- and polyfluoroalkyl substances (PFAS) are classed as Endocrine Disrupting Compounds (EDCs) but continue to be used in many products. This includes firefighting foams, flame retardants, utensil coatings and waterproofing of food packaging. PFAS exposure aberrantly modulates lipid, metabolite and bile acid (BA) levels, increasing susceptibility to onset and severity of metabolic diseases, such as diabetes and non-alcoholic fatty liver disease (NAFLD). To date, most studies in humans have focused on PFAS-exposure effects in adults. In this study we now demonstrate that PFAS are present in the human fetal liver and that they have metabolic consequences for the human fetus. Methods Human fetal livers from elective termination of pregnancies between 11-19 weeks of gestation (n = 78) were analysed by both targeted and untargeted metabolomic analyses of lipids, polar metabolites, BAs and PFAS. Stringent bioinformatic and statistical methods were applied to this data to generate a network of interacting molecules. Findings Metabolites associated with PFAS were identified in the fetal liver and these varied with gestational age . Conjugated BAs were markedly positively associated with fetal age. Several amino acids, fatty acids and sugar derivatives in fetal livers were inversely associated with PFAS exposure, while the BA glycolithocholic acid (GLCA) was markedly positively associated with all quantified PFAS. Furthermore, 7Ī±-hydroxy-4-cholesten-3-one (C4), a marker of BA synthesis rate, was strongly positively associated with PFAS levels and was detectable as early as gestational week 12. Interpretation The data show direct evidence for in-utero effects of PFAS exposure on specific key hepatic products. Our results provide evidence that PFAS exposure, with potential future consequences, manifests in the human fetus as early as the first trimester of gestation. Furthermore, the profiles of metabolic changes resemble those observed in perinatal PFAS exposures. Such exposures are already linked with susceptibility, initiation, progression and/or exacerbation of a wide range of metabolic diseases.
INSTITUTE
Ćrebro University
LAST_NAME
McGlinchey
FIRST_NAME
Aidan
ADDRESS
School of Medical Sciences, Ćrebro, Ćrebro, 70281, Sweden
Urolithin A (UroA) acts at CYP1A1/1B1 substrates leading to enhanced aryl hydrocarbon receptor (AHR) activity in vivo
STUDY_SUMMARY
Many AHR ligands are CYP1A1/1B1 substrates, which can result in the rapid clearance within the intestinal tract and other tissues, limiting both the level and duration of AHR activation. This leads to the hypothesis that there are dietary constituents capable of inhibiting CYP1A1/1B1 increasing the half-live of potent AHR ligands. To test this hypothesis, we examined the ability of urolithin A (UroA) to act at CYP1A1/1B1 substrates leading to enhanced AHR activity in vivo.
INSTITUTE
Pennsylvania State University
LAST_NAME
DONG
FIRST_NAME
FANGCONG
ADDRESS
309 Life Sciences Building, State college, PA, 16802, USA
Non-targeted screening of natural products from 288 fungal endophytes from Canadian fruit crops
STUDY_SUMMARY
Fungal endophytes often live in symbiotic relationships with various plant hosts, conferring positive effects to their host organism. These endophytes frequently produce a wide variety of secondary metabolites with bioactivities that are often responsible for the beneficial effects seen in the host, such as antifungal or anti-insectan activity. A large group of fungal endophytes isolated from Canadian fruit crops including blueberries, raspberries, cranberries, grapes, and pears, was analyzed using molecular networking by GNPS in an effort to simplify the process of examining a large dataset. Molecular networking increased the speed and efficiency of examining this dataset, permitting the dereplication of 60 known compounds and the discovery of seven putative novel compounds, which will be purified, characterized, and tested for bioactivity in future studies.
Metabolomic analysis of maternal mid-gestation plasma and cord blood: primary metabolism
STUDY_SUMMARY
Metabolomic analysis of maternal mid-gestation plasma and cord blood reveals evidence in autism spectrum disorder of inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. The discovery of prenatal and neonatal molecular biomarkers has the potential to yield insights into autism spectrum disorder (ASD) and facilitate early diagnosis. We characterized metabolomic profiles in ASD using plasma samples collected in the Norwegian Autism Birth Cohort from mothers at weeks 17-21 gestation (maternal mid-gestation, MMG, n=408) and from children on the day of birth (cord blood, CB, n=418). We analyzed associations using sex-stratified adjusted logistic regression models with Bayesian analyses. Chemical enrichment analyses (ChemRICH) were performed to determine altered chemical clusters. We also employed machine learning algorithms to assess the utility of metabolomics as ASD biomarkers. We identified ASD associations with a variety of chemical compounds including arachidonic acid, glutamate, and glutamine, and metabolite clusters including hydroxy eicospentaenoic acids, phosphatidylcholines, and ceramides in MMG and CB plasma that are consistent with inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. Girls with ASD have disruption of ether/non-ether phospholipid balance in the MMG plasma that is similar to that found in other neurodevelopmental disorders. ASD boys in the CB analyses had the highest number of dysregulated chemical clusters. Machine learning classifiers distinguished ASD cases from controls with AUC values ranging from 0.710 to 0.853. Predictive performance was better in CB analyses than in MMG. These findings may provide new insights into the sex-specific differences in ASD and have implications for discovery of biomarkers that may enable early diagnosis and intervention.
Simultaneous targeting of PD-1 and IL2RĪ²Ī³ with radiation therapy to inhibit pancreatic cancer growth and metastasis - mouse serum metabolomics
STUDY_SUMMARY
Mice were orthotopically implanted with PK5L1940 cells then radiation therapy (8 Gy) administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. Serum was collected at time of sacrifice and profiled by mass spectrometry-based metabolomics to visualize changes in systemic metabolism in the mouse model.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Simultaneous targeting of PD-1 and IL2RĪ²Ī³ with radiation therapy to inhibit pancreatic cancer growth and metastasis - mouse blood T-cell metabolomics
STUDY_SUMMARY
Mice were orthotopically implanted with PK5L1940 cells then radiation therapy (8 Gy) administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. Blood was collected at time of sacrifice and sorted for CD8+ T-cells. The obtained T-cells were profiled by mass spectrometry-based metabolomics.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Simultaneous targeting of PD-1 and IL2RĪ²Ī³ with radiation therapy to inhibit pancreatic cancer growth and metastasis - mouse spleen and lymph node T-cell metabolomics
STUDY_SUMMARY
Mice were orthotopically implanted with PK5L1940 cells then radiation therapy (8 Gy) administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. Spleen and associated lymph nodes were collected at time of sacrifice and sorted for CD8+ T-cells. The obtained T-cells were profiled by mass spectrometry-based metabolomics.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Simultaneous targeting of PD-1 and IL2RĪ²Ī³ with radiation therapy to inhibit pancreatic cancer growth and metastasis - mouse serum metabolomics
STUDY_SUMMARY
C57BL/6 mice were orthotopically implanted with PK5L1940 cells. 8 Gy RT was administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. Spleens and lymph nodes were harvested 17 days post implantation and the sorted T-cells cultured in the presence of 13C6 glucose for 6h. This study contains longitudinal sampling of the cell culture supernatants.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Simultaneous targeting of PD-1 and IL2RĪ²Ī³ with radiation therapy to inhibit pancreatic cancer growth and metastasis
STUDY_SUMMARY
Mice were orthotopically implanted with PK5L1940 cells then radiation therapy (8 Gy) administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week beginning day 7 post-implantation. T-cells from spleen were obtained through sorting, cultured in the presence of 13C6 glucose for 6h, and glucose fluxes profiled via mass spectrometry-based metabolomics.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Simultaneous targeting of PD-1 and IL2RĪ²Ī³ with radiation therapy to inhibit pancreatic cancer growth and metastasis - human plasma metabolomics
STUDY_SUMMARY
Patient plasma samples were collected as part of a Phase I radiation dose-escalation clinical trial (NCT02873598) before, during (6 hours post), and post (6 weeks) SBRT. Samples were collected and analyzed per COMIRB19ā0328 for branched chain amino acids and kynurenine.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
We establish that six tryptophan metabolites are essentially not CYP1A1 substrates and the level of these metabolite in mouse serum are not affected by a lack of CYP1A1 or any other Aryl hydrocarbon Receptor (AHR) dependent metabolic pathway expression in vivo. These results establish that tryptophan metabolites that circulate at significant levels in vivo are not subject to an autoregulatory feedback loop between the AHR and CYP1A1.
INSTITUTE
Pennsylvania State University
LAST_NAME
Dong
FIRST_NAME
Fancong
ADDRESS
309 Life Sciences Building, University Park, PA 16802
The goal of this study was to use metabolomics as a platform to elucidate the chemical composition of plants in order to increase their resolution and in turn use the identified chemicals to reveal potential health impacts. 20 plant foods were studied: apple, banana, tomato, lettuce, strawberry, carrot, peach, onion, spinach, pepper, corn, garlic, basil, potato, soybean, black bean, olive, chickpea, sugarbeet, and pear.
INSTITUTE
Northeastern University; Massachusets Institute of Technology
DEPARTMENT
Department of Physics
LABORATORY
BarabasiLab
LAST_NAME
Barabasi
FIRST_NAME
Albert-Laszlo
ADDRESS
177 Huntington Ave, 11th Floor, Boston, MA, 02115, USA
Composition of raw plant-based food items Pilot Study
STUDY_TYPE
Composition of food
STUDY_SUMMARY
The goal of this pilot study was to be a preliminary metabolomics study on the platforms used to elucidate the chemical composition of plants in order to increase their resolution and in turn use the identified chemicals to reveal potential health impacts. In this pilot we focused on 6 food items: apple, basil, lettuce, strawberry, tomato, and garlic.
INSTITUTE
Northeastern University; Massachusets Institute of Technology
DEPARTMENT
Department of Physics
LABORATORY
BarabasiLab
LAST_NAME
Barabasi
FIRST_NAME
Albert-Laszlo
ADDRESS
177 Huntington Ave, 11th Floor, Boston, MA, 02115, USA
Disrupted intestinal microbiota contributes to the pathogenesis of anorexia nervosa (Part 1)
STUDY_SUMMARY
Anorexia nervosa (AN) is an eating disorder with a high mortality affecting about 1% of women, where no evidence-based effective treatment exists. The pathogenesis likely involves genetic and environmental alterations. We hypothesized that a disrupted gut microbiota contributes to AN pathogenesis. In analyses comparing 70 AN with 77 healthy females, we found multiple taxa, functional modules, structural variants and growth rates of bacterial gut microbiota, and viral gut microbiota that were altered in AN with parts of these perturbations linked to estimates of eating behavior and mental health. In silico, causal inference analyses implied serum bacterial metabolites mediated parts of the impact of altered gut microbiota on AN behavior, and in vivo, three independent fecal microbiota transplantation from AN cases to germ-free mice under energy restricted feeding mirroring AN eating behavior consistently induced a lower body weight gain and hypothalamic and adipose tissue gene expressions related to aberrant energy metabolism and eating and mental behavior.
Disrupted intestinal microbiota contributes to the pathogenesis of anorexia nervosa (Part 2)
STUDY_SUMMARY
Anorexia nervosa (AN) is an eating disorder with a high mortality affecting about 0.5% of women, where no evidence-based effective treatment exists. The pathogenesis likely involves genetic and environmental alterations. We hypothesized that a disrupted gut microbiota contributes to AN pathology. In analyses comparing 70 AN with 77 healthy females, we found multiple taxa, functional modules, structural variants and growth rates of bacterial gut microbiota, and viral gut microbiota that were altered in AN with parts of these perturbations linked to estimates of eating behavior and mental health. In silico, causal inference analyses implied bacterial metabolites mediated parts of the impact of altered gut microbiota on AN behavior, and in vivo, fecal microbiota transplantation from AN cases to germ-free mice induced a lower body weight and hypothalamic and adipose tissue gene expressions related to aberrant energy metabolism and eating and mental behavior.
Study of environmental toxicants and gut microbiome in relation to obesity and insulin resistance
STUDY_SUMMARY
Background & Aims: Environmental toxicants (ETs) associate with various adverse health outcomes. Here, we hypothesized that exposures to ETs are associated with obesity and insulin resistance via a dysbiotic gut microbiota and derived alterations in microbiome-mediated bile acid (BA) synthesis. Methods: Serum BAs, per- and polyfluoroalkyl substances (PFAS) and additional twenty-seven ETs were measured by mass spectrometry in 264 Danes (121 women and 143 men, age 56.6 Ā± 7.3 years, BMI 29.7 Ā± 6.0 kg/m2). Bacterial species were identified based on whole-genome shotgun (WGS) sequencing of DNA extracted from purified stool samples. Personalized genome-scale metabolic models (GEMs) of gut microbial communities were developed to elucidate regulation of BA pathways. Subsequently, we compared findings in the human study with metabolic implications of perfluorooctanoic acid (PFOA) exposure in a PPARļ”-humanized murine model. Results: Fasting serum concentrations of twelve ETs associated directly with measures of obesity and insulin resistance. Several bacterial species including Dorea longicatena, Dorea formicigenerans, Subdoligranulum spp., Veillonella spp., and Roseburia intestinalis associated positively and in a sex-dimorphic manner, particularly in women, with high exposure to ETs. Moreover, high serum concentrations of ETs were linked with higher fasting serum levels of microbiome-synthesized secondary BAs such as lithocholic acid (LCA) and ursodeoxycholic acid (UDCA). These findings were substantiated by the outcome of a murine exposure study. Conclusion: Serum concentrations of ETs, particularly in women, were associated with an altered gut microbiome-mediated secondary BA biosynthesis, linked with obesity and insulin resistance.
INSTITUTE
Ćrebro University
DEPARTMENT
Department of Medical Sciences
LABORATORY
Systems Medicine
LAST_NAME
OreÅ”iÄ
FIRST_NAME
Matej
ADDRESS
School of Medical Sciences, Ćrebro, Ćrebro, 70281, Sweden
Postnatal hyperglycemia alters amino acid profile in retinas
STUDY_SUMMARY
Nutritional deprivation occurring in most preterm infants postnatally, can induce hyperglycemia, a significant and independent risk factor for suppressing physiological retinal vascularization (Phase I retinopathy of prematurity (ROP)), leading to compensatory but pathological neovascularization. Amino acid supplementation reduces retinal neovascularization in mice. Little is known about amino acid contribution to Phase I ROP. Significant changes in retinal amino acids (including most decreased L-leucine, L-isoleucine and L-valine) were found in mice modeling hyperglycemia-associated Phase I ROP, and parenteral (i.p.) L-isoleucine suppressed physiological retinal vascularization. In premature infants, severe ROP was associated with a higher mean intake of parenteral versus enteral amino acids in the first two weeks of life after adjustment for treatment group, gestational age at birth, birth weight and sex. The number of days with parenteral amino acids support independently predicted severe ROP. Further understanding and modulating amino acids may help improve nutritional intervention and prevent Phase I ROP
Plasma Metabolomics Profiling of 580 Patients from the Weill Cornell Medicine Early Detection Research Network Prostate Cancer Cohort
STUDY_SUMMARY
Prostate cancer is the second most common cancer in men and affects 1 in 9 men in the United States. Early screening for prostate cancer often involves monitoring levels of prostate-specific antigen (PSA) and performing digital rectal exams. However, a prostate biopsy is always required for definitive cancer diagnosis. The Early Detection Research Network (EDRN) is a consortium within the National Cancer Institute aimed at improving screening approaches and early detection of cancers. As part of this effort, the Weill Cornell EDRN Prostate Cancer has collected and biobanked specimens from men undergoing a prostate biopsy between 2008 and 2017. In this report, we describe blood metabolomics measurements for a subset of this population. The dataset includes detailed clinical and prospective records for 580 patients who underwent prostate biopsy, 287 of which were subsequentially diagnosed with prostate cancer, combined with profiling of 1,482 metabolites from plasma samples collected at the time of biopsy. We expect this dataset to provide a valuable resource for scientists investigating prostate cancer metabolism.
Plasma metabolomic signatures from patients following high-dose total body irradiation
STUDY_SUMMARY
Plasma metabolic characteristics were investigated from patients of hematopoietic stem cell transplantation following high-dose TBI pretreatment utilizing gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS).
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (ReCHARGE Cohort)
STUDY_TYPE
Case-Control Study
STUDY_SUMMARY
The ReCHARGE study is a case control study of children with Autism Spectrum Disorder, Developmental Delay, and who are typically developing. They were initially recruited into the study between the ages and 2 and 5 years of age, and are being seen again during middle childhood and adolescence. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. ReCHARGE is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
University of California, Davis
DEPARTMENT
School of Medicine, Public Health Sciences
LAST_NAME
Deborah
FIRST_NAME
Bennett
ADDRESS
Public Health Sciences Medical Sciences 1-C Davis, CA 95616
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (IKIDS Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
IKIDS is a prospective birth cohort in Central Illinois. Pregnant women were recruited at two obstetrical clinics in Urbana Champaign IL from 2014-2021. Women were enrolled between 8-14 weeks of pregnancy. Eligibility criteria included 18-40 years of age, English speaking, not in a high risk pregnancy, planning to stay in the area until their child's first birth day and living within 30 minutes of the University of Illinois campus. Health, demographic and life style data and biospecimens were collected at 5 time points across pregnancy and children were followed prospectively from birth including assessments at birth, 1-5 weeks 4.5 months, 7.5 months, 2 years, 3 years, 4 years and 7.5 years of age. Physical and neurobehavioral development were assessed, with a particular focus on the impacts of prenatal chemical exposures (phthalates, phenols, PFAS) or prenatal maternal stress on child health outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. IKIDS is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
University of Illinois Urbana-Champaign
DEPARTMENT
Beckman Institute for Advanced Science and Technology
LAST_NAME
Schantz
FIRST_NAME
Susan
ADDRESS
2325/21 Beckman Institute 405 North Mathews Avenue Urbana, IL 61801
Endothelial Cell CD36 Regulates Membrane Ceramide Formation, Exosome Fatty Acid Delivery to Tissues and Circulating Fatty Acid Levels
STUDY_TYPE
Membrane ceramide and Fatty Acid Uptake
STUDY_SUMMARY
Endothelial cell (EC) CD36 controls tissue fatty acid (FA) uptake. Here we visualized FA transfer by ECs using click chemistry. Apical FA interaction with EC CD36 induces actin reorganization, caveolin-1 tyrosine 14 (Cav-1Y14) phosphorylation, and ceramide generation in caveolae, culminating in caveolae internalization into FA/CD36/ceramide vesicles that are secreted basolaterally as small extracellular vesicles, sEVs (exosomes). In transwells, ECs expressing fluorescent CD63 (exosomal marker) transfer FAs to underlying myotubes in CD63-labeled sEVs. In vivo FA-CD63-CD36 transfer from ECs to muscle is shown using a mouse with EC-specific expression of emGFP-CD63. CD36 depletion, actin-binding latrunculin-B, Src inhibition, Cav-1Y14 mutation, and membrane neutral sphingomyelinase inhibition by GW4869 helped map the FA-sEV pathway. Injecting GW4869 to mice reduced muscle FA uptake, raised plasma FAs and lowered glucose, mimicking prominent Cd36-/- phenotypes. The FA-sEV pathway we describe contributes to crosstalk between ECs and underlying cells and links regulation of membrane ceramide to blood FAs.
INSTITUTE
Washington University in St. Louis
DEPARTMENT
IM-Nutitrional Science
LABORATORY
Abumrad Lab
LAST_NAME
Palacios
FIRST_NAME
Hector
ADDRESS
West Building 00201, St. Louis, Missouri, 63110, USA
Lipid droplets and peroxisomes are co-regulated to drive lifespan extension in response to mono-unsaturated fatty acids
STUDY_SUMMARY
Dietary mono-unsaturated fatty acids (MUFAs) are linked to human longevity and extend lifespan in several species. But the mechanisms by which MUFAs extend lifespan remain unclear. Here we show that an organelle network involving lipid droplets and peroxisomes is critical for lifespan extension by MUFAs in C. elegans. MUFA accumulation increases lipid droplet number in fat storage tissues, and this is necessary for MUFA-induced longevity. Lipid droplet number in young or middle-aged individuals can predict remaining lifespan, consistent with a beneficial effect of lipid droplets on lifespan. Lipidomics datasets reveal that MUFA accumulation also modifies the ratio of membrane lipids and ether lipids, a signature predictive of decreased lipid oxidation. We validate that MUFAs decrease lipid oxidation in middle-aged individuals, and that this is important for MUFA-induced longevity. Intriguingly, the increase in lipid droplet number in response to MUFAs is accompanied by a concomitant increase in peroxisome number. Using a targeted screen, we identify genes involved in the co-regulation or uncoupling of this lipid droplet-peroxisome network. We find that induction of both organelles is optimal for lifespan extension. Our study uncovers an organelle network involved in lipid homeostasis and lifespan regulation and identifies a mechanism of action for MUFAs to extend lifespan, opening new avenues for lipid-based interventions to delay aging. For the manuscript only the conditions ācontrolā and āash-2 RNAiā are plotted
A Mammalian Conserved Circular RNA CircLARP2 Regulates Hepatocellular Carcinoma Metastasis and Lipid Metabolism (Part 1)
STUDY_SUMMARY
Circular RNAs (circRNAs) have emerged as crucial regulators in physiology and human diseases. However, evolutionarily conserved circRNAs with potent functions in cancers are rarely reported. Here, we identified a mammalian conserved circRNA circLARP2 that played critical roles in hepatocellular carcinoma (HCC). With clinical specimens, we found that patients with high circLARP2 levels in HCC had advanced prognostic stage and poor overall survival. CircLARP2 facilitated HCC metastasis and lipid accumulation in HCC cell lines. CircLARP2 was one of the rare ones that were identified in HCC metastasis and conserved in mammals, which enabled further studies with animal models. CircLARP2-deficient mice exhibited reduced metastasis and less lipid accumulation in an induced HCC model. We provided lines of evidence at molecular, cellular, and whole organismal levels, to support that circLARP2 functioned as a protein sponge of AUF1. CircLARP2 sequestered AUF1 from binding to LKB1 mRNA, which led to decreased LKB1 mRNA stability and lower LKB1 protein levels. LKB1 as a kinase promoted the phosphorylation of AMPK and then the phosphorylation of ACC, the rate limiting enzyme of fatty acid synthesis. Knockdown of Lkb1 with AAV8-shLkb1 in mice HCC model also proved that Lkb1 was a key element in the regulation. Through this AUF1-LKB1-AMPK-ACC pathway, circLARP2 promoted HCC metastasis and lipid accumulation.
Natural abundance of isotopic metabolite detection in mouse eye orgnaoids.
STUDY_SUMMARY
Because the natural abundance of isotopic metabolites in the early eye organoids has not yet been reported, we have performed LC-MS/MS without exogenous isotopic labeling to show the natural abundance of isotopic carbons. Cell Name AES0145 : Rx-GFP K/I EB5 (RIKEN Cell Bank): Organoid method (PMID: 21475194).
Time-course analysis of C13 labeling in mouse eye organoids.
STUDY_SUMMARY
The direct quantification of glucose consumption using 13C glucose time-course tracing was performed in cultured eye organoids and measured by LC-MS/MS analysis. The incubation was performed from 15 minutes to 2 hours. Cell Name AES0145 : Rx-GFP K/I EB5 (RIKEN Cell Bank): Organoid method (PMID: 21475194).
13C-isotopic labeling of mouse eye organoids under three conditions.
STUDY_SUMMARY
To examine the potential involvement of metabolites, we performed 13C-glucose labeling under three conditions: 13C-glucose = control, 13C-glucose + GNE-140 = LHDi, 13C-lactate in glucose free media mimicking the lack of glucose. Cell Name AES0145 : Rx-GFP K/I EB5 (RIKEN Cell Bank): Organoid method (PMID: 21475194).
INSTITUTE
Northwestern University
LAST_NAME
TAKATA
FIRST_NAME
NOZOMU
ADDRESS
303 East Superior Street, 10-220, Chicago, Illinois, 60611, USA
Time course 1: Growth of Eggerthella lenta in defined media
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of supernatants from 3 strains of Eggerthella lenta grown in defined EDM1 media with varying acetate concentrations.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Strain supernatants: Strain diversity of Eggerthella lenta metabolites in defined media
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of supernatants from stationary phase of a collection of 30 strains of Eggerthella lenta grown in defined EDM1 media.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Gnotobiotic mice: Metabolites in intestinal contents of germ-free mice colonized with strains of gut bacterium Eggerthella lenta
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of intestinal contents of gnotobiotic mice either colonized with different strains of Eggerthella lenta for 2 weeks, or germ-free controls.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Gnotobiotic mice: Metabolites in serum of germ-free mice colonized with strains of gut bacterium Eggerthella lenta
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of serum of gnotobiotic mice either colonized with different strains of Eggerthella lenta for 2 weeks, or germ-free controls.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
The investigation of the role of dietary inulin in NASH progression with mouse fecal metabolites
STUDY_TYPE
non-targeted LC-Mass analysis
STUDY_SUMMARY
To investigate the role of dietary fiber in non-alcoholic steatohepatitis (NASH) progression, male C57 mice was randomly assigned into four groups that received normal chow diet (NCD), choline deficient high fat diet (CDHFD), CDHFD + 10% inulin (CDHFD-I), CDHFD + 10% Cellulose (CDHFD-C). Additionally, some mice received inulin or cellulose was treated with 13C labelled fiber for 36 hours. Fecal metabolites were analyzed by non-targeted metabolomics.Results shown that the fecal metabolites from Inulin treatment group signficantly distinguished from that from CDHFD only group, while only negligible alterations was induced by cellulose treatment, indicating that inulin not cellulose could be well fermented by gut microbiota. The 13C tracing results shown that nineteen 13C-inulin labelled metabolites were also captured including pantothenate, phosphoethanolamine and adenosine. These metabolites are reported to play a protective role in reducing fat accumulation and ameliorating cellular oxidative stress and inflammation, indicating that the mechanism inulin suppresses NASH may through mediating modulating gut metabolites.
Time course 2: Acetate quantification from growth of Eggerthella lenta in defined media
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains targeted metabolomics analysis of carboxylic acids in supernatants from 3 strains of Eggerthella lenta grown in defined EDM1 media with varying acetate concentrations.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Time course 2: Growth of Eggerthella lenta in defined media with some samples receiving 13C2 stable isotope labeled acetate
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of supernatants from 3 strains of Eggerthella lenta grown in defined EDM1 media with varying acetate concentrations. One set of samples grew in EDM1 containing 13C2 stable isotope labeled acetate.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Time course 2: Growth of Eggerthella lenta in defined media with some samples receiving 13C2 stable isotope labeled acetate (intracellular samples)
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of supernatants from 3 strains of Eggerthella lenta grown in defined EDM1 media with varying acetate concentrations. One set of samples grew in EDM1 containing 13C2 stable isotope labeled acetate. Samples were collected at a subset of time points for extraction of intracellular metabolites.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Time course 3: Growth of Eggerthella lenta in defined media with some samples receiving 13C6 stable isotope labeled arginine
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of supernatants from Eggerthella lenta DSM 2243 grown in defined EDM1 media. One set of samples grew in EDM1 containing 13C6 stable isotope labeled arginine.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
Time course 3: Growth of Eggerthella lenta in defined media with some samples receiving 13C6 stable isotope labeled arginine (intracellular samples)
STUDY_TYPE
Untargeted LC-MS
STUDY_SUMMARY
This dataset contains untargeted metabolomics analysis of supernatants from Eggerthella lenta grown in defined EDM1 media. One set of samples grew in EDM1 containing 13C6 stable isotope labeled arginine. Samples were collected at a subset of time points for extraction of intracellular metabolites.
INSTITUTE
University of California, San Francisco
LAST_NAME
Noecker
FIRST_NAME
Cecilia
ADDRESS
513 Parnassus Ave HSW1501, San Francisco, CA 94143
The investigation of the role of dietary inulin in Non-alcoholic Steatohepatitis (NASH) progression with portal vein serum metabolites
STUDY_SUMMARY
To investigate the role of dietary fiber in non-alcoholic steatohepatitis (NASH) progression, male C57 mice was randomly assigned into four groups that received normal chow diet (NCD), choline deficient high fat diet (CDHFD), CDHFD + 10% inulin (CDHFD-I), CDHFD + 10% Cellulose (CDHFD-C). Mice portal vein serum was collected.Results shown that the fecal metabolites from Inulin treatment group signficantly distinguished from that from CDHFD only group, while only negligible alterations was induced by cellulose treatment, indicating that inulin not cellulose could be well fermented by gut microbiota. By overlaping with fecal metabolites, the inulin derived pentadecaonic acid was consistently enriched in portal vein serum, which was reported as a beneficial metabolite.The results indicate that inulin suppresses NASH may through direct modulating gut metabolites.
Wide-Coverage Serum Metabolomic Profiling Reveals a Comprehensive Lipidome Signature of Ovarian Cancer.
STUDY_SUMMARY
Distinguishing ovarian cancer (OC) from other benign or cancerous gynecological malignancies remains a critical unmet medical need with significant implications on patient survival. Substantially better results are observed when women with OC are correctly diagnosed and ensured the right treatment. However, non-specific symptoms along with our lack of understanding of OC pathogenesis hinder its diagnosis, consequently leading to a very low survival rate. Accumulating evidence suggests the link between OC and deregulated lipid metabolism. Most studies, however, are limited by small sample sizes and metabolite coverage, thereby constraining the robustness of the results. Here, we performed a comprehensive serum lipidome profiling of OC and various other gynecological malignancies (non-OC). A relatively large patient cohort with 208 OC and 137 non-OC patients, including 93 OC patients with early-stage OC, was recruited from two independent clinical sites in South Korea. Samples were analyzed with high-coverage liquid chromatography high-resolution mass spectrometry, providing extensive lipidome coverage with 994 successfully annotated lipid features. Lipidome differences between OC and other gynecological malignancies were investigated via statistical and machine learning approaches. Our data suggest that lipidome alterations unique to OC can be detected as early as when the cancer is localized, and those changes amplify as the diseases progresses. Comparison of the relative lipid abundances revealed specific patterns based on lipid class with most lipid classes showing decreased abundance in ovarian cancer. This study provides a systemic analysis of lipidome alterations in OC, emphasizing the potential of circulating lipids as a complementary class of blood-based biomarkers for OC diagnosis.
Lipidomics study on the effect of LBP protein on hepatic lipid composition in mice
STUDY_SUMMARY
Stress elevates the formation of ROS and lipid peroxidation, which induce lipid droplets (LDs) accumulation and adverse metabolic disturbance. Here, we explored the novel role of Lipopolysaccharide-binding protein (LBP) as an anti-oxidant, which can capture unsaturated triglyceride (TG) into LDs to avoid lipid peroxidation. Oxidative stress upregulates LBP level and promotes LDs growth via the LBP/TG phase transition. Upon N-Acetyl-L-cysteine (NAC) elimination of ROS, LBP is exported from LD along with PRDX4, resulting in an increase in phospholipid synthesis. Chronic stress causes LBP upregulation and leads to obesity, which can be rescued by NAC treatment in vivo. These results support that LBP maintains homeostasis by coupling lipid metabolism and redox signal, which provides insights into redox medicine that mitigate stress-induced metabolic dysfunction. Hepatic lipidomics in overexpressed LBP and WT mice treated with NAC after 24h fasting
INSTITUTE
University of Science and Technology of China
DEPARTMENT
Department of Endocrinology and Laboratory for Diabetes
LABORATORY
The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine
Candida expansion in the human gut is associated with an ecological signature that supports growth under dysbiotic conditions
STUDY_SUMMARY
The overgrowth of Candida species in the human gut is considered a prerequisite for invasive candidiasis. However, our understanding of how gut bacteria promote or restrict overgrowth of Candida species in the human gut is still limited. By integrating mycobiome and shotgun metagenomics data from stool of 75 patients at risk but with no systemic candidiasis, we revealed that bacterial communities from high Candida samples had greater metabolic potential whereas communities from low Candida had greater functional redundancy. In addition, we developed machine learning models that used only bacterial taxa or functional relative abundances to predict the levels of Candida genus and species in an external validation cohort with an area under the curve of 78.6-81.1%. Last, we proposed an intriguing mechanism for Candida species overgrowth based on a decrease in short-chain fatty acid producing-bacteria resulting in increased oxygen levels. These conditions create a metabolic niche for Candida species to use lactate as a carbon source and overtake their fungal competitors in the human gut.
INSTITUTE
Leibniz Institute for Natural Product Research and Infection Biology Hans KnĆĀ¶ll Institute
In situ microwave fixation provides an instantaneous snapshot of the brain metabolome - Part 1
STUDY_SUMMARY
We demonstrate exhaustion of glycogen and glucose and an increase in lactate production during conventional rapid brain resection prior to preservation by liquid nitrogen that is not observed with microwave fixation. Next, microwave fixation was employed to define the impact of brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple regions of the mouse brain, evidenced by reduced 13C enrichment into glycogen, glycolysis, and the TCA cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation to study terminal brain metabolism in rodent models.
In situ microwave fixation provides an instantaneous snapshot of the brain metabolome - Part 2
STUDY_SUMMARY
We demonstrate exhaustion of glycogen and glucose and an increase in lactate production during conventional rapid brain resection prior to preservation by liquid nitrogen that is not observed with microwave fixation. Next, microwave fixation was employed to define the impact of brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple regions of the mouse brain, evidenced by reduced 13C enrichment into glycogen, glycolysis, and the TCA cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation to study terminal brain metabolism in rodent models.
Stable isotope tracing of 15N2-glutamine in orthotopic pancreatic ductal adenocarcinoma tumor bearing mice and non tumor-bearing controls
STUDY_SUMMARY
Stable isotope tracing by bolus intravenous injections of 15N2-glutamine in orthotopic PDAC tumor bearing mice and non tumor-bearing controls followed by plasma sampling and tumor extraction for analysis of intratumoral metabolite labeling during the period of kinetic labeling
HPLC-MS-MS analysis amino acid levels in PDAC IF samples upon arginase inhibition
STUDY_SUMMARY
To test the hypothesis that myeloid arginase-1 activity could be responsible for pancreatic ductal adenocarninoma microenvironmental arginine starvation (PMID: 30990168), we generated orthotopic allograft mPDAC tumors in a mouse model with myeloid specific Arg1 knockout (LysM-Cre+/+-; Arg1fl/fl) and control animals (Arg1fl/fl). We also tested this with pharmacological inhibition of arginase-1 with the small-molecule inhibitor CB-1158. We then isolated IF from these tumors at end-stage and measured the levels of amino acids including arginine and ornithine in these samples.
Concentrations of amino acids in interstitial fluid and whole tumor samples of a murine PDAC orthotopic tumor model, measured by LC-MS
STUDY_SUMMARY
We extracted polar metabolites from the interstitial fluid and whole tumor samples of orthotopic murine PDAC tumors. Weused LC-MS to measure the concentration of amino acids in the interstitial fluid of orthotopic murine PDAC tumors and compared this to the intratumoral concentrations.
LCMS analysis of amino acid levels in PDAC interstitial fluid samples upon ass1 cancer cell knock out
STUDY_SUMMARY
We used CRISPR-Cas9 to knockout (KO) Ass1 in mPDAC cells. We tested if inhibiting arginine de novo synthesis would impair PDAC tumor growth. To test this, we generated orthotopic PDAC tumors with mPDAC-Ass1 KO cells and control cells where Ass1 was re-expressed (Ass1KO; mASS1).
Cellular consumption/release of polar metabolites by mPDAC cells cultured in different culture media conditions
STUDY_SUMMARY
We used quantitative LC-MS metabolite profiling to perform analysis of 108 metabolites that mPDAC cells consume or release while growing in RPMI or TIFM media.
Nontargeted metabolomics analysis on kidney tissue treated with cisplatin
STUDY_TYPE
Nontargeted metabolomics analysis
STUDY_SUMMARY
Cisplatin-induced acute kidney injury (AKI) is a severe clinical complication with no satisfactory therapies in the clinic. Tumor necrosis factor receptor (TNFR)-associated factor 1 (TRAF1) plays an important role in both inflammation and metabolism. However, the role of TRAF1 in cisplatin-induced AKI needs to be evaluated. In this study, TRAF1 expression was decreased in cisplatin-treated mice and mouse proximal tubular cells (mPTCs), suggesting a potential role of TRAF1 in cisplatin-associated kidney injury. Thus, TRAF1 plasmids were introduced into male C57BL/6J mice by a tail vein high-pressure injection method to overexpress TRAF1. Then, cisplatin was administrated by a single intraperitoneal (i.p.) injection (20 mg/kg). Mice were sacrificed 72 h after cisplatin administration. Serum and kidney tissues were collected for further analysis. In vitro, mPTCs were transfected with TRAF1 plasmids before treatment with cisplatin (5 Āµg/mL) for 24 h. Western blotting, Massonās trichrome and hematoxylin-eosin (HE) staining and tandem mass spectrometry (LCāMS/MS) analysis were employed to evaluate kidney injury. Following cisplatin treatment, we observed a marked downregulation of TRAF1 in mouse kidneys and mPTCs treated with cisplatin. In mice, TRAF1 overexpression attenuated cisplatin-induced AKI, as evidenced by decreased levels of serum creatinine (Scr) and blood urea nitrogen (BUN). Moreover, TRAF1 delivery obviously ameliorated cisplatin-induced renal tubular injury, as shown by the improved histological damage and blocked upregulation of NGAL and KIM-1. Moreover, the NF-ĪŗB activation and inflammatory cytokine production enhanced by cisplatin were significantly blunted by TRAF1. In line with the attenuated inflammatory response, the increased number of apoptotic cells (TUNEL staining) and enhanced expression of BAX and cleaved Caspase-3 were markedly decreased by TRAF1 overexpression. In vitro, TRAF1 also attenuated renal tubular cell inflammation and apoptosis induced by cisplatin. In addition, disordered cellular metabolism, which was reported as an important pathogenic factor of AKI, was examined by metabolomics analysis. Interestingly, a significant correction of the metabolic disturbance, including perturbations in energy generation and lipid and amino acid metabolism, was observed in the kidneys of cisplatin-treated mice. In conclusion, TRAF1 overexpression significantly attenuated cisplatin-induced nephrotoxicity, possibly by correcting the impaired metabolism, inhibiting inflammation, and blocking apoptosis in renal tubular cells.
Untargeted Metabolomics on First Trimester Serum Implicates Metabolic Perturbations Associated with BMI in Development of Hypertensive Disorders: A Discovery Study
STUDY_SUMMARY
Body mass index (BMI) in early pregnancy is a critical risk factor for hypertensive disorders of pregnancy (HDP). The pathobiology of the interplay between BMI and HDP is not fully understood and represents the focus of this investigation. BMI and 1st-trimester serum samples were obtained from the Global Alliance to Prevent Prematurity and Stillbirth repository for 154 women (105 without HDP and 49 with HDP). Metabotyping was conducted using ultra-high-performance liquid-chromatography high-resolution mass spectrometry (UHPLC HR-MS). Regression models were used to determine metabolites and pathway perturbations associated with BMI in women with and without HDP, and to determine metabolites and pathway perturbations associated with HDP for women in categories of obese, overweight, and normal weight based on the 1st trimester BMI. This study will lay the groundwork for larger studies of predictive markers and biological pathways leading to infant morbidity and mortality.
Using Mass Spectrometry Imaging to Map Fluxes Quantitatively in the Tumor Ecosystem
STUDY_SUMMARY
Tumors are comprised of a multitude of cell types spanning different microenvironments. Mass spectrometry imaging (MSI) has the potential to identify metabolic patterns within the tumor ecosystem and surrounding tissues, but conventional workflows have not yet fully integrated the breadth of experimental techniques in metabolomics. Here, we combine MSI, stable isotope labeling, and a spatial variant of Isotopologue Spectral Analysis to map distributions of metabolite abundances, nutrient contributions, and metabolic turnover fluxes across the brains of mice harboring GL261 glioma, a widely used model for glioblastoma. When integrated with MSI, the combination of ion mobility, Desorption Electrospray Ionization, and Matrix Assisted Laser Desorption revealed disruption in multiple anabolic pathways. De novo fatty acid synthesis flux was determined to be increased by approximately 3-fold in glioma relative to surrounding healthy tissue. Fatty acid elongation flux was elevated even higher at 8-fold and highlights the importance of elongase activity in glioma. The fluxes we examined were uniformly increased throughout the entire tumor region, revealing a high degree of metabolic homogeneity in our model of glioblastoma.
INSTITUTE
Washington University in St. Louis
DEPARTMENT
Chemistry
LABORATORY
Patti
LAST_NAME
Stancliffe
FIRST_NAME
Ethan
ADDRESS
1 Brookings Dr. Campus Box 1134, St. Louis, MO 63105
Relationships between the gut microbiome and cognitive impairment in residents of long-term aged care.
STUDY_SUMMARY
Ageing-associated cognitive decline affects more than half of those in long-term residential aged care. Emerging evidence suggests that gut microbiome-host interactions influence the effects of modifiable risk factors. We investigated the relationship between gut microbiome characteristics and severity of cognitive impairment (CI) in 159 residents of long-term aged care. Severe CI was associated with a significantly increased abundance of proinflammatory bacterial species, including Methanobrevibacter smithii and Alistipes finegoldii, and decreased relative abundance of beneficial bacterial clades. Severe CI was associated with increased microbial capacity for methanogenesis, and reduced capacity for synthesis of short-chain fatty acids, neurotransmitters glutamate and gamma-aminobutyric acid, and amino acids required for neuro-protective lysosomal activity. These relationships were independent of age, sex, antibiotic exposure, and diet. Our findings implicate multiple gut microbiome-brain pathways in ageing-associated cognitive decline, including inflammation, neurotransmission, and autophagy, and highlight the potential to predict and prevent cognitive decline through microbiome-targeted strategies.
INSTITUTE
South Australian Health and Medical Research Institute
LAST_NAME
Shoubridge
FIRST_NAME
Andrew
ADDRESS
North Terrace, Adelaide, South Australia, 5000, Australia
Effectors enabling adaptation to mitochondrial complex I loss in HĆ¼rthle cell carcinoma
STUDY_TYPE
comparison of tumor versus normal tissue
STUDY_SUMMARY
With the goal of performing RNA-seq and metabolomic profiling, a cohort of fresh frozen oncocytic (HĆ¼rthle cell) thyroid carcinoma (HCC) samples was established with confirmation of mtDNA mutation status and chromosomal copy number. This cohort contained 24 oncocytic (HĆ¼rthle cell) tumors with 18 cases having matched normal thyroid tissue. Tumor samples included 21 primary tumors comprised of 19 HCC (8 widely invasive, 11 minimally invasive) and 2 oncocytic (HĆ¼rthle cell) adenomas as well as 2 locoregional recurrences (LR) and 1 distant metastasis (DM). HCC samples were collected and stored as part of the Mass General Brigham Institutional Review Board (protocol number 2008P001466). Frozen tissue was accessed to create the cohort used in the study.
INSTITUTE
Broad Institute of MIT and Harvard
DEPARTMENT
Metabolomics Platform
LAST_NAME
Clish
FIRST_NAME
Clary
ADDRESS
415 Main Street, Cambridge, MA, 02142, USA
EMAIL
clary@broadinstitute.org
PHONE
617-714-7654
NUM_GROUPS
2
TOTAL_SUBJECTS
tumors from 24 subjects and matched normal tissue from a subset of 18 subjects
Osmoprotectants play a major role in the Portulaca oleracea resistance to high levels of salinity stress - Insights from a metabolomics and proteomics integrated approach
STUDY_SUMMARY
Purslane is an invasive plant and is considered the eighth most common weed in the world. Because of that, its outdoor production in extensive areas faces several concerns. Kong & Zheng (2014) evaluated the potential of producing purslane in a hydroponic system, generating approximately 5.75 kg of fresh matter per m2 per month, which might yield 57.5 tons/hectare/year if cultivated in a bimestrial regime. The high productivity of purslane, when grown in controlled-environment agriculture, can open many opportunities for the purslane industry, even in the context of biosaline agriculture. Building up a robust multi-omics database on the response of purslane to salt stress and the subsequent study of it via an MOI analysis can create the basis for a future system biology approach to decode the genetics behind its resistance to salinity stress. The present study is a second step in building a robust database on the morpho-physiological and molecular responses of Portulaca oleracea L. to salinity stress and its subsequent use in attempting to decode the genetics behind its resistance to this abiotic stress. After reporting on the characterization of the morpho-physiological responses of young purslane plants to such stress using a robust salinization protocol, here we report a study on adult purslane plants through the characterization of the untargeted metabolome and proteome profiles on the leaves and roots of this halophyte species submitted to very high salinity stress, and the consequent use of single- and multi-omics analysis strategies to study it.
INSTITUTE
Embrapa Agroenergy
LAST_NAME
Souza JĆŗnior
FIRST_NAME
Manoel Teixeira
ADDRESS
Parque Estacao Biologica final Asa Norte BrasĆlia DF 70770-901 BR, PQEB, sn - Asa Norte, DF
Microbial metabolomic responses to changes in temperature and salinity along the western Antarctic Peninsula.
STUDY_TYPE
Study of particulate metabolites in phytoplankton and sea-ice algae along the Western Antarctic Peninsula
STUDY_SUMMARY
Seasonal cycles within the marginal ice zones in polar regions include large shifts in temperature and salinity that strongly influence microbial abundance and physiology. However, the combined effects of concurrent temperature and salinity change on microbial community structure and biochemical composition during transitions between seawater and sea ice are not well understood. Coastal marine communities along the western Antarctic Peninsula were sampled and surface seawater was incubated at combinations of temperature and salinity mimicking the formation (cold, salty) and melting (warm, fresh) of sea ice to evaluate how these factors may shape community composition and particulate metabolite pools during seasonal transitions. Bacterial and algal community structures were tightly coupled to each other and distinct across sea-ice, seawater, and sea-ice-meltwater field samples, with unique metabolite profiles in each habitat. During short-term (approximately 10-day) incubations of seawater microbial communities under different temperature and salinity conditions, community compositions changed minimally while metabolite pools shifted greatly, strongly accumulating compatible solutes like proline and glycine betaine under cold and salty conditions. Lower salinities reduced total metabolite concentrations in particulate matter, which may indicate a release of metabolites into the labile dissolved organic matter pool. Low salinity also increased acylcarnitine concentrations in particulate matter, suggesting a potential for fatty acid degradation and reduced nutritional value at the base of the food web during freshening. Our findings have consequences for food web dynamics, microbial interactions, and carbon cycling as polar regions undergo rapid climate change.
Deriving Schwann Cells from hPSCs Enables Disease Modeling and Drug Discovery for Diabetic Peripheral Neuropathy
STUDY_SUMMARY
Schwann cells (SCs) are the major glia of the peripheral nervous system (PNS) and are essential for its function. Defects in SCs are associated with many PNS disorders including diabetic peripheral neuropathy (DPN), a condition affecting millions of patients. We have developed a strategy for deriving SCs from human pluripotent stem cells (hPSCs) which recapitulate the molecular features of primary SCs and are capable of engrafting efficiently and producing myelin in vitro and in injured sciatic nerves in rats. We further established an hPSC-based model of DPN that revealed the selective vulnerability of human SCs to hyperglycemia-induced cytotoxicity. By high-throughput screening we found bupropion counteracts glucose-mediated cytotoxicity in SCs and normalizes glucose-induced transcriptional and metabolic abnormalities in SCs. Treatment of hyperglycemic mice with bupropion rescued sensory function, prevented SC death, and counteracted myelin damage in sciatic nerves. Our retrospective analysis of patient health records revealed that bupropion treatment was associated with a lower incidence of neuropathy among diabetic patients that receive antidepressant medications.
Methionine restriction constrains lipoylation and activates mitochondria for nitrogenic synthesis of amino acids (Part 1)
STUDY_SUMMARY
Methionine restriction (MR) provides metabolic benefits in many organisms. However, mechanisms underlying the MR-induced effect remain incompletely understood. Here, we show in the budding yeast S. cerevisiae that MR relays a signal of S-adenosylmethionine (SAM) deprivation to adapt bioenergetic mitochondria to nitrogenic anabolism. In particular, decreases in cellular SAM constrain lipoate metabolism and protein lipoylation required for the operation of the tricarboxylic acid (TCA) cycle in the mitochondria, leading to incomplete glucose oxidation with an exit of acetyl-CoA and alpha-ketoglutarate from the TCA cycle to the syntheses of amino acids, such as arginine and leucine. This mitochondrial SAM-induced response, namely mitoSIR, promotes cell fitness through the coordination of mitochondrial fuel metabolism with the nitrogenic synthesis of amino acids.
Methionine restriction constrains lipoylation and activates mitochondria for nitrogenic synthesis of amino acids (Part 2)
STUDY_SUMMARY
Methionine restriction (MR) provides metabolic benefits in many organisms. However, mechanisms underlying the MR-induced effect remain incompletely understood. Here, we show in the budding yeast S. cerevisiae that MR relays a signal of S-adenosylmethionine (SAM) deprivation to adapt bioenergetic mitochondria to nitrogenic anabolism. In particular, decreases in cellular SAM constrain lipoate metabolism and protein lipoylation required for the operation of the tricarboxylic acid (TCA) cycle in the mitochondria, leading to incomplete glucose oxidation with an exit of acetyl-CoA and alpha-ketoglutarate from the TCA cycle to the syntheses of amino acids, such as arginine and leucine. This mitochondrial SAM-induced response, namely mitoSIR, promotes cell fitness through the coordination of mitochondrial fuel metabolism with the nitrogenic synthesis of amino acids.
GC/MS analysis of hypoxic volatile metabolic markers in the MDA-MB-231 breast cancer cell line
STUDY_SUMMARY
Hypoxia in disease describes persistent low oxygen conditions, observed in a range of pathologies, including cancer. In the discovery of biomarkers in biological models, pathophysiological traits present a source of translatable metabolic products for the diagnosis of disease in humans. Part of the metabolome is represented by its volatile, gaseous fraction; the volatilome. Human volatile profiles, such as those found in breath, are able to diagnose disease, however accurate volatile biomarker discovery is required to target reliable biomarkers to develop new diagnostic tools. Using custom chambers to control oxygen levels and facilitate headspace sampling, the MDA-MB-231 breast cancer cell line was exposed to hypoxia (1% oxygen) for 24 hours. The maintenance of hypoxic conditions in the system was successfully validated over this time period. Targeted and untargeted gas chromatography mass spectrometry approaches revealed four significantly altered volatile organic compounds when compared to control cells. Three compounds were actively consumed by cells: methyl chloride, acetone and n-Hexane. Cells under hypoxia also produced significant amounts of styrene. This work presents a novel methodology for identification of volatile metabolisms under controlled gas conditions with novel observations of volatile metabolisms by breast cancer cells.
INSTITUTE
University of York
LAST_NAME
Issitt
FIRST_NAME
Theo
ADDRESS
Biology Dept. University of York, Personal
EMAIL
ti538@york.ac.uk
NUM_GROUPS
4
PUBLICATIONS
T. Issitt et al., Volatile compounds in human breath: critical review and meta-analysis Journal of Breath Research, Volume 16, Number 2 (2022) https://iopscience.iop.org/article/10.1088/1752-7163/ac5230#jbrac5230s2
Metabolomic study on the chronic Toxoplasma gondii infection in mice.
STUDY_SUMMARY
Cachexia is a life-threatening disease characterized by chronic, inflammatory muscle wasting and systemic metabolic impairment. Despite its high prevalence, there are no efficacious therapies for cachexia. Mice chronically infected with the protozoan parasite Toxoplasma gondii represent a novel animal model recapitulating the chronic kinetics of cachexia. To understand how perturbations to metabolic tissue homeostasis influence circulating metabolite availability we used mass spectrometry analysis. Despite the significant reduction in circulating triacylglycerides, nonesterified fatty acids, and glycerol, sphingolipid long-chain bases and a subset of phosphatidylcholines (PCs) were significantly increased in the sera of mice with T. gondii infection-induced cachexia. In addition, the TCA cycle intermediates -ketoglutarate, 2- hydroxyglutarate, succinate, fumarate, and malate were highly depleted in cachectic mouse sera. Sphingolipids and their de novo synthesis precursors PCs are the major components of the mitochondrial membrane and regulate mitochondrial function consistent with a causal relationship in the energy imbalance driving T. gondii-induced chronic cachexia.
Lipidomic profile of Toxoplasma gondii-infected mice (Positive mode MS)
STUDY_SUMMARY
Cachexia is a life-threatening disease characterized by chronic, inflammatory muscle wasting and systemic metabolic impairment. Despite its high prevalence, there are no efficacious therapies for cachexia. Mice chronically infected with the protozoan parasite Toxoplasma gondii represent a novel animal model recapitulating the chronic kinetics of cachexia. To understand how perturbations to metabolic tissue homeostasis influence circulating metabolite availability we used mass spectrometry analysis. Despite the significant reduction in circulating triacylglycerides, nonesterified fatty acids, and glycerol, sphingolipid long-chain bases and a subset of phosphatidylcholines (PCs) were significantly increased in the sera of mice with T. gondii infection-induced cachexia. In addition, the TCA cycle intermediates -ketoglutarate, 2- hydroxyglutarate, succinate, fumarate, and malate were highly depleted in cachectic mouse sera. Sphingolipids and their de novo synthesis precursors PCs are the major components of the mitochondrial membrane and regulate mitochondrial function consistent with a causal relationship in the energy imbalance driving T. gondii-induced chronic cachexia.
Lipidomic profile of Toxplasma gondii-infected mice (Negative mode MS)
STUDY_SUMMARY
Cachexia is a life-threatening disease characterized by chronic, inflammatory muscle wasting and systemic metabolic impairment. Despite its high prevalence, there are no efficacious therapies for cachexia. Mice chronically infected with the protozoan parasite Toxoplasma gondii represent a novel animal model recapitulating the chronic kinetics of cachexia. To understand how perturbations to metabolic tissue homeostasis influence circulating metabolite availability we used mass spectrometry analysis. Despite the significant reduction in circulating triacylglycerides, nonesterified fatty acids, and glycerol, sphingolipid long-chain bases and a subset of phosphatidylcholines (PCs) were significantly increased in the sera of mice with T. gondii infection-induced cachexia. In addition, the TCA cycle intermediates -ketoglutarate, 2- hydroxyglutarate, succinate, fumarate, and malate were highly depleted in cachectic mouse sera. Sphingolipids and their de novo synthesis precursors PCs are the major components of the mitochondrial membrane and regulate mitochondrial function consistent with a causal relationship in the energy imbalance driving T. gondii-induced chronic cachexia.
C57bl/6 mice subjected to either PBS or bleomycin treatment to develop fibrosis
STUDY_SUMMARY
C57bl/6 mice subjected to either PBS or bleomycin treatment. By day 21, lung fibrosis will develop. We resected the lung and performed LC-QTOF analysis on small molecule metabolites between the two groups.
INSTITUTE
University of Florida
LAST_NAME
Sun
FIRST_NAME
Ramon
ADDRESS
1200 Newell Drive, ARB, Gainesville, FL, 32610, USA
To study the global metabolic impact of myeloperoxidase-derived oxidants on airway epithelial cells, we developed an in vitro oxidant exposure model. Experiments were conducted with BEAS-2B cells that were supplemented with 100pg/ml recombinant epidermal growth factor and 0.1% FBS prior to oxidant exposure. Measurements were taken at 2, and 6 hours for: (i) H2O2, (ii) HOCl, (iii) HOBr, (iv) HOSCN, and (v) control. Additional measurements were taken at 24 hours for HOSCN and control only.
Lipidomic analysis of serum from mice with Toxoplasma gondii infection
STUDY_SUMMARY
Cachexia is a life-threatening disease characterized by chronic, inflammatory muscle wasting and systemic metabolic impairment. Despite its high prevalence, there are no efficacious therapies for cachexia. Mice chronically infected with the protozoan parasite Toxoplasma gondii represent a novel animal model recapitulating the chronic kinetics of cachexia. To understand how perturbations to metabolic tissue homeostasis influence circulating metabolite availability we used mass spectrometry analysis. Despite the significant reduction in circulating triacylglycerides, nonesterified fatty acids, and glycerol, sphingolipid long-chain bases and a subset of phosphatidylcholines (PCs) were significantly increased in the sera of mice with T. gondii infection-induced cachexia. In addition, the TCA cycle intermediates -ketoglutarate, 2- hydroxyglutarate, succinate, fumarate, and malate were highly depleted in cachectic mouse sera. Sphingolipids and their de novo synthesis precursors PCs are the major components of the mitochondrial membrane and regulate mitochondrial function consistent with a causal relationship in the energy imbalance driving T. gondii-induced chronic cachexia.
Acute respiratory distress syndrome (ARDS) is a heterogeneous disease in its etiology and clinical aspects, and it has been an important interest in how to diagnose it and classify its subtypes, and apply them to treatment. Metabolomics is becoming important for identifying ARDS biology and distinguishing subtypes. The aim of this study is to identify metabolites distinguishing sepsis-induced ARDS patients from healthy controls using a targeted metabolomics approach, and to find out whether direct and indirect ARDS are metabolically distinct groups and confirm their metabolites and associated pathways. Targeted metabolomics was performed to explore metabolome changes between pARDS (pediatric ARDS) and eARDS.
Metabolomics dataset of CNTF induced axon regeneration in mice post optic nerve crush
STUDY_SUMMARY
Axons are processes or extensions of a neuron that help connect one neuron with the next. In the eye all retinal ganglion cells (RGCs) reside within the retina but their axons travel a very long distance traversing through the optic nerve they connect with other neurons in the lateral geniculate nucleus in the brain. Loss of axons results in blindness in glaucoma and traumatic optic neuropathies. Optic nerve crush (ONC) is mouse is an assay system that enable pharmacological induction of axon regeneration from existing RGCs. Lipids form the outer boundary of axons, their synthesis or alterations are associated with metabolite changes. Our motivation was to understand what metabolite changes occurred when ONC axons regenerated due to ciliary neurotrophic factor (CNTF) treatment. We found metabolite profile changes associated with regeneration after crush induced by CNTF. This metabolite dataset was collected from C57Bl/6 mice expressing either AAV2-CNTF to promote regeneration or AAV2-Green Lantern as a control. Animals were subjected to optic nerve crush injury and allowed to recover for either 7 days or 14 days. At the respective time points, animals were euthanized and optic nerves were collected. Nerves underwent two rounds of extraction using a Precellys 24 Touch Homogenizer and a two solvent system of 1:1 Methanol/Water and 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) using a Vanquish Horizon Binary HPLC coupled to a Q Exactive Orbitrap mass spectrometer. Metabolites were identified using Compound Discoverer 3.3 and quantified using isotopic internal metabolite standards.
Biomarker discovery in galactosemia: Metabolomics with UPLC/HRMS in dried blood spots
STUDY_TYPE
Newborn screening
STUDY_SUMMARY
Galactosemia (GAL) is an autosomal recessive genetic disorder characterized by galactose metabolism disturbances. GAL develops non-preventable life-threatening complications even with a reduced content of galactose and lactose patientās diet. Thus, the underlying pathophysiology of long-term complications in GAL remains poorly understood. The current study used a metabolomics approach using ultra-performance liquid chromatography coupled with high-resolution mass spectrometry to investigate the metabolomic changes in the dried blood spots of 15 patients with GAL and 39 healthy individuals. Compared to the control group, 2,819 metabolites underwent significant changes in patients with GAL. In all, 480 human endogenous metabolites were identified, of which 209 and 271 were upregulated and downregulated, respectively. PA (8:0/LTE4) and ganglioside GT1c (d18:0/20:0) metabolites showed the most significant difference between GAL and the healthy group, with an area under the curve of 1 and 0.995, respectively. Additionally, our findings showed novel potential biomarkers for GAL, such as 17-alpha-estradiol-3-glucuronide and 16-alpha-hydroxy DHEA 3-sulfatediphosphate. In conclusion, this metabolomics study deepened the understanding of the pathophysiology of GAL and presented metabolites that might serve as potential prognostic biomarkers to monitor the progression or support the clinical diagnosis of GAL.
Exploring the Impact of Oral Arabic Gum Consumption on Sphingolipid Metabolism and human metabolites in Chronic Kidney Disease: A Mass Spectrometry Analysis
STUDY_SUMMARY
Globally, the incidence of chronic kidney disease is increasing, raising serious concerns about its impact on public health. It also poses significant difficulty in finding novel early diagnostics, understanding biochemical pathways, monitoring patients, and prognosis. Any metabolite found in a biofluid, or tissue may act as a driver, signal, or both in the emergence or spread of the disease. As a result, metabolomics is a very useful strategy in this therapeutic setting. Broad metabolite coverage is essential since it strives to offer a representative image of a biological system. An untargeted metabolomics-based method was used in this cross-sectional study to identify metabolomic changes and their relationship to pathways in the Arabic gum patient group and control participants. Plasma samples were collected from 88 participants who met the inclusion criteria, of whom 43 control patients were treated with a placebo and 45 intervention patients were treated with Arabic gum. Highly sensitive ultra-high-performance liquid chromatography with electrospray ionization and quadrupole time-of-flight mass spectrometry was used to analyze the plasma samples (UHPLC-ESI-QTOF-MS). We investigated the effect of Arabic gum on individual metabolites using a two-tailed independent student t-test. The results showed that 31 out of 92 identified metabolites were found to be statistically significant (p < 0.05). L-Leucine and 5'-Methylthioadenosine were the significantly increased metabolites in the Arabic gum group. Conversely, triethylamine, D-limonene, 4-methylphenylacetic acid, and sphingosine levels were significantly lower in the Arabic gum group compared to the control. Arabic gum primarily affected multiple metabolic pathways, including glycine and serine, arginine and proline, valine, leucine, and isoleucine degradation, phenylalanine and tyrosine, urea cycle, and sphingolipid. The results from this study provide insights into the potential diagnostic significance of different metabolites in chronic kidney disease and their impact on specific metabolic pathways. However, further research involving larger cohorts is necessary to validate the observed metabolite changes following Arabic gum intake and their diagnostic value for chronic kidney disease.
INSTITUTE
Sharjah Institute for Medical Research
DEPARTMENT
Sharjah Institute for Medical Research
LABORATORY
Biomarker Discovery Group
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Untargeted lipidomic analysis of blood plasma samples from drug-naĆÆve patients with bipolar disorder and schizophrenia
STUDY_TYPE
Untargeted lipidomic analysis
STUDY_SUMMARY
In this study, we obtained a lipidomic profile of plasma samples from patients with schizophrenia (SZ) and bipolar disorder (BD) in comparison to healthy controls (CT). The sample cohort consisted of 60 drug-naĆÆve patients and 30 control individuals. Untargeted lipidomics strategy using liquid chromatography coupled to high resolution mass spectrometry was employed to obtain the data, and univariate and multivariate statistical tools were applied to evaluate the results. Metabolic pathway networks were constructed and our results demonstrated alterations in different lipid pathways, such as glycerophospholipids, sphingolipids, and prostaglandins between schizophrenia and bipolar disorder patients. The differential diagnosis is crucial for effective treatment and improving the quality of life of patients with psychotic disorders.
INSTITUTE
University of Campinas
DEPARTMENT
Institute of Chemistry
LABORATORY
LaBIOmics - Laboratory of Bioanalytics and Integrated Omics
Ethnicity-Specific Differences in Ovarian Cancer Metabolic Signatures
STUDY_TYPE
Cultured cells
STUDY_SUMMARY
Ovarian cancer is a leading cause of cancer-related deaths among women worldwide. Cancer cell metabolism plays a critical role in tumor growth and progression, and metabolic alterations in cancer cells have been implicated in treatment resistance. In this study, we performed metabolomic analysis using ovarian cancer cells derived from patients in the United States and Korea. Our results reveal significant ethnic-specific differences in the metabolic signatures of ovarian cancer cells, with differential regulation of metabolites derived from glycolytic pathways, lipid metabolism, and microbiome modified metabolites. These findings have important therapeutic implications, as differences in ovarian cancer metabolism between ethnic groups may influence treatment response and resistance. Targeting the unique metabolic signatures of ovarian cancer cells based on ethnic specificity may improve the effectiveness of precision medicine approaches in the treatment of ovarian cancer. This study highlights the potential for personalized and targeted therapeutic options based on the tumor metabolome and ethnic background of the patient. Overall, our results suggest that investigating ethnic-specific differences in cancer metabolism is critical for developing effective and personalized cancer therapies. The identification of unique metabolic signatures in ovarian cancer cells based on ethnic specificity provides a promising avenue for improving treatment outcomes and advancing the field of precision medicine in ovarian cancer.
Blood metabolomics and impacted cellular mechanisms during transition into lactation in dairy cows that develop metritis
STUDY_TYPE
Case-Control Study
STUDY_SUMMARY
The objective of this study was to identify metabolites associated with metritis and use them for identification of cellular mechanisms affected during transition into lactation. Holstein cows (n = 104) had blood collected in the prepartum period (d-14 Ā± 6), at calving (d0), and at the day of metritis diagnosis (d7 Ā± 2). Cows with reddish or brownish, watery, and fetid discharge were diagnosed with metritis (n = 52). Cows with metritis were paired with herdmates without metritis (n = 52) based on DIM. The metabolome of plasma samples was evaluated using untargeted gas chromatography time-of-flight mass spectrometry. Univariate analyses included t-tests and fold change analyses. Metabolites with false discovery rate (FDR) adjusted P ā¤ 0.10 on t-tests were used for partial least squares ā discriminant analysis PLS-DA coupled with permutational analysis using 2,000 permutations. Metabolites with FDR adjusted P ā¤ 0.10 on t-tests were also used for enriched pathway analyses and identification of cellular processes. Cows that developed metritis had affected cellular processes associated with lower amino acid metabolism in the prepartum period, greater lipolysis, cell death, and oxidative stress at calving and at metritis diagnosis, and greater leukocyte activation at calving, but lower immune cell activation at metritis diagnosis. In summary, cows that developed metritis had plasma metabolomic changes associated with greater lipolysis, oxidative stress, and a dysregulated immune response which may predispose cows to metritis development.
Untargeted Metabolomics Identifies Biomarkers for MCADD Neonates in Dried Blood Spots
STUDY_TYPE
Newborn screening
STUDY_SUMMARY
Medium-chain acyl-CoA dehydrogenase deficiency (MCADD) is the most common inherited mitochondrial metabolic disease of fatty acid Ī²-oxidation, especially in newborns. MCADD is clinically diagnosed using Newborn Bloodspot Screening (NBS) and genetic testing. Still, these methods have limitations, such as false negatives or positives in NBS and variants of uncertain significance in genetic testing. Thus, complementary diagnostic approaches for MCADD are needed. Recently, untargeted metabolomics has been proposed as a diagnostic approach for inherited metabolic diseases (IMDs) due to its ability to detect a wide range of metabolic alterations. We performed untargeted metabolic profiling of dried blood spots (DBS) from MCADD newborns (n=14) and healthy controls (n=14) to discover potential metabolic biomarkers/pathways associated with MCADD. Extracted metabolites from DBS samples were analyzed using UPLC-QToF-MS for untargeted metabolomics analyses. Multivariate and univariate analyses were used to analyze the metabolomics data, and pathway and biomarker analyses were also performed on the significantly endogenous identified metabolites. MCADD newborns had 1034 significantly dysregulated metabolites compared to healthy newborns (Moderated t-test, no correction, p-value ā¤ 0.05, FC 1.5). 23 endogenous metabolites were upregulated, while 84 endogenous metabolites were downregulated. Pathway analyses showed phenylalanine, tyrosine, and tryptophan biosynthesis as the most affected pathway. Potential metabolic biomarkers for MCADD were PGP (a21:0/PG/F1alpha) and glutathione with an area under the curve (AUC) of 0.949 and 0.898, respectively. PGP (a21:0/PG/F1alpha) was the only oxidized lipid in the top-15 biomarker list with the highest p-value and FC. Also, glutathione was chosen to indicate oxidative stress events that could happen during fatty acid oxidation defects. Our findings suggest that MCADD newborns may have oxidative stress events as signs of the disease. However, further validations of these biomarkers are needed in future studies to ensure their accuracy and reliability as complementary markers with established MCADD markers for clinical diagnosis.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
LAST_NAME
AlMalki
FIRST_NAME
Reem
ADDRESS
Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia
Extraction and Untargeted Analysis of Metabolome from Undemineralised Cortical Bone Matrix for Forensic Application
STUDY_SUMMARY
Untargeted metabolomics has become the gold standard for the profiling of low-molecular-weight compounds. Recently, metabolomics has shown great potential in forensic science in the field of toxicology and postmortem interval estimation. The current study aims to evaluate three extraction protocol and four liquid chromatography coupled with mass spectrometry assays that could offer a valuable tool to identify biomarkers for PMI estimation. One fragment for anterior human skeletal tibia from a 82 years old male individual belonging to the Forensic Anthropology Center - Texas State University collection was powdered and extracted in five replicates to be extracted according to a the biphasic chloroform/methanol/water protocol and two single phase protocols based on methanol/water and methanol/acetonitrile/water. Formal analysis was carried out ThermoFisher Ultimate 3000 HPLC in hydrophilic interaction (HILIC) and reverse phase (RP) liquid chromatography coupled with SCIEX 6600 TripleTOF Q-TOF mass spectrometer.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (VCSIP Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
Maternal smoking during pregnancy negatively effects fetal lung development and causes lifelong decreases in offspring pulmonary function and offspring respiratory health including increased risk of childhood asthma development. The VCSIP cohort was created from 2 RCTs to determine if vitamin C supplementation to pregnant smokers could ameliorate some of the negative effects of maternal smoking during pregnancy on offspring lung health (JAMA 311:2074-2082, 2014; JAMA Pediatr 177:16-24, 2023). Both pre- and postnatal tobacco exposures have been carefully monitored and extensive respiratory phenotyping on the offspring have been performed. Please contact Cindy McEvoy at Mcevoyc@ohsu.edu for questions related to subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. VCSIP is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
Oregon Health & Science University
DEPARTMENT
Division of Neuroscience
LAST_NAME
Spindel
FIRST_NAME
Eliot
ADDRESS
3181 SW Sam Jackson Park Road Portland, OR 97239-3098
Hydroxylated acylcarnitines as potential biomarkers for VLCADD newborn patients in Saudi Arabia
STUDY_SUMMARY
Very long acylcarnitine dehydrogenase deficiency (VLCADD) is an inherited metabolic disorder related to fatty acid Ī²-oxidation. It is characterized by genetic mutations in ACADVL gene and accumulations of acylcarnitines. VLCADD can be developed in the neonatal period or during adulthood. Certain diagnostic approaches are used to confirm the diagnosis of VLCADD including genetic sequencing and newborn bloodspot screening (NBS). The last two approaches have shown some limitations such as VUS with genetic sequencing and false positive or negative results in NBS. Therefore, there are demands for additional diagnostic tools for VLCADD. Since VLCADD is associated with disrupted metabolism, untargeted metabolomics, which is an analytical technique used to detect a large-scale profiling of metabolites in biological samples, could be a useful tool for diagnosis. We hypothesized that VLCADD newborns patients may exhibit a unique metabolic profile and biomarkers compared to healthy newborns. Untargeted metabolomics approach was conducted using liquid chromatography-mass spectrometry (LC-MS) to measure the global metabolites in DBS cards collected from VLCADD newborns (n=15) and healthy controls (n=15). Metabolite extraction was performed and followed by LC-MS analysis. Multivariate and univariate analyses were used to analyze the metabolomics data, and pathway and biomarker analyses were also performed on the significantly endogenous identified metabolites. A moderate T-test was used for statistical analysis with no correction, and the cutoff was (p-value ā¤ 0.05 and Fold Change 1.5). VLCADD newborns had 2012 significantly dysregulated metabolites compared to healthy newborns. 58 endogenous metabolites were upregulated while 148 endogenous metabolites were downregulated. Pathway analyses showed phenylalanine, tyrosine, and tryptophan biosynthesis as the most affected pathway. Potential metabolic biomarker for VLCADD was 3,4-dihydroxytetradecanoylcarnitine with an area under the curve (AUC) of 1, was in the top-15 biomarker list with the highest p-value and FC, suggesting its high possibility to be used for diagnosis. However, validation experiments of the biomarker is needed in following-up studies to ensure its accuracy and reliability to be used as a VLCADD marker in the clinical practice.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
LAST_NAME
AlMalki
FIRST_NAME
Reem
ADDRESS
Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia
Metabolomic profiling of PMM2-CDG zebrafish in presence and absence of epalrestat
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Targeting AR with epalrestat decreases polyol levels and increases GDP-mannose in vivo in pmm2 mutant zebrafish.
Metabolomic profiling of PMM2-CDG patient fibroblasts in presence and absence of epalrestat
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Targeting AR with epalrestat decreases polyol levels and increases GDP-mannose both in vitro in patient-derived fibroblasts.
Metabolomic profiling of PMM2-CDG after siRNA mediated KD of AKR1b1
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Ssing tracer glucose studies, we demonstrate that AR inhibition diverts glucose flux away from polyol production towards the synthesis of sugar nucleotides.
Metabolomic profiling of PMM2-CDG after siRNA mediated KD of AKR1b1 and neuraminidase treatment
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Ssing tracer glucose studies, we demonstrate that AR inhibition diverts glucose flux away from polyol production towards the synthesis of sugar nucleotides, which results in increase in glucose flux towards glycans.
Metabolomic profiling of PMM2-CDG after siRNA mediated KD of AKR1b1 - 13C6 glucose and fructose study
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Ssing tracer glucose studies, we demonstrate that AR inhibition diverts glucose flux away from polyol production towards the synthesis of sugar nucleotides.
Metabolomic profiling of PMM2-CDG patient fibroblasts
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols.
Metabolomics of Human islets treated with DHT and high-glucose challenge
STUDY_SUMMARY
Dihydrotestosterone (DHT) driven Androgen receptor activation in pancreatic beta-cells leads to the amplification of GLP-1R-mediated insulin exocytosis. Here we study the impact of DHT on the metabolic architecture of human pancreatic islets upon stimulation with DHT at high (16.7mM) and low (2.8mM) Glucose.
Assessment of metabolic changes in different animal models of maladaptive right ventricular hypertrophy in chronic pulmonary hypertension by lipidomics and HILIC
STUDY_SUMMARY
We analyzed plasma samples from 33 Yucatan pigs belonging to four different experimental models of pulmonary hypertension (PH): M1 chronic postcapillary PH by pulmonary vein banding; M2 chronic PH by aorto-pulmonary shunting; M3: right ventricular pressure overload by pulmonary artery banding, thus without PH; and M0 sham procedure. Briefly, blood samples were collected 8 months after surgery and untargeted lipidomics and HILIC metabolomic analyses were performed for negative and positive polarities. For quality control, 5 QC samples were included in each analysis, iterative MSMS was performed for metabolite annotation. Plasma metabolomic patterns differed among groups, displaying arginine-nitric oxide and histidine deficiency in both PH models (M1 and M2), altered taurine and purine pathways in M2, and lipidomic changes in all three models of pressure overload.
Evaluation of Extraction Parameters for the Analysis of Lipid Classes in Plants
STUDY_SUMMARY
The aim of this study was to preliminary evaluate the lipid profile alterations on Pitcairnia flammea leaves based on variations in solvent proportion and ultrasonic ice bath extraction time, followed by a lipid class-enriched analysis employing chemometric techniques. In the plant extraction method employed, the sample was separated in 3 phases 2 : organic, aqueous and protein phases. The parameters evaluated in our study were different solvent proportion (1:2, 1:3 or 1:4 v/v) and time under ultrasonic in ice-cold bath (10, 20 or 30 min). The extraction solvents were methanol (MeOH) and methyl tert-butyl ether (MTBE). For each condition, experiments were prepared in triplicate resulting in 27 samples. Samples extracted in MeOH:MTBE (1:2 v/v) for 10 min and 20 min ultrasonic bath did not separate in three phases. Then, their organic phase chromatographic analyses were not performed resulting on 42 experimental samples chromatograms. Ultra-high performance liquid chromatography coupled to electrospray ionization mass spectrometry (UHPLC-ESI-MS) was used to acquire raw data and MS-DIAL and MetaboAnalyst platforms were used to perform data preprocessing and statistical analysis. The statistical analysis of UHPLC-ESI-MS data in both ionization modes enabled the visualization of a trend distribution based on extraction time. Furthermore, we were able to establish that the solvent proportion had a greater impact on group separation in data samples extracted for 30 min versus 10 and 20 min. Moreover, diacylglycerol or/and lysophophatidylcholine are lipid subclasses that can be favored depending on the extraction time in the MS analyses using positive ESI mode.
Quantification of cytokinins in ap1 cal AP1-GR and drmy1 ap1 cal AP1-GR inflorescences using LC-MS
STUDY_TYPE
Quantification using mass spectrometry
STUDY_SUMMARY
Robustness is the invariant development of phenotype despite environmental changes and genetic perturbations. In the Arabidopsis flower bud, four sepals initiate at robust positions and times and grow to equal size to enclose and protect the inner floral organs. We previously characterized the mutant development related myb-like1 (drmy1), where 3-5 sepals initiate at irregular positions and variable times and grow to different sizes, compromising their protective function. This loss of robustness was caused by a uniform increase in cytokinin signaling, as revealed by the TCS::GFP reporter, in the floral meristem before sepal initiation. We hypothesized that the increase in cytokinin signaling in drmy1 was due to an increase in the level of cytokinins. To test this idea, we extracted cytokinins from induced inflorescences of wild-type (5 bio-reps) and drmy1 (6 bio-reps) in ap1 cal AP1-GR background. We measured the level of three cytokinin bases, trans-Zeatin (tZ), cis-Zeatin (cZ), and isopentenyladenine (iP), and their corresponding nucleosides (tZR, cZR, and iPR), using liquid chromatography-mass spectrometry. We found that there was no statistically significant differences in cytokinin levels between these genotypes, indicating that the increase in cytokinin signaling in the drmy1 floral meristem is not due to increased cytokinin levels.
Steady State and Cysteine Flux Metabolomics Study in PTEN WT and PTEN KO MEFs
STUDY_TYPE
Quantitative Targeted Mass Spec
STUDY_SUMMARY
In earlier studies we had indication that the tumor suppressor PTEN was downregulating the cystine glutamate antiporter, xCT; therefore to probe whether this effect on xCT was altering cystine uptake and downstream cystine metabolism, we performed steady state metabolomics via targeted LC-MS/MS on PTEN WT and PTEN KO MEFs. Steady state metabolomics revealed that Pten KO MEFs have a sixfold and fourfold increase in intracellular cystine and cysteine abundance, respectively, as well as a higher abundance of glutathione and the glutathione synthesis intermediate gamma-glutamylcysteine, compared to Pten WT MEFs. In addition to cystine import by xCT as a source of cysteine, cysteine can also be funneled into or recycled from the transsulfuration and choline oxidation pathways. Pten KO MEFs were also found to have increased abundance of transsulfuration pathway metabolites, as well as choline oxidation pathway metabolites. Collectively, this suggests that PTEN regulates cysteine and glutathione metabolism and that PTEN KO cells have more glutathione compared to PTEN WT cells. Next to determine if the increased glutathione in the Pten KO MEFs was being synthesized from increased cystine being brought into the cell by xCT, we performed cystine flux metabolomics via targeted LC-MS/MS on PTEN WT and PTEN KO MEFs and using the heavy isotope 13C2-cystine. Cystine Flux metabolomics revealed Pten KO MEFs were found to have a fourfold and threefold higher accumulation of 13C into intracellular cystine and cysteine, respectively, than Pten WT MEFs, indicating that more extracellular cystine is being brought into the cell by xCT. This result seems plausible given these cells were observed to have higher levels of xCT transporters compared to Pten WT MEFs. Furthermore, there was more cystine flux into glutathione synthesis in Pten KO MEFs, indicated by the sevenfold higher accumulation of heavy isotope labeled glutathione and higher accumulation of its preceding intermediate -glutamylcysteine. Together these findings suggest that PTEN loss heightened the cellās ability to import cystine via xCT and as a result increased glutathione pools.
Steady State and Cystine Flux Metabolomics Study in PTEN WT and PTEN KO MEFs
STUDY_TYPE
Quantitative Targeted Mass Spec
STUDY_SUMMARY
In earlier studies we had indication that the tumor suppressor PTEN was downregulating the cystine glutamate antiporter, xCT; therefore to probe whether this effect on xCT was altering cystine uptake and downstream cystine metabolism, we performed steady state metabolomics via targeted LC-MS/MS on PTEN WT and PTEN KO MEFs. Steady state metabolomics revealed that Pten KO MEFs have a sixfold and fourfold increase in intracellular cystine and cysteine abundance, respectively, as well as a higher abundance of glutathione and the glutathione synthesis intermediate gamma-glutamylcysteine, compared to Pten WT MEFs. In addition to cystine import by xCT as a source of cysteine, cysteine can also be funneled into or recycled from the transsulfuration and choline oxidation pathways. Pten KO MEFs were also found to have increased abundance of transsulfuration pathway metabolites, as well as choline oxidation pathway metabolites. Collectively, this suggests that PTEN regulates cysteine and glutathione metabolism and that PTEN KO cells have more glutathione compared to PTEN WT cells. Next to determine if the increased glutathione in the Pten KO MEFs was being synthesized from increased cystine being brought into the cell by xCT, we performed cystine flux metabolomics via targeted LC-MS/MS on PTEN WT and PTEN KO MEFs and using the heavy isotope 13C2-cystine. Cystine Flux metabolomics revealed Pten KO MEFs were found to have a fourfold and threefold higher accumulation of 13C into intracellular cystine and cysteine, respectively, than Pten WT MEFs, indicating that more extracellular cystine is being brought into the cell by xCT. This result seems plausible given these cells were observed to have higher levels of xCT transporters compared to Pten WT MEFs. Furthermore, there was more cystine flux into glutathione synthesis in Pten KO MEFs, indicated by the sevenfold higher accumulation of heavy isotope labeled glutathione and higher accumulation of its preceding intermediate -glutamylcysteine. Together these findings suggest that PTEN loss heightened the cellās ability to import cystine via xCT and as a result increased glutathione pools.
Metabolomic profiling of PMM2-CDG patient fibroblasts by GC/MS
STUDY_SUMMARY
Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols such as sorbitol and mannitol.
Hydroxyproline Modulates Adaptive PD-L1 Expression and Autophagy
STUDY_SUMMARY
The immune checkpoint protein PD-L1 plays critical roles in both immune system homeostasis and tumor progression. Impaired PD-1/PD-L1 function promotes autoimmunity and PD-L1 expression within tumors promotes immune evasion. If and how changes in metabolism or defined metabolites regulate PD-L1 expression is not fully understood. Here, using a metabolomics activity screening-based approach, we have determined that hydroxyproline (Hyp) significantly and directly enhances adaptive (i.e., IFN-Ī³ induced) PD-L1 expression in multiple relevant myeloid and cancer cell types. Mechanistic studies reveal that Hyp acts as an inhibitor of IFN-Ī³-induced autophagic flux, which allows it to regulate this negative feedback mechanism, thereby contributing to its overall effect on PD-L1 expression. Due to its prevalence in fibrotic tumors, these findings suggest that hydroxyproline could contribute to the establishment of an immunosuppressive tumor microenvironment and that Hyp metabolism could be targeted to pharmacologically control PD-L1 expression for the treatment of cancer or autoimmune diseases.
INSTITUTE
Scripps Research Institute/University of California, Los Angeles
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Plasma - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Targeted Keto Acids
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
Metabolomics analysis of ALDH1L1-expressing HuH7 cell lines.
STUDY_SUMMARY
One-carbon metabolism is utilized for the de novo synthesis of purines. Given that tumor cells require large amounts of nucleotides, they are highly dependent on one-carbon metabolism. Among the one-carbon-metabolizing enzymes, the aldehyde dehydrogenase 1 family member L1 (ALDH1L1), which catalyzes 10-formyltetrahydrofolate to trahydrofolate, is considered a tumor suppressor gene, as its expression is reported to be attenuated or diminished in hepatocellular carcinoma. To clarify the effect of ALDH1L1 expression, we generated control and ALDH1L1-expressing HuH-7 (H7-1L1) cells, and measured the amounts of intracellular metabolites in both control and ALDH1L1-expressing by CE-TOFMS analysis.
INSTITUTE
Tohoku Medical and Pharmaceutical University
LAST_NAME
Sasaki
FIRST_NAME
Masato
ADDRESS
4-4-1 Komatsushima, Aoba-ku, Sendai, Miyagi, 981-8558, Japan
Zinc treatment reverses and anti-Zn-regulated miRs suppress esophageal carcinomas in vivo
STUDY_SUMMARY
Using gas chromatography time-of-flight mass spectrometry GC-TOF MS (3), we performed untargeted metabolomic profiling on esophageal mucosa of Zn-treated vs Zn-untreated rats (n = 10 rats per cohort). Thirty-eight significantly altered metabolites (24 down-, 14 up-regulated, P < 0.05) were identified in the Zn-treated vs Zn-untreated esophagus. Of the 24 down-regulated metabolites, 15 (63%) were involved in anabolic/biosynthetic pathways, including amino acid/pyrimidine/purine metabolism and polyamine biosynthesis. Putrescine (intermediate in polyamine biosynthesis), shown to be up 6.4-fold in ESCC-bearing ZD esophagus (3), was down-regulated -3.96-fold in Zn-treated esophagus. Ornithine decarboxylase (ODC) is the rate liming enzyme in the polyamine biosynthetic pathway to form putrescine, which is converted into spermidine and spermine. Polyamines are indispensable for cell proliferation/tumor growth, and depletion of polyamines inhibits tumor growth (6). Of the 14 metabolites that were significantly up-regulated in the Zn-treated esophagus, five (37%) were carbohydrates, including glucose, which is up-regulated 3.4-fold, pointing to a decreased uptake of glucose and a reversal of the Warburg effect after Zn treatment. Critical to cancer aerobic glycolysis is the metabolic enzyme hexokinase 2 (HK2) that catalyzes the first committed step in glucose metabolism where glucose is phosphorylated to form glucose-6-phosphate. HK2 overexpression accounts for the high glycolytic rate in cancer cells (7). In summary, Zn treatment that significantly reduced ESCC incidence reversed classic cancer cell metabolic phenotypes such as increased glycolysis and nucleoside intermediates, with decrease in putrescine, increase in glucose, accompanied by down-regulation of metabolite enzymes ODC and HK2.
Stable isotope tracer analysis (SITA) using 13C6-glucose and 13C5-glutamine measured by GC/MS in BIN-67 cells Ā± SMARCA4 restoration
STUDY_SUMMARY
SMARCA4/2-loss drives the metabolic shift preferring glutamine than glucose to sustain the TCA cycle. SITA using 13C6-glucose confirmed that glucose uptake was increased in BIN-67 cells upon SMARCA4-restoration, leading to overall increased metabolic flux through glucose (lactate) and TCA cycle intermediates (citrate, fumarate, malate, aspartate) via pyruvate carboxylation.Conversely, the 13C5-glutamine SITA in BIN-67 cells revealed that SMARCA4-restoration decreased glutamine uptake and utilization via glutaminolysis to fuel the TCA cycle as indicated by reduced glutamine metabolites (glutamate, Ī±-KG, m+5 metabolites) and TCA cycle intermediates (succinate, fumarate, malate, aspartate, m+4 metabolites)
INSTITUTE
McGill University
DEPARTMENT
Biochemistry
LABORATORY
Sidong Huang Lab
LAST_NAME
Fu
FIRST_NAME
Zheng
ADDRESS
McIntyre Medical Sciences Building, 3655 promenade Sir-William-Osler
Stable isotope tracer analysis (SITA) using 13C6-glucose and 13C5-glutamine measured by GC/MS in H1703 cells Ā± SMARCA4/A2 restoration
STUDY_SUMMARY
SMARCA4/2-loss drives the metabolic shift preferring glutamine than glucose to sustain the TCA cycle. SITA using 13C6-glucose confirmed that glucose uptake was increased in H1703 cells upon SMARCA4/A2-restoration. Conversely, the 13C5-glutamine SITA in H1703 cells revealed that SMARCA4/A2-restoration decreased glutamine uptake and utilization via glutaminolysis to fuel the TCA cycle as indicated by reduced glutamine metabolites (glutamate, Ī±-KG, metabolites) and TCA cycle intermediates (succinate, fumarate, malate, aspartate, metabolites)
INSTITUTE
McGill University
DEPARTMENT
Biochemistry
LABORATORY
Sidong Huang Lab
LAST_NAME
Fu
FIRST_NAME
Zheng
ADDRESS
McIntyre Medical Sciences Building, 3655 promenade Sir-William-Osler
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Keto Acids
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Nucleotides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Heart - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Targeted Keto Acids
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Targeted Nucleotides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Liver - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Targeted Keto Acids
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Targeted Nucleotides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Amines
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Ethanolamides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Keto Acids
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Nucleotides
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Oxylipins
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Tricarboxylic Acid Cycle
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Plasma - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Plasma - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Plasma - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Plasma - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Cortex Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Hypothalamus Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Vastus Lateralis Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Adrenal Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Gastrocnemius Powder - Targeted Acyl-CoA
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Targeted Acyl-CoA
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Targeted Acyl-CoA
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Targeted Acyl-CoA
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Colon Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Spleen Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Testes Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Ovaries Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:Lung - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Lung Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Small Intestine Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Liver Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Brown Adipose Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Untargeted Ion-Pair Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Tissue:White Adipose - Untargeted Lipidomics, Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Untargeted Lipidomics, Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
Per- and poly-fluoroalkyl substances (PFAS) Exposures and Child Health (PEACH) Study: Using targeted exposure assessment and untargeted metabolic profiling to characterize molecular pathways and mechanisms underlying PFAS toxicity on adverse birth and child health outcomes
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The overarching goal of the Per- and poly-fluoroalkyl substances Exposures And Child Health (PEACH) Study is to apply an advanced untargeted metabolomics workflow to investigate associations between PFAS levels, perturbations in maternal and newborn metabolome and adverse birth outcomes. The Emory ECHO team has established a socio-economically diverse, exceptionally phenotyped African American (AA) maternal-child cohort that enrolls pregnant women in the early prenatal period and extends dyad follow-up through age five. PEACH draws from repeated metabolic profiling on a subset of 320 AA pregnant people within the Atlanta ECHO cohort, PFAS assessment, and untargeted metabolomics from newborn blood spots (n=279). Please contact Drs. Donghai Liang (Donghai.liang@emory.edu) and Anne Dunlop (amlang@emory.edu) via email for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) OIF program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. PEACH is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat White Adipose Powder - Untargeted Reversed-Phase Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Vena Cava Powder - Untargeted HILIC-Positive
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
Deficiency of the lipid flippase ATP10A causes diet-induced dyslipidemia in female mice
STUDY_TYPE
MS Untargeted Lipidomics
STUDY_SUMMARY
Genetic association studies have linked ATP10A and closely related type IV P-type ATPases (P4-ATPases) to insulin resistance and vascular complications, such as atherosclerosis. In addition, prior studies of mice harboring large, overlapping chromosomal deletions implicated Atp10A in the development of diet-induced obesity and insulin resistance. Here, we generated gene-specific Atp10A knockout mice and show that Atp10A-/- mice fed a high-fat diet did not gain excess weight relative to wild-type littermates. However, Atp10A-/- mice displayed female-specific dyslipidemia characterized by elevated plasma triglycerides, free fatty acids and cholesterol, as well as altered VLDL and HDL properties. We also observed increased circulating levels of several sphingolipid species along with reduced levels of eicosanoids and bile acids. The Atp10A-/- mice also displayed hepatic insulin resistance without perturbations to whole-body glucose homeostasis. Thus, ATP10A has a sex-specific role in regulating plasma lipid composition and maintaining hepatic liver insulin sensitivity in mice.
Employing Hindlimb Unloading Model for The Identification of Serum Biomarkers Associated with Cardiovascular and Skeletal Muscle Deconditioning.
STUDY_SUMMARY
Microgravity and prolonged periods of inactivity cause a variety of diseases, including skeletal muscle mass loss and weakening as well as cardiovascular deconditioning. The primary causes of the inadequate preventative measures for these deconditionings are the lack of biomarkers and unknown underlying mechanisms of cardiovascular and skeletal muscle deconditioning in these conditions. Here, we used a hindlimb unloading (HU) mouse model that replicates astronauts in space and bedridden patients to first evaluate cardiovascular and skeletal muscle performance. Serum samples from these mice were used toĀ identify new biomarkers using metabolomic and proteomic approaches. Three weeks of unloading resulted in alterations in cardiovascular system function in C57/Bl6 mice, as measured by changes in mean arterial pressure and heart weight. Unloading for three weeks also altered skeletal muscle function, resulting in a decrease of grip strength in HU mice, as well as skeletal muscle atrophy, as shown by a drop in muscle mass. A two-week recovery time from the unloading condition partially reversed these alterations, stressing the importance of the recovery process.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Systemic host inflammation induces stage-specific transcriptomic modification and slower maturation in malaria parasites
STUDY_TYPE
Biomedical research
STUDY_SUMMARY
We found that host inflammation altered the plasma environment surrounding Plasmodium falciparum parasites in vivo, and that this altered plasma environment contained inhibitory factors that directly impaired maturation of early trophozoite stages. We demonstrated with LPS-conditioning that systemic host inflammation alone, in the absence of confounding factors such as ongoing infection, slowed the rate at which parasites transited from one generation of RBC to the next. While this is consistent with the idea that host inflammatory responses can impair parasite maturation, other TLR agonists, CpG and Poly I:C, did not elicit such a response. Metabolomics also identified 1-methylhypoxanthine as elevated in both LPS conditioned and acutely-infected plasma. Plasmodium survival depends on host hypoxanthine, inosine and xanthine for purine synthesis. 1-Methylhypoxanthine can bind effectively to and possibly limit the action of hypoxanthine-guanine phosphoribosyl transferase (HGPRTase)25, an enzyme critical for purine synthesis. Interestingly, hypoxanthine, inosine and xanthine were also all reduced in the plasma of LPS-conditioned and acutely infected mice supporting the possibility that inhibition of purine synthesis by 1-methylhypoxanthine might have been partly aided by the lack of substrates for this pathway.
INSTITUTE
Peter Doherty Institute for Infection and Immunity
DEPARTMENT
Department of Microbiology and Immunology
LABORATORY
Ashraful Haque lab
LAST_NAME
Skinner
FIRST_NAME
Oliver
ADDRESS
792 Elizabeth Street, The University of Melbourne, Victoria 3000 Australia
Spatially resolved metabolomics and isotope tracing reveal dynamic metabolic responses of dentate granule neurons with acute stimulation
STUDY_SUMMARY
To define the metabolic adaptations of the neuron-enriched dentate granule cell layer within the native brain environment, we utilize acute hippocampal brain slices and perform fast metabolite preservation followed by mass spectrometry imaging (MALDI-MSI) to obtain spatially resolved metabolomics and isotope tracing data.
Metabolomic analysis of maternal mid-gestation plasma and cord blood: lipidomics
STUDY_SUMMARY
Metabolomic analysis of maternal mid-gestation plasma and cord blood reveals evidence in autism spectrum disorder of inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. The discovery of prenatal and neonatal molecular biomarkers has the potential to yield insights into autism spectrum disorder (ASD) and facilitate early diagnosis. We characterized metabolomic profiles in ASD using plasma samples collected in the Norwegian Autism Birth Cohort from mothers at weeks 17-21 gestation (maternal mid-gestation, MMG, n=408) and from children on the day of birth (cord blood, CB, n=418). We analyzed associations using sex-stratified adjusted logistic regression models with Bayesian analyses. Chemical enrichment analyses (ChemRICH) were performed to determine altered chemical clusters. We also employed machine learning algorithms to assess the utility of metabolomics as ASD biomarkers. We identified ASD associations with a variety of chemical compounds including arachidonic acid, glutamate, and glutamine, and metabolite clusters including hydroxy eicospentaenoic acids, phosphatidylcholines, and ceramides in MMG and CB plasma that are consistent with inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. Girls with ASD have disruption of ether/non-ether phospholipid balance in the MMG plasma that is similar to that found in other neurodevelopmental disorders. ASD boys in the CB analyses had the highest number of dysregulated chemical clusters. Machine learning classifiers distinguished ASD cases from controls with AUC values ranging from 0.710 to 0.853. Predictive performance was better in CB analyses than in MMG. These findings may provide new insights into the sex-specific differences in ASD and have implications for discovery of biomarkers that may enable early diagnosis and intervention.
The brain and gut are intricately connected in response to various stressful stimuli. Here we describe a brain-to-gut pathway in mice that instructs intestinal stem cells (ISCs) lineage commitment via bacterial metabolic signals. Psychological stress signals from the brain trigger a sympathetic pathway to enrich gut commensal Lactobacillus, which contributes to a transferrable loss of intestinal secretory cell subtypes. Indole-3-acetate (IAA) production by Lactobacillus murinus disrupts mitochondrial bioenergetics of ISCs and blocks secretory lineage commitment. In patients with mental stress, we observe similar enrichment of IAA and Lactobacillus species associated with gut dysfunction. These findings uncover a stress-responsive brain-gut signalling mechanism that skews ISCs fate decision and could be targeted for stress-driven gut-brain comorbidities.
A targeted metabolomics approach for sepsis-induced ARDS and its subphenotypes
STUDY_SUMMARY
Acute respiratory distress syndrome (ARDS) is etiologically and clinically a heterogeneous disease. Its diagnostic characteristics and subtype classification, and the application of these features to treatment, have been of considerable interest. Metabolomics is becoming important for identifying ARDS biology and distinguishing its subtypes. This study aimed to identify metabolites that could distinguish sepsis-induced ARDS patients from non-ARDS controls, using a targeted metabolomics approach, and to identify whether sepsis-induced direct and sepsis-induced indirect ARDS are metabolically distinct groups, and if so, confirm their metabolites and associated pathways.
Multi-Omics Analysis Revealed a Significant Molecular Changes in Doxorubicin-Resistant Lung Cancer Cells.
STUDY_TYPE
LC/MS/MS
STUDY_SUMMARY
Lung cancer is the second most common cancer and the leading cause of cancer-related deaths worldwide. Chemotherapy resistance in lung cancer is one of the major characteristics of an aggressive phenotype with poor prognosis. Therefore, there is a critical need to explore the significant molecular changes associated with resistance to conventional chemotherapy, and identify potential therapeutic targets for treatment of resistant lung cancer. In this study, we have performed comprehensive quantitative proteomics and metabolomics analysis of non-small cell lung cancer cells (A549-P) and doxorubicin resistant A549 cells (A549-R), using state-of-the-art Trapped Ion Mobility Spectroscopy, Time-of-Flight Mass Spectrometry (TIMS-TOF-MS). The results revealed 30 dysregulated proteins and 37 significantly altered metabolites in A549-R cells compared to A549-P cells. Among the significantly upregulated proteins are liver carboxylesterase 1, anterior gradient protein 2 homolog and nicotinamide phosphoribosyltransferase. A group of the upregulated proteins are endogenous and xenobiotic-metabolizing enzymes, including UDP-glucuronosyltransferase 1-6, CES1, and epoxide hydrolase 1. While Importin, ATP-citrate synthase and CTP synthase are downregulated. The significantly altered metabolites include sepiapterin, glutathione, glycine, pyridine and niacinamide. The performed multi-omics integrated analysis revealed the involvement of purine and glutathione metabolism, ABC transporters, citric acid cycle in the development of resistance in A549 cells, besides the involvement of energy metabolism, pathways related to cancer progression, invasion and migration, and redox homeostasis. Collectively, this exploratory study effectively revealed the significantly dysregulated proteins and metabolites in doxorubicin resistant A549 cells and shed the light on potential biomarkers for chemotherapy resistant non-small cell lung cancer. In addition, multi-omics integrated analysis elucidates the involved pathways in resistance including pathways related to progression and invasion which would improve prognosis and open the door for new potential therapeutic targets.
INSTITUTE
University of Sharjah
DEPARTMENT
Research institute of medical and health science
LABORATORY
Biomarker Discovery Group
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (UCP Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The Utah Children's Project is a prospetive cohort study of children and parents recruited from population-based and community-based samples. The purpose of the study is to study the influence of genetics and environment on child health and development. Eligibility required prior participation in a preconception or prenatal cohort study based at the University of Utah and/or Utah State University, with data and biospecimens available to carry forward into the Utah Children's project. The index child enrolled was a result of the pregnancy in the prior study, and up to one sibling was also eligible to enroll. At least one biologic parent enrolled, and both biologic parents were invited to enroll. For further information, contact ucp@hsc.utah.edu, or 801-587-7400. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. UCP is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
University of Utah
DEPARTMENT
School of Medicine
LAST_NAME
Stanford
FIRST_NAME
Joseph
ADDRESS
DFPM UU, 375 Chipeta Way #A, Salt Lake City, UT 84108
Lipidomic Analysis of subventricular zone in adult mice.
STUDY_SUMMARY
Lipid metabolism plays an important role in neurogenesis. The present study was performed to investigate the role of serine racemase in lipid metabolism and adult neurogenesis. Conditional deletion of serine racemase in nestin precursor cells (nestin cre+) showed significant alterations in the different lipid classes in the subventricular zone of adult nestin-cre+ mice based on a lipidomics study. The raw data for the lipidomics study are presented.
Disruption of the circadian clock in skeletal muscle worsens local and systemic health, leading to decreased muscle strength, metabolic dysfunction, and aging-like phenotypes. Whole-body knockout mice that lack Bmal1, a key component of the molecular clock, display premature aging. Here, by using adeno-associated viruses, we rescued Bmal1 expression specifically in the skeletal muscle of Bmal1-KO mice and found that this improves their healthspan and lifespan. Plasma samples from 40-week-old KO and KO+AAV male mice were collected at ZT1 to characterize the systemic effects of muscle-specific expression of Bmal1. Overall, our findings highlight the critical role of skeletal muscle in systemic health.
INSTITUTE
University of Florida
LAST_NAME
Esser
FIRST_NAME
Karyn
ADDRESS
1345 Center Drive, M552, Gainesville, Florida, 32610, USA
Levels of T3 (triiodothyronine) and T4 (thyroxine) in CSF (cerebrospinal fluid) and plasma as part of natural diurnal variation
STUDY_SUMMARY
This study employs targeted LC-MS analysis of CSF and plasma to assess relative changes in the levels of thyroid hormone (T3: triiodothyronine and T4: thyroxine) at two time points in the diurnal cycle. For this purpose, wild-type CD1 mice were kept in a circadian cabinet housing with 12-hour light cycle (7 a.m. on/ 7 p.m. off) and tissues were collected consistently at an a.m and a p.m time point (9 a.m. and 9 p.m). CSF was collected from adult (3 months old). Internal standards (13C-labelled T3 and T4) as well as calibration curves were used to estimate the respective T3 and T4 concentration in the examined biofluids.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Levels of central carbon metabolites in choroid plexus as part of natural diurnal variation
STUDY_SUMMARY
This study employs targeted LC-MS analysis of choroid plexus (ChP) tissue to assess relative changes in the levels of intermediates from the central carbon metabolism, with additional attention to mitochondrial energy precursors (ATP/ADP, NAD(P) and NAD(P)H) at two time points in the diurnal cycle. For this purpose, mice were kept in a circadian cabinet housing with 12-hour light cycle (7 a.m. on/ 7 p.m. off) and tissues were collected consistently at an a.m and a p.m time point (9 a.m. and 9 p.m). Two different ChP tissues were collected for analysis (LV - left ventricle and 4V - 4th ventricle). Also two separate extractions were performed: 80% MetOH based ("MetOH" study factor) and FB (MetOH with Na-ascorbate and Na-acetate additives, "FB" study factor). The two extractions are optimal for central carbon metabolites or NAD(P)/H respectively.
FH variant pathogenicity promotes purine salvage pathway dependence in kidney cancer
STUDY_SUMMARY
The tricarboxylic citric acid cycle enzyme fumarate hydratase (FH) is a tumor suppressor. When lost in cells, its substrate fumarate accumulates to mM levels and drives oncogenic signaling and transformation. Germline alterations lead to an autosomal dominant condition known as hereditary leiomyomatosis and renal cell cancer (HLRCC) where patients are predisposed to various benign tumors and an aggressive form of kidney cancer. FH alterations of unclear significance are frequently observed with germline testing; thus, there is an unmet need to classify FH variants by their cancer-associated risk, allowing for screening, early diagnosis and treatment. Here we quantify catalytic efficiency of 74 FH variants of uncertain significance. Over half were enzymatically inactive which is strong evidence of pathogenicity. We generated a panel of HLRCC cell lines expressing FH variants with a range of catalytic activities, then correlated fumarate levels with metabolic features. We found that fumarate accumulation blocks purine biosynthesis, rendering FH-deficient cells more sensitive to the purine salvage pathway inhibitor 6-mercaptopurine. Together, these findings suggest pathogenicity of many patientassociated FH variants and reveal nucleotide salvage as a targetable vulnerability in FHdeficient cancer cells.
Uncoupled glycerol-3-phosphate shuttle in kidney cancer reveals that cytosolic GPD is essential to support lipid synthesis
STUDY_SUMMARY
The glycerol-3-phosphate shuttle (G3PS) is a major NADH shuttle that regenerates reducing equivalents in the cytosol and produces energy in the mitochondria. Here, we demonstrate that G3PS is uncoupled in kidney cancer cells where the cytosolic reaction is 4.5 times faster than the mitochondrial reaction. The high flux through cytosolic glycerol-3-phosphate dehydrogenase (GPD) is required to maintain redox balance and support lipid synthesis. Interestingly, inhibition of G3PS by knocking down mitochondrial GPD (GPD2) has no effect on mitochondrial respiration. Instead, loss of GPD2 upregulates cytosolic GPD on a transcriptional level and promotes cancer cell proliferation by increasing glycerol-3-phosphate supply. The proliferative advantage of GPD2 knockdown tumor can be abolished by pharmacologic inhibition of lipid synthesis. Taken together, our results suggest that G3PS is not required to run as an intact NADH shuttle but is instead truncated to support complex lipid synthesis in kidney cancer.
INSTITUTE
Harvard Medical School
LAST_NAME
Yao
FIRST_NAME
Conghui
ADDRESS
LHRRB RM301, 240 Longwood Ave, Boston, MA, 02115, USA
Metabolomic analysis of maternal mid-gestation plasma and cord blood: biogenic amines
STUDY_SUMMARY
Metabolomic analysis of maternal mid-gestation plasma and cord blood reveals evidence in autism spectrum disorder of inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. The discovery of prenatal and neonatal molecular biomarkers has the potential to yield insights into autism spectrum disorder (ASD) and facilitate early diagnosis. We characterized metabolomic profiles in ASD using plasma samples collected in the Norwegian Autism Birth Cohort from mothers at weeks 17-21 gestation (maternal mid-gestation, MMG, n=408) and from children on the day of birth (cord blood, CB, n=418). We analyzed associations using sex-stratified adjusted logistic regression models with Bayesian analyses. Chemical enrichment analyses (ChemRICH) were performed to determine altered chemical clusters. We also employed machine learning algorithms to assess the utility of metabolomics as ASD biomarkers. We identified ASD associations with a variety of chemical compounds including arachidonic acid, glutamate, and glutamine, and metabolite clusters including hydroxy eicospentaenoic acids, phosphatidylcholines, and ceramides in MMG and CB plasma that are consistent with inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. Girls with ASD have disruption of ether/non-ether phospholipid balance in the MMG plasma that is similar to that found in other neurodevelopmental disorders. ASD boys in the CB analyses had the highest number of dysregulated chemical clusters. Machine learning classifiers distinguished ASD cases from controls with AUC values ranging from 0.710 to 0.853. Predictive performance was better in CB analyses than in MMG. These findings may provide new insights into the sex-specific differences in ASD and have implications for discovery of biomarkers that may enable early diagnosis and intervention.
Ranolazine induced metabolic rewiring improves melanoma responses to targeted therapy and immunotherapy - metabolomics
STUDY_SUMMARY
Metabolomics analysis was performed on A375 melanoma cells resistant to BRAF inhibitor vemurafenib (VR) and cells resistant to VR and treated with fatty acid oxidation inhibitor ranolazine.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Ranolazine induced metabolic rewiring improves melanoma responses to targeted therapy and immunotherapy - lipidomics
STUDY_SUMMARY
Lipidomics analysis was performed on A375 melanoma cells resistant to BRAF inhibitor vemurafenib (VR) and cells resistant to VR and treated with fatty acid oxidation inhibitor ranolazine.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Loss of microglial MCT4 leads to defective synaptic pruning and anxiety-like behavior in mice
STUDY_SUMMARY
Microglia, the innate immune cells of the central nervous system, actively participate in brain development by supporting neuronal maturation and refining synaptic connections. These cells are emerging as highly metabolically flexible, able to oxidize different energetic substrates to meet their energy demand. Lactate is particularly abundant in the brain, but whether microglia use it as a metabolic fuel has been poorly explored. Here we show that microglia can import lactate, and this is coupled with increased lysosomal acidification. In vitro, loss of the monocarboxylate transporter MCT4 in microglia prevents lactate-induced lysosomal modulation and leads to defective cargo degradation. Microglial depletion of MCT4 in vivo leads to impaired synaptic pruning, associated with increased excitation in hippocampal neurons, enhanced E/I ratio, vulnerability to seizures and anxiety-like phenotype. Overall, these findings show that selective disruption of the MCT4 transporter in microglia is sufficient to alter synapse refinement and to induce defects in brain development and adult behavior.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Metabolic alteration of MCF-7 cells upon indirect exposure to E. coli secretome: A model of studying the microbiota effect on human breast tissue
STUDY_SUMMARY
Cancer is a challenging disease that requires a comprehensive approach for effective treatment. Various bacterial species, including clostridia, bifidobacteria, and salmonellae, have been investigated in numerous animal tumor models, cell lines, and clinical trials as gene carriers for anti-cancerous genes, including tumor suppressor genes, suicide genes, or tumor-associated antigens. Therefore, they render cell cancer more sensitive to treatment, and they can be used as drug/gene delivery vehicles. E. coli, as one of the breast tissue microbiomes, secretes metabolites that could influence the metabolism of MCF-7 cells to ensure their survival. This in vitro investigation concentrated primarily on the role of E. coli secretome modulation on the MCF-7 cells metabolism. The intra- and extracellular metabolomes of the E. coli secretome and secretome exposed MCF-7 cells were profiled using the liquid chromatography-mass spectrometry (LC-MS) metabolomics approach. Secretome-exposed MCF-7 cells were compared to unexposed controls; a total of 31 and 56 metabolites were significantly altered intra- and extracellularly, respectively. The most common metabolic pathways dysregulated after exposure were aminoacyl-tRNA biosynthesis, purine metabolism, and energy metabolism. The decrease in some purine metabolites would suggest that altering nucleotide metabolism is one of the ways the bacterial secretome kills cancer cells. The maximum discrimination between the two groups was found in lactate levels, which plays a crucial role in cancer progression. The Warburg effect causes cancer tissue to have an acidic microenvironment, which impacts cancer cell metastasis and proliferation, inflammation, immune cell function, and blood vessel development; the decrease in lactate content may also be a method by which the secretome affects cancer. Finally, some microbial metabolites from bacterial secretome have shown promising anticancer effects and can be employed as innovative ways for cancer treatment, either alone or in combination with other medicines.
Ventricle-specific myocardial protein and metabolite characterisation in healthy humans, with differential regulation in end-stage cardiomyopathies (Part 1)
STUDY_SUMMARY
The left and right ventricles of the human heart are functionally and developmentally distinct such that genetic or acquired insults can cause dysfunction in one or both ventricles resulting in heart failure. First, we performed unbiased quantitative mass spectrometry on the myocardium of 25-27 pre-mortem cryopreserved non-diseased human hearts to compare the metabolome and proteome between the normal left and right ventricles. Constituents of gluconeogenesis, glycolysis, lipogenesis, lipolysis, fatty acid catabolism, the citrate cycle and oxidative phosphorylation were down-regulated in the left ventricle, while glycogenesis, pyruvate and ketone metabolism were up-regulated. Inter-ventricular significance of these metabolic pathways was then found to be diminished within end-stage dilated cardiomyopathy and ischaemic cardiomyopathy (n = 30-33), while heart failure-associated pathways were increased in the left ventricle relative to the right within ischaemic cardiomyopathy, such as fluid sheer-stress, increased glutamine to glutamate ratio, and down-regulation of contractile proteins indicating a left ventricular pathological bias.
Ventricle-specific myocardial protein and metabolite characterisation in healthy humans, with differential regulation in end-stage cardiomyopathies (Part 2)
STUDY_SUMMARY
The left and right ventricles of the human heart are functionally and developmentally distinct such that genetic or acquired insults can cause dysfunction in one or both ventricles resulting in heart failure. First, we performed unbiased quantitative mass spectrometry on the myocardium of 25-27 pre-mortem cryopreserved non-diseased human hearts to compare the metabolome and proteome between the normal left and right ventricles. Constituents of gluconeogenesis, glycolysis, lipogenesis, lipolysis, fatty acid catabolism, the citrate cycle and oxidative phosphorylation were down-regulated in the left ventricle, while glycogenesis, pyruvate and ketone metabolism were up-regulated. Inter-ventricular significance of these metabolic pathways was then found to be diminished within end-stage dilated cardiomyopathy and ischaemic cardiomyopathy (n = 30-33), while heart failure-associated pathways were increased in the left ventricle relative to the right within ischaemic cardiomyopathy, such as fluid sheer-stress, increased glutamine to glutamate ratio, and down-regulation of contractile proteins indicating a left ventricular pathological bias.
SAND: automated time-domain modeling of NMR spectra applied to metabolic quantification
STUDY_TYPE
spike-in urine sample
STUDY_SUMMARY
New developments in untargeted nuclear magnetic resonance (NMR) metabolomics enable the profiling of hundreds to thousands of biological samples in biomedical studies, with great potential in drug discovery and diagnostics. The exploitation of this rich information requires detailed quantification of spectral features. However, the development of a consistent and automatic workflow for NMR feature quantification has been a long-standing challenge because of the difficulties of extensive spectral overlap. To address this challenge, we introduce the software SAND (Spectral Automated NMR Deconvolution), for automated feature quantification in the time domain. SAND follows upon the previous success of time-domain modeling and provides automated quantification of entire spectra without the need for manual interaction. SAND employs subsampling, global optimization, and statistic model selection, which are readily expandable to higher dimensional NMR and non-uniform sampling applications. Here, we demonstrate the accuracy of the SAND approach (a correlation around 0.9) using highly overlapped simulated datasets, a two-compound mixture, and a urine spectral series spiked with differing amounts of a four-compound mixture. We further demonstrate automated annotation using correlation networks derived from SAND deconvoluted peaks, and on average 74% of peaks for each compound can be recovered in a single correlation network cluster. SAND is currently integrated with NMRbox and the Network for Advanced NMR (NAN).
INSTITUTE
University of Georgia
DEPARTMENT
Genetics; Biochemistry and Molecular Biology; Institute of Bioinformatics; College of Engineering; Complex Carbohydrate Research Center
Comparison of metabolic of A549 cells before and after Gossypol acetate (GAA) treatment
STUDY_SUMMARY
GAA is a natural product with anti-cancer application prospect, but its anti-tumor molecular mechanism is still controversial. Previous studies have showed that GAA can suppress the expression of mitochondrial DNA encoded proteins by targeting LRPPRC, suggesting that GAA may affect mitochondrial metabolism. Here, metabonomics was applied to study of effect of GAA on central carbon metabolism in A549 cells. The metabolomic data showed that GAA significantly decreased tricarboxylic acid cycle ralated metabolites and significantly increased glycolysis-related metabolites. These results indicated that GAA could inhibite oxidative phosphorylation in A549 cells.
INSTITUTE
Hangzhou Institute of Medicine, Chinese Academy of Sciences
LABORATORY
University of Chinese Academy of Sciences (Zhejiang Cancer Hospital)
Metabolic characterization of the polar endometabolome of Triple-Negative Breast Cancer parental and cDDP-resistant cells
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Metabolomics can measure the metabolic response of drug-exposed cells, unveiling insight into drug mechanisms and metabolic markers of drug efficacy, toxicity and resistance. The present 1H NMR metabolomics study aims to describe the polar endometabolome of both MDA-MB-231 parental and cDDP-resistant cells (MDA-MB-231R), which are representative of Triple-Negative Breast Cancer, aiding the current knowledge about the resistant cells metabolism rewiring and disclosing metabolic hotspots as possible targets to counteract the therapy resistance.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LABORATORY
Metabolomics from Ana M. Gil
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Repeated Exposure to Wood Smoke Alters Pulmonary Gene and Metabolic Profiles
STUDY_SUMMARY
This project aims to evaluate the association of eucalyptus wood smoke exposure with perturbations to lung metabolism in a pre-clinical rat model. This cross-sectional study of male Long-Evans rats were exposed whole body to eucalyptus smoke for 4 consecutive days on week 1, followed by a 3-day break, and 3 subsequent days of smoke exposure on week 2. The smoke exposures were set to a target CO of 10ppm for the low smoke group and 20ppm for the high smoke group. Animal were sacrificed and lungs were extracted for metabolomics analysis. Exposures to eucalyptus smoke and biological perturbations to lung metabolism will be associated with known metabolic dysregulation associated with smoke exposures.
There are limited data on the diagnostic accuracy of gut microbial signatures for predicting hepatic decompensation in patients with cirrhosis. The aim of this study is to determine whether a stool genomic and metabolic signature accurately detects hepatic decompensation and mortality risk in cirrhosis secondary to nonalcoholic fatty liver disease (NAFLD). Shotgun metagenomic sequencing was performed on fecal samples collected from a prospective cohort of adults with NAFLD-related cirrhosis. The signatures were further validated with a metabolomic study on serum samples. Finally, we developed a Random Forest machine learning algorithm to make predictions on hepatic decompensation and mortality in NAFLD-related cirrhosis. Here we uploaded the metabolomics study data from LC-MS/MS.
Ceramides profile in HeLa overexpressing nSMase2 (SMPD3)
STUDY_TYPE
Lipidomics LC-MS/MS
STUDY_SUMMARY
This study aim to determine which species of ceramides are modified by overexpression of the protein neutral sphingomyelinase 2 (nSMase2, gene SMPD3). HeLa cells were transfected with a plasmid containing V5-tagged nSMase2. Lipids were extracted after 24h after transfection and the sphingolipid profile was determined by LC-MS/MS.
Metabolic and Proteomic Divergence is Present in Circulating Monocytes and Tissue Resident Macrophages from Berkeley Sickle Cell Anemia and B-thalassemia mice (PBMC's)
STUDY_SUMMARY
Sickle cell disease and Beta-thalassemia represent hemoglobinopathies arising from dysfunctional or under produced beta-globin chains, respectively. In both diseases, red blood cell injury and anemia are the impetus for end organ injury. Because persistent erythrophagocytosis is a hallmark of these genetic maladies it is critical to understand how macrophage phenotype polarizations in tissue compartments can inform on disease progression. Murine models of sickle cell disease and Beta-thalassemia allow for a basic understanding of mechanisms and provide for translation to human disease. A multi-omics approach to understanding macrophage metabolism and protein changes in two murine models of beta-globinopathy was performed on peripheral blood mononuclear cells as well as spleen and liver macrophages isolated from Berkley sickle cell disease (Berk-ss) and heterozygous B1/B2 globin gene deletion (Hbbth3/+) mice. Results from these experiments revealed the metabolome and proteome of macrophages are polarized to a distinct phenotype in Berk-ss and Hbbth3/+ compared each other and their common background mice (C57BL6/J). Further, spleen and liver macrophages revealed distinct disease specific phenotypes, suggesting macrophages become differentially polarized and reprogrammed within tissue compartments. We conclude that tissue recruitment, polarization, metabolic and proteomic reprogramming of macrophages in Berk-ss and Hbbth3/+ mice may be relevant to disease to progression in other tissue.
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboratation with David Irwin
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
Metabolic and Proteomic Divergence is Present in Circulating Monocytes and Tissue Resident Macrophages from Berkeley Sickle Cell Anemia and B-thalassemia mice (Spleen)
STUDY_SUMMARY
Sickle cell disease and Beta-thalassemia represent hemoglobinopathies arising from dysfunctional or under produced beta-globin chains, respectively. In both diseases, red blood cell injury and anemia are the impetus for end organ injury. Because persistent erythrophagocytosis is a hallmark of these genetic maladies it is critical to understand how macrophage phenotype polarizations in tissue compartments can inform on disease progression. Murine models of sickle cell disease and Beta-thalassemia allow for a basic understanding of mechanisms and provide for translation to human disease. A multi-omics approach to understanding macrophage metabolism and protein changes in two murine models of beta-globinopathy was performed on peripheral blood mononuclear cells as well as spleen and liver macrophages isolated from Berkley sickle cell disease (Berk-ss) and heterozygous B1/B2 globin gene deletion (Hbbth3/+) mice. Results from these experiments revealed the metabolome and proteome of macrophages are polarized to a distinct phenotype in Berk-ss and Hbbth3/+ compared each other and their common background mice (C57BL6/J). Further, spleen and liver macrophages revealed distinct disease specific phenotypes, suggesting macrophages become differentially polarized and reprogrammed within tissue compartments. We conclude that tissue recruitment, polarization, metabolic and proteomic reprogramming of macrophages in Berk-ss and Hbbth3/+ mice may be relevant to disease to progression in other tissue.
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboratation with David Irwin
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
Metabolic and Proteomic Divergence is Present in Circulating Monocytes and Tissue Resident Macrophages from Berkeley Sickle Cell Anemia and B-thalassemia mice (Liver)
STUDY_SUMMARY
Sickle cell disease and Beta-thalassemia represent hemoglobinopathies arising from dysfunctional or under produced beta-globin chains, respectively. In both diseases, red blood cell injury and anemia are the impetus for end organ injury. Because persistent erythrophagocytosis is a hallmark of these genetic maladies it is critical to understand how macrophage phenotype polarizations in tissue compartments can inform on disease progression. Murine models of sickle cell disease and Beta-thalassemia allow for a basic understanding of mechanisms and provide for translation to human disease. A multi-omics approach to understanding macrophage metabolism and protein changes in two murine models of beta-globinopathy was performed on peripheral blood mononuclear cells as well as spleen and liver macrophages isolated from Berkley sickle cell disease (Berk-ss) and heterozygous B1/B2 globin gene deletion (Hbbth3/+) mice. Results from these experiments revealed the metabolome and proteome of macrophages are polarized to a distinct phenotype in Berk-ss and Hbbth3/+ compared each other and their common background mice (C57BL6/J). Further, spleen and liver macrophages revealed distinct disease specific phenotypes, suggesting macrophages become differentially polarized and reprogrammed within tissue compartments. We conclude that tissue recruitment, polarization, metabolic and proteomic reprogramming of macrophages in Berk-ss and Hbbth3/+ mice may be relevant to disease to progression in other tissue.
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboratation with David Irwin
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (CHES Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The aims of the NYU Childrenās Health and Environment Study (CHES) are to evaluate influences of prenatal non-persistent chemical exposures on fetal and postnatal growth and pool our data with the US National Institutes of Health Environmental influences on Child Health Outcomes (ECHO) Program to answer collaborative research questions on the impact of the preconceptual, prenatal, and postnatal environment on childhood obesity, neurodevelopment, pre/peri/postnatal outcomes, upper and lower airway outcomes, and positive health. Eligible women were ā„ 18 years old, < 18 weeks pregnant, had a pregnancy that is not medically threatened, and planned to deliver at NYU Langone HospitalāManhattan, Bellevue Hospital, or NYU Langone HospitalāBrooklyn. Data collection in pregnancy was nested in clinical care, with questionnaire and specimen collection conducted during routine prenatal visits at < 18, 18ā25, and > 25 weeks gestation. These have been followed by questionnaire and specimen collection at birth and regular postpartum intervals. Please contact Dr. Leonardo Trasande at Leonardo.Trasande@nyulangone.org for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. CHES is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
NYU Grossman School of Medicine
DEPARTMENT
Department of Pediatrics
LABORATORY
Division of Environmental Pediatrics
LAST_NAME
Trasande
FIRST_NAME
Leonardo
ADDRESS
NYU Langone Health 227 E 30th St, 8th Floor New York, NY 10016
Improved Endurance Capacity of Diabetic Mice during SGLT2 Inhibition: Potential Role of AICARP, an Endogenous AMPK Activator.
STUDY_SUMMARY
Diabetes is associated with an increased risk of deleterious changes in muscle mass and function or sarcopenia, leading to physical inactivity and worsening glycemic control. Given the negative energy balance during sodium-glucose cotransporter 2 (SGLT2) inhibition, whether SGLT2 inhibitors affect skeletal muscle mass and function is a matter of concern. However, how SGLT2 inhibition affects the skeletal muscle function in patients with diabetes remains insufficiently explored. We aimed to explore the effects of canagliflozin (CANA), an SGLT2 inhibitor, on skeletal muscles in genetically diabetic db/db mice.
INSTITUTE
Medical Institute of Bioregulation, Kyushu University
LAST_NAME
Takahashi
FIRST_NAME
Masatomo
ADDRESS
Maidashi 3-1-1, Higashi-ku, Fukuoka, Fukuoka, 8128582, Japan
Multi-Omics profiling of Candida albicans from agar plate and suspension media
STUDY_SUMMARY
Candida albicans is an opportunistic pathogen that is a significant challenge to healthcare facilities worldwide, commonly found in the human gastrointestinal, respiratory, and genitourinary systems. Morphological transition allows yeast cells to diffuse through bloodstream to colonize internal organs, whilst filamentous forms is related to penetration of host mucosa and epidermal surfaces. With the help of novel analytical techniques and instruments developed in the past years, which enabled accurate, simultaneous detection and quantification of proteins and metabolites. We investigated and compared the proteome and metabolome of C. albicans grown on agar plate verses suspension culture to gain insight into the different environmental adaptation and response to stress. Multi-omics (proteomics & metabolomics) analyses were performed using a high-resolution timsTOF mass spectrometer. From the findings reported in this experiment it is worth highlighting that ease of nutritional access in suspension media favours core growth metabolism and increased translation, while impeded access in solid media favours more diverse metabolic pathways. Core growth and replication machinery are enhanced in suspension media, with several terms related to protein translation and core metabolism increased in this media. In contrast, pathogenic cell wall proteins and proteins related to cell surface were increased in cells grown on solid media.
INSTITUTE
University of Sharjah
DEPARTMENT
Research institute of medical and health science
LABORATORY
Biomarker Discovery Group
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Untargeted Multi-Omics of LNCaP Cell-line Treated with Novel DNA Minor Groove Binder and /or Doxorubicin Using Mass-Spectrometry
STUDY_SUMMARY
Prostate cancer poses a significant health risk, ranking as the second most common cancer among men in the United States. However, the effectiveness of current anti-prostate cancer drugs is limited due to increasing drug resistance and side effects. Consequently, there is a pressing need to develop new compounds and identify novel drug targets that can surpass these limitations. Due to their targeted mechanism, DNA minor groove binders (MGBs) are becoming more popular as a relatively safe and effective alternative. In our research, we employed multi-omics techniques to investigate the mechanism of action of a novel MGB compound (MGB4) through LC-MS/MS-based untargeted metabolomics combined with discovery proteomics analysis performed on LNCaP cells, which were treated with MGB4, doxorubicin, or a combination of both compounds. Through a one-way ANOVA test with a significance level of p-value < 0.05, we identified 99 metabolites and 1143 proteins associated with the treatments. Our findings indicate that treating LNCaP cells with doxorubicin or the MGB4 lead compound yielded similar effects, albeit not identical, on the cells. Both compounds deactivated the translation pathway in the cells. Furthermore, we observed alterations in sphingolipid and amino acid metabolic pathways, potentially contributing to the suppression of prostate cancer cell proliferation and division. Additionally, doxorubicin and combined treatments resulted in reduced metabolism of spermine and spermidine, likely stemming from decreased protein synthesis of key enzymes involved in their pathways. Moreover, the combined treatment exhibited a synergistic interaction between the two compounds, leading to altered purine metabolism and a more pronounced reduction in metabolite abundance compared to individual treatments. Overall, our study demonstrates the robustness of the multi-omics approach in elucidating the mechanism of action of promising drug candidates. It also suggests that MGB4 shows potential as a candidate for prostate cancer treatment.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Impaired metabolism predicts coronary artery calcification in women with systemic lupus erythematosus
STUDY_SUMMARY
Background. Patients with systemic lupus erythematosus (SLE) exhibit a high risk for cardiovascular diseases which is not fully explained by the classical Framingham risk factors. SLE is characterized by major metabolic alterations which could contribute to the elevated prevalence of CVD. In order to address this hypothesis, a comprehensive analysis of the circulating metabolome and lipidome was conducted in a large cohort of 211 women with SLE who underwent a multi-detector computed tomography (CT) scan for quantification of coronary artery calcium (CAC), a robust predictor of coronary heart disease (CHD). Results. Beyond traditional risk factors, including age and hypertension, disease activity and duration were independent risk factor for developing CAC in women with SLE. The presence of coronary calcium was associated with major alterations of circulating lipidome dominated by a high abundance of circulating ceramides with very long chain fatty acids. Alteration in multiple metabolic pathways, including purine metabolism, arginine and proline metabolism, and microbiota-derived metabolites, were also associated with CAC in women with SLE. Backward stepwise logistic regression models of lipidomic and metabolomic variables were used to develop prognostic scores. Strikingly, combining metabolic and lipidomic variables to clinical and biological parameters markedly improved the prediction (Area under the curve: 0.887, P<0.001) of the presence of coronary calcium in women with SLE. Conclusion. The present study uncovers the contribution of disturbed metabolism in the presence of coronary artery calcium and the prediction of CHD in SLE. The identification of these novel lipid and metabolite biomarkers may help to stratify patients for reducing CVD morbidity and mortality in SLE.
Plasma metabolic fingerprints for large-scale screening and personalized risk stratification of metabolic syndrome
STUDY_SUMMARY
Direct diagnosis and accurate assessment of metabolic syndrome (MetS) would allow for prompt clinical interventions. However, diagnostic strategies use only traditional risk factors, without considering the complex heterogeneity of MetS. Here, we performed an advanced ferric particle-assisted laser desorption/ionization mass spectrometry (LDI-MS)-based metabolomic analysis of 100 nL of plasma per participant collected from the largest general community cohort (n=13,554) reported to date and extracted a set of 26 hub plasma metabolic fingerprints (PMFs) for MetS and its early identification (pre-MetS). We develop machine learning-based diagnostic models for pre-MetS and MetS with convincing performance through independent validation. These PMFs were applied to assess the contributions of four MetS risk factors in the general population as follows, from large to small contribution: hyperglycemia, hypertension, dyslipidemia, and obesity. We devised a personalized three-dimensional plasma metabolic risk (PMR) stratification to decode the individual metabolic risk into three patterns. During the 4-year follow-up period of 13,554 participants, the accumulation analysis of all-cause death events showed that patients with medium and high risk had HRs of 1.54 (95% CI 1.05-2.28, p = 0.029) and 1.85 (95% CI 1.22-2.79, p = 0.004), respectively, compared to those with low risk. Overall, we provided efficient screening tools to identify patients with pre-MetS and MetS who require treatments in the general community and defined the heterogeneous risk stratification of metabolic phenotypes in real-world settings.
INSTITUTE
Shanghai Jiao Tong University affiliated Renji Hospital
Integrative multi-omics analysis of oncogenic EZH2 mutants: from epigenetic reprogramming to molecular signatures
STUDY_SUMMARY
Metabolic study of Enhancer of zeste homolog 2 (EZH2) isogenic mutants with gain-of-function (GOF) and loss-of-function (LOF) enzymatic activity in HEK-293T cell lines.
Untargeted metabolomics revealed multiple metabolic perturbations in plasma of T2D patients in response to Liraglutide
STUDY_SUMMARY
Despite the global efforts put into the clinical research and studies in order to protect against Type-2 diabetes mellitus (T2DM), the incidence of T2DM remains high causing a major health problem and impacting the health and care systems. Therefore, T2DM-related treatments and therapies are continuously invented for the clinical use, including Liraglutide. The last is a GLP-1 analogue and shows its beneficial health outcomes e.g., improved glycemic control, lower body weight, and reduced cardiovascular disease risks. The intrinsic mechanisms of these beneficial effects are not fully understood; however, our research group has previously published proteomics work demonstrating the involvement of certain important proteins in part in the beneficial health outcomes of Liraglutide. Since proteomics and metabolomics are complementary to each other in the context of the biological pathways, studying the metabolic impacts of Liraglutide on T2DM patients would add further information about the beneficial health outcomes of Liraglutide. Thus, herein, we performed an untargeted metabolomics approach for identifying metabolic pathways impacted by the treatment of Liraglutide in T2DM patients. Methods: Untargeted liquid chromatography coupled with mass spectrometry was used for metabolomics analysis of plasma samples collected from T2DM patients (n=20) before and after receiving Liraglutide treatment. Metabolic profiling and related pathway and network analyses were conducted. Results: The metabolic profiling analyses identified 93 endogenous metabolites were significantly affected by the Liraglutide treatments, which 49 metabolites up-regulated and 44 metabolites down-regulated. Moreover, the metabolic pathway analyses revealed that the most pronounced metabolite and metabolic pathways that were affected by the Liraglutide treatment was Pentose and glucuronate interconversion, suggesting the last may be a potential target of the Liraglutide treatment could be involved in part in the beneficial effects seen in T2DM patients, specially, we found that glucuronate interconversion pathway which is known by its role in eliminating toxic and undesirable substances from the human body, impacted in Liraglutide treated patients. The last findings ar consistence with our previous proteomics findings. Conclusion: These findings, taken together with our previous results, provide a deeper understanding of the underlying mechanisms involved in the beneficial effects of Liraglutide at the proteomic and metabolic levels in T2DM patients.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
LAST_NAME
Al Mogren
FIRST_NAME
Maha
ADDRESS
Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia
Assessing mitochondrial bioenergetics in coronary artery disease: A translational multiomic tissue study in humans (The AMBITION study).
STUDY_SUMMARY
Background: Severe or recurrent myocardial ischemia can lead to chronic left ventricular (LV) dysfunction and heart failure in patients with coronary artery disease (CAD). Objectives: To assess the multiomic profile of LV myocardium in patients with stable CAD. Methods: Patients undergoing coronary artery bypass grafting (CABG) had preoperative quantitative stress perfusion cardiovascular magnetic resonance. During surgery, paired transmural LV biopsies were acquired on the beating heart from a region of inducible ischemia, and a remote LV segment. LV samples from human organ donors were used as controls. Myocardial biopsies underwent high-energy phosphate quantification, liquid chromatography-mass spectrometry and single-nuclei ribonucleic acid sequencing. Results: From 33 patients, 63 LV biopsies were acquired on the beating heart during CABG (mean age 60Ā±9 years, median LV ejection fraction 67% [IQR: 61-71%]). Analysis of LV samples from 11 essentially healthy donor hearts were included. The global myocardial ATP/ADP ratio was reduced in CAD patients as compared to donor LV tissue (median [IQR]: 2.2 [1.5-2.8] versus 7.4 [6.8-8.6], P=0.001), with increased expression of oxidative phosphorylation (OXPHOS) genes encoding the electron transport chain complexes across multiple cell types. Paired analyses of biopsies obtained during CABG from LV segments with or without inducible ischemia revealed no significant difference in the ATP/ADP ratio (P=0.36), broader metabolic profile or expression of ventricular cardiomyocyte genes implicated in OXPHOS. Conclusions: Our results suggest that viable human myocardium in patients with stable CAD has global alterations in bioenergetic and transcriptional profile without large regional differences between areas with or without inducible ischemia.
Human milk-derived 2ā-fucosyllactose (2ā-FL) consumption is associated with health benefits in infancy that extend into adulthood. However, the exact biological functions of 2ā-FL and corresponding mechanisms of action remain largely unknown. Here, we investigated the impact of 2ā-FL on gut microbial metabolism for the prevention of colitis in adulthood. The gut microbiota from adult mice treated with 2ā-FL showed an increase in abundance of several health-associated genera, including Bifidobacterium, and exhibited preventive effects on colitis. Microbial metabolic analysis demonstrated that 26 pathways that are significantly different between non-inflammatory bowel disease individuals and patients with ulcerative colitis (UC) are significantly regulated by 2ā-FL in mice, indicating that 2ā-FL has the potential to directly regulate dysregulated microbial metabolism in UC. Exploratory metabolomics of Bifidobacterium infantis identified novel secreted metabolites significantly enriched by 2ā-FL consumption, including pantothenol. Remarkably, pantothenate significantly protects mucosal barrier and mitigates colitis in adult mice. Thus 2ā-FL-modulated gut microbial metabolism may contribute to the prevention of intestinal inflammation in adulthood.
Metabolic effect of Lamin A/C in oligodendrocyte on brain function
STUDY_TYPE
LC-MS/MS metabolomics of cell type specific Lmna conditional knockout and wildtype mice brains at 26 weeks
STUDY_SUMMARY
Oligodendrocytes are specialized cells which insulate and support axons with their myelin membrane, allowing proper brain function. Here, we identify Lamin A/C (LMNA/C) as essential for transcriptional and functional stability of myelinating oligodendrocytes. We show that LMNA/C levels increase with differentiation of progenitors and that loss of Lmna in differentiated oligodendrocytes profoundly alters their chromatin accessibility and transcriptional signature. Lmna deletion in myelinating glia is compatible with normal developmental myelination. However, altered chromatin accessibility is detected in fully differentiated oligodendrocytes together with increased expression of progenitor genes and decreased levels of lipid-related transcription factors and inner mitochondrial membrane transcripts. As mice age, they start to develop myelin-thinning and progressively worsening motor phenotype. To address the metabolic effect of LMNA/C in oligodendrocyte on brain function, we carried out LC-MS/MS metabolomic study of myelinating glia cell specific Lmna conditional knockout and wildtype mice brains at 26 weeks. Each LC-MS/MS experiment was performed with 3 biological replicates and 4 technical replicates per genotype. Overall, our data identify LMNA/C as essential for maintaining the transcriptional and functional stability of myelinating oligodendrocytes.
INSTITUTE
Advanced Science Research Center - CUNY
DEPARTMENT
Neuroscience
LABORATORY
Casaccia lab, He lab, MALDI and MS core.
LAST_NAME
He
FIRST_NAME
Ye
ADDRESS
85 St. Nicholas Terrace, New York, New York, 10031, USA
Non-targeted metabolomics screen comparing 13C2-acetate labeling of metabolites in CD8+ T cells and NK cells from mouse spleens.
STUDY_SUMMARY
Non-targeted metabolomics screen comparing 13C2-acetate labeling of metabolites in CD8+ T cells and NK cells from mouse spleens (wild type vs ACSS2 knockout C57Bl/6 mice). Metabolites were analyzed using a high-resolution, high-performance LC-MS analysis.
Integration of Meta-Multi-Omics Data Using Probabilistic Graphs and External Knowledge
STUDY_SUMMARY
Multi-omics has the promise to provide a detailed molecular picture for biological systems. Although obtaining multi-omics data is relatively easy, methods that analyze such data have been lagging. In this paper, we present an algorithm that uses probabilistic graph representations and external knowledge to perform optimum structure learning and deduce a multifarious interaction network for multi-omics data from a bacterial community. Kefir grain, a microbial community that ferments milk and creates kefir, represents a self-renewing, stable, natural microbial community. Kefir has been shown to associate with a wide range of health benefits. We obtained a controlled bacterial community using the two most abundant and well-studied species in kefir grains: Lentilactobacillus kefiri and Lactobacillus kefiranofaciens. We applied growth temperatures of 30Ā°C and 37Ā°C, and obtained transcriptomic, metabolomic, and proteomic data for the same 20 samples (10 samples per temperature). We obtained a multi-omics interaction network, which generated insights that would not have been possible with single-omics analysis. We identified interactions among transcripts, proteins, and metabolites suggesting active toxin/antitoxin systems. We also observed multifarious interactions that involved the shikimate pathway. These observations helped explain bacterial adaptation to different stress conditions, co-aggregation, and increased activation of L. kefiranofaciens at 37Ā°C.
Differentiation of ayahuasca samples according to the origin, religious groups, and botanical varieties using multivariate statistical analysis of UHPLC-MS qualitative data
STUDY_SUMMARY
Ayahuasca is a brew used by Amazonian natives for medicinal, spiritual, and cultural purposes; it is prepared by Psychotria viridis Ruiz & Pav. (Rubiaceae) leaves and Banisteriopsis caapi (Spruce ex Griseb.) Morton (Malpighiaceae) liana decoction, mainly. When considering the potential of ayahuasca as a new phytomedicine to treat depression, it is important to establish which parameters can significantly modify the composition of the brew. Therefore, we report herein statistical analyses regarding ayahuasca chemical composition from different geographic origins, religious groups, and botanical varieties of the sample. Also, we analyzed which biological pathway could be associated to the Banisteriopsis caapi varieties' emergence. Ayahuasqueiro group has more influence on sample differentiation than geographical origin. The most important identified compounds for ayahuasqueiro group brew differentiation are glycosylated and/or phenolic. The metabolic pathway with significant variation related to the Banisteriopsios caapi (Spruce ex Griseb.) Morton variety was Arginine and proline metabolism.
Metabolomics comparison of lung fibroblasts from Pteropus alecto and Homo sapiens
STUDY_TYPE
MS Quantitative analysis
STUDY_SUMMARY
Targeted Metabolomics was performed to identify differences in central carbon metabolism between lung fibroblasts from black flying fox fruit bat Pteropus alecto (PaLung cells) and Humans (WI-38 cells). For both PaLung and WI-38 cells, metabolomics measurements were carried out for n = 3 biological replicates.
INSTITUTE
Duke-NUS Medical School
LAST_NAME
Jagannathan
FIRST_NAME
Narendra Suhas
ADDRESS
8 College Road, Singapore, Singapore, 169857, Singapore
Proteins and RNA are able to phase separate from the aqueous cellular environment to form sub-cellular compartments called condensates. This process results in a protein-RNA mixture that is chemically distinct from the surrounding aqueous phase. Here we use mass spectrometry to characterize the metabolomes of condensates. To test this, we prepared mixtures of phase-separated proteins and cellular metabolites and identified metabolites enriched in the condensate phase. Here, we confirmed the presence of lipids in MED1 samples and condensates using reversed-phase LC-MS.
Proteins and RNA are able to phase separate from the aqueous cellular environment to form sub-cellular compartments called condensates. This process results in a protein-RNA mixture that is chemically distinct from the surrounding aqueous phase. Here we use mass spectrometry to characterize the metabolomes of condensates. To test this, we prepared mixtures of phase-separated proteins and cellular metabolites and identified metabolites enriched in the condensate phase. Here, we quantified the concentration of a select set of phospholipids in the aqueous and condensate phase of condensates formed from the low complexity domain of MED1 by comparison with isotopic-labeled phospholipid standards.
Evolutionary genomics identifies host-directed therapeutics to treat intracellular bacterial infections
STUDY_SUMMARY
Obligate intracellular bacteria from the Rickettsiaceae family have shed essential biosynthetic pathways during their evolution towards host dependency. By systematically comparing this cytosolic family of bacteria to the related vacuolar Anaplasmataceae family using a novel computational pipeline called PoMeLo, we identified 20 metabolic pathways that may have been lost since the divergence of Anaplasmataceae and Rickettsiaceae, corresponding to the latterās change to a cytosolic niche. We hypothesized that drug inhibition of these host metabolic pathways would reduce the levels of metabolic products available to the bacteria, thereby inhibiting bacterial growth. We tested 22 commercially available inhibitors for 14 of the identified pathways and found that 59% of the inhibitors reduced bacterial growth at concentrations that did not contribute to host cell cytotoxicity. Of these, 5 inhibitors with an IC50 under 5 ĀµM were tested to determine whether their mode of inhibition was bactericidal or bacteriostatic. Both mycophenolate mofetil, an inhibitor of inosine-5'-monophosphate dehydrogenase in the purine biosynthesis pathway, and roseoflavin, an analog of riboflavin, displayed bactericidal activity. We then took an unbiased metabolomics approach to Rickettsia-infected cells to determine whether there was any overlap between our predicted host pathways and depletion of metabolite levels in infected cells, as measured by mass spectrometry. Our results show that 13 pathways were identified as metabolic gaps in both our computational predictions and our metabolomics analysis. These in vitro validation studies support the feasibility of a novel evolutionary genomics-guided approach for antibiotic drug development against obligate pathogens.
INSTITUTE
CZ Biohub
LAST_NAME
DeFelice
FIRST_NAME
Brian
ADDRESS
1291 Welch Rd., Rm. G0821 (SIM1), Stanford CA, California, 94305, USA
HER2 overexpression initiates breast tumorigenesis non-cell autonomously by inducing oxidative stress in the tissue microenvironment
STUDY_SUMMARY
HER2 is a driver oncogene overexpressed in the majority of premalignant breast tumors known as ductal carcinoma in situ (DCIS). Due to their stemness features, breast cancer stem cells (BCSC) are considered the main drivers of breast tumor initiation and progression. Here, we used clinical samples and mouse models of HER2+ breast tumorigenesis to demonstrate that neither BCSCs nor their cell-of-origin express HER2/Neu in early-stage breast tumors. Instead, our results demonstrate that Neu overexpression results in the transformation of BCSCs in a non-cell autonomous manner via triggering DNA damage and somatic mutagenesis in their Neu-negative cell-of-origin. This is caused by the increased oxidative stress in the tissue microenvironment generated by altered energy metabolism and increased reactive oxygen species level in Neu-overexpressing mammary ducts. Therefore, our findings illustrate a previously unrecognized mechanism of HER2-induced breast tumor initiation, which may have an impact on future preventive treatments for patients with HER2+ DCIS.
Untargeted analysis on Bromeliades leaf samples. The aim of this study was to evaluate the lipid profile alterations on Pitcairnia flammea leaves based on the different altitudes where they were collected. A lipidomic approach was applied to the samples. Ultra-high performance liquid chromatography coupled to electrospray ionization mass spectrometry (UHPLC-ESI-MS) was used to acquire raw data and MS-DIAL was used to perform data preprocessing. The statistical analysis of UHPLC-ESI-MS data in both ionization modes enabled the visualization of a trend distribution based on the altitude. Our study grouped the individuals into three major groups: one with individuals from the lower elevations (UBA, RAN, and COR), another with individuals from the highest elevations (PAP and MAR), and another with individuals from mixed localities from intermediate elevations. Higher elevation population showed an increase in very long chain fatty acids and monounsaturated lipids compared to lower elevation as a response to cold higher elevations affecting the leaves' membrane fluidity. In addition, the higher elevation population showed a higher abundance of hexosylceramides and phosphatidylglycerol compared to lower ones.
A Comprehensive Metabolomics Profile for Newborns with Maple syrup urine disease
STUDY_TYPE
Untargeted LCMS
STUDY_SUMMARY
Background: Maple syrup urine disease (MSUD) is a genetic inherited disorder caused by a defect in the branched-chain alpha-ketoacid dehydrogenase (BCKAD) complex function. This complex usually breaks down three amino acids: leucine, isoleucine, and valine. Therefore, abnormal activity in this process, can impact important bodily functions and lead to metabolic dysregulation related to the disease complications. A wide range of studied endogenous metabolites and dysregulated biomarkers and pathways provide a huge core support for the treatment and follow-up of newborn MSUD patients. Objectives: In this study, we aim to investigate MSUDās distinctive profile in newborn MSUD patients using untargeted metabolomics to contribute to the growing knowledge surrounding MSUD and pathways involved for improving patient outcomes. Methods: In this study, untargeted metabolomics analyses via liquid chromatographyāmass spectrometry was used to investigate metabolic changes in dry blood spot (DBS) of 22 MSUD newborns and 22 healthy newborns. Results: The metabolomics results revealed 1040 significantly dysregulated metabolites, where 303 and 737 were up- and down-regulated, respectively. 480 metabolites were annotated and 210 were identified as endogenous metabolites. The study identified potential biomarkers for MSUD such as L-Alloisoleucine and Methionine sulfoxide were upregulated in MSUD newborn compared to healthy newborns, while LysoPI was downregulated in MSUD newborns. In addition, the most affected pathways in MSUD Newborns were ascorbate and aldarate, Pentose and glucuronate interconversions and pyrimidine metabolism. Conclusion: Our results demonstrate metabolomics as a noninvasive strategy to understand the pathophysiology of the disease and is a promising tool to evaluate the potential biomarkers in the early diagnosis of newborn MSUD. Future studies are needed to correlate these dysregulated metabolites with defective mechanisms.
In this study we used LC-MS and MS/MS to characterize the metabolomes of the input mouse liver metabolites used in the first two studies of this submission.
Proteins and RNA are able to phase separate from the aqueous cellular environment to form sub-cellular compartments called condensates. This process results in a protein-RNA mixture that is chemically distinct from the surrounding aqueous phase. In this project we used mass spectrometry to characterize the metabolomes of condensates. To test this, we prepared mixtures of phase-separated proteins and cellular metabolites and identified metabolites enriched in the condensate phase. These proteins included SARS-CoV-2 nucleocapsid, as well as low complexity domains of MED1 and HNRNPA1. In this sub-study, we examined the metabolomes of the mouse liver samples that were used to conduct the condensate metabolome experiment described above.
Comparative analyses of metabolome for the intact mammary ducts of 8-week-old MMTV-Neu-IRES-Cre mice versus their wildtype littermates
STUDY_TYPE
Comparative
STUDY_SUMMARY
Transgenic overexpression of HER2/Neu oncogene in murine mammary glands result in mammary tumor formation. To elucidate the potential changes in the metabolic activities of HER2/Neu-overexpressing mammary epithelia prior to any obvious tumorigenic growth, we isolated intact mammary ducts from the No:4 inguinal mammary glands of 8-week-old MMTV-Neu-IRES-Cre and WT littermate mice via mild collagenase treatment and performed a LC/MS based untargeted metabolomics assay.
INSTITUTE
Manchester Institute of Biotechnology, University of Manchester
LAST_NAME
Ucar
FIRST_NAME
Ahmet
ADDRESS
Michael Smith building, Oxford Road, Manchester, M13 9PT, UK
100Ī¼l serum of 11-12 weeks old male mice with a background from various diet groups were processed in Metabolon (https://www.metabolon.com) for metabolics analysis.Diet groups are CDm CDl CD, CDm CDl HFD, CDmHFDl HFD, HFDm CDl CD, HFDm CDl HFD, HFDm HFDl CD and HFDm CDl CD. (m: maternal diet, l: lactation)
Metabolomics Profiling of the Antiproliferative, Anti-migratory and Anti-invasive Potential of Amlodipine in Lung Cancer Cells
STUDY_TYPE
LC/MS/MS
STUDY_SUMMARY
Lung cancer is still among the most leading causes of cancer-related deaths across the world. Although chemotherapy is considered as a critical choice to manage/limit cancer growth in lung cancer patients with early-stage and advanced cancer stages, it has many limitations including, at least, the severe side effects and chemoresistance. The latter is one of the considerable challenges to lung cancer treatment. Therefore, identification of new alternative therapies with lesser cytotoxic effects when compared to the currently used chemotherapeutics is one of the current research approaches. Calcium channel blockers (CCBs) are emerging as anti-cancer agents in several cancer types. Our objective is to determine the cytotoxic effect of amlodipine on non-small cell lung cancer (NSCLC) cells. Colorimetric MTT cell proliferation assay was used to analyze cell viability following treatments with amlodipine in A549 and H1299 NSCLC cell lines. ANOVA and Tukeyās multiple comparison test were used to detect statistical significance. Half maximal (50%) inhibitory concentration (IC50) values were obtained by applying nonlinear regression curve fit analysis. To assess the effect of amlodipine on A549 and H1299 NSCLC cells migration and invasion scratch wound-healing assay and cell invasion assay were used. Our study revealed that amlodipine significantly reduced proliferation of cancer cells in a dose-dependent fashion with half maximal (50%) inhibitory concentration (IC50) values of 23 and 25.66 ĀµM in A549 and H1299 cells, respectively. Furthermore, amlodipine was able to reduce the invasiveness and migration of cancer cells, both of which are hallmarks in the pathogenesis of cancer, in both cell lines in a dose-dependent manner. Accordingly, our study provides empirical evidence that amlodipine expresses anti-cancer effect to NSCLC cells. However, additional investigations are required to further confirm our results on a larger scale at the clinical level.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
In this study, we tested the hypothesis that the dose-dependent disruption of propionyl-CoA metabolism produces toxic intermediates that contribute to TCDD hepatotoxicity and progression of steatosis to steatohepatitis with fibrosis. Our results suggest TCDD dose-dependently reduced cobalamin (Cbl aka vitamin B12) levels compromising methylmalonyl-CoA mutase (MUT) activity and limiting the metabolism of propionyl-CoA to succinyl-CoA using the canonical Cbl-dependent carboxylation pathway. Consequently, accumulating propionyl-CoA was redirected to the alternate Cbl-independent beta-oxidation-like pathway resulting in the dose-dependent accumulation of acrylyl-CoA, as indicated by the increase in S-(2-carboxyethyl)-L-cysteine, a conjugate produced following the spontaneous reaction between the sulfhydryl group of cysteine and highly reactive acrylyl-CoA.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (CIOB Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
CIOB is a prospective prenatal cohort study measuring maternal exposures to endocrine disrupting chemicals and place-based, perceptual, and biological markers of chronic psychosocial stressors and their relationships to adverse pregnancy outcomes and neurodevelopment in children. We have recruited nearly 800 pregnant participants from two locations in San Francisco; Zuckerberg San Francisco General Hospital (ZSFG), which serves primarily low income and Latinx patients, and Mission Bay Hospital (MB), which serves higher income and primarily white and Asian patients. We recruited women during their 2nd trimester (13 ā 27 weeks) and collected biospecimens (urine, blood, hair, nails) and questionnaires during in-person visits, and umbilical cord and maternal blood and urine at delivery. We measured multiple classes of chemical exposures in our participants including flame retardants, per- and poly-fluoroalkyl substances (PFAS), phthalates, pesticides, phenols, aromatic amines, and we collaborated on development and implementation of shared standardized core data elements, questionnaires at multiple time points during pregnancy and childhood, and collection of biospecimens for chemical and mechanistic analyses during pregnancy and childhood. Please contact Erin DeMicco at erin.demicco@ucsf.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. CIOB is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Metabolic responses of normal rat kidneys to a high salt intake (Plasma)
STUDY_SUMMARY
In this study, novel methods were developed which allowed continuous (24/7) measurement of arterial blood pressure and renal blood flow in freely moving rats and the intermittent collection of arterial and renal venous blood to estimate kidney metabolic fluxes of O2 and metabolites. Specifically, the study determined the effects of a high salt (HS; 4.0% NaCl) diet upon whole kidney O2 consumption and arterial and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. A separate group of rats was studied to determine changes in the cortex and outer medulla tissue metabolomic profiles before and following the switch from a 0.4% to 4.0% NaCl diet.
Metabolic responses of normal rat kidneys to a high salt intake (Kidney cortex)
STUDY_TYPE
Time-course metabolomics experiment
STUDY_SUMMARY
In this study, novel methods were developed which allowed continuous (24/7) measurement of arterial blood pressure and renal blood flow in freely moving rats and the intermittent collection of arterial and renal venous blood to estimate kidney metabolic fluxes of O2 and metabolites. Specifically, the study determined the effects of a high salt (HS; 4.0% NaCl) diet upon whole kidney O2 consumption and arterial and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. A separate group of rats was studied to determine changes in the cortex and outer medulla tissue metabolomic profiles before and following the switch from a 0.4% to 4.0% NaCl diet.
Metabolic responses of normal rat kidneys to a high salt intake (Kidney outer medulla)
STUDY_TYPE
Time-course metabolomics experiment
STUDY_SUMMARY
In this study, novel methods were developed which allowed continuous (24/7) measurement of arterial blood pressure and renal blood flow in freely moving rats and the intermittent collection of arterial and renal venous blood to estimate kidney metabolic fluxes of O2 and metabolites. Specifically, the study determined the effects of a high salt (HS; 4.0% NaCl) diet upon whole kidney O2 consumption and arterial and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. A separate group of rats was studied to determine changes in the cortex and outer medulla tissue metabolomic profiles before and following the switch from a 0.4% to 4.0% NaCl diet.
Metabolic responses of normal rat kidneys to a high salt intake (Urine)
STUDY_TYPE
Time-course metabolomics experiment
STUDY_SUMMARY
In this study, novel methods were developed which allowed continuous (24/7) measurement of arterial blood pressure and renal blood flow in freely moving rats and the intermittent collection of arterial and renal venous blood to estimate kidney metabolic fluxes of O2 and metabolites. Specifically, the study determined the effects of a high salt (HS; 4.0% NaCl) diet upon whole kidney O2 consumption and arterial and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. A separate group of rats was studied to determine changes in the cortex and outer medulla tissue metabolomic profiles before and following the switch from a 0.4% to 4.0% NaCl diet.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (ECHO-NOVI Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The Neonatal Neurobehavior and Outcomes in Very Preterm Infants cohort is a prospective study of infants born <30 weeks postmenstrual age who through ECHO are being followed from age 2 to 7 to understand the neurobehavioral and other health outcomes of very preterm infants. The goal is to identify environmental factors and behaviors that can place these children at greater risk for or provide protection from adverse developmental outcomes. Please contact Carmen Marsit at carmen.j.marsit@emory.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. ECHO-NOVI is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO-NOVI Cohort Award, UH3OD023347 to mPIs Barry Lester, Women and Infants Hospital, Providence, RI and Carmen Marsit, Emory University, Atlanta, GA. ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
Emory University
DEPARTMENT
Rollins School of Public Health
LAST_NAME
Marsit
FIRST_NAME
Carmen
ADDRESS
1518 Clifton Road, NE 8008 Claudia Nance Rollins Building Atlanta, GA 30322
Metabolomic and lipidomic characterization of Haemophilus influenzae Rd KW20
STUDY_SUMMARY
This study aims to comprehensively characterize the polar metabolome and the phosphlipidome of Haemophilus influenzae Rd Kw20, to provide an improved understanding of this bacterial metabolome by comparison with data inferred from genome annotation data.
INSTITUTE
Universidad CEU San Pablo
LAST_NAME
GarcĆa
FIRST_NAME
Antonia
ADDRESS
Centro de MetabolĆ³mica y BioanĆ”lisis (CEMBIO), Facultad de Farmacia, Universidad San Pablo-CEU, UrbanizaciĆ³n MonteprĆncipe, 28660, Boadilla del Monte, EspaƱa
Identification of pre-diagnostic lipid sets associated with liver cancer risk using untargeted lipidomics and chemical set analysis ā a nested case-control study within the ATBC cohort
STUDY_SUMMARY
In pre-disposed individuals, a reprogramming of the hepatic lipid metabolism may support liver cancer initiation. We conducted a high-resolution mass spectrometry based untargeted lipidomics analysis of pre-diagnostic serum samples from a nested case-control study (219 liver cancer cases and 219 controls) within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. Out of 462 annotated lipids, 158 (34.2%) were associated with liver cancer risk in a conditional logistic regression analysis at a false discovery rate (FDR) < 0.05. A chemical set enrichment analysis (ChemRICH) and co-regulatory set analysis suggested that 22/28 lipid classes and 47/83 correlation modules were significantly associated with liver cancer risk (FDR <0.05). Strong positive associations were observed for monounsaturated fatty acids (MUFA), triacylglycerols (TAGs), and phosphatidylcholines (PCs) having MUFA acyl chains. Negative associations were observed for sphingolipids (ceramides and sphingomyelins), lysophosphatidylcholines, cholesterol esters and polyunsaturated fatty acids (PUFA) containing TAGs and PCs. Stearoyl-CoA desaturase enzyme 1 (SCD1), a rate limiting enzyme in fatty acid metabolism and ceramidases seems to be critical in this reprogramming. In conclusion, our study reports pre-diagnostic lipid changes that provide novel insights into hepatic lipid metabolism reprogramming may contribute to a pro-cell growth and anti-apoptotic tissue environment and, in turn, support liver cancer initiation. Study
INSTITUTE
Icahn School of Medicine at Mount Sinai
DEPARTMENT
Environmental Medicine and Public Health
LABORATORY
Integrated Data Science Laboratory for Metabolomics and Exposomics
Disorder of Lipids Induced by Gestational Asthma and its Effect on the Development of Fetal Lung Function and Fetal Health
STUDY_SUMMARY
Maternal asthma during pregnancy is highly correlated with fetal growth and development, and can cause damage to both the mother and fetus, but the underlying mechanisms are not yet clear. Amniotic fluid, as the environment for fetal growth and development, may be affected by lipid metabolism disorders, which can impact fetal lung function development. A rat model of asthma during pregnancy induced by common allergen house dust mite (HDM) was used to investigate changes in lipid composition in amniotic fluid and bronchoalveolar lavage fluid (BALF) by ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS), revealing the impact of maternal asthma during pregnancy on fetal lipid metabolism. In this study, maternal asthma aggravated inflammatory indicators and pathological manifestations after fetal allergen exposure, creating a high oxidative stress growth environment for the fetus, and causing metabolic differences in various lipid groups, including phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and fatty acids (FA), indicating significant lipid metabolism disorders. Improving lipid metabolism may help asthmatic pregnant women maintain healthy fetal development.
INSTITUTE
Nanjing University of Chinese Medicine
LAST_NAME
Fang
FIRST_NAME
Huafeng
ADDRESS
No.138 xianlin road, nanjing city, Nanjing, China, 210046, China
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (GAPPS Cohort)
STUDY_SUMMARY
The GAPPS study was designed to establish a large biorepository during the pregnancy and early postnatal period that could be used for research on reducing adverse birth outcomes. Pregnant women were eligible if they were older than 18 years of age, English-speaking and had plans to deliver at the hospital in which they enrolled. Eligibility for ECHO-PATHWAYS included the following: recruitment at a Washington study site, consent to contact for future research, at least one pregnancy urine sample available, pregnancy questionnaire data collected and child age between 4ā8 years old. GAPPS-PW has two study sites, Seattle, Washington, and Yakima, Washington. GAPPS is conducting two middle childhood visits ā one at age 5-6 years and another at 8-9 years. The research is supported by NIH (UG3/UH3OD023271, P30ES007033, UL1 TR002319). This research was conducted using specimens and data collected and stored on behalf of the Global Alliance to Prevent Prematurity and Stillbirth (GAPPS) Repository. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. GAPPS is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539 Citation: LeWinn KZ, Karr CJ, Hazlehurst M, Carroll K, Loftus C, Nguyen R, Barrett E, Swan SH, Szpiro AA, Paquette A, Moore P, Spalt E, Younglove L, Sullivan A, Colburn T, Byington N, Sims Taylor L, Moe S, Wang S, Cordeiro A, Mattias A, Powell J, Johnson T, Norona-Zhou A, Mason A, Bush NR, Sathyanarayana S. Cohort profile: the ECHO prenatal and early childhood pathways to health consortium (ECHO-PATHWAYS). BMJ Open. 2022 Oct 21;12(10):e064288. doi: 10.1136/bmjopen-2022-064288. PMID: 36270755; PMCID: PMC9594508.
Metabolomic analysis of maternal mid-gestation plasma and cord blood: oxylipins
STUDY_SUMMARY
Metabolomic analysis of maternal mid-gestation plasma and cord blood reveals evidence in autism spectrum disorder of inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. The discovery of prenatal and neonatal molecular biomarkers has the potential to yield insights into autism spectrum disorder (ASD) and facilitate early diagnosis. We characterized metabolomic profiles in ASD using plasma samples collected in the Norwegian Autism Birth Cohort from mothers at weeks 17-21 gestation (maternal mid-gestation, MMG, n=408) and from children on the day of birth (cord blood, CB, n=418). We analyzed associations using sex-stratified adjusted logistic regression models with Bayesian analyses. Chemical enrichment analyses (ChemRICH) were performed to determine altered chemical clusters. We also employed machine learning algorithms to assess the utility of metabolomics as ASD biomarkers. We identified ASD associations with a variety of chemical compounds including arachidonic acid, glutamate, and glutamine, and metabolite clusters including hydroxy eicospentaenoic acids, phosphatidylcholines, and ceramides in MMG and CB plasma that are consistent with inflammation, disruption of membrane integrity, and impaired neurotransmission and neurotoxicity. Girls with ASD have disruption of ether/non-ether phospholipid balance in the MMG plasma that is similar to that found in other neurodevelopmental disorders. ASD boys in the CB analyses had the highest number of dysregulated chemical clusters. Machine learning classifiers distinguished ASD cases from controls with AUC values ranging from 0.710 to 0.853. Predictive performance was better in CB analyses than in MMG. These findings may provide new insights into the sex-specific differences in ASD and have implications for discovery of biomarkers that may enable early diagnosis and intervention.
Dysregulation of neural activity and microglia function following exposure to the global environmental contaminant perfluorooctane sulfonate (PFOS)
STUDY_SUMMARY
Humans are chronically exposed to complex chemical mixtures and, correspondingly, researchers are disentangling the contribution of different contaminants to human neuropathologies. Per- and polyfluoroalkyl substances (PFAS) are biopersistent pollutants and, due to their diverse applications, have become global contaminants. Perfluorooctane sulfonate (PFOS), a prevalent PFAS congener, impairs humoral immunity; however, its impact on innate immunity is unclear. Given the critical roles of innate immune cells, namely microglia, in brain development and homeostasis, we asked whether exposure adversely affects microglial function. Herein, we demonstrate developmental PFOS exposure produces microglial activation and upregulation of the microglia activation gene p2ry12. PFOS-induced microglial activation heightened microglial responses to brain injury, in the absence of increased cell death or inflammation. Use of the photoconvertible calcium indicator CaMPARI revealed PFOS exposure heightened neural activity, while optogenetic silencing of neurons was sufficient to normalize microglial responses to injury. Through an untargeted metabolome wide association study (MWAS), we further determined that PFOS-exposed larvae exhibit significant neurochemical imbalances. Exposure to the perfluorooctanoic acid, an immunotoxic PFAS, did not alter neuronal activity or microglial behavior, further supporting a role for neural activity as a critical modifier of microglial function. Together, this study reveals how contaminant-induced changes in brain activity can shape brain health.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (INSPIRE Cohort)
STUDY_TYPE
Prospective Birth Cohort Study
STUDY_SUMMARY
The INSPIRE (Infant Susceptibility to Pulmonary Infections and Asthma Following RSV Exposure Study) is a population-based birth cohort of 1952 term healthy infants who will be followed to at least 11 years of age. The INSPIRE cohort was established under NIAID U19 funding in 2012 and is one of the larger ECHO cohorts. The cohort was designed such that infants were born June through December and were, on average, 6 months of age or less during their first RSV season. Biweekly surveillance was done November through April of the first year of life, with in-person respiratory illness visits conducted for those who met criteria. Viral pathogens were identified by PCR, and RSV serology was done at one year. Children are followed annually, in some years by survey, and in some years by in-person visits which include skin prick testing to aeroallergens, spirometry with test of reversibility and FeNO. Intermediate outcomes include infant respiratory morbidity, recurrent wheezing, and atopy. Asthma is defined annually using standardized questionnaires. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. INSPIRE is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
Vanderbilt University Medical Center
DEPARTMENT
Vanderbilt University Medical Center, Department of Medicine, Department of Pediatrics
LAST_NAME
Hartert
FIRST_NAME
Tina
ADDRESS
Center for Asthma Research 2525 West End Ave, Suite 450 Nashville, TN 37203
Metabolomics and Molecular Networking-Guided Screening of Bacillus-Derived Bioactive Compounds Against a Highly Lethal Vibrio Species
STUDY_SUMMARY
During our search for active substances capable of inhibiting a newly discovered highly lethal Vibrio strain vp-HL, we found that the fermentation broth of multiple Bacillus strains exhibited antibacterial activity. However, the substances responsible for the activity remained unclear. Metabolomics was employed in conjunction with bioactivity screening to identify the antibacterial compounds from Bacillus strains. The Ethyl Acetate extracts of the broth of 20 Bacillus strains were analyzed using UPLC-MS/MS. Principal Component Analysis (PCA) and Orthogonal Partial Least Square - Discriminant Analysis (OPLS-DA) was used for pattern recognition. The strains fall into two groups in the scores plot from PCA. The S-plot from OPLS-DA offered information on biomarkers.
INSTITUTE
Third Institute of Oceanography, Ministry of Natural Resources
Day-night fluctuations in Cerebrospinal fluid metabolomics
STUDY_TYPE
Circadian (NaĆÆve animals)
STUDY_SUMMARY
This study was designed to investigate whether the molecular drivers underlying the diurnal fluctuation of CSF secretion rate could be found in either the choroid plexus tissue itself and/or in the CSF surrounding it. Male Sprague-Dawley rats, kept in light-controlled 12:12 conditions, presented with diurnally modulated CSF metabolite composition. This change was accompanied with differential expression of 2778 genes within the choroid plexus, with several plasma membrane transporters and CLOCK-associated genes diurnally modulated.
INSTITUTE
University of Copenhagen
DEPARTMENT
Department of Neuroscience
LABORATORY
MacAulay Group
LAST_NAME
Edelbo
FIRST_NAME
Beatriche
ADDRESS
Blegdamsvej 3B
EMAIL
beatriche.henriksen@sund.ku.dk
PHONE
+4527697585
NUM_GROUPS
2
TOTAL_SUBJECTS
25
NUM_MALES
25
STUDY_COMMENTS
2 night animals were excluded due to comments regarding sampling/behavior (N4 and N11)
PUBLICATIONS
Day-night fluctuations in choroid plexus transcriptomics and cerebrospinal fluid metabolomics
A nested case-control study of untargeted plasma metabolomics and lung cancer risk among never-smoking women in Shanghai Womenās Health Study
STUDY_SUMMARY
Background: The etiology of lung cancer among never smokers has not been fully elucidated despite 15% of cases in men and 53% in women worldwide are not smoking-related. Metabolomics provides a snapshot of dynamic biochemical activities, including those found to be driving tumor formation and progression. This study used untargeted metabolomics with network analysis to agnostically identify network modules and independent metabolites in pre-diagnostic blood samples among never-smokers to further understand the pathogenesis of lung cancer. Methods and Findings: Within the prospective Shanghai Womenās Health Study, we conducted a nested case-control study of 395 never-smoking incident lung cancer cases and 395 never-smoking controls matched on age. We performed liquid chromatography high-resolution mass spectrometry to quantify 20,348 metabolic features in plasma. We agnostically constructed 28 network modules using a weighted correlation network analysis approach and assessed associations for network modules and individual metabolites with lung cancer using conditional logistic regression models, adjusting for covariates. We accounted for multiple testing using a false discovery rate (FDR) < 0.20. We identified a network module of 122 metabolic features enriched in lysophosphatidylethanolamines that was associated with all lung cancer combined (p = 0.001, FDR = 0.028) and lung adenocarcinoma (p = 0.002, FDR = 0.056) and another network module of 440 metabolic features that was associated with lung adenocarcinoma (p = 0.014, FDR = 0.196). Metabolic features were enriched in pathways associated with cell growth and proliferation, including oxidative stress, bile acid biosynthesis, and metabolism of nucleic acids, carbohydrates, and amino acids, including 1-carbon compounds. Conclusions: Our prospective study suggests that untargeted plasma metabolomics in pre-diagnostic samples could provide new insights into the etiology of lung cancer in never-smokers. Replication and further characterization of these associations are warranted.
INSTITUTE
Emory University
DEPARTMENT
Gangarosa Department of Environmental Health
LABORATORY
Comprehensive Laboratory for Untargeted Exposome Science
LAST_NAME
Walker
FIRST_NAME
Douglas
ADDRESS
1518 Clifton Rd, CNR 7025, Atlanta, GA 30322
EMAIL
douglas.walker@emory.edu
PHONE
(404) 727-6123
NUM_GROUPS
2
TOTAL_SUBJECTS
790
NUM_FEMALES
790
STUDY_COMMENTS
Samples were collected from participants enrolled in the Shanghai Women's Health Study
Metabolomics studies on L4-5 Dorsal Root Ganglia of Ctrl and cKO mice
STUDY_SUMMARY
Metabolomics studies on Dorsal Root Ganglia (DRGs) (Ctrl and cKO).The specific knockout of PP2Cm in the DRG was achieved by intracellular injection of AAV9-Pirt-Cre (1.0Ć1013 vg/ml) or AAV9-Pirt (1.0Ć1013 vg/ml)) as control. Mice were used for experiments at 4 weeks after the injection.
INSTITUTE
West China Hospital of Sichuan University
LAST_NAME
Li
FIRST_NAME
Tao
ADDRESS
No. 37 Guoxue Road, Wuhou District, Chendu 610041, Sichuan, China
Zebrafish Retina Regeneration Metabolomics - 3 Days Post Crush
STUDY_SUMMARY
Retinal regeneration has been at the forefront of optic research. Regenerative model organisms provide key information regarding treatment for optic nerve and retinal degeneration in mammalian species; specifically, Zebrafish (Danio Rerio) have the capacity for successful adult retinal regeneration. Mammals, however, lack this intrinsic ability and undergo irreversible neurodegeneration seen in glaucoma, diabetes and other optic neuropathies. Optic nerve and retinal regeneration are often studied using the retina obtained via optic nerve crush, a mechanical neurodegenerative model. Untargeted metabolomic studies within successful regenerative models are deficient. Evaluation of tissue metabolomic changes in active zebrafish retinal regeneration can elucidate prioritized metabolite pathways that can be targeted in mammalian systems for therapeutic development. Female and male (6 month to 1 year old) right Zebrafish (Tg(gap43:GFP)) optic nerves were crushed and the retinas were collected three days after. Contralateral, uninjured optic nerve retinas were collected as controls. The tissue was dissected from euthanized fish and frozen on dry ice. Samples were pooled for each category (female crush, female control, male crush, male control) and pooled at n = 10-12 to obtain sufficient metabolite concentrations for analysis. Retinal regeneration was verified by microscope visualization of GFP fluorescence. Metabolites were extracted using a Precellys Homogenizer and a serial extraction method: (1) 1:1 Methanol/Water and (2) 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. Metabolites were identified and quantified using Compound Discoverer 3.3 and isotopic internal metabolites standards.
Zebrafish Optic Nerve Regeneration, Tectum Metabolomics - 3 Days Post Crush
STUDY_SUMMARY
Optic nerve crush provides insight into the metabolome of the tectum. Regenerative model organisms provide key information regarding treatment degeneration in mammalian species; specifically, Zebrafish (Danio Rerio) have the capacity for successful adult optic nerve regeneration. Mammals, however, lack this intrinsic ability and undergo irreversible neurodegeneration seen in glaucoma, diabetes and other optic neuropathies. Optic nerve regeneration as well as the tectum metabolome are often studied to enhance regenerative research. Untargeted metabolomic studies within successful regenerative models are deficient. Evaluation of tectum tissue metabolomic changes in active zebrafish regeneration can elucidate prioritized metabolite pathways that can be targeted in mammalian systems for therapeutic development. Female and male (6 month to 1 year old) right Zebrafish (Tg(gap43:GFP)) optic nerves were crushed and the tecta were collected three days after. Contralateral, uninjured optic nerve tecta were collected as controls. The tissue was dissected from euthanized fish and frozen on dry ice. Samples were pooled for each category (female crush, female control, male crush, male control) and pooled at n = 10-12 to obtain sufficient metabolite concentrations for analysis. Optic nerve regeneration was verified by microscope visualization of GFP fluorescence. Metabolites were extracted using a Precellys Homogenizer and a serial extraction method: (1) 1:1 Methanol/Water and (2) 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. Metabolites were identified and quantified using Compound Discoverer 3.3 and isotopic internal metabolites standards.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (Project Viva)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
Project Viva is a ground breaking longitudinal research study of women and their children. The initial goal of Project Viva was to find ways to improve the health of mothers and children by looking at the effects of mother's diet as well as other factors during pregnancy and after birth. Over the past two decades, Project Viva has expanded its focus to include a wider range of experiences that influence health extending into midlife for the mothers, and young adulthood for their children. Please see projectviva.org or contact Emily Oken at Project_Viva@point32health.org for more information or questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Project Viva is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Cell Lineage-Guided Microanalytical Mass Spectrometry Reveals Increased Energy Metabolism and Reactive Oxygen Species in the Vertebrate Organizer
STUDY_SUMMARY
We performed targeted metabolomic analysis on the Spemann-Mangold Organizer (SMO) tissue in the frog (Xenopus laevis) and the remainder of dissected embryos (RE). Metabolites were extracted from the dissected tissues, reconstituted, and analyzed using liquid chromatography (LC) electrospray ionization (ESI) mass spectrometry (MS). The targeted metabolite measurements were performed on a trapped ion mobility time-of-flight mass spectrometer (timsTOF PRO, Bruker).
Contribution of sugar transporters to spatial microbiota colonization along the longitudinal root axis of Arabidopsis
STUDY_SUMMARY
Longitudinal gut-axes of animals show spatial heterogeneity of microbiota related to physiological differentiation. Plant roots show functional heterogeneity in cellular architecture, transcriptome, metabolic states, and immunity. We hypothesized that axial differentiation impacts spatial colonization by rhizobiota along the root axis. We developed two growth systems, ArtSoil and CD-Rhizotrons, to dissect Arabidopsis thaliana roots into three segments. We identified distinct rhizobiota communities in the segments, supporting spatial microbiota differentiation along the axis. Root metabolite profiling revealed differential enrichment and specificity. Correlation analyses point to strong reliance of rhizobiota on carbohydrate supply from the host. Bioinformatic analyses and GUS histochemistry indicate sugar and/or microbe-induced accumulation of SWEET2, 4, and 12 sugar uniporters. Profiling of root-segments in sweet mutants showed impaired spatial rhizobiota arrangement. Correlation analysis revealed complex interconnected metabolite-rhizobiota networks. This work uncovers interdependency between root physiology and microbiota colonization and a contribution of SWEETs to shaping local adaptation of root microbiota.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (CANDLE Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The CANDLE study is a contemporary birth cohort representing a racially and socioeconomically diverse southern metropolitan population of the US. It was designed to investigate the effects of different factors, such as prenatal characteristics, home environment, childhood experiences, genetics, and exposure to potentially harmful substances on the neurocognitive development of children from birth to age four years. The CANDLE study enrolled a total of 1,503 healthy women during their 2nd trimester of pregnancy at four hospitals in Memphis, TN. The sample accrued between 2006 and 2011, demographically representing Shelby County TN, a highly disadvantaged urban population. Study participants are primarily African American and White. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. CANDLE is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier.D U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Lipidomics analysis of maternal obesity model - wild type
STUDY_SUMMARY
10Ī¼l liver tissue of 11-12 weeks old male mice from various diet groups with various genetic background were homogenized and lipids were extracted for shotgun lipidomics experiments. Raw files were converted to .mzml files and imported into and analyzed by LipidXplorer software using custom mfql files to identify sample lipids and internal standards. For further data processing, absolute amounts were calculated using the internal standard intensities followed by the calculated mol% of the identified lipids. For wild-type C57BL/6JRcc mice, diet groups are CDm CDl CD, CDm CDl HFD, CDm HFDl HFD, HFDm CDl CD, HFDm CDl HFD, HFDm HFDl CD and HFDm CDl CD. m: maternal diet, l: lactation phase diet.
Lipidomics analysis of maternal obesity model - knock out
STUDY_SUMMARY
10Ī¼l liver tissue of 11-12 weeks old male mice from various diet groups with various genetic background were homogenized and lipids were extracted for shotgun lipidomics experiments. Raw files were converted to .mzml files and imported into and analyzed by LipidXplorer software using custom mfql files to identify sample lipids and internal standards. For further data processing, absolute amounts were calculated using the internal standard intensities followed by the calculated mol% of the identified lipids. For Hif1a flox/flox or Hif1a flox/flox;LysMCre/+ mice with a C57BL/6JRcc background, diet groups contain CDm CD, CDm HFD, HFDm CD and HFDm HFD. m: maternal diet
Metabolic changes of the pig kidney during isolated normothermic perfusion with autologous whole blood - perfusate
STUDY_SUMMARY
This study investigates how glucose, lactate and 20 amino acids in the perfusate of normothermically perfused pig kidneys changes over time. It also studies how type and severity of ischemia before perfusion (cold or warm ischemia) change this profile.
Metabolic changes of the pig kidney during isolated normothermic perfusion with red blood cell based perfusate - perfusate
STUDY_SUMMARY
This study investigates how glucose, lactate and 20 amino acids in the perfusate of normothermically perfused pig kidneys changes over time. It also studies how type and severity of ischemia before perfusion (cold or warm ischemia) change this profile.
Metabolic changes of the human kidney during isolated normothermic perfusion with red blood cell based perfusate - perfusate
STUDY_TYPE
Experimental
STUDY_SUMMARY
This study investigates how glucose, lactate and 20 amino acids in the perfusate of normothermically perfused human kidneys that were not suitable for transplantation changes over time.
Metabolic changes of the pig kidney during isolated normothermic perfusion with autologous whole blood - urine
STUDY_SUMMARY
This study investigates how glucose, lactate and 20 amino acids in the urine of normothermically perfused pig kidneys changes over time. It also studies how type and severity of ischemia before perfusion (cold or warm ischemia) change this profile.
Metabolomic analysis of gut metabolites in colorectal cancer patients: correlation with disease development and outcome
STUDY_SUMMARY
In this study, targeted metabolomic sequencing was performed on fecal samples from 35 colorectal cancer (CRC) patients, 37 colorectal adenoma patients (CRA), and 30 healthy controls (HC) to identify metabolite biomarkers. Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify metabolomic features distinguishing the three groups. ROC analysis found that 9,10-diHOME, cholesterol CE (18:2), and lipoxinA4 distinguished CRC from HC with an AUC of 0.969. The study highlights the advantages and potential applications of using LC-MS for targeted metabolomic analysis.
INSTITUTE
Wuhan University of Science and Technology
DEPARTMENT
School of Medicine
LABORATORY
Hubei Province Key Laboratory of Occupational Hazard Identification and Control
LAST_NAME
Xie
FIRST_NAME
Zhufu
ADDRESS
No.2 Huangjiahu Road, Hongshan District, Wuhan City, Hubei Province, China
Metabolomic analysis of particulate matter in the NPSG during a 2018 cruise on the R/V Falkor
STUDY_SUMMARY
Targeted and untargeted analysis of metabolomics samples from the North Pacific Subtropical Gyre taken during the 2018 March/April SCOPE cruise on the R/V Falkor (FK180310) across a mesoscale eddy dipole. Particulate matter was collected on 0.2um filters and extracted using a modified Bligh & Dyer before analysis on a QE Orbitrap. Results show significant changes in the absolute quantity and relative composition of particles in the gyre between anticyclonic and cyclonic eddies.
Metabolomic analysis of particulate matter in the NPSG during a 2017 cruise on the R/V Kilo Moana as part of the MESOSCOPE program
STUDY_SUMMARY
Targeted and untargeted analysis of metabolomics samples from the North Pacific Subtropical Gyre taken during the 2017 June/July SCOPE cruise on the R/V Kilo Moana (KM1709) across a mesoscale eddy dipole, with high-resolution depth profile sampling across the DCM in the center of each eddy. Particulate matter was collected on 0.2um filters and extracted using a modified Bligh & Dyer before analysis on a QE Orbitrap. Results show significant changes in the absolute quantity and relative composition of particles in the gyre between anticyclonic and cyclonic eddies.
Community metabolomes reflect taxon-specific fingerprints of phytoplankton and heterotrophic bacteria in the ocean (expanded)
STUDY_TYPE
Metabolomic survey of 46 phytoplankton and heterotrophic bacteria species
STUDY_SUMMARY
This study is an updated version of the prior study in this project, ST001514. With this update we introduce additional marine species, in particular the heterotrophic bacteria, and include our expanded library of authentic standards (available at https://github.com/IngallsLabUW/Ingalls_Standards/blob/430992e10c0464a817c69d22e3905d73f2ea196f/Ingalls_Lab_Standards.csv) for improved targeted coverage.
The Experiment analyzes the cancer cell metabolism in two pancreatic cancer cell lines, (i.e. Panc02 and KPC FC 1245) after the knockdown of the protein cytidine deaminase (CDA). Murine Panc02 and KPC FC1245 pancreatic cancer cell lines were genetically engineered using a doxycycline inducible CRISPR/Cas9 platform with a target specific gRNA for CDA and a control non-targeting NT gRNA. CDA knockdown cells show a significant decrease of intracellular uridine levels and the accumulation of intracellular cytidine. Consistent with the decrease in intracellular uridine, sgCda cells show reduced intracellular levels of UMP, UDP and UTP compared to sgNT cells while we see no change in adenine and cytosine nucleotides (i.e., AMP, ADP, ATP and CMP, CDP, CTP, respectively) or in UDP-hexose.
Chemoproteomics validates selective targeting of Plasmodium M1 alanyl aminopeptidase as a cross-species strategy to treat malaria
STUDY_SUMMARY
All current treatments for malaria are threatened by drug resistance, and new drug candidates that act on novel pathways are urgently needed. Here, we describe MIPS2673, a selective inhibitor of the Plasmodium M1 alanyl metalloaminopeptidase, which displays excellent in vitro antimalarial activity with no significant host cell toxicity. Biochemical assays revealed potent inhibition of recombinant Plasmodium falciparum (PfA-M1) and Plasmodium vivax (Pv-M1) M1 metalloaminopeptidases, with selectivity over other Plasmodium and human aminopeptidases. Orthogonal chemoproteomic methods based on thermal stability and limited proteolysis reproducibly identified PfA-M1 as the sole target of MIPS2673 in parasites from approximately 2,000 detected proteins. Furthermore, the limited proteolysis approach enabled estimation of the binding site on PfA-M1 to within ~5 Ć of that determined by X-ray crystallography. Functional investigation by untargeted metabolomics further demonstrated that MIPS2673 inhibits the key role of PfA-M1 in haemoglobin digestion. Combined, our proteomics and metabolomics target deconvolution strategies provided unbiased confirmation of the on-target activity of a PfA-M1 inhibitor, and validated selective inhibition of this enzyme as a promising multi-stage and cross-species antimalarial strategy.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Interspecies metabolite transfer fuels methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production
STUDY_SUMMARY
The major oral odor compound methyl mercaptan (CH3SH) is strongly associated with halitosis and periodontitis. CH3SH production stems from metabolism of polymicrobial communities in periodontal pockets and on the tongue dorsum. However, understanding of CH3SH-producing oral bacteria and their interactions is limited. This study aimed to investigate CH3SH production by major oral bacteria and the impact of interspecies interactions on its generation. Using a newly constructed large-volume anaerobic non-contact coculture system, Fusobacterium nucleatum was found to be a potent producer of CH3SH, with that production stimulated by metabolic interactions with Streptococcus gordonii, an early dental plaque colonizer. Furthermore, analysis of extracellular amino acids using an S. gordonii arginine-ornithine antiporter (ArcD) mutant demonstrated that ornithine excreted from S. gordonii is a key contributor to increased CH3SH production by F. nucleatum. Further study with 13C, 15N-methionine, as well as gene expression analysis, revealed that ornithine secreted by S. gordonii increased the demand for methionine through accelerated polyamine synthesis by F. nucleatum, leading to elevated methionine pathway activity and CH3SH production. Collectively, these findings suggest that interaction between S. gordonii and F. nucleatum plays a key role in CH3SH production, providing new insight into the mechanism of CH3SH generation in oral microbial communities. Better understanding of the underlying interactions among oral bacteria involved in CH3SH generation can lead to development of more appropriate prophylactic approaches to treat halitosis and periodontitis.
Metabolomics Study on Plasma and Lung Tissue of Rats Exposed to Whole Thorax Irradiation.
STUDY_SUMMARY
Although some progress has been made in the study of radiation injury, there are still no effective prevention and treatment methods for severe acute radiation syndrome or sickness (ARS). Accordingly, a thorough understanding of biological characteristics associated with high-dose radiation is essential for revealing the mechanisms underlying the varied biological processes following high dose radiation and the development of novel potent radioprotective agents. In present study, plasma metabolic characteristics were investigated from patients of hematopoietic stem cell transplantation following high-dose TBI pretreatment utilizing gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS). The most potential panel of four metabolic markers for radiation injury was selected and the metabolic disorders involved were explored. The metabolic disorders implied the dysregulation of gut microflora, shift of energy supply from aerobic respiration to ketogenesis, protein synthesis and metabolism in response to TBI. Although similar metabolic alternation patterns exist between male and female following high-dose irradiation, specific changes are observed in either male or female patients. These findings provide valuable information for further selecting biomarkers, clues of pathogenic mechanisms involved in high-dose radiation exposure.
INSTITUTE
Soochow University
LAST_NAME
Wang
FIRST_NAME
Chang
ADDRESS
Suzhou, No. 199, Renai Road, Suzhou Industrial Park
Functional divergence of CYP76AKs shapes the chemodiversity of abietane-type diterpenoids in genus Salvia
STUDY_SUMMARY
The genus Salvia L. (Lamiaceae) comprises myriad distinct medicinal herbs, with terpenoids as one of their major active chemical groups. Abietane-type diterpenoids (ATDs), such as tanshinones and carnosic acids, are specific to Salvia and exhibit taxonomic chemical diversity among lineages. To elucidate how ATD chemical diversity evolved, we carried out large-scale metabolic and phylogenetic analyses of 71 Salvia species, combined with enzyme function, ancestral sequence and chemical trait reconstruction, and comparative genomics experiments. This integrated approach showed that the lineage-wide ATD diversities in Salvia were induced by differences in the oxidation of the terpenoid skeleton at C-20, which was caused by the functional divergence of the cytochrome P450 subfamily CYP76AK. These findings present a unique pattern of chemical diversity in plants that was shaped by the loss of enzyme activity and associated catalytic pathways.
INSTITUTE
Shanghai University of Traditional Chinese Medicine
Comparison of different blood samples (whole blood, serum and plasma)
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
To evaluate the performance of compounds annotation in real biological samples, we conducted LC-MS/MS based metabolomics including both DDA and SWATH-DIA on paired blood samples of different types (plasma, serum and whole blood) and compared their chemical differences.
INSTITUTE
McGill University
DEPARTMENT
Faculty of Agricultural and Environmental Sciences
LABORATORY
Xia Lab, Institute of Parasitology
LAST_NAME
Xia
FIRST_NAME
Jeff
ADDRESS
Macdonald Campus, McGill University 21,111 Lakeshore Road
Fetal metabolic adaptations to cardiovascular stress in twin-twin transfusion syndrome
STUDY_SUMMARY
Monochorionic-diamniotic twin pregnancies comprise 70% of identical twin pregnancies and are susceptible to unique complications arising from a single placenta shared by two fetuses. Twin-twin transfusion syndrome (TTTS) is a constellation of disturbances caused by unequal blood flow within the shared placenta giving rise to a major hemodynamic imbalance between the twins. If untreated, it leads to fetal cardiac failure and death. Here, we applied TTTS as a model to uncover fetal metabolic adaptations to cardiovascular stress. We compared untargeted mass spectrometry-based metabolomic analyses of amniotic fluid samples from a cohort of severe TTTS cases showing sonographic evidence of increased afterload and heart failure vs. uncomplicated singleton controls. Amniotic fluid metabolites demonstrated footprints of changes in fatty acid, glucose, and steroid hormone metabolism in TTTS. Among TTTS cases, unsupervised principal component analysis revealed two distinct clusters of disease defined by levels of glucose metabolites, amino acids, urea, and redox status. Our results suggest that the human fetal heart can adapt to hemodynamic stress by modulating its glucose metabolism. Furthermore, we have uncovered heterogeneity among cases of severe TTTS suggesting potential differences in the ability of individual fetuses to respond to cardiovascular stress.
INSTITUTE
University of Texas Health Science Center at Houston
Evaluation of quantitative performance with serial dilutions
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
To benchmark the quantitative performance of MetaboAnalystR, a dilution series was prepared by mixing serum and urine in a cross-gradient manner (0:100, 1:1024, 1:256, 1:64, 1:16, 1:4, 1:1, 4:1, 16:1, 64:1, 256:1, 1024:1, 100:0).
INSTITUTE
McGill University
DEPARTMENT
Faculty of Agricultural and Environmental Sciences
LABORATORY
Xia Lab, Institute of Parasitology
LAST_NAME
Xia
FIRST_NAME
Jeff
ADDRESS
Macdonald Campus, McGill University 21,111 Lakeshore Road
Gestational asthma causes lipid disturbance in fetal alveolar lavage fluid
STUDY_SUMMARY
Maternal asthma during pregnancy is highly correlated with fetal growth and development, and can cause damage to both the mother and fetus, but the underlying mechanisms are not yet clear. Amniotic fluid, as the environment for fetal growth and development, may be affected by lipid metabolism disorders, which can impact fetal lung function development. A rat model of asthma during pregnancy induced by common allergen house dust mite (HDM) was used to investigate changes in lipid composition in amniotic fluid and bronchoalveolar lavage fluid (BALF) by ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS), revealing the impact of maternal asthma during pregnancy on fetal lipid metabolism. In this study, maternal asthma aggravated inflammatory indicators and pathological manifestations after fetal allergen exposure, creating a high oxidative stress growth environment for the fetus, and causing metabolic differences in various lipid groups, including phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and fatty acids (FA), indicating significant lipid metabolism disorders. Improving lipid metabolism may help asthmatic pregnant women maintain healthy fetal development.
INSTITUTE
Nanjing University of Chinese Medicine
LAST_NAME
Fang
FIRST_NAME
Huafeng
ADDRESS
No.138 xianlin road, nanjing city, Nanjing, China, 210046, China
Hepatic oxylipin profiles in mouse models of Wilson disease: new insights into early hepatic manifestations.
STUDY_SUMMARY
Hepatic inflammation is commonly identified in Wilson disease (WD), a genetic disease of hepatic and brain copper accumulation. Copper accumulation is associated with increased reactive oxygen species and activation of the non-enzymatic oxidation of membrane-bound polyunsaturated fatty acids (PUFA), with impairment of cellular structures and function. Products of PUFA oxidation are collectively known as oxylipins (OXL), which can also be produced via enzymatic pathways including lipoxygenases (LOXs), cyclooxygenases (COXs), and cytochrome P450 monooxygenases (CYPs). These bioactive lipids modulate hepatic inflammation. We aimed to examine hepatic OXLs profile at early stages of WD in mouse model of Wilson disease, the toxic milk from The Jackson Laboratory (tx-j) compared to mice with normal copper metabolism (C3H). Targeted lipidomic profiling of OXLs was performed by ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) in livers from 16 weeks old male and female mice. Hepatic OXL profiles were altered, with higher levels of PUFA alcohols, diols, and ketones. Markers of oxidative stress, 9-HETE and 9-HEPE were increased in tx-j mice. Hepatic prostaglandin and thromboxane levels in the COX pathway were increased in tx-j mice. tx-j showed altered PUFA-epoxides, suggesting altered CYP(s) activities. Our findings suggest that both non-enzymatic ROS-dependent and enzymatic PUFAs oxidation via COX and LOX pathways are associated with early stages liver disease in WD. It also indicates altered CYPs activities in animal model of WD. These novel pathways could be the target for future therapies.
INSTITUTE
University of California, Davis
LAST_NAME
Sarode
FIRST_NAME
Gaurav
ADDRESS
Genome & Biomedical Sciences Facility, 451 E. Health Sciences Dr., Davis, CA, 95616, USA
Investigation of metabolism in hypertrophic cardiomyopathy
STUDY_SUMMARY
We propose the use of targeted metabolomics to define the metabolic and molecular pathways altered in 2 mouse models (R92W-TnT and R403Q-MHC) of hypertrophic cardiomyopathy (HCM) that span the spectrum of human disease (heart failure and sudden death), with the goal of identifying treatment targets. Parallel targeted metabolomics studies will be performed in heart tissue and plasma at rest and following inotrope stimulation. Since energy compromise is expected to be most marked when the heart is subject to increased workload, as is the case during high-intensity exercise or inotropic stimulation, we propose metabolomics studies in heart tissue and plasma, at rest and following inotrope stimulation. We anticipate that the results of these studies will allow us to move forward with further investigations into specific metabolites of interest as biomarkers, to be tested in HCM patients in future studies.
INSTITUTE
University of California, San Francisco
LAST_NAME
Abraham
FIRST_NAME
Maria Roselle
ADDRESS
555 Mission Bay Boulevard South, San Francisco, CA 94158
EMAIL
Roselle.Abraham@ucsf.edu
PHONE
415-502-3911
NUM_GROUPS
2 groups of 2 (i.e., mutant vs control of two mouse models of HCM (R402Q-MyHC, R92W-TnT)
Early life metabolic abnormalities in the bone marrow of the offspring of obese mothers
STUDY_SUMMARY
Obese mothers predispose their babies to develop metabolic disorders and obesity before they are even born. The study used a mouse model of maternal high-fat diet-induced obesity that recapitulates cardiovascular and metabolic abnormalities seen in humans born to obese mothers. to compare the metabolic characteristics of bone marrow cells in newly weaned mice. The results showed that the bone marrow of offspring born to mothers on a high-fat diet primarily used glucose through OXPHOS and presented decreased amino acid metabolism. These metabolic changes were observed in the bone marrow of the offspring at a very young age before any symptomatic metabolic disease was present.
INSTITUTE
Oregon Health and Science University
DEPARTMENT
Knight Cardiovascular Institute
LABORATORY
Maloyan Lab
LAST_NAME
Maloyan
FIRST_NAME
Alina
ADDRESS
3181 S.W. Sam Jackson Park Road, Portland, Oregon, 97239-3098, USA
Metabolic Adaptations of Microbacterium sediminis YLB-01 for Survival in the Challenging High-Pressure of the Deep Sea
STUDY_SUMMARY
In this study, we investigated the metabolic adaptations of Microbacterium sediminis YLB-01, a cold and stress-tolerant microorganism isolated from deep-sea sediments, in response to high-pressure conditions. By using NMR-based metabolomic analysis and proteomic analysis, we conducted a comprehensive examination of the significantly altered metabolic pathways involved in the adaptation of YLB-01 cells to high-pressure conditions.
INSTITUTE
Xiamen University
LAST_NAME
Qiu
FIRST_NAME
Xu
ADDRESS
No. 422, Siming South Road, Xiamen, Fujian, China.
IL-1Ī²-mediated adaptive re-programming of endogenous human cardiac fibroblasts to cells with immune features during fibrotic remodeling
STUDY_SUMMARY
The source and roles of fibroblasts and CD4 helper T-cells during maladaptive remodeling and myocardial fibrosis in pulmonary arterial hypertension (PAH) have been long debated. We demonstrate, using single-cell mass cytometry, a sub-population of endogenous human cardiac fibroblasts expressing increased levels of CD4, a helper T-cell marker, in addition to myofibroblast markers distributed in human fibrotic RV tissue, interstitial/perivascular lesions of SUGEN/Hypoxia (SuHx) rats and fibroblasts labelled with pdgfrĪ± CreERt2/+ in R26R-tdTomato mice. Recombinant IL-1Ī² increases IL-1R, CCR2 receptor expression, modifies the secretome, and differentiates cardiac fibroblasts to form CD68 positive cell clusters. IL-1Ī² also activates stemness markers such as NANOG and SOX2 and genes involved in de-differentiation, lymphoid cell function and metabolic reprogramming. IL-1Ī² induction of lineage traced primary mouse cardiac fibroblasts causes these cells to lose their fibroblast identity and acquire an immune phenotype. Our results identify IL-1Ī² induced immune-competency in human cardiac fibroblasts and suggest that fibroblast secretome modulation may constitute a therapeutic approach to PAH and other diseases typified by inflammation and fibrotic remodeling.
INSTITUTE
Brown University
DEPARTMENT
Molecular Pharmacology, Physiology and Biotechnology
Comprehensive Metabolic Profiling of MYC-Amplified Medulloblastoma Tumors Reveals Key Dependencies on Amino Acid, Tricarboxylic Acid and Hexosamine Pathways
STUDY_SUMMARY
Reprogramming of cellular metabolism is a hallmark of cancer. Altering metabolism allows cancer cells to overcome unfavorable microenvironment conditions and to increase and invade. Medulloblastoma is the most common malignant brain tumor in children. Genomic amplification of MYC defines a subset of poor-prognosis medulloblastoma. We performed comprehensive metabolic studies of human MYC-amplified medulloblastoma by comparing the metabolic profiles of tumor cells in three different conditionsāin vitro, in flank xenografts, and orthotopic xenografts in the cerebellum. Principal component analysis showed that the metabolic profiles of brain and flank high-MYC medulloblastoma tumors clustered closely together and separated away from the normal brain and in vitro MYC-amplified cells. Compared to typical brains, MYC-amplified medulloblastoma orthotopic xenograft tumors showed upregulation of the TCA cycle and the synthesis of nucleotides, hexosamines, amino acids, and glutathione. There was significantly higher glucose uptake and usage in orthotopic xenograft tumors compared to flank xenograft tumors and cells in culture. In orthotopic tumors, glucose was the primary carbon source for the de novo synthesis of glutamate, glutamine, and glutathione through the TCA cycle. In vivo, the glutaminase II pathway was the main pathway utilizing glutamine. Glutathione was the most abundant upregulated metabolite in orthotopic tumors compared to normal brains. Glutamine-derived glutathione was synthesized through the glutamine transaminase K (GTK) enzyme in vivo. In conclusion, high MYC medulloblastoma cells have different metabolic profiles in vitro compared to in vivo; critical vulnerabilities may be missed by not performing in vivo metabolic analyses.
INSTITUTE
Johns Hopkins
LAST_NAME
Pham
FIRST_NAME
Khoa
ADDRESS
600 N. Wolfe Street, Pathology Bldg., Rm. 401, Baltimore, Maryland, 21287, USA
Metabolites are low molecular-weight molecules produced during cellular metabolism. The global expression of the (meta-)metabolome could thus potentially be used to characterize the exposure of an organism or a community to a specific stressor. Here, the meta-metabolomic fingerprints of mature biofilms were examined after 1, 3 and 7 days of exposure to five concentrations of cobalt (0, 10-7, 10-6, 5 x 10-6 and 10-5 M) in aquatic microcosms. The global changes in meta-metabolomic fingerprints were in plain agreement with those of the other biological parameters studied (cobalt bioaccumulation, biomass, chlorophyll). To better understand the dose-responses of the biofilm meta-metabolome, the untargeted LC-HRMS metabolomic data were further processed using DRomics to build dose-response model curves and to calculate benchmark doses to be aggregated into an empirical cumulative density function. A trend analysis of the dose-response curves suggests the presence of homeostasis (0 - 4.7 x 10-7 M), defense (4.7 x 10-7 ā 2.7 x 10-6 M) and damage (higher than 2.7 x 10-6 M) response ranges in biofilms exposed to various cobalt concentrations. The present study demonstrates that the use of untargerted meta-metabolomics represents a promising approach for environmental risk assessment of metals by relating molecular defense and damage mechanisms to contaminant concentration.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (DINE Cohorts)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
Developmental Impact of NICU Exposures (DINE) as part of ECHO has been designed to determine the impact of and interactions among early childhood phthalate, manganese, lead, various other environmental chemicals and stress exposures on neurodevelopmental outcomes in preterm children. This research was supported by the Environmental Influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. DINE is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
Zucker School of Medicine at Hofstra / Northwell
DEPARTMENT
Department of Pediatrics and Department of Occupational Medicine, Epidemiology & Prevention
LAST_NAME
Stroustrup
FIRST_NAME
Annemarie
ADDRESS
Cohen Childrenās Medical Center 269-01 76th Avenue NICU, CH346 New Hyde Park, NY 11040
Autosomal dominant polycystic kidney disease (ADPKD), the most common potentially lethal genetic disease in humans and the fourth leading cause of kidney disease, exhibits features of both a ciliary and metabolic disorder. Our previous research revealed that cells overexpressing Exoc5 with elongated cilia demonstrate enhanced recovery from oxidative stress. To investigate the connection between primary cilia and metabolism, we conducted an unbiased metabolomics screen. Global metabolic profiling was performed on canine MDCK cells (Control, Exoc5 ciliary targeting sequence mutation (CTS-mut), Exoc5 knockdown (KD), Exoc5 overexpression (OE)) and murine cells (Ift88 knockout (KO), Ift88 rescue). Knockdown (KD) or ciliary targeting sequence mutation (CTS-mut) in Exoc5, a central exocyst component, resulted in cilia loss. Similarly, Ift88 knockout (KO) resulted in cilia loss. For each experimental group, we cultivated six independent replicates of Exoc5 OE, KD, CTS-mut, and control MDCK cells, as well as six independent replicates of murine Ift88 KO and rescue cells. Cell pellets were obtained from the cultures, and we analyzed the global metabolic profiles for all 36 cell pellets. The most significant findings from the metabolomics screen indicated defects in tryptophan metabolism. This discovery suggests a potential link between primary cilia function and tryptophan-related metabolic pathways. Further exploration of these findings may shed light on the underlying mechanisms and implications for ADPKD pathogenesis and metabolic disturbances.
GAS2 encodes a 2-oxoglutarate dependent dioxygenase involved in ABA catabolism
STUDY_SUMMARY
Liu et al. [1] recently reported the characterization of Arabidopsis thaliana GAS2 (Gain of Function in ABA-modulated Seed Germination 2), which was described as an enzyme that catalyzes the stereospeciļ¬c hydration of GA12 to produce GA12 16, 17-dihydro-16Ī±-ol (DHGA12). A second paper describes the conversion of GA12 to an unidentified product by GAS2 and also reports that this enzyme does not convert ABA [2]. However, as previously reported [3], we did not find any conversion of [17-14C]-labeled or [1-,7-,12-,18-14C4]-labeled GA12 by GAS2. Instead, we present here data showing that the recombinant GAS2 enzyme is capable of catabolising abscisic acid (ABA) to phaseic acid (PA) and further to a second product, putative 8ā-carboxy-ABA (compound A; Fig. 1a) [4]. References: [1] Liu, H. et al. Biosynthesis of DHGA12 and its roles in Arabidopsis seedling establishment. Nat. Commun. 10, 1768 (2019). [2] Xiong, W. et al. The dioxygenase GIM2 functions in seed germination by altering gibberellin production in Arabidopsis. J. Integr. Plant Biol. 60, 276-291 (2018). [3] Lange, T. & Pimenta Lange, M. J. The Multifunctional Dioxygenases of Gibberellin Synthesis. Plant Cell Physiol. 61, 1869-1879 (2020). [4] Lange, T., Atiq, N., Pimenta Lange. GAS2 encodes a 2-oxoglutarate dependent dioxygenase involved in ABA catabolism. bioRxiv, doi: 10.1101/2022.11.16.516706 (2022).
Metabolomics panel associated with cystic fibrosis-related diabetes towards biomarker discovery
STUDY_SUMMARY
Background: The most prevalent comorbidity among cystic fibrosis (CF) patients is cystic fibrosis-related diabetes (CFRD). CFRD has been linked to one of the worse clinical outcomes and higher mortality. Improved clinical results have been related to earlier diagnosis and treatment of CFRD. Therefore, the present study aimed to investigate the metabolome of human serum of patients with CFRD. This might aid in identifying novel biomarkers linked with the pathophysiology of CFRD and its diagnosis. Methods: The liquid chromatographyāhigh-resolution mass spectrometry (LCāHRMS) metabolomics approach was utilized for serum samples from patients with CF (n= 36) and healthy control (n=36). Among the CF group, nine patients were with CFRD and 27 were non-CFRD (nCFRD). Results: A total of 2328 metabolites were significantly altered in CF compared to the healthy control. Among those, 799 significantly dysregulated metabolites were identified between CFRD and nCFRD. Arachidonic acid (AA), ascorbate, and aldarate metabolism were the most common metabolic pathways dysregulated in CF. L-homocysteic acid (L-HCA) levels were significantly reduced in CF and CFRD compared to control and nCFRD, respectively. In addition, gamma-glutamylglycine and L-5-hydroxytryptophan (5-HTP) had the highest discrimination between CFRD and nCFRD with AUC (0.716 and 0.683, respectively). Conclusions: These biomarkers might serve as diagnostic biomarkers and aid in understanding potential metabolic changes linked to CF and CFRD.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
LAST_NAME
Al Mogren
FIRST_NAME
Maha
ADDRESS
Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia
Metabolomic profiling of PMM2-CDG brain organoids - Part 1
STUDY_SUMMARY
PMM2-CDG is a rare inborn error of metabolism caused by deficiency of the PMM enzyme, which leads to impaired protein glycosylation. While the disorder has primarily neurological presentation, there is limited knowledge about the specific brain-related changes that result from PMM deficiency. Utilizing 3D brain organoids derived from individuals with PMM2-CDG, we identified abnormal glucose metabolism in PMM2-CDG organoids, indicating disturbances in metabolic utilization. In this experiment, day 160 organoids were used.
Effects of acute cold exposure on mouse metabolome
STUDY_SUMMARY
Cold-induced thermogenesis is widely studied as a potential avenue to treat obesity, but a thorough understanding of the metabolic changes driving thermogenesis is lacking. We analyzed mouse plasma and tissue samples from fasted mice housed at room temperature or acutely exposed to 4Ā°C. First, C57BL/6 mice (n=41) were divided into 2 groups (n=20-21 per group) and underwent surgery to implant an indwelling arterial catheter. Mice originally housed at room temperature were housed without food for 6 hours at room temperature or 4Ā°C, after which blood was collected from the arterial catheter and centrifuged, and plasma was collected for metabolomic analyses. Second, C57BL/6 mice (n=12) were divided into 2 groups (n=6 per group) and housed without food for 6 hours at room temperature or 4Ā°C. Mice were euthanized, and tissues (brown adipose tissue, heart, liver, quadricepts muscle, diaphragm, and gonadal-white adipose tissue) were collected, freeze-clamped in liquid nitrogen, and used for metabolomic analyses. Overall our data detail a coordinated and broad metabolic response governing the thermogenic response to acute cold exposure in mice.
Atlas of fetal metabolism during mid-to-late gestation and diabetic pregnancy!
STUDY_SUMMARY
Mounting evidence supports an instructive role for metabolism in stem cell fate decisions. However, much is yet unknown about how fetal metabolism changes during mammalian development and how altered maternal metabolism shapes fetal metabolism. Here, we present a descriptive atlas of in vivo fetal murine metabolism during mid-to-late gestation in normal and diabetic pregnancy. Using 13C-glucose and LC-MS, we profiled the metabolism of fetal brains, hearts, livers, and placentas harvested from pregnant dams between embryonic days (E)10.5 and 18.5. Comparative analysis of our large metabolomics dataset revealed metabolic features specific to fetal tissues developed under a hyperglycemic environment as well as metabolic signatures that may denote developmental transitions during euglycemic development. We observed sorbitol accumulation in fetal tissues and altered neurotransmitter levels in fetal brains isolated from dams with maternal hyperglycemia. Tracing 13C-glucose revealed disparate nutrient sourcing in fetuses depending on maternal glycemic states. Regardless of glycemic state, histidine-derived metabolites accumulated during late development in fetal tissues and maternal plasma. Our rich dataset presents a comprehensive overview of in vivo fetal tissue metabolism and alterations occurring as a result of maternal hyperglycemia.
INSTITUTE
University of California, Los Angeles
DEPARTMENT
Biological Chemistry
LABORATORY
Heather Christofk
LAST_NAME
Matulionis
FIRST_NAME
Nedas
ADDRESS
615 Charles E Young Drive South Los Angeles, CA, 90095
Investigation of metabolism in hypertrophic cardiomyopathy - HILIC
STUDY_SUMMARY
We propose the use of targeted metabolomics to define the metabolic and molecular pathways altered in 2 mouse models (R92W-TnT and R403Q-MHC) of hypertrophic cardiomyopathy (HCM) that span the spectrum of human disease (heart failure and sudden death), with the goal of identifying treatment targets. Parallel targeted metabolomics studies will be performed in heart tissue and plasma at rest and following inotrope stimulation. Since energy compromise is expected to be most marked when the heart is subject to increased workload, as is the case during high-intensity exercise or inotropic stimulation, we propose metabolomics studies in heart tissue and plasma, at rest and following inotrope stimulation. We anticipate that the results of these studies will allow us to move forward with further investigations into specific metabolites of interest as biomarkers, to be tested in HCM patients in future studies.
INSTITUTE
University of California, San Francisco
DEPARTMENT
Hypertrophic Cardiomyopathy Center of Excellence
LABORATORY
UCSF Smith Cardiovascular Research Building
LAST_NAME
Abraham
FIRST_NAME
Maria Roselle
ADDRESS
555 Mission Bay Boulevard South, San Francisco, CA 94158
EMAIL
Roselle.Abraham@ucsf.edu
NUM_GROUPS
2 groups of 2 (i.e., mutant vs control of two mouse models of HCM (R402Q-MyHC, R92W-TnT)
Metabolomic profiling of PMM2-CDG brain organoids - Part 2
STUDY_SUMMARY
PMM2-CDG is a rare inborn error of metabolism caused by deficiency of the PMM enzyme, which leads to impaired protein glycosylation. While the disorder has primarily neurological presentation, there is limited knowledge about the specific brain-related changes that result from PMM deficiency. Utilizing 3D brain organoids derived from individuals with PMM2-CDG, we identified abnormal glucose metabolism in PMM2-CDG organoids, indicating disturbances in metabolic utilization. In this experiment, day 40 organoids were used.
Investigation of metabolism in hypertrophic cardiomyopathy
STUDY_SUMMARY
We propose the use of targeted metabolomics to define the metabolic and molecular pathways altered in 2 mouse models (R92W-TnT and R403Q-MHC) of hypertrophic cardiomyopathy (HCM) that span the spectrum of human disease (heart failure and sudden death), with the goal of identifying treatment targets. Parallel targeted metabolomics studies will be performed in heart tissue and plasma at rest and following inotrope stimulation. Since energy compromise is expected to be most marked when the heart is subject to increased workload, as is the case during high-intensity exercise or inotropic stimulation, we propose metabolomics studies in heart tissue and plasma, at rest and following inotrope stimulation. We anticipate that the results of these studies will allow us to move forward with further investigations into specific metabolites of interest as biomarkers, to be tested in HCM patients in future studies.
INSTITUTE
University of California, San Francisco
DEPARTMENT
Hypertrophic Cardiomyopathy Center of Excellence
LABORATORY
UCSF Smith Cardiovascular Research Building
LAST_NAME
Abraham
FIRST_NAME
Maria Roselle
ADDRESS
555 Mission Bay Boulevard South, San Francisco, CA 94158
EMAIL
Roselle.Abraham@ucsf.edu
PHONE
415-502-3911
NUM_GROUPS
2 groups of 2 (i.e., mutant vs control of two mouse models of HCM (R402Q-MyHC, R92W-TnT)
Evaluation of Novel Candidate Filtration Markers from a Global Metabolomics Discovery for Glomerular Filtration Rate Estimation (ALTOLD)
STUDY_SUMMARY
Background: Creatinine and cystatin-C are recommended for estimating glomerular filtration rate (eGFR) but accuracy is suboptimal. Using untargeted metabolomics data, we sought to identify candidate filtration markers using a novel approach based on their maximal joint association with measured GFR (mGFR) with flexibility to consider their biological and chemical properties later. Methods: We analyzed metabolites measured in seven diverse studies of 2,851 participants on the Metabolon H4 platform that had Pearson correlations with log mGFR <-0.5. We used a stepwise approach to develop models to estimate mGFR including two to 15 metabolites with and without inclusion of creatinine and demographics. We then selected candidate filtration markers from those metabolites found >20% in models that did not demonstrate substantial overfitting in cross-validation and with small (<0.1 in absolute value) coefficients for demographics. Results: In total, 456 named metabolites were present in all studies, and 36 had correlations <-0.5 with mGFR. We developed 2,225 models including these metabolites; all had lower RMSEs and smaller coefficients for demographic variables compared to estimates using untargeted creatinine. Cross-validated RMSEs (0.187-0.213) were similar to original RMSEs for models with ā¤ 10 metabolites. Our criteria identified 17 metabolites, including 12 new candidate filtration markers. Conclusion: We identified candidate metabolites with maximal joint association with mGFR and minimal association with demographic variables across varied clinical settings. Future analyses will assess metabolite biological and chemical characteristics in the path towards development of a panel eGFR that is more accurate and less reliant on demographic variables than current eGFR. ACRONYMS AASKG1: African American Study of Kidney (patient data at G1 visit). ALTOLD: Assessing Long Term Outcomes in Living Kidney Donors study. MDRD: The Modification of Diet in Renal Disease study.
Evaluation of Novel Candidate Filtration Markers from a Global Metabolomics Discovery for Glomerular Filtration Rate Estimation (MDRD)
STUDY_SUMMARY
Background: Creatinine and cystatin-C are recommended for estimating glomerular filtration rate (eGFR) but accuracy is suboptimal. Using untargeted metabolomics data, we sought to identify candidate filtration markers using a novel approach based on their maximal joint association with measured GFR (mGFR) with flexibility to consider their biological and chemical properties later. Methods: We analyzed metabolites measured in seven diverse studies of 2,851 participants on the Metabolon H4 platform that had Pearson correlations with log mGFR <-0.5. We used a stepwise approach to develop models to estimate mGFR including two to 15 metabolites with and without inclusion of creatinine and demographics. We then selected candidate filtration markers from those metabolites found >20% in models that did not demonstrate substantial overfitting in cross-validation and with small (<0.1 in absolute value) coefficients for demographics. Results: In total, 456 named metabolites were present in all studies, and 36 had correlations <-0.5 with mGFR. We developed 2,225 models including these metabolites; all had lower RMSEs and smaller coefficients for demographic variables compared to estimates using untargeted creatinine. Cross-validated RMSEs (0.187-0.213) were similar to original RMSEs for models with ā¤ 10 metabolites. Our criteria identified 17 metabolites, including 12 new candidate filtration markers. Conclusion: We identified candidate metabolites with maximal joint association with mGFR and minimal association with demographic variables across varied clinical settings. Future analyses will assess metabolite biological and chemical characteristics in the path towards development of a panel eGFR that is more accurate and less reliant on demographic variables than current eGFR. ACRONYMS AASKG1: African American Study of Kidney (patient data at G1 visit). ALTOLD: Assessing Long Term Outcomes in Living Kidney Donors study. MDRD: The Modification of Diet in Renal Disease study.
Evaluation of Novel Candidate Filtration Markers from a Global Metabolomics Discovery for Glomerular Filtration Rate Estimation (AASKG1)
STUDY_SUMMARY
Background: Creatinine and cystatin-C are recommended for estimating glomerular filtration rate (eGFR) but accuracy is suboptimal. Using untargeted metabolomics data, we sought to identify candidate filtration markers using a novel approach based on their maximal joint association with measured GFR (mGFR) with flexibility to consider their biological and chemical properties later. Methods: We analyzed metabolites measured in seven diverse studies of 2,851 participants on the Metabolon H4 platform that had Pearson correlations with log mGFR <-0.5. We used a stepwise approach to develop models to estimate mGFR including two to 15 metabolites with and without inclusion of creatinine and demographics. We then selected candidate filtration markers from those metabolites found >20% in models that did not demonstrate substantial overfitting in cross-validation and with small (<0.1 in absolute value) coefficients for demographics. Results: In total, 456 named metabolites were present in all studies, and 36 had correlations <-0.5 with mGFR. We developed 2,225 models including these metabolites; all had lower RMSEs and smaller coefficients for demographic variables compared to estimates using untargeted creatinine. Cross-validated RMSEs (0.187-0.213) were similar to original RMSEs for models with ā¤ 10 metabolites. Our criteria identified 17 metabolites, including 12 new candidate filtration markers. Conclusion: We identified candidate metabolites with maximal joint association with mGFR and minimal association with demographic variables across varied clinical settings. Future analyses will assess metabolite biological and chemical characteristics in the path towards development of a panel eGFR that is more accurate and less reliant on demographic variables than current eGFR. ACRONYMS AASKG1: African American Study of Kidney (patient data at G1 visit). ALTOLD: Assessing Long Term Outcomes in Living Kidney Donors study. MDRD: The Modification of Diet in Renal Disease study.
The ECHO Cohort Exposome: First Steps using HHEAR Analysis ā An Opportunity for ALL ECHO Cohorts to Contribute Type A Samples ā Untargeted Analysis (PRISM Cohort)
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The PRogramming of Intergenerational Stress Mechanisms (PRISM) study is a prospective pregnancy cohort of mother-child dyads originally designed to examine the effects of prenatal and early life psychosocial and environmental exposures on child developmental outcomes. Pregnant women receiving prenatal care at health centers in the Northeastern United States were recruited at 23.6 + 9.4 weeksā gestation. Women were eligible if they were English or Spanish speaking, 18 years or older, did not have definite plans to move out of the area within a year after consent, were planning to give birth at the hospital affiliated with the recruitment site, had a singleton pregnancy, endorsed drinking <7 alcoholic drinks per week prior to pregnancy recognition and none following pregnancy recognition, had an HIV negative status, and delivered an infant without congenital abnormalities that would impede ongoing participation. Written informed consent was obtained from women in their preferred language. In 2016, funding was obtained to support ongoing prenatal recruitment and follow-up as part of the Environmental influences on Child Health Outcomes (ECHO) program through UH3OD023337. Please contact Rosalind Wright, MD MPH at rosalind.wright@mssm.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental Influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. PRISM is an ECHO cohort which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 North Carolina Human Health Exposure Analysis Resource Hub: Research Triangle Institute: Fennell T, University of North Carolina at Chapel Hill: Sumner S, University of North Carolina at Charlotte: Du X; U2C ES030857 Human Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
Effect of ERR Agonist in Mouse Heart Post Pressure Overload
STUDY_SUMMARY
The present heart dataset comprises a total of 855 biochemicals, 787 compounds of known identity (named biochemicals) and 68 compounds of unknown structural identity (unnamed biochemicals). The muscle dataset comprises a total of 711 biochemicals, 663 compounds of known identity (named biochemicals) and 48 compounds of unknown structural identity (unnamed biochemicals). Following normalization to mass of sample extracted for heart tissue (equal mass was extracted for muscle tissue samples, so no additional normalization was performed on muscle samples prior to statistical analysis), log transformation and imputation of missing values, if any, with the minimum observed value for each compound, 2-way ANOVA or Welchās two-sample t-test analyses were used to identify biochemicals that differed significantly between experimental groups. A summary of the numbers of biochemicals that achieved statistical significance (pā¤0.05), as well as those approaching significance (0.05<p<0.10), is shown below.
Smoking Induced Gut Microbial Dysbiosis Mediates Cancer Progression Through Adaptive Immune System Modulation
STUDY_SUMMARY
Cigarette smoke exposure (CSE), either through active smoking or secondhand smoke, increases the risk for a plethora of cancers. Studies have estimated that one in three cancer deaths is associated with cigarette smoke exposure. However, despite ongoing research on numerous carcinogens, the underlying mechanism(s) remain poorly understood. Recent evidence indicates that the gut microbiome can influence cancer progression by immune system modulation. Since CSE alters the gut microbiome, we hypothesized that the gut microbiome serves as a causative link between smoking and cancer growth. Through a combination of rigorous syngeneic animal models and fecal microbiome transplantation studies, we establish an essential role for smoke-induced dysbiosis in cancer growth. Using Flow cytometric analysis of tumor specimens and experiments in Rag 1 KO and CD8 KO, we demonstrate that smoke induced tumor growth requires functional adaptive immunity. We further characterized the unique gut microbial and metabolomic signatures induced by CSE using high throughput 16s rRNA sequencing and mass spectrometric techniques. Finally, utilizing gut microbial ablation strategies with broad and narrow-spectrum antibiotics, we demonstrate the reversal of phenotypic effects of CSE and present a novel actionable target to mitigate CSE-induced tumor promotion.
INSTITUTE
University of Alabama, Birmingham
DEPARTMENT
Department of Surgery
LAST_NAME
Vikas
FIRST_NAME
Dudeja
ADDRESS
1808 7th Avenue South Boshell Building- Suite 573 Birmingham, AL 35294
Leukemia inhibitory factor suppresses hepatic de novo lipogenesis and induces cachexia
STUDY_SUMMARY
We performed metabolomic analysis to measure polar and lipid metabolites in the serum from TgLC and TgL mice with Tamoxifen (TAM) injection. Mice under both fed and fasted conditions were included.
Brain Metabolomics in Fragile X-Associated Tremor/Ataxia Syndrome (FXTAS)
STUDY_SUMMARY
The course of pathophysiological mechanisms involved in fragile X-associated tremor/ataxia syndrome (FXTAS) remains largely unknown. Previous proteomics and metabolomics studies conducted in blood samples collected from FMR1 premutation carriers with FXTAS reported abnormalities in energy metabolism, and precursors of gluconeogenesis showed significant changes in plasma expression levels in FMR1 premutation carriers who developed FXTAS. We conducted an analysis of postmortem human brain tissues from 44 donors, 25 brains with FXTAS, and 19 matched controls. We quantified the metabolite relative abundance in the inferior temporal gyrus and the cerebellum using untargeted mass spectrometry (MS)-based metabolomics. We investigated how the metabolite type and abundance relate to the number of cytosine-guanine-guanine (CGG) repeats, to markers of neurodegeneration, and to the symptoms of FXTAS.
INSTITUTE
UC Davis
LAST_NAME
MartĆnez-CerdeƱo
FIRST_NAME
VerĆ³nica
ADDRESS
2425 Stockton Blvd, 6th Floor, Sacramento, CA 95817
Cold-stimulated brown adipose tissue activation is related to changes in serum metabolites relevant to NAD+ metabolism in humans
STUDY_SUMMARY
Cold-induced brown adipose tissue (BAT) activation is considered to improve metabolic health. In murine BAT, cold increases the fundamental molecule for mitochondrial function, nicotinamide adenine dinucleotide (NAD+), but limited knowledge on NAD+ metabolism during cold in human BAT metabolism exists. We show that cold increases the serum metabolites of the NAD+ salvage pathway (nicotinamide and 1-methylnicotinamide) in humans. Additionally, individuals with coldstimulated BAT activation have decreased levels of metabolites from the de novo NAD+ biosynthesis pathway (tryptophan, kynurenine). Serum nicotinamide correlates positively with cold-stimulated BAT activation, whereas tryptophan and kynurenine correlates negatively. Furthermore, the expression of genes involved in NAD+ biosynthesis in BAT are related to markers of metabolic health. Our data indicate that cold increases serum tryptophan conversion to nicotinamide to be further utilized by BAT. We conclude that NAD+ metabolism is activated upon cold in humans and probably regulated in a coordinated fashion by several tissues.
INSTITUTE
University of Turku
LAST_NAME
Virtanen
FIRST_NAME
Kirsi
ADDRESS
Turku PET Centre, Turku University Hospital, Turku, Finland
Multi-assay nutritional metabolomics profiling of low vitamin A status versus adequacy is characterized by reduced plasma lipid mediators among lactating women in the Philippines: A pilot study.
STUDY_TYPE
Case-control
STUDY_SUMMARY
Low vitamin A (VA) status is common among lactating women in low-income countries. Lactation has substantial effects on motherās metabolism and VA is required in multiple biological processes, including growth, vision, immunity, and reproduction. The objective of this pilot study was to utilize metabolomics profiling to conduct a broad, exploratory assessment of differences in plasma metabolites associated with low VA status versus VA adequacy in lactating women. Plasma samples from lactating women who participated in a survey in Samar, Philippines, were selected from a cross-sectional study based on plasma retinol concentrations indicating low (VA-; n=5) or adequate (VA+; n=5) VA status (plasma retinol <0.8 or >1.05 Āµmol/L). The plasma results collected from six metabolomics assays (oxylipins, endocannabinoids, bile acids, primary metabolomics, biogenic amines, and lipidomics) were compared by group using liquid chromatography mass spectrometry. Twenty-eight metabolites were altered in the VA- versus VA+ status groups, with 24 being lipid mediators (p<0.05). These lipid mediators included lower concentrations of arachidonic acid- and eicosapentaenoic acid-derived oxylipins, as well as lysophospholipids and sphingolipids, in the VA- group (p<0.05). Chemical similarity enrichment analysis identified HETEs, HEPEs, and DiHETEs as significantly altered oxylipin clusters (p<0.0001, false discovery rate (FDR) p<0.0001), as well as sphingomyelins, saturated lysophosphatidylcholines, phosphatidylcholines, and phosphatidylethanolamines (p<0.001, FDR p<0.01). The multi-assay nutritional metabolomics profiling of low VA status compared with adequacy in lactating women was characterized by reduced lipid mediator concentrations. Future studies with stronger study designs and larger sample size are needed to confirm and validate these preliminary results.
INSTITUTE
California Polytechnic State University, San Luis Obispo
DEPARTMENT
Food Science and Nutrition
LABORATORY
Cal Poly Metabolomics Service Center
LAST_NAME
La Frano
FIRST_NAME
Michael
ADDRESS
Attn: Dr. Michael La Frano Bldg 11 Room 239 Cal Poly State University 1 Grand Avenue San Luis Obispo, CA 93407
Role of the Afferent Lymph as an Immunological Conduit to Analyze Tissue Antigenic and Inflammatory Load - metabolomics
STUDY_SUMMARY
The intestinal barrier, and the gut-associated lymphoid tissue, are the interface of the host-microbiome/pathogens interactions and, its disruption, has been associated with a series of inflammatory, autoimmune, and degenerative diseases. Herein we performed the analysis of the pre-nodal mesenteric lymph at steady state and following disruption of the intestinal barrier to map the antigenic and pro-inflammatory load in both mice and human subjects with inflammatory bowel disease. The antigenic signature of gastrointestinal tissue inflammation was reflected in the mesenteric nodal dendritic-cells (DC)-MHCII-eluted immunopeptidome, in a tissue specific manner, when compared to the DC-eluted-MHCII-peptidome from cervical lymph nodes. Pro-inflammatory and microbiome-derived by-products, such as amino acids, deoxy sugars, component of the bacterial wall and other immunomodulators of the gut-brain axis, were found in the afferent lymph following damage to the gut epithelium. Our data points to the relevance of the lymphatic fluid to probe the tissue-specific antigenic and inflammatory load transported to the draining lymph node.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Nucleotide, phospholipid, and kynurenine metabolites are robustly associated with COVID-19 severity and time of plasma sample collection in a prospective cohort study
STUDY_SUMMARY
Introduction: A deep understanding of the molecular underpinnings of disease severity and progression in large human studies is necessary to develop metabolism-related preventive strategies of severe disease outcomes, particularly in viral pandemics like that of COVID-19. The use of samples collected before disease diagnosis, however, is limited and thus metabolites and metabolic pathways that predispose to severe disease are not well understood. Further, current studies are limited in sample size, number of metabolites evaluated, and/or do not adjust for comorbidities. Methods: We generated comprehensive plasma metabolomic profiles in more than 600 patients from the Longitudinal EMR and Omics COVID-19 Cohort (LEOCC). Samples were collected before (n = 441), during (n = 86), and after (n = 82) COVID-19 diagnosis. Regression models were used to determine (1) metabolites associated with predisposition to and/or persistent effects of COVID-19 severity within each time of sample collection, using logistic regression and (2) metabolites associated with time of sample collection, using linear regression, to better understand transient or lingering metabolic alterations over the disease course. All models were controlled for demographic (age, sex, race, ethnicity), risk (smoking status, BMI), and comorbidities (Charlson Index). Metabolites with an FDR-adjusted p-value < 0.05 were considered significant. Results: Of the 1,546 metabolites measured, 506 were associated with disease severity or time of sample collection. Among these, sphingolipids and phospholipids were negatively associated with severity and exhibited lingering elevations after disease, while modified nucleotides were positively associated with severity and had lingering decreases after disease. Cytidine and uridine metabolites, which were positively and negatively associated with COVID-19 severity, respectively, were transiently elevated in active disease, reflecting particular importance of pyrimidine metabolism in active COVID-19. Conclusions: We identified novel metabolites reflecting predisposition to severe disease and changes to global metabolism from before to during and after COVID-19 diagnosis. This is the first large metabolomics study using COVID-19 plasma samples before, during, and/or after disease. This study lays the groundwork for identifying putative clinical biomarkers and identifying preventative strategies for severe disease outcomes.
INSTITUTE
National Institutes of Health
DEPARTMENT
Division of Preclinical Innovation - National Center for Advancing Translational Sciences
LABORATORY
Informatics Core - Division of Preclinical Innovation
Summary Reactive oxygen species (ROS) are by-products of metabolism of oxygen and they play an important role in normal homeostasis and cell signaling, as well as in the initiation of diseases including cancer when their production is upregulated. Thus, it is imperative to understand the cellular and molecular basis by which ROS impact on various biological and pathological processes. Here, we identified 2-oxindole, a tryptophan derivative, was a major catabolic product in hydrogen peroxide-treated cell culture medium. We used 2-oxindole to study its role in regulating AhR signaling and tryptophan metabolic pathways. We found that 2-oxindole significantly increased the activity of AhR, leading to enhanced expression of its downstream targets including cytochrome P450 genes.
Resource competition predicts assembly of in vitro gut bacterial communities- HILIC
STUDY_SUMMARY
Microbiota dynamics arise from a plethora of interspecies interactions, including resource competition, cross-feeding, and pH modulation. The individual contributions of these mechanisms are challenging to untangle, especially in natural or complex laboratory environments where the landscape of resource competition is unclear. Here, we developed a framework to estimate the extent of multi-species niche overlaps by combining metabolomics data of individual species, growth measurements in pairwise spent media, and mathematical models. When applied to an in vitro model system of human gut commensals in complex media, our framework revealed that a simple model of resource competition described most pairwise interactions. By grouping metabolomic features depleted by the same set of species, we constructed a coarse-grained consumer-resource model that predicted assembly compositions to reasonable accuracy. Moreover, deviations from model predictions enabled us to identify and incorporate into the model additional interactions, including pH-mediated effects and cross-feeding, which improved model performance. In sum, our work provides an experimental and theoretical framework to dissect microbial interactions in complex in vitro environments.
Resource competition predicts assembly of in vitro gut bacterial communities- 2022-C18
STUDY_SUMMARY
Microbiota dynamics arise from a plethora of interspecies interactions, including resource competition, cross-feeding, and pH modulation. The individual contributions of these mechanisms are challenging to untangle, especially in natural or complex laboratory environments where the landscape of resource competition is unclear. Here, we developed a framework to estimate the extent of multi-species niche overlaps by combining metabolomics data of individual species, growth measurements in pairwise spent media, and mathematical models. When applied to an in vitro model system of human gut commensals in complex media, our framework revealed that a simple model of resource competition described most pairwise interactions. By grouping metabolomic features depleted by the same set of species, we constructed a coarse-grained consumer-resource model that predicted assembly compositions to reasonable accuracy. Moreover, deviations from model predictions enabled us to identify and incorporate into the model additional interactions, including pH-mediated effects and cross-feeding, which improved model performance. In sum, our work provides an experimental and theoretical framework to dissect microbial interactions in complex in vitro environments.
Resource competition predicts assembly of in vitro gut bacterial communities- 2021-C18
STUDY_SUMMARY
Microbiota dynamics arise from a plethora of interspecies interactions, including resource competition, cross-feeding, and pH modulation. The individual contributions of these mechanisms are challenging to untangle, especially in natural or complex laboratory environments where the landscape of resource competition is unclear. Here, we developed a framework to estimate the extent of multi-species niche overlaps by combining metabolomics data of individual species, growth measurements in pairwise spent media, and mathematical models. When applied to an in vitro model system of human gut commensals in complex media, our framework revealed that a simple model of resource competition described most pairwise interactions. By grouping metabolomic features depleted by the same set of species, we constructed a coarse-grained consumer-resource model that predicted assembly compositions to reasonable accuracy. Moreover, deviations from model predictions enabled us to identify and incorporate into the model additional interactions, including pH-mediated effects and cross-feeding, which improved model performance. In sum, our work provides an experimental and theoretical framework to dissect microbial interactions in complex in vitro environments.
Investigation of FGFR signaling controlled metabolism in FGFR2-fusion+ intrahepatic cholangiocarcinoma
STUDY_SUMMARY
Genomic alterations that activate FGFR2 are common in intrahepatic cholangiocarcinoma (ICC) and confer sensitivity to treatment with FGFR inhibitors. However, the depth and duration of responses are often limited. Elucidating the FGFR2-driven oncogenic program and the adaptions to FGFR inhibition is needed to gain insight into the biology of these tumors and inform future therapeutic development. Here, we conducted metabolomic analysis of patient-derived models to define the pathways that fuel tumor growth downstream of oncogenic FGFR2 signaling in ICC and to uncover compensatory mechanisms associated with pathway inhibition. We find FGFR2-fusion+ ICC maintains a highly glycolytic phenotype in FGFR2+ ICC. Conversely, FGFR inhibition blocks glucose uptake and glycolysis, while inciting a series of adaptive changes. Thus, we show that glycolysis is a key downstream effector of oncogenic FGFR2 signaling in ICC, and that pronounced metabolic reprogramming in response to FGFR inhibition confers new targetable vulnerabilities.
Bacterial tryptophan metabolites increased by prebiotic galactooligosaccharide reduce microglial reactivity and are associated with lower anxiety-like behavior (Intestine)
STUDY_SUMMARY
Prebiotic galactooligosaccharides (GOS) reduce anxiety-like behaviors in mice and humans. However, the biological pathways behind these behavioral changes are not well understood. To begin to study these pathways, C57BL/6 mice were fed a standard diet with or without GOS supplementation for 3 weeks prior to testing on the open field. After behavioral testing, colonic contents and serum were collected for bacteriome (16S rRNA gene sequencing, colonic contents only) and metabolome (UPLC-MS, colonic contents and serum data) analyses. As expected, GOS significantly reduced anxiety-like behavior (i.e., increased time in the center (p < 0.05)). Time in the center was significantly correlated with serum methyl-indole-3-acetate (mI3A). This metabolite, which is a methylated form of indole-3-acetate, is derived from bacterial metabolism of tryptophan. Sequencing analyses showed that GOS significantly increased Akkermansia, which is known to metabolize both GOS and tryptophan. To test the hypothesis that mI3A can reduce anxiety-like behavior and affect microglial activity, we first tested mI3A effects on LPS-stimulated BV2 microglia. Cells treated with mI3A produced significantly less CCL2 and TNF-Ī± than vehicle-treated cells (p<.05). We then treated mice with an intraperitoneal injection of mI3A or vehicle control, and found that mice given mI3A had lower CCL2 in the prefrontal cortex and hippocampus, as well as a reduction in a composite behavioral score in the open field. Together, these data support a novel pathway through which GOS reduces anxiety-like behaviors in mice, and suggests that the bacterial metabolite mI3A, which is elevated by GOS, reduces microglial CCL2, which in turn reduces anxiety-like behavior.
INSTITUTE
National Center for Advancing Translational Sciences
Bacterial tryptophan metabolites increased by prebiotic galactooligosaccharide reduce microglial reactivity and are associated with lower anxiety-like behavior (Blood)
STUDY_SUMMARY
Prebiotic galactooligosaccharides (GOS) reduce anxiety-like behaviors in mice and humans. However, the biological pathways behind these behavioral changes are not well understood. To begin to study these pathways, C57BL/6 mice were fed a standard diet with or without GOS supplementation for 3 weeks prior to testing on the open field. After behavioral testing, colonic contents and serum were collected for bacteriome (16S rRNA gene sequencing, colonic contents only) and metabolome (UPLC-MS, colonic contents and serum data) analyses. As expected, GOS significantly reduced anxiety-like behavior (i.e., increased time in the center (p < 0.05)). Time in the center was significantly correlated with serum methyl-indole-3-acetate (mI3A). This metabolite, which is a methylated form of indole-3-acetate, is derived from bacterial metabolism of tryptophan. Sequencing analyses showed that GOS significantly increased Akkermansia, which is known to metabolize both GOS and tryptophan. To test the hypothesis that mI3A can reduce anxiety-like behavior and affect microglial activity, we first tested mI3A effects on LPS-stimulated BV2 microglia. Cells treated with mI3A produced significantly less CCL2 and TNF-Ī± than vehicle-treated cells (p<.05). We then treated mice with an intraperitoneal injection of mI3A or vehicle control, and found that mice given mI3A had lower CCL2 in the prefrontal cortex and hippocampus, as well as a reduction in a composite behavioral score in the open field. Together, these data support a novel pathway through which GOS reduces anxiety-like behaviors in mice, and suggests that the bacterial metabolite mI3A, which is elevated by GOS, reduces microglial CCL2, which in turn reduces anxiety-like behavior.
INSTITUTE
National Center for Advancing Translational Sciences
Metabolic alteration during ferroptotic process in cancer cells.
STUDY_SUMMARY
Targeting ferroptosis, an iron-dependent form of regulated cell death triggered by the lethal overload of lipid peroxides, in cancer therapy is impeded by our limited understanding of the intersection of tumourās metabolic feature and ferroptosis vulnerability. In this study, we performed metabolomics in cancer cell lines pretreated with ferroptosis inducer RSL3 and control DMSO. We noted that the levels of a series of metabolites were significantly impacted by the RSL3 treatment, such as upregulated polyamines including putrescine, spermidine, and spermine. According to our previous study, we proved the pro-ferroptotic feature of polyamines in tumor cells, which was derived from the H2O2 produced during the polyamine metabolic process. This finding is consistent with our RNA-Seq data indicating upregulated ODC1 in the ferroptotic process. Therefore, it could be speculated that the RSL3-induced polyamine production leads to a positive-feedback loop that amplifies ferroptosis in tumor cells.
Collaborative role of YqgC and superoxide dismutase (MnSOD) in manganese intoxication: Replicate Experiment 1
STUDY_TYPE
Targeted MS Analysis
STUDY_SUMMARY
In this study, we are validating the mutual impact of yqgC and superoxide dismutase (MnSOD) deletion (YS strain) and resultant hypersensitivity to Manganese (Mn). Our genetic and physiological studies show that Mn intoxication is a result of distinct enzymatic vulnerabilities via alleviated expression of Mg-dependent, chorismate-utilizing enzymes of the menaquinone, siderophore, and tryptophan (MST) family (due to mismetalation).
Collaborative role of YqgC and superoxide dismutase (MnSOD) in manganese intoxication: Replicate Experiment 2
STUDY_TYPE
Targeted MS Analysis
STUDY_SUMMARY
In this study, we are validating the mutual impact of yqgC and superoxide dismutase (MnSOD) deletion (YS strain) and resultant hypersensitivity to Manganese (Mn). Our genetic and physiological studies show that Mn intoxication is a result of distinct enzymatic vulnerabilities via alleviated expression of Mg-dependent, chorismate-utilizing enzymes of the menaquinone, siderophore, and tryptophan (MST) family (due to mismetalation).
Leukemia inhibitory factor suppresses hepatic de novo lipogenesis and induces cachexia (Fenofibrate)
STUDY_SUMMARY
To investigate whether fenofibrate can restore hepatic lipid homeostasis, TgLC mice were fed with a regular chow or fenofibrate diet. Lipid metabolites in liver tissues were analyzed after Tamoxifen injection. A similar effect of fenofibrate on lipid metabolites was observed in C26 tumor-bearing mice.
Leukemia inhibitory factor suppresses hepatic de novo lipogenesis and induces cachexia (Tamoxifen)
STUDY_SUMMARY
We compared the levels of lipid metabolites in the liver from TgLC and TgL mice with Tamoxifen injection. The lipid metabolites levels in the liver from Balb/c mice bearing C26 or C26-LIF KO tumors were also measured. Mice under both fed and fasted conditions were included.
Methylprednisolone therapy induces differential metabolic trajectories in severe COVID-19 patients
STUDY_SUMMARY
Corticosteroids have become a choice for managing severe COVID-19, but the molecular mechanisms behind the response after corticosteroid administration remain incompletely understood. In order to unravel this, comparisons between temporal metabolic profiles in the plasma samples of methylprednisolone (MP) - and placebo-treated COVID-19 patients were performed at different time points. The patient plasma samples used were obtained from a double blind, randomized, placebo-controlled Phase IIb clinical trial performed on severe COVID-19 patients in the Brazilian Amazon where the patients received placebo or 0.5 mg/kg MP intravenously twice daily for five days. The MP treatment reduced the number of metabolites in the plasma of patients during follow-up. The longitudinal changes in the MP-group was in eight metabolic pathways related to steroid hormones and eicosanoids. Direct comparison between the two groups, revealed differences at baseline, which peaked five days after initiation of MP treatment. The metabolic pathways differing between the two groups over time included galactose metabolism, glucose and gluconeogenesis, N-glycan metabolism, and prostaglandin formation from arachidonate. Deoxy-galactose, prostaglandin H2, sphingosine, and sphinganine exhibited differential trajectories by day 14 after initiating the MP treatment. Survival of MP-treated COVID-19 patients was associated with modulation of tryptophan metabolism. Network analysis revealed that MP treatment is highly associated with alterations in pathways reflecting eicosanoid metabolism, such as arachidonic acid and prostaglandins. Curiously, there is crosstalk between metabolomics, biochemistry and cytokine components. Treatment of systemic and inflammatory conditions induced by SARS-CoV-2 viral infections with methylprednisolone modulates metabolic activity associated with tryptophan and inflammatory lipids.
INSTITUTE
Federal University of GoiƔs
LAST_NAME
Gardinassi
FIRST_NAME
Luiz Gustavo
ADDRESS
R. 235 s/n - Institute of Tropical Pathology and Public Health - Federal University of GoiƔs
Apolipoprotein E suppresses hyperlipidemia-driven hematopoiesis & inflammation by controlling mitochondrial metabolism
STUDY_SUMMARY
Apolipoprotein E (ApoE) is recognized for its pleiotropic properties that suppress inflammation. We report that ApoE serves as a metabolic rheostat that regulates microRNA-control of glycolytic and mitochondrial activity in myeloid cells and hematopoietic stem & progenitor cells (HSPCs). ApoE expression in myeloid cells increases microRNA-146a, which reduces NF-ĪŗB-driven GLUT1 expression and glycolytic activity. In contrast, ApoE expression reduces microRNA-142a, which increases CPT1A expression, fatty acid oxidation, and oxidative phosphorylation. Improved mitochondrial metabolism by ApoE expression causes an enrichment of TCA cycle metabolites and NAD+ in macrophages. The study of mice with conditional ApoE expression supports the capacity for ApoE to foster microRNA-controlled immunometabolism. Modulation of microRNA-146a & -142a in the hematopoietic system of hyperlipidemic mice using RNA mimics & antagonists, respectively, improves mitochondrial metabolism, which suppresses inflammation and hematopoiesis. Our findings unveil an RNA regulatory network, controlled by ApoE, which exerts metabolic control over hematopoiesis and inflammation in hyperlipidemia.
Targeting Pancreatic Cancer Metabolic Dependencies through Glutamine Antagonism.
STUDY_SUMMARY
Pancreatic ductal adenocarcinoma (PDAC) cells utilize glutamine (Gln) to support proliferation and redox balance. Earlier attempts to inhibit Gln metabolism using glutaminase inhibitors resulted in rapid metabolic reprogramming and therapeutic resistance. Here, we demonstrated that treating PDAC cells with a Gln antagonist, 6-Diazo-5-oxo-L-norleucine (DON), led to a metabolic crisis in vitro. In addition, we observed a profound decrease in tumor growth in various in vivo models using DRP-104 (sirpiglenastat), a pro-drug version of DON that was designed to circumvent DON associated toxicity. We found that ERK signaling is increased as a compensatory mechanism. Combinatorial treatment of DRP-104 and Trametinib led to a significant increase in survival in a syngeneic model PDAC. These proof-of-concept studies suggested that broadly targeting Gln metabolism could provide a therapeutic avenue for PDAC. The combination with an ERK signaling pathway inhibitor could further improve the therapeutic outcome.
INSTITUTE
New York University
LAST_NAME
Encarnacion Rosado
FIRST_NAME
Joel
ADDRESS
Smilow Research Building Room 907G New York, NY 10016
The cellular CD1 system binds lipid antigens for display to T cells. Here we developed a lipidomics platform that detected > 2000 distinct lipids in cellular CD1 complexes, demonstrating a broad display of self-sphingolipids and phospholipids to T cells. The four types of human CD1 antigen-presenting molecules show differing lipid capture motifs based on the length and chemical structures of lipids bound, pointing to general self-lipid capture mechanisms. For CD1a and CD1d, lipid size matches CD1 cleft volume, whereas CD1b shows a nearly universal size mismatch with its ligands, which results from the uniform seating of two small lipids within its large cleft. Further, the list of compounds that comprise the assembled CD1 lipidomes provides a resource for matching to bioactive lipids from other fields of research and supports the ongoing discovery of lipid blockers and antigens for T cells.
The cellular CD1 system binds lipid antigens for display to T cells. Here we developed a lipidomics platform that detected > 2000 distinct lipids in cellular CD1 complexes, demonstrating a broad display of self-sphingolipids and phospholipids to T cells. The four types of human CD1 antigen presenting molecules show differing lipid capture motifs based on the length and chemical structures of lipids bound, pointing to general self-lipid capture mechanisms. For CD1a and CD1d, lipid size matches CD1 cleft volume, whereas CD1b shows a nearly universal size mismatch with its ligands, which results from the uniform seating of two small lipids within its large cleft. Further, the list of compounds that comprise the assembled CD1 lipidomes provides a resource for matching to bioactive lipids from other fields of research and supports the ongoing discovery of lipid blockers and antigens for T cells.
Bap1 Promotes Osteoclast Function by Metabolic Reprogramming
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
Treatment of osteoporosis most commonly diminishes osteoclast number which suppresses bone formation thus compromising fracture prevention. Bone formation is not suppressed, however, when bone degradation is reduced by retarding osteoclast functional resorptive capacity, rather than differentiation. We find deletion of deubiquitinase, BRCA1-associated protein 1 (Bap1), in myeloid cells (Bap1āLysM), arrests osteoclast function but not formation. Bap1āLysM osteoclasts fail to organize their cytoskeleton which is essential for bone degradation. Consequently, bone mass increases in the mutant mice. We find the deubiquitinase activity of Bap1 regulates osteoclast function by metabolic reprogramming. Bap1 deficient osteoclast lineage cells upregulate the cystine transporter, Slc7a11, by enhanced H2Aub occupancy of its promoter. SLC7A11 regulates cellular ROS levels and redirects the mitochondrial metabolites away from the TCA cycle, both of which are necessary for osteoclast function. Thus in osteoclasts, Bap1 appears to regulate epigenetic-metabolic axis and is a potential target to reduce bone degradation while maintaining osteogenesis in osteoporotic patients.
INSTITUTE
Washington University in St. Louis
DEPARTMENT
Pathology and Immunology, Medicine, Chemistry
LABORATORY
Teitelbaum and Patti Laboratories
LAST_NAME
Cho
FIRST_NAME
Kevin
ADDRESS
1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA
Metabolic caracterization of liver metastasis organotropism
STUDY_SUMMARY
Transcriptomic and metabolomic analyses in animals revealed distinct metabolic adaptations, particularly related to the TCA cycle and OxPhos, specific to liver metastases compared to concurrent lung metastases. This finding was substantiated by analyzing RNA-seq data from a considerable number of patient metastases across various cancer types. Further analysis of exome/RNA-seq data from melanoma patients indicated more frequent PIK3CA mutations, lower transcript levels of PIP4K2C, and enrichment of TCA cycle and OxPhos pathways in liver metastases.
INSTITUTE
Columbia University
DEPARTMENT
Department of Medicine
LABORATORY
Division of Hematology/Oncology
LAST_NAME
Izar
FIRST_NAME
Benjamin
ADDRESS
Columbia University Irving Medical Center, New York, NY, USA
MYC is a regulator of androgen receptor inhibition-induced metabolic requirements in prostate cancer
STUDY_SUMMARY
Advanced prostate cancers are treated with therapies targeting the androgen receptor (AR) signaling pathway. While many tumors initially respond to AR inhibition, nearly all develop resistance. It is critical to understand how prostate tumor cells respond to AR inhibition in order to exploit therapy-induced phenotypes prior to the outgrowth of treatment-resistant disease. Here, we comprehensively characterize the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics and bioenergetics approaches. The metabolic response to AR inhibition is defined by reduced glycolysis, robust elongation of mitochondria, and increased reliance on mitochondrial oxidative metabolism. We establish DRP1 activity and MYC signaling as mediators of AR blockade-induced metabolic phenotypes. Rescuing DRP1 phosphorylation after AR inhibition restores mitochondrial fission, while rescuing MYC restores glycolytic activity and prevents sensitivity to complex I inhibition. Our study provides new insight into the regulation of treatment-induced metabolic phenotypes and vulnerabilities in prostate cancer.
Targeting Pancreatic Cancer Metabolic Dependencies through Glutamine Antagonism (Lipidomics-human)
STUDY_SUMMARY
Pancreatic ductal adenocarcinoma (PDAC) cells utilize glutamine (Gln) to support proliferation and redox balance. Earlier attempts to inhibit Gln metabolism using glutaminase inhibitors resulted in rapid metabolic reprogramming and therapeutic resistance. Here, we demonstrated that treating PDAC cells with a Gln antagonist, 6-Diazo-5-oxo-L-norleucine (DON), led to a metabolic crisis in vitro. In addition, we observed a profound decrease in tumor growth in various in vivo models using DRP-104 (sirpiglenastat), a pro-drug version of DON that was designed to circumvent DON associated toxicity. We found that ERK signaling is increased as a compensatory mechanism. Combinatorial treatment of DRP-104 and Trametinib led to a significant increase in survival in a syngeneic model PDAC. These proof-of-concept studies suggested that broadly targeting Gln metabolism could provide a therapeutic avenue for PDAC. The combination with an ERK signaling pathway inhibitor could further improve the therapeutic outcome.
INSTITUTE
New York University
LAST_NAME
Encarnacion Rosado
FIRST_NAME
Joel
ADDRESS
Smilow Research Building Room 907G New York, NY 10016
HILIC-IM-MS for Simultaneous Lipid and Metabolite Profiling of Microorganisms
STUDY_SUMMARY
Progress in the ion mobility mass spectrometry (IM-MS) field has significantly increased our ability to make small molecule and lipid identifications, making it an attractive approach for untargeted multi-omics experiments. The dimensionality of collision cross section (CCS) coupled with tandem mass spectrometry (MS/MS) for feature annotation has become a useful tool for high confidence structural elucidation in complex mixtures in the absence of authentic standards. A comprehensive method for feature identification of small organisms has remained limited to exploring genetic markers and protein signatures, however these methods for identification only scratch the surface of effective methods for bacterial classification. Multi-omic methods that include the metabolome and lipidome have grown in popularity due to the increased capacity for organism specific information. We have achieved species-level identification of Enterococcus faecium, Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa using a modern single-phase extraction method with hydrophilic interaction liquid chromatography (HILIC) coupled to traveling wave ion mobility mass spectrometry (TWIMS). To test the robustness of this optimized workflow, we included internal standards as a metric for efficiency of the extraction, and well known calibrants for validation for our CCS calibration method. We observed significant differences in metabolite profiles at the strain level using multi-variate statistics, primarily including quorum sensing metabolites in Gram-negative strains, and energy production metabolites in the Gram-positive strains. Lipid profiles showed staggering differences in acyl tail compositions that effectively categorized the microbes, including several classes of phospholipids and glycolipids. We have demonstrated a powerful workflow using multi-dimensional techniques for bacterial speciation in a single injection.
Targeting Pancreatic Cancer Metabolic Dependencies through Glutamine Antagonism (Lipidomics mus musculus)
STUDY_SUMMARY
Pancreatic ductal adenocarcinoma (PDAC) cells utilize glutamine (Gln) to support proliferation and redox balance. Earlier attempts to inhibit Gln metabolism using glutaminase inhibitors resulted in rapid metabolic reprogramming and therapeutic resistance. Here, we demonstrated that treating PDAC cells with a Gln antagonist, 6-Diazo-5-oxo-L-norleucine (DON), led to a metabolic crisis in vitro. In addition, we observed a profound decrease in tumor growth in various in vivo models using DRP-104 (sirpiglenastat), a pro-drug version of DON that was designed to circumvent DON associated toxicity. We found that ERK signaling is increased as a compensatory mechanism. Combinatorial treatment of DRP-104 and Trametinib led to a significant increase in survival in a syngeneic model PDAC. These proof-of-concept studies suggested that broadly targeting Gln metabolism could provide a therapeutic avenue for PDAC. The combination with an ERK signaling pathway inhibitor could further improve the therapeutic outcome.
INSTITUTE
New York University
LAST_NAME
Encarnacion Rosado
FIRST_NAME
Joel
ADDRESS
Smilow Research Building Room 907G New York, NY 10016
Validation of on-target effect of IACS-010759 in vehicle- and Enzalutamide-treated 16D cells
STUDY_SUMMARY
In this study, we sought to validate that 30nM IACS-010759, a complex I inhibitor, had an on-target effect in vehicle- and Enzalutamide-treated 16D prostate cancer cells.
INSTITUTE
UCLA
LABORATORY
Goldstein Lab
LAST_NAME
Giafaglione
FIRST_NAME
Jenna
ADDRESS
610 Charles E Young Dr S, Los Angeles, CA, 90024, USA
Intracerebroventricular Transplantation of Foetal Allogeneic Neural Stem Cells in Patients with Secondary Progressive Multiple Sclerosis (hNSC-SPMS): a phase I dose-escalation clinical trial - Metabolomics Analysis of Human CSF
STUDY_SUMMARY
This is an open-label, first-in-human, dose-escalation phase I study (NCT03282760, EudraCT2015ā004855ā37) to determine the feasibility, safety, and tolerability of the transplantation of allogeneic human neural stem/progenitor cells (hNSCs) for the treatment of progressive multiple sclerosis. We report the analysis of 1 year of data from the first cohort of 15 patients from two trial sites that received increasing numbers of allogeneic hNSCs delivered via intracerebroventricular injection in combination with an immunosuppressive regimen. No treatment-related deaths nor serious adverse events (AEs) were observed over the 12-month follow-up. Participants displayed stability of clinical and laboratory parameters, as well as lesion load and activity at the brain MRIs, compared to study entry. Longitudinal metabolomics and lipidomics analyses of cerebrospinal fluid and serum from these patients identified time and dose-dependent responses, with increased levels of free fatty acids and acylcarnitines in the CSF, especially at the highest dose of injected hNSCs at the one-year follow-up time point. Finally, a significant inverse correlation was found between the highest dose of injected hNSCs and the smaller parenchymal brain volume change (PBVC; Spearmanās rho= -0.7, p= 0.03), clinical covariates that correlated with CSF levels of free fatty acids, acyl-carnitines, oxylipins, conjugated bile acids and purine breakdown and deamination products, such as hypoxanthine. The absence of AEs and the stability of functional and structural outcomes is reassuring in terms of risks and represent a main milestone to rigorously address the challenges for the safe translation of key principles of stem cell biology into effective regenerative medicines.
INSTITUTE
University of Colorado
DEPARTMENT
Department of Biochemistry and Molecular Genetics
LABORATORY
Angelo DāAlessandro
LAST_NAME
Stephenson
FIRST_NAME
Daniel
ADDRESS
Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA
Intracerebroventricular Transplantation of Foetal Allogeneic Neural Stem Cells in Patients with Secondary Progressive Multiple Sclerosis (hNSC-SPMS): a phase I dose-escalation clinical trial - Metabolomics Analysis of Human Serum
STUDY_SUMMARY
This is an open-label, first-in-human, dose-escalation phase I study (NCT03282760, EudraCT2015ā004855ā37) to determine the feasibility, safety, and tolerability of the transplantation of allogeneic human neural stem/progenitor cells (hNSCs) for the treatment of progressive multiple sclerosis. We report the analysis of 1 year of data from the first cohort of 15 patients from two trial sites that received increasing numbers of allogeneic hNSCs delivered via intracerebroventricular injection in combination with an immunosuppressive regimen. No treatment-related deaths nor serious adverse events (AEs) were observed over the 12-month follow-up. Participants displayed stability of clinical and laboratory parameters, as well as lesion load and activity at the brain MRIs, compared to study entry. Longitudinal metabolomics and lipidomics analyses of cerebrospinal fluid and serum from these patients identified time and dose-dependent responses, with increased levels of free fatty acids and acylcarnitines in the CSF, especially at the highest dose of injected hNSCs at the one-year follow-up time point. Finally, a significant inverse correlation was found between the highest dose of injected hNSCs and the smaller parenchymal brain volume change (PBVC; Spearmanās rho= -0.7, p= 0.03), clinical covariates that correlated with CSF levels of free fatty acids, acyl-carnitines, oxylipins, conjugated bile acids and purine breakdown and deamination products, such as hypoxanthine. The absence of AEs and the stability of functional and structural outcomes is reassuring in terms of risks and represent a main milestone to rigorously address the challenges for the safe translation of key principles of stem cell biology into effective regenerative medicines.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Stephenson
FIRST_NAME
Daniel
ADDRESS
Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA
Gut Microbiota-associated Metabolites Affected the Susceptibility to Heart Health Abnormality in Young Migrants at High-altitude-Human Serum Metabolomics
STUDY_SUMMARY
Background: Young migrants in plateau are susceptible to heart health abnormality and even high-altitude heart disease (HAHD). Though the gut microbial community was found to be drastically affected when exposed to a hypobaric hypoxia environment, there is limited knowledge about the roles of gut microbiota and gut microbiota-associated serum metabolites (GMSMs) in high-altitude associated heart diseases. Hence, we performed multi-omics integration analysis of 230 graduates from the same university in this study (163 who migrated to Tibet Plateau and 67 matched controls currently living in Chengdu Plain) to explore how the gut microbiota affect the development of high-altitude associated heart health abnormality. Results: Here, we found 206 differential metabolites (82 from serum and 124 from feces) and 369 differential species among the plateau migrants and plain controls. Of these, 27 differential microbial species and 4 differential metabolites (L-Asp, betaine, 3-GUA, and Ī±-KG) that both existed in serum and feces were related to the plateau migrants with undermined heart health (HH-A), which were diagnosed by biomedical detection, electrocardiography (ECG), frequency-domain Cardiogram (FCG) and ultrasonic cardiogram (UCG). Moreover, the abundances of Streptococcus rubneri and Veillonella rogosae were related with the serum levels of L-Asp, betaine, and Ī±-KG in HH-A individuals. And lower these microbiome biomarkers and GMSMs abundances were validated in an independent cohort, both of which together had an excellent discernibility efficacy of heart health abnormality in plateau migrants, with a higher AUC value of 0.7857. Besides, supplement with the two species or each of GMSMs were confirmed to effectively attenuate hypobaric hypoxia-induced higher serum lactic acid, glycolysis, myocardial damage and cardiac hypertrophy. Integrated analysis revealed significant shift in gut microbiome exerted negative regulations in Malate-Aspartate (MA) shuttle, Tricarboxylic acid cycle (TCA) and oxidative phosphorylation in HH-A individuals. Conclusion: Plateau migration altered profoundly the signatures of gut microbiome and metabolome in young migrants. Hypobaric hypoxia-induced lower abundances of Veillonella rogosae, Streptococcus rubneri, and related GMSMs promoted the remodeling of metabolic processes, resulting in higher susceptibility to heart health abnormality in high-altitude. Our findings not only presented elaborate microbial mechanisms, but also provided potential early risk prediction and therapeutic interventions for HAHD.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Liu
FIRST_NAME
Jingjing
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Gut Microbiota-associated Metabolites Affected the Susceptibility to Heart Health Abnormality in Young Migrants at High-altitude-Human Faeces Metabolomics
STUDY_SUMMARY
Background: Young migrants in plateau are susceptible to heart health abnormality and even high-altitude heart disease (HAHD). Though the gut microbial community was found to be drastically affected when exposed to a hypobaric hypoxia environment, there is limited knowledge about the roles of gut microbiota and gut microbiota-associated serum metabolites (GMSMs) in high-altitude associated heart diseases. Hence, we performed multi-omics integration analysis of 230 graduates from the same university in this study (163 who migrated to Tibet Plateau and 67 matched controls currently living in Chengdu Plain) to explore how the gut microbiota affect the development of high-altitude associated heart health abnormality. Results: Here, we found 206 differential metabolites (82 from serum and 124 from feces) and 369 differential species among the plateau migrants and plain controls. Of these, 27 differential microbial species and 4 differential metabolites (L-Asp, betaine, 3-GUA, and Ī±-KG) that both existed in serum and feces were related to the plateau migrants with undermined heart health (HH-A), which were diagnosed by biomedical detection, electrocardiography (ECG), frequency-domain Cardiogram (FCG) and ultrasonic cardiogram (UCG). Moreover, the abundances of Streptococcus rubneri and Veillonella rogosae were related with the serum levels of L-Asp, betaine, and Ī±-KG in HH-A individuals. And lower these microbiome biomarkers and GMSMs abundances were validated in an independent cohort, both of which together had an excellent discernibility efficacy of heart health abnormality in plateau migrants, with a higher AUC value of 0.7857. Besides, supplement with the two species or each of GMSMs were confirmed to effectively attenuate hypobaric hypoxia-induced higher serum lactic acid, glycolysis, myocardial damage and cardiac hypertrophy. Integrated analysis revealed significant shift in gut microbiome exerted negative regulations in Malate-Aspartate (MA) shuttle, Tricarboxylic acid cycle (TCA) and oxidative phosphorylation in HH-A individuals. Conclusion: Plateau migration altered profoundly the signatures of gut microbiome and metabolome in young migrants. Hypobaric hypoxia-induced lower abundances of Veillonella rogosae, Streptococcus rubneri, and related GMSMs promoted the remodeling of metabolic processes, resulting in higher susceptibility to heart health abnormality in high-altitude. Our findings not only presented elaborate microbial mechanisms, but also provided potential early risk prediction and therapeutic interventions for HAHD.
INSTITUTE
Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University
DEPARTMENT
Shanghai Center for Systems Biomedicine
LABORATORY
Lu Group
LAST_NAME
Liu
FIRST_NAME
Jingjing
ADDRESS
800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China
Metabolic profiling and glucose tracing in naive and Enzalutamide-treated 16D prostate cancer cells
STUDY_SUMMARY
Advanced prostate cancers are treated with therapies targeting the androgen receptor (AR) signaling pathway. While many tumors initially respond to AR inhibition, nearly all develop resistance. It is critical to understand how prostate tumor cells respond to AR inhibition in order to exploit therapy-induced phenotypes prior to the outgrowth of treatment-resistant disease. Here, we comprehensively characterize the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics and bioenergetics approaches. The metabolic response to AR inhibition is defined by reduced glycolysis, robust elongation of mitochondria, and increased reliance on mitochondrial oxidative metabolism. We establish DRP1 activity and MYC signaling as mediators of AR blockade-induced metabolic phenotypes. Rescuing DRP1 phosphorylation after AR inhibition restores mitochondrial fission, while rescuing MYC restores glycolytic activity and prevents sensitivity to complex I inhibition. Our study provides new insight into the regulation of treatment-induced metabolic phenotypes and vulnerabilities in prostate cancer.
INSTITUTE
University of California, Los Angeles
DEPARTMENT
Molecular, Cell and Developmental Biology
LABORATORY
Andrew Goldstein
LAST_NAME
Goldstein
FIRST_NAME
Andrew
ADDRESS
610 Charles E Young Dr East, Goldstein Lab 3141 Terasaki Life Sci Bld, Los Angeles, CA, 90095, USA
Metabolic profiling, glucose tracing and glutamine tracing in naive and Enzalutamide-treated 16D prostate cancer cells expressing RFP or MYC
STUDY_SUMMARY
Advanced prostate cancers are treated with therapies targeting the androgen receptor (AR) signaling pathway. While many tumors initially respond to AR inhibition, nearly all develop resistance. It is critical to understand how prostate tumor cells respond to AR inhibition in order to exploit therapy-induced phenotypes prior to the outgrowth of treatment-resistant disease. Here, we comprehensively characterize the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics and bioenergetics approaches. The metabolic response to AR inhibition is defined by reduced glycolysis, robust elongation of mitochondria, and increased reliance on mitochondrial oxidative metabolism. We establish DRP1 activity and MYC signaling as mediators of AR blockade-induced metabolic phenotypes. Rescuing DRP1 phosphorylation after AR inhibition restores mitochondrial fission, while rescuing MYC restores glycolytic activity and prevents sensitivity to complex I inhibition. Our study provides new insight into the regulation of treatment-induced metabolic phenotypes and vulnerabilities in prostate cancer.
INSTITUTE
University of California, Los Angeles
DEPARTMENT
Molecular, Cell and Developmental Biology
LABORATORY
Andrew Goldstein
LAST_NAME
Goldstein
FIRST_NAME
Andrew
ADDRESS
610 Charles E Young Dr East, Goldstein Lab 3141 Terasaki Life Sci Bld, Los Angeles, CA, 90095, USA
Metabolic profiling, glucose tracing and glutamine tracing in 16D prostate cancer cells treated with vehicle, AR inhibitor Enzalutamide, AR inhibitor Apalutamide, or AR degrader/PROTAC ARCC-4
STUDY_SUMMARY
Advanced prostate cancers are treated with therapies targeting the androgen receptor (AR) signaling pathway. While many tumors initially respond to AR inhibition, nearly all develop resistance. It is critical to understand how prostate tumor cells respond to AR inhibition in order to exploit therapy-induced phenotypes prior to the outgrowth of treatment-resistant disease. Here, we comprehensively characterize the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics and bioenergetics approaches. The metabolic response to AR inhibition is defined by reduced glycolysis, robust elongation of mitochondria, and increased reliance on mitochondrial oxidative metabolism. We establish DRP1 activity and MYC signaling as mediators of AR blockade-induced metabolic phenotypes. Rescuing DRP1 phosphorylation after AR inhibition restores mitochondrial fission, while rescuing MYC restores glycolytic activity and prevents sensitivity to complex I inhibition. Our study provides new insight into the regulation of treatment-induced metabolic phenotypes and vulnerabilities in prostate cancer.
INSTITUTE
University of California, Los Angeles
DEPARTMENT
Molecular, Cell and Developmental Biology
LABORATORY
Andrew Goldstein
LAST_NAME
Goldstein
FIRST_NAME
Andrew
ADDRESS
610 Charles E Young Dr East, Goldstein Lab 3141 Terasaki Life Sci Bld, Los Angeles, CA, 90095, USA
The role of PFAS exposures in nonalcoholic fatty liver disease and hepatocellular carcinoma in the Multiethnic Cohort
STUDY_SUMMARY
Funding was approved by the NIH HHEAR Program (Study #2020-0500), and analysis was performed within the Mount Sinai HHEAR Laboratories (U2C ES030859 and U2CES026561). 2,952 plasma samples collected from the Multiethnic Cohort (MEC) were provided by the PI (Veronica Setiawan, PhD) and analyzed using liquid chromatography with high-resolution mass spectrometry (LC-HRMS) using a hybrid targeted and untargeted method that provides quantitation of known poly- and per-fluoroalkyl substances (PFAS), as well as untargeted detection of small molecules. Results were reported in terms of untargeted mass spectral signals (Level 5), identified small molecules (Level 1), and computational annotations (Level 3 or 4) according to the MSI Annotation Schemes
INSTITUTE
University of Southern California
LAST_NAME
Veronica
FIRST_NAME
Setiawan
ADDRESS
Genetic Epidemiology NRT 1502 1450 Biggy Street Health Sciences Campus Los Angeles
This study provides a comprehensive list of root VOCs detected from the rhizosphere of Arabidopsis thaliana and the mutants belonging to their biosynthetic pathways.
Pathogenic Staphylococcus epidermidis ICE25 response to skin and blood pH
STUDY_SUMMARY
Staphylococcus epidermidis (SE) is one of the most common bacteria of the human skin microbiota. Despite its role as a commensal, SE has emerged as an opportunistic pathogen, associated with 80% of medical devices related infections. Moreover, these bacteria are extremely difficult to treat due to their ability to form biofilms and accumulate resistance to almost all classes of antimicrobials developed so far. Thus new preventive and therapeutic strategies are urgently needed. In spite of its clinical importance, the molecular mechanisms associated with SE colonisation and disease are still poorly understood. A deeper understanding of the metabolic and cellular processes associated with response to environmental factors characteristic of SE ecological niches in health and disease might provide new clues on colonisation and disease processes. Here we studied the impact of pH conditions, mimicking the skin pH (5.5) and blood pH (7.4), in a S. epidermidis pathogenic strain, belonging to the A/C clonal lineage, by means of next-generation proteomics and 1H NMR-based metabolomics. Moreover, we evaluated the metabolic changes occurring when a sudden pH change arise, simulating the skin barrier break produced by a catheter.
Identifying Biodegradation Pathways of Cetrimonium Bromide (CTAB) Using Metagenome, Metatranscriptome, and Metabolome Tri-omics Integration
STUDY_SUMMARY
Traditional research on biodegradation of emerging organic pollutants involves slow and labor-intensive experimentation. Currently, fast-developing metagenome, metatranscriptome, and metabolome technologies promise to expedite mechanistic research on biodegradation of emerging organic pollutants. Integrating the metagenome, metatranscriptome, and metabolome (i.e., tri-omics) makes it possible to link gene abundance and expression with the biotransformation of the contaminant and the formation of metabolites from this biotransformation. In this study, we used this tri-omics approach to study the biotransformation pathways for cetyltrimethylammonium bromide (CTAB) under aerobic conditions. The tri-omics analysis showed that CTAB undergoes three parallel first-step mono-/di-oxygenations ; intermediate metabolites and expressed enzymes were identified for all three pathways, and the beta-carbon mono-/di-oxygenation is a novel pathway. Four metabolites ā palmitic acid, trimethylamine N-oxide (TMAO), myristic acid, and betaine ā were the key identified biodegradation intermediates of CTAB, and they were associated with first-step mono-/di-oxygenations This tri-omics approach with CTAB demonstrates its power for identifying promising paths for future research on the biodegradation of complex organics by microbial communities.
A carbon-nitrogen negative feedback loop underlies the repeated evolution of cnidarian-Symbiodiniaceae symbioses across >700 Myr
STUDY_SUMMARY
Symbiotic associations with Symbiodiniaceae have evolved independently across a diverse range of cnidarian taxa including reef-building corals, sea anemones, and jellyfish, yet the molecular mechanisms underlying their regulation and repeated evolution are still elusive. Here we show that despite their independent evolution, cnidarian hosts employ the same carbon-nitrogen negative feedback loop to control symbiont proliferation. Symbiont-derived photosynthates are used to assimilate nitrogenous waste via GS/GOGAT mediated amino acid biosynthesis in a carbon dependent manner, which regulates the availability of nitrogen to the symbionts. Using nutrient supplementation experiments, we show that the provision of additional carbohydrates significantly reduces symbiont density while ammonium promotes symbiont proliferation. UHPLC-HR-MS analysis confirmed that all hosts co-incorporated glucose-derived 13C and ammonium-derived 15N via GS/GOGAT mediated amino acid biosynthesis. Our results reveal a general carbon-nitrogen negative feedback loop underlying these symbioses and provide a parsimonious explanation for their repeated evolution.
INSTITUTE
King Abdullah University of Science and Technology
Comparative multi-omics analyses of cardiac mitochondrial stress in three mouse models of frataxin deficiency
STUDY_SUMMARY
Cardiomyopathy is often fatal in Friedreich Ataxia (FA). However, the FA heart maintains adequate function until disease end stage, suggesting that it can initially adapt to the loss of frataxin (FXN). Conditional knockout mouse models with no Fxn expression show transcriptional and metabolic profiles of cardiomyopathy and mitochondrial integrated stress response (ISRmt). However, ISRmt has not been investigated in models with disease-relevant, partial decrease of FXN. We characterized the heart transcriptomes and metabolomes of three mouse models of partial FXN loss, YG8-800, KIKO-700, and FxnG127V. Few metabolites were significantly changed in YG8-800 mice and did not provide a signature of cardiomyopathy or ISRmt. Instead, several metabolites were altered in FxnG127V and KIKO-700 hearts. Transcriptional changes were found in all models, but differentially expressed genes consistent with cardiomyopathy and ISRmt were only identified in FxnG127V hearts. However, these changes were surprisingly mild even at an advanced age (18-months), despite a severe decrease in FXN levels to 1% of WT. These findings indicate that the mouse heart has extremely low reliance on FXN, highlighting the difficulty in modeling genetically relevant FA cardiomyopathy.
The ganglioside GM3 protects against abdominal aortic aneurysm (AAA) by suppressing ferroptosis
STUDY_SUMMARY
Background: Abdominal aortic aneurysm (AAA) is a potentially life-threatening condition, but approved medical therapies to prevent AAA progression and rupture are currently lacking. Sphingolipids metabolism disorders are associated with the occurrence and development of AAA. It has been discovered that ganglioside GM3, a sialic acid-containing type of glycosphingolipid, plays a protective role in atherosclerosis which is an important risk factor for AAA, but the potential contribution of GM3 to AAA development has not been investigated. Methods: We performed a metabolomics study to evaluated GM3 level in plasma of human AAA patients. We profiled GM3 synthase (ST3GAL5) expression in the mouse model of aneurysm and human AAA tissues through western blotting and immunofluorescence staining. RNA sequencing, proteomic analysis, affinity purification and mass spectrometry, surface plasmon resonance (SPR) analysis, and functional studies were used to dissect the molecular mechanism of GM3-regulating ferroptosis. We conditionally deleted and overexpressed St3gal5 in smooth muscle cells (SMCs) in vivo to investigate its role in AAA. Results: We found significantly reduced plasma levels of the GM3 in human AAA patients. GM3 content and ST3GAL5 expression were all decreased in abdominal aortic VSMCs in AAA patients and mouse model. RNA-sequencing analysis showed that ST3GAL5 silencing in human aortic SMCs (HASMCs) induced ferroptosis. Importantly, we showed that GM3 interacted directly with the extracellular domain of transferrin receptor 1 (TFR1), a cell membrane protein critical for cellular iron uptake, disrupted its interaction with holo-transferrin. SMC-specific St3gal5 knockout exacerbated iron accumulation at lesion sites and significantly promoted AAA development, while GM3 supplementation suppressed lipid peroxidation, reduced iron deposition in aortic VSMCs and markedly decreased AAA incidence. Conclusions: Together, these results suggest that GM3 dysregulation promotes ferroptosis of VSMCs in AAA. Furthermore, GM3 may constitute a new therapeutic target for the treatment of AAA.
INSTITUTE
Tianjin Medical University
LAST_NAME
Li
FIRST_NAME
Kan
ADDRESS
Tianjin Medical University, Tianjin, China., Tianjin, Tianjin, 300070, China
Quantification of metabolites in kynurenine pathway
STUDY_SUMMARY
Urine samples were collected from two independent cohorts. Patients with LN were classified into proliferative (class III/IV) and membranous (class V) by kidney histopathology. Quantification of metabolites in kynurenine pathway was performed using LC-MS/MS.
INSTITUTE
Mahidol University
DEPARTMENT
Siriraj Metabolomics and Phenomics Center
LABORATORY
Siriraj Center of Research Excellence in Metabolomics and Systems Biology
The LAT1-4F2hc complex (SLC7A5-SLC3A2) facilitates uptake of essential amino acids, hormones and drugs. Its dysfunction is associated with many cancers and immune/neurological disorders. Here, we apply native mass spectrometry (MS)-based approaches to provide evidence of super-dimer formation (LAT1-4F2hc)2. When combined with lipidomics, and site-directed mutagenesis, we discover four endogenous phosphatidylethanolamine (PE) molecules at the interface and C-terminus of both LAT1 subunits. We find that interfacial PE binding is regulated by 4F2hc-R183 and is critical for regulation of palmitoylation on neighbouring LAT1-C187. Combining native MS with mass photometry (MP) we reveal that super-dimerization is sensitive to pH, and modulated by complex N-glycans on the 4F2hc subunit. Free LAT1 is also present in the lysosome after interferon-Ī³ (IFN-Ī³) stimulation suggesting that the assembly of LAT1-4F2hc is regulated by redox dynamics in vivo. Together our results link PTM and lipid binding with regulation and trafficking of the LAT1-4F2hc super dimer.
INSTITUTE
University of Oxford
DEPARTMENT
Kavli Institute for Nanoscience Discovery
LABORATORY
Prof. Carol Robinson Group
LAST_NAME
Wu
FIRST_NAME
Di
ADDRESS
Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, OX1 3QU, UK
High Level Expression of NSD2 Creates a Metabolic Dependency in Multiple Myeloma
STUDY_SUMMARY
Multiple myeloma (MM) is a malignancy of plasma cells with several molecular subtypes and variable prognosis. Despite therapeutic advances, most patients ultimately relapse due to drug resistance. Frontline treatments for MM target malignant cells based on their differentiated B cell nature, but not the underlying genetic lesions. Chromosomal translocation t(4;14), observed in 15% of MM patients, results in overexpression of the histone methyltransferase NSD2, which contributes to MM pathogenesis by promoting an oncogenic transcriptional program and is associated with a worse prognosis. A genome-wide CRISPR based functional screen in isogenic MM cell lines with high and low NSD2 expression identified the mitochondrial enzyme adenylate kinase 2 (AK2) as a NSD2-driven MM cell dependency. AK2 loss in t(4;14) MM cells induced apoptosis and inhibited cell growth in vitro and in vivo. Consistent with a defect in shuttling ATP from the mitochondria to intracellular utilization sites, AK2 depletion impaired ATP-dependent protein folding in the ER and increased MM cell sensitivity to the proteasome inhibitor bortezomib. Furthermore, AK2 suppression decreased intracellular NAD(H) phosphorylation resulting in lower NADP(H) levels. Cytosolic NADPH is necessary for reducing thioredoxin, an essential cofactor for ribonucleotide reductase which is critical for deoxyribonucleotides (dNTP) synthesis. Consequently, AK2 deficiency in MM cells resulted in dNTP pool depletion and induced DNA replication stress. Creatine phosphorylation by mitochondrial creatine kinase is an alternative route for shuttling ATP from the mitochondria to the cytosol. Metabolomics analysis revealed decreased levels of creatine and accumulation of its precursor guanadoacetate in NSD2 high cells, in association with elevated levels of S-adenosyl homocysteine (SAH) indicating consumption of the carbon donor S-Adenosyl methionine (SAM). This along with the 6-fold increase in genome wide H3K36me2 levels and 40% increase in DNA methylation levels in NSD2 high cells suggested that overexpression of NSD2 redirected one-carbon metabolism to the epigenome and away from the SAM-dependent creatine synthesis. Therefore, decreased creatine levels in NSD2 overexpressing cells underlie the increased reliance on AK2. Correspondingly, supplementation with exogenous creatine restored NADP(H) levels and rescued AK2 deficient cells from apoptosis. These findings revealed a novel metabolic susceptibility in t(4;14) MM and provided insight into a novel therapeutic strategy to improve patient prognosis. We performed metabolomic analysis of multiple myeloma cell lines with either high or low NSD2 levels with or without NSD2 knockdown.
Metabolic Profiling of Raw264.7 Mouse Macrophage Cells Cultured with Alanine
STUDY_SUMMARY
To identify the catabolites of L-Alanine on promoting phagocytosis, GC-MS based metabolomics analysis was adopted to explore L-Alanine-reprogrammed metabolome. The metabolic flow of the TCA cycle was dysregulated. Meanwhile, six metabolites (oleate, palmitate, stearate, myristate, arachidonate and linoleate) in biosynthesis of saturated and unsaturated fatty acids were increased upon L-Alanine treatment, where palmitate was the biggest absolute increment in abundance. Thus, L-Alanine promotes the biosynthesis of fatty acids.
INSTITUTE
Sun Yat-sen University
LAST_NAME
jiang
FIRST_NAME
ming
ADDRESS
No. 135, Xingang Xi Road, Guangzhou, 510275, P. R. China, guangzhou, guangdong, 510006, China
Atlas of fetal metabolism during mid-to-late gestation and diabetic pregnancy. Dynamic Labelling experiment.
STUDY_SUMMARY
Mounting evidence supports an instructive role for metabolism in stem cell fate decisions. However, much is yet unknown about how fetal metabolism changes during mammalian development and how altered maternal metabolism shapes fetal metabolism. Here, we present a descriptive atlas of in vivo fetal murine metabolism during mid-to-late gestation in normal and diabetic pregnancy. Using 13C-glucose and LC-MS, we profiled the metabolism of fetal brains, hearts, livers, and placentas harvested from pregnant dams between embryonic days (E)10.5 and 18.5. Comparative analysis of our large metabolomics dataset revealed metabolic features specific to fetal tissues developed under a hyperglycemic environment as well as metabolic signatures that may denote developmental transitions during euglycemic development. We observed sorbitol accumulation in fetal tissues and altered neurotransmitter levels in fetal brains isolated from dams with maternal hyperglycemia. Tracing 13C-glucose revealed disparate nutrient sourcing in fetuses depending on maternal glycemic states. Regardless of glycemic state, histidine-derived metabolites accumulated during late development in fetal tissues and maternal plasma. Our rich dataset presents a comprehensive overview of in vivo fetal tissue metabolism and alterations occurring as a result of maternal hyperglycemia.
INSTITUTE
University of California, Los Angeles
DEPARTMENT
Biological Chemistry
LABORATORY
Heather Christofk
LAST_NAME
Matulionis
FIRST_NAME
Nedas
ADDRESS
615 Charles E Young Drive South Los Angeles, CA, 90095
A distinct, hypoxia-related lipid composition of Solanum lycopersicum root tissue was observed. Out of 965 lipid species, 33 were exclusively detected in this condition. Among the lipid classes observed, glycerolipids and glycerophospholipids dominated by far (77%). Significantly, the abundance of triacylglycerols increased with hypoxic stress, while sitosterol esters, digalactosyldiacylglycerols, and phosphatidylcholine decreased. Alongside, an increased level of polyunsaturation was observed in the fatty acid chains, with 18:2, 18:3 residues showing a significant increase. Of note, hexadecatetraenoic acid (16:4) was identified in hypoxia condition samples.
INSTITUTE
Leibniz Institute for Plasma Science and Technology
Characterization of the in vivo deuteration of native phospholipids by mass spectrometry yields guidelines for their regiospecific customization
STUDY_SUMMARY
Customization of deuterated biomolecules is vital for many advanced biological experiments, including neutron scattering. However, because it is challenging to control the proportion and regiospecificity of deuterium incorporation in live systems, often only two or three synthetic lipids are mixed together to form simplistic model membranes. This limits the applicability and biological accuracy of the results generated with these synthetic membranes. Despite some limited prior examination of deuterating E. coli lipids in vivo, this approach has not been widely implemented. Here, an extensive mass spectrometry-based profiling of E. coli phospholipid deuteration states with several different growth media was performed and a computational method to describe deuterium distributions with a one-number summary is introduced. The deuteration states of thirty-six lipid species were quantitatively profiled in fifteen different growth conditions and tandem mass spectrometry was used to reveal deuterium localization. Regressions were employed to enable the prediction of lipid deuteration for untested conditions. Small-angle neutron scattering was performed on select deuterated lipid samples, which validated the deuteration states calculated from the mass spectral data. Based on these experiments, guidelines for the design of specifically deuterated phospholipids are described. This unlocks even greater capabilities from neutron-based techniques, enabling experiments that were formerly impossible.
INSTITUTE
University of Tennessee
DEPARTMENT
Genome Science and Technology (Bredesen Center)
LAST_NAME
Matthew
FIRST_NAME
Keller
ADDRESS
The Bredesen Center for Interdisciplinary Research and Graduate Education 444 Greve Hall, 821 Volunteer Blvd. Knoxville, TN 37996-3394
Glutamine metabolism improves left ventricular function but not macrophage-mediated inflammation following myocardial infarction
STUDY_SUMMARY
Glutamine is a critical amino acid that serves as an energy source, building block, and signaling molecule for both the heart tissue and the immune system. However, the role of glutamine metabolism in regulating cardiac remodeling following myocardial infarction (MI) is unknown. In this study, we show that glutamine metabolism is altered both in the remote (contractile) area and in infiltrating macrophages in the infarct area after MI in adult male mice by permanent left anterior descending artery occlusion. Using untargeted metabolomics in extracted LV macrophages, we found that metabolites related to glutamine metabolism were differentially altered at days 1, 3, and 7 after MI. Glutamine metabolism in live cells was found to be increased after MI relative to no MI controls. Gene expression in the remote area of the heart indicated a loss of glutamine metabolism. Glutamine administration improved LV function at days 1, 3, and 7, which was associated with improved contractile and metabolic gene expression.
INSTITUTE
Vanderbilt University
DEPARTMENT
Chemistry
LABORATORY
Center for Innovative Technology
LAST_NAME
Codreanu
FIRST_NAME
Simona
ADDRESS
1234 STEVENSON CENTER LANE, NASHVILLE, TN, 37235, USA
Folate levels in K562 cells following folate depletion
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 100 nM folic acid for up to 8 days followed my metabolomics targeting folate metabolites. All samples were harvested at the same time and the study was set up as a reverse time-course. Day 0 samples were cultured in 2,000 nM folate for the full 8 days.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Folate depletion time-course in MEL cells with analysis for porphyrin metabolites
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 100 nM folic acid for 0, 1, 2, 4, 6, or 8 days followed my LC-MS targeting porphyrin metabolites. This is a reverse timecourse where all samples are harvested on the same day. Day 0 in 100 nM folic acid indicates 8 days culture in 2,000 nM folic acid.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Folate levels in MEL cells following folate depletion
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 100 nM folic acid for up to 8 days followed my metabolomics targeting folate metabolites. All samples were harvested at the same time and the study was set up as a reverse time-course. Day 0 samples were cultured in 2,000 nM folate for the full 8 days.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Polar metabolite levels in K562 cells following folate depletion and inosine supplementation
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 100 nM folic acid for 48hr in the presence or absence of 100 uM inosine followed my metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Polar metabolite levels in MEL cells following folate depletion and inosine supplementation
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 100 nM folic acid for 48hr in the presence or absence of 100 uM inosine followed my metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Polar metabolite levels in K562 cells following folate depletion, SHIN1 or AICAR treatment
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 2000 nM folic acid for 48hr in the presence or absence of SHIN1 (1.25 uM) or AICAR (500 nM). In addition, K562 cells were cultured in 100 nM folic acid. These samples were followed up by metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Polar metabolite levels in MEL cells following folate depletion, SHIN1 or AICAR treatment
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 2000 nM folic acid for 48hr in the presence or absence of SHIN1 (1.25 uM) or AICAR (500 nM). In addition, K562 cells were cultured in 100 nM folic acid. These samples were followed up by metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Polar metabolite levels in erythroid progenitor cells following SHIN1 and inosine treatment
STUDY_SUMMARY
Culture of erythroid progenitor cells cultured in SFEM II media supplemented with SHIN1 and/or inosine for 48 hours followed up by metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Folate metabolite levels in erythroid progenitor cells following SHIN1 and inosine treatment
STUDY_SUMMARY
Culture of erythroid progenitor cells cultured in SFEM II media supplemented with SHIN1 and/or inosine for 48 hours followed up by metabolomics targeting folate metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Potential serum metabolite markers and predictive features of depressive-like behavior and effective fluoxetine treatment in chronically socially isolated rats
STUDY_SUMMARY
Metabolic perturbation has been associated with depression. To examine peripheral metabolome changes, untargeted metabolic profiling was employed in the serum of chronic social isolation (CSIS) rats, an animal model of depression, and/or chronic fluoxetine (Flx) treatment using liquid chromatography-high resolution mass spectrometry.
Quantitative analysis of fatty acid compositions of retail cowās milk and milk alternatives in Thailand using GC-MS
STUDY_SUMMARY
All 36 brands of commercial milk samples available in retail stores in Thailand were collected in July 2019. They included 17 brands of whole cowās milk, 6 brands of lactose-free cowās milk, 9 brands of soymilk, and 4 brands of almond milk. Of all the cowās milk samples, there were 6, 2, and 9 ultra-high temperature processing (UHT), sterilized, pasteurized milk samples, respectively. Quantification of fatty acids was performed using GC-MS.
INSTITUTE
Mahidol University
DEPARTMENT
Siriraj Metabolomics and Phenomics Center
LABORATORY
Siriraj Center of Research Excellence in Metabolomics and Systems Biology
Identification and targeting of microbial putrescine acetylation in bloodstream infections
STUDY_TYPE
comparison of septic shock versus control plasma
STUDY_SUMMARY
To identify bacterial metabolites elevated in human plasma during infection, we performed metabolomics on an existing cohort of patient plasma samples from 21 septic shock patients admitted to the intensive care unit (ICU) with culture positive gram-negative BSIs (Escherichia coli, Klebsiella spp., Pseudomonas spp.) who had banked blood samples drawn contemporaneously or near-contemporaneously with their positive blood cultures as well from 22 controls admitted to the ICU for other reasons.
Porphyrin analysis of K562 cells following C13-Serine and C13-Glycine tracing during folate depletion.
STUDY_SUMMARY
Culture of K562 cells for 7 days in RPMI media containing 2000 nM folic acid or 100 nM folic acid. At day 8, media was changed to 2000 nM or 100 nM folic acid with unlabeled serine and glycine, or 2-C13-Serine and 2-C13-Glycine at RPMI levels. Amino acid tracing was performed for 24 hours. These samples were followed up by metabolomics targeting porphyrin metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Plasma metabolomics reveals distinct biological and diagnostic signatures for melioidosis
STUDY_SUMMARY
Rationale: The global burden of sepsis is greatest in low-resource settings. Melioidosis, infection with the Gram-negative bacterium Burkholderia pseudomallei, is a frequent cause of fatal sepsis in endemic tropical regions such as Southeast Asia. Objectives: To investigate whether plasma metabolomics would identify biological pathways specific to melioidosis and yield clinically meaningful biomarkers. Methods: Using a comprehensive approach, differential enrichment of plasma metabolites and pathways was systematically evaluated in individuals selected from a prospective cohort of patients hospitalized in rural Thailand with infection. Statistical and bioinformatics methods were used to distinguish metabolomic features and processes specific to melioidosis patients, and between fatal and non-fatal cases. Measurements and Main Results: Metabolomic profiling and pathway enrichment analysis of plasma samples of melioidosis (n=175) and non-melioidosis infections (n=75) revealed a distinct immuno-metabolic state among patients with melioidosis, as suggested by excessive tryptophan catabolism in the kynurenine pathway and significantly increased levels of sphingomyelins and ceramide species. We derived a 12-metabolite classifier to distinguish melioidosis from other infections, yielding an area under the receiver operating characteristic curve of 0.87 in a second validation set of patients. Melioidosis non-survivors (n=94) had a significantly disturbed metabolome compared to survivors (n=81) with increased leucine, isoleucine and valine metabolism, and elevated circulating free fatty acids and acylcarnitines. A limited 8-metabolite panel showed promise as an early prognosticator of mortality in melioidosis. Conclusions: Melioidosis induces a distinct metabolomic state that can be examined to distinguish underlying pathophysiological mechanisms associated with death. A twelve-metabolite signature accurately differentiates melioidosis from other infections and may have diagnostic applications.
INSTITUTE
University of Washington
LAST_NAME
Gharib
FIRST_NAME
Sina
ADDRESS
Center for Lung Biology, 850 Republican St. Seattle WA 98109
Identifying the impact of RelA overexpression in triple negative breast cancer cells using mass spectroscopy-based proteomics and metabolomics analysis
STUDY_SUMMARY
Introduction: The incidence of chemotherapeutic resistance among breast cancer patients continues to rise steadily. The expression of RelA, a prominent member of the Rel family, is closely associated with the aggressiveness of triple-negative breast cancer (TNBC) and a poor prognosis for patients. Consequently, it is crucial to deeply investigate the molecular changes that underlie breast cancer progression and chemotherapy resistance associated with RelA overexpression. Materials and Methods: In this study, we performed a comprehensive quantitative analysis of proteomics and metabolomics in triple negative breast cancer (TNBC) cells overexpressing RelA, compared to TNBC cells with basal levels. State-of-the-art Trapped Ion Mobility Spectroscopy, Time-of-Flight Mass Spectrometry (TIMS-TOF-MS) was employed to achieve high-resolution analysis. Results: The results unveiled 27 significantly dysregulated proteins and 21 altered metabolites in MDA-MB-231 RelA cells relative to MDA-MB-231 cells. The upregulated proteins in MDA-MB-231 RelA cells include interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and cytochrome b5, which are involved in tumor migration and progression and regulation of the cellular redox system, respectively. Conversely, the adhesion molecule, integrin alpha-2, was downregulated in MDA-MB-231 RelA cells. In addition, metabolomics analysis revealed significant upregulation in the signal transducer, cyclic AMP, and downregulation in multiple nucleotides such as pyridine, adenine, and thymidine. The integrated analysis of multi-omics data highlighted the highest impacted pathways, including ABC transporters, arginine biosynthesis, purine, pyruvate, pyrimidine, glutathione, and phenylalanine metabolism. Conclusion: This study effectively recognized significantly dysregulated proteins and metabolites in MDA-MB-231 RelA cells, providing valuable insights into potential proteins, metabolites, and signaling pathways that mediate the aggressiveness of TNBC through RelA. Moreover, the multi-omics integrated analysis elucidated RelA role in chemotherapy resistance, tumor progression, migration, and invasion, which would propose potential biomarkers and novel therapeutic targets.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
LiLA: Lipid Lung-based ATLAS built Through a Comprehensive Workflow Designed for an Accurate Lipid Annotation
STUDY_SUMMARY
Accurate lipid annotation is crucial for understanding the role of lipids in health and disease and identifying therapeutic targets. However, annotating the wide variety of lipid species in biological samples remains challenging in untargeted lipidomic studies. In this work, we present an optimized lipid annotation workflow based on the combination of LC-MS and MS/MS strategies, four bioinformatic tools, and a decision-tree-based approach to support the accurate annotation and semi-quantification of the lipid species present in lung tissue from control mice. The developed workflow allowed us to generate a lipid lung-based ATLAS (LiLA), which was then employed to unveil the lipidomic signatures of the Mycobacterium tuberculosis (Mtb) infection at two different time points for a deeper understanding of the disease progression. This workflow, combined with manual inspection strategies of MS/MS data, can enhance the annotation process for lipidomic studies and guide the generation of sample-specific lipidome maps. LiLA serves as a freely available data resource that can be employed in future studies to address lipidomic alterations in mice lung tissue.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
Centro de MEtabolĆ³mica y BioanĆ”lisis (CEMBIO)
LAST_NAME
GonzƔlez
FIRST_NAME
Carolina
ADDRESS
km 0, Universidad CEU-San Pablo UrbanizaciĆ³n MonteprĆncipe. M-501
Polar metabolite levels in MEL cells following folate depletion
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 100 nM folic acid for up to 8 days followed my metabolomics targeting polar metabolites. All samples were harvested at the same time and the study was set up as a reverse time-course. Day 0 samples were cultured in 2,000 nM folate for the full 8 days.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Polar metabolite levels in K562 cells following folate depletion
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 100 nM folic acid for up to 8 days followed my metabolomics targeting polar metabolites. All samples were harvested at the same time and the study was set up as a reverse time-course. Day 0 samples were cultured in 2,000 nM folate for the full 8 days.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Untangling the Dynamics of Lysine Acetylation and Phosphorylation in Adipogenesis in the Established Human and Mouse Adipocyte Cell Lines SGBS and 3T3L1
STUDY_SUMMARY
Obesity is one of the most pressing global public health challenges of our time with yet increasing prevalence. Characterized by enlarged and dysfunctional adipose tissue, it is often associated with the development of metabolic or cardiovascular comorbidities. A sound understanding of the processes underlying adipogenesis, the differentiation of a preadipocyte into a mature and lipid laden adipocyte, provide the basic knowledge for future research on the causes and consequences of obesity. The tricarboxylic acid cycle represents the central metabolic hub, that provides energy equivalents, metabolic building blocks and critical intermediates such as acetyl-CoA as precursor for protein acetylation and de novo lipogenesis. Stable cell lines are an integral part of research into the development and physiology of adipocytes in health and disease and various models have been used to date. In this study we demonstrated the vivid temporal dynamics of central carbon metabolites during the differentiation of SGBS and 3T3-L1 adipocytes. Detected metabolite levels showed distinct temporal profiles, partly with cell-line specificities.
MoTrPAC: Endurance exercise training study in young adult rats, Rat Kidney Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
MoTrPAC: Endurance exercise training study in young adult rats, Rat Heart Powder - Untargeted Reversed-Phase Negative
STUDY_SUMMARY
The goal of the endurance exercise training study in young adult rats (internal code: PASS1B-06) was to perform exercise training studies in adult (6 month) F344 rats, and from these rats collect as many tissues as feasible in order to provide high quality samples for detailed analysis by chemical analysis sites. Tissues were collected from 10-12 rats sedentary control rats concurrent with the collection of the 8-week training groups. The 8-week training group and controls were from the same cohort and same age at euthanasia (either 8). For the older age group, an additional set of controls (n=5-6) were collected with the 1-2 week training group. Rats were either sedentary or underwent an exercise training program. Rats were exercised on the rodent treadmill 5 days per week using a progressive training protocol designed to exercise the rats at approximately 70% of VO2max as outlined in the Table on the next page. Training was performed no earlier than 10:00 am and no later than 5:00 pm over 5 consecutive days per week. Training was initiated with the treadmill set at 70% of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of exercise was increased by one minute each day until day 31 of training (start of week 7), when a duration of 50 min was reached. Speed and grade of each training session increased in larger increments due to treadmill parameters. The highest intensity and duration of training began on day 31. This intensity was maintained for the final 10 days of the protocol to ensure steady state had been achieved. If any rats were unable to perform at least 4 days of training per week they were removed from the study and euthanized. It is important to note that the starting treadmill speed varied depending on the sex and age of the rat. The initial and maximum speeds were based on VO2max measurements obtained during the pre-training testing of the compliant rats. Rats assigned to the control group followed a schedule similar to the training group. They were placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The treadmill was set at 0 m/min at an incline that corresponded to the incline being used by the training group.
INSTITUTE
University of Michigan
DEPARTMENT
Biomedical Research Core Facilities
LABORATORY
Metabolomics core
LAST_NAME
Burant
FIRST_NAME
Charles
ADDRESS
6120 Brehm Tower,1000 Wall St., Ann Arbor, MI, 48105-5714
Transporter-mediated depletion of apoplastic proline directly contributes to plant pattern-triggered immunity against a bacterial pathogen
STUDY_SUMMARY
GC-MS analysis of apoplastic fluid extracted from arabidopsis plants treated with 100 nM flg22 or a mock treatment for 8 hours. Col-0 is wild type arabidopsis plants, QKO is a quadruple knockout mutant in the Col-0 background with knockouts in dde2-2, ein2-1, pad4-1, and sid2-2.
INSTITUTE
Oregon State University
DEPARTMENT
Botany and Plant Pathology
LABORATORY
Jeff C Anderson
LAST_NAME
Rogan
FIRST_NAME
Conner
ADDRESS
Cordley Hall, 2701 SW Campus Way, Corvallis, OR 97331
Metabolic Profiling in mouse Infected with Vibrio parahaemolyticus
STUDY_SUMMARY
mice were subject to intraperitoneal (i.p.) injection with the V. parahaemolyticus, and control mice were injected with saline. Data were collected and analyzed using unsupervised hierarchical clustering. In general, the abundance of metabolites increased more often than it decreased in animals with higher resistance to infection. Load weight analysis of biological pathways suggested that alanine, aspartate, glutamate metabolism could play roles in the capacity of mice to survive infection with V. parahaemolyticus.
INSTITUTE
Sun Yat-sen University
LAST_NAME
jiang
FIRST_NAME
ming
ADDRESS
No. 135, Xingang Xi Road, Guangzhou, 510275, P. R. China
Short-term metabolic insulin response of DINCH- and MINCH-treated cells
STUDY_SUMMARY
In the second part of the project, we examined the short-term metabolic insulin response of DINAH- and MINCH-treated cells and compared them with rosiglitazone-differentiated cells. For this purpose, the human SGBS preadipocyte cell line was exposed to differentiation media conditioned with DINCH (10 nM or 10 ĀµM), MINCH (10 nM or 10 ĀµM), or rosiglitazone, and the insulin response was measured by analyzing the changes in glycolysis and PPP between insulin-stimulated and non-insulin-stimulated cells. In conclusion, the insulin response of glycolysis and PPP of cells treated with 10 ĀµM MINCH, but not with 10 nM MINCH or DINCH, was similar to cells differentiated with rosiglitazone.
Possible PPARG-independent effects of DINCH and MINCH on central carbon metabolism
STUDY_SUMMARY
In the third part of the project, we investigated the PPARG-independent effects of DINCH and MINCH on the central carbon metabolism of human SGBS cells. SGBS preadipocytes were treated for 12 days with 10 ĀµM MINCH, 10 ĀµM DINCH, or rosiglitazone in the presence of the PPARG inhibitor GW9662. Irreversible blocking of PPARG was achieved by incubating cells with 10 ĀµM GW9662 1 hour before treatment and maintaining a regular treatment interval of 2 days. In conclusion, although the PPARG inhibitor GW9662 greatly reduced the effects of MINCH and rosiglitazone, slightly increased lipid accumulation along with increased lactate secretion and increased concentrations of pyruvate cycle metabolites were consistently induced by MINCH treatment even after PPARG inhibition. This suggests that MINCH exerts its effects on lipid accumulation and central carbon metabolism at least in part via a PPARG-independent mechanism.
Metabolomic Characteristics of Nontuberculous Mycobacterial Pulmonary Disease
STUDY_SUMMARY
While the burden of nontuberculous mycobacterial pulmonary disease (NTM-PD) continues to increase, knowledge of biomarkers for NTM-PD remains insufficient. Furthermore, metabolic changes in NTM-PD have not yet been investigated. The identification of specific metabolites and associated metabolic pathways unique to NTM-PD will provide a deeper understanding of its pathogenesis. In this study, we aimed to discover specific metabolic biomarkers for NTM-PD using a metabolomics approach. In this study, we underwent untargeted metabolomic profiling using a liquid chromatography system coupled with the quadrupole-orbitrap mass spectrometer to analyze serum samples from patients with NTM-PD (n = 50), patients with non-NTM bronchiectasis (n = 50), and HC subjects (n = 60). To validate the results, an additional 86 serum samples for each group were analyzed using the same analytical methods. We identified several NTM-PD significant metabolites that differentiate patients with NTM-PD from healthy individuals. The machine learning-based classification model demonstrated the proficiency of the selected metabolic features in distinguishing between patients with NTM-PD and healthy individuals. These findings may contribute to a better understanding of the pathogenesis of NTM-PD and provide insights for the development of novel treatment approaches.
INSTITUTE
Seoul National University College of Medicine and Hospital
Effects of DINCH and MINCH on adipocyte metabolism of human SGBS cells.
STUDY_SUMMARY
In the first part of the project, we investigated the effects of DINCH and MINCH on central carbon metabolism. For this purpose, the human SGBS preadipocyte cell line (Wabitsch et al., 2001) was exposed to DINCH and MINCH at concentrations ranging from 10 nM to 10 ĀµM and compared with cells differentiated with rosiglitazone (adipogenic reference) and without rosiglitazone (undifferentiated control). Analysis of central carbon metabolism showed that MINCH, similar to rosiglitazone, induces lipid accumulation mainly through PPARG-mediated upregulation of the pyruvate cycle. In addition, increased lactate production suggests altered glucose homeostasis induced by MINCH-treatment. Our results suggest that MINCH could potentially lead to a weight-promoting effect, as observed with thiazolidinediones, because of the similarity of the observed changes to the effects of the thiazolidinedione rosiglitazone.
Gut sphingolipid metabolites in infants with atopic dermatitis associated with food allergy - Part 1
STUDY_SUMMARY
Food allergy (FA) may be present in the range of 20ā80% in atopic dermatitis (AD). Food sensitization through the skin can cause FA due to damage to the skin barrier, and failure to acquire tolerance to food allergens in the gut can equally cause the development of FA. Gut metabolites can influence the physical gut barrier and intestinal homeostasis. Therefore, it is possible that gut metabolites related to gut immunity play an important role in the development of FA. Sphingolipids are key factors in cell inflammatory response and affect gut epithelial cells and skin barrier integrity and function. FA is associated with a marked decrease in serum sphingolipid levels. However, there are no reports of FA-associated gut sphingolipid metabolites in infants by targeted metabolomics. In our previous study, we showed that when FA is present in various phenotypes of AD in early life, it might be associated with the later development of asthma. The discovery of a biomarker that can distinguish the phenotypes that accompany AD and FA from other AD phenotypes is therefore expedient. Consequently, we aimed to investigate whether FA in AD infants. can be classified as gut sphingolipid metabolites using targeted metabolomics.
Gut sphingolipid metabolites in infants with atopic dermatitis associated with food allergy - Part 2
STUDY_SUMMARY
This study determines sphingolipids and diacylglycerols from infant feces to explore the lipid changes with food allergy in atopic dermatitis. Food allergy (FA) may be present in the range of 20ā80% in atopic dermatitis (AD). Food sensitization through the skin can cause FA due to damage to the skin barrier, and failure to acquire tolerance to food allergens in the gut can equally cause the development of FA. Gut metabolites can influence the physical gut barrier and intestinal homeostasis. Therefore, it is possible that gut metabolites related to gut immunity play an important role in the development of FA. Sphingolipids are key factors in cell inflammatory response and affect gut epithelial cells and skin barrier integrity and function. FA is associated with a marked decrease in serum sphingolipid levels. However, there are no reports of FA-associated gut sphingolipid metabolites in infants by targeted metabolomics. In our previous study, we showed that when FA is present in various phenotypes of AD in early life, it might be associated with the later development of asthma. The discovery of a biomarker that can distinguish the phenotypes that accompany AD and FA from other AD phenotypes is therefore expedient. Consequently, we aimed to investigate whether FA in AD infants. can be classified as gut sphingolipid metabolites using targeted metabolomics.
INSTITUTE
Asan Medical Center
LAST_NAME
Yoo
FIRST_NAME
Hyun Ju
ADDRESS
88, Olympic-ro, 43-gil, Songpa-gu, Seoul, Seoul, 05505, Korea, South
Metabolite analysis of hepatic Pptc7-ko mice under fed or fasting conditions
STUDY_SUMMARY
Hepatic Pptc7 knockout leads to mitochondrial loss under fed conditions in adult mice, which is exacerbated upon fasting. Under fasting conditions, WT liver upregulates Pptc7 levels to repress mitophagy and maintain mitochondrial mass. However, Pptc7-KO liver accumulated higher levels of Bnip3, resulting in mitophagy activation and further mitochondrial loss. To explore the metabolic changes upon Pptc7 knockout under both fed and fasting conditions, we performed a metabolite analysis of wild-type or Pptc7-KO mouse liver samples in this study.
INSTITUTE
National Institute of Biological Sciences, Beijing
Multi-āomicsā analysis reveals the orphan P. falciparum protein kinase PfPK8 regulates multi-gene family expression
STUDY_SUMMARY
Protein kinases are a large group of proteins that serve regulatory and signalling functions in eukaryotic cells. Whilst kinases can be readily identified by highly conserved kinase domains, the downstream function of many protein kinases remains unknown, even more so for kinases of divergent eukaryotes, such as the Plasmodium parasites that cause malaria. PfPK8 (PF3D7_0203100) is an orphan kinase in Plasmodium falciparum that bears some homology to STE kinases but has no known function. To reveal the function of PfPK8 we investigated PfPK8 knockout parasites with an untargeted multi-omics workflow. Phosphoproteomics analysis identified six putative phosphorylation targets in the parasite nucleus, including another kinase, a histone acetyltransferase, and three transcription-associated proteins, including the transcription factor AP2-12 (PF3D7_1239200). Untargeted metabolomics and proteomics analysis demonstrated no impact of the PfPK8 knockout on metabolism but revealed differential regulation of exported surface proteins from multi-gene families. Transcriptomics analysis confirmed differential expression of these multi-gene family proteins, particularly de-repression of var genes encoding PfEMP1 variants. DAP-seq analysis of genes bound to the AP2-12 transcription factor also identified significant enrichment of var genes, with significant overlap between the group B/C and C var genes enriched in both the PfPK8-KO transcriptomics and AP2-12 DAP-seq analysis. Overall, this study revealed that the primary function of PfPK8 is to regulate transcription of antigenic variant genes via phosphorylation of nuclear targets including the AP2-12 transcription factor.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
O-GlcNAcase activity maintains stress granules for proximity-enhanced ATP production to ensure recovery from stress
STUDY_SUMMARY
Accurate disassembly of stress granules (SGs) after environmental stimuli release is essential for cells to maintain homeostasis , which requires ATP-consuming processes. However, the molecular mechanism whereby regulation of SGs programmatically disassembly and ATP restoration remain poorly understood in mammalian cells. Here we found that defect of OGA in cells leads to aggregates formation, severe autophagy and eventually apoptosis during stress recovery. OGA, which localized in SGs, had no effect on SGs formation but could protect SGs from rapid disassembly during stress recovery stage. Then the SGs localized glycolysis-related enzymes were reserved and concentrated in SGs during stress release for ATP production in a proximity manner, which was vital to guarantee cells resistant to stress and survival during recovery. Finally, supplementation of ATP to OGA knockdown cells during stress recovery significantly rescue cell from aggregates, autophagy and apoptosis. Together, these results describe a brand new mechanism on how OGA regulates the programmed disassembly of stress granules and restoration of ATP to safeguard cell viability in a very precisely programmed process, whose rate is rigorous regulated.
INSTITUTE
Zhejiang University
DEPARTMENT
Life Sciences Institute
LABORATORY
Shixian Lin
LAST_NAME
Chen
FIRST_NAME
Yulin
ADDRESS
Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang province, China
The major TMEM106B dementia risk allele affects TMEM106B protein levels, fibril formation, and myelin lipid homeostasis in the ageing human hippocampus
STUDY_SUMMARY
Background: The risk for dementia increases exponentially from the seventh decade of life. Identifying and understanding the biochemical changes that sensitize the ageing brain to neurodegeneration will provide new opportunities for dementia prevention and treatment. This study aimed to determine how ageing and major genetic risk factors for dementia affect the hippocampal proteome and lipidome of neurologically-normal humans over the age of 65. The hippocampus was chosen as it is highly susceptible to atrophy with ageing and in several neurodegenerative diseases. Methods: Mass spectrometry-based proteomic and lipidomic analysis of CA1 hippocampus samples from 74 neurologically normal human donors, aged 66 ā 104, was used in combination with multiple regression models and gene set enrichment analysis to identify age-dependent changes in the proteome and lipidome. ANOVA was used to test the effect of major dementia risk alleles in the TMEM106B and APOE genes on the hippocampal proteome and lipidome, adjusting for age, gender, and post-mortem interval. Fibrillar C-terminal TMEM106B fragments were isolated using sarkosyl fractionation and quantified by immunoblotting. Results: Forty proteins were associated with age at false discovery rate-corrected P<0.05, including proteins that regulate cell adhesion, the cytoskeleton, amino acid and lipid metabolism, and ribosomal subunits. TMEM106B, a regulator of lysosomal and oligodendrocyte function, was regulated with greatest effect size. The increase in TMEM106B levels with ageing was specific to carriers of the rs1990622-A allele in the TMEM106B gene that increases risk for frontotemporal dementia, Alzheimerās disease, Parkinsonās disease, and hippocampal sclerosis with ageing. Rs1990622-A was also associated with higher TMEM106B fibril content. Hippocampal lipids were not significantly affected by APOE genotype, however levels of myelin-enriched sulfatides and hexosylceramides were significantly lower, and polyunsaturated phospholipids were higher, in rs1990622-A carriers after controlling for APOE genotype. Conclusions: Our study demonstrates that TMEM106B protein abundance is increased with brain ageing in humans, establishes that dementia risk allele rs1990622-A predisposes to TMEM106B fibril formation in the hippocampus, and provides the first evidence that rs1990622-A affects brain lipid homeostasis, particularly myelin lipids. Our data suggests that TMEM106B is one of a growing list of major dementia risk genes that affect glial lipid metabolism.
Hypoxia-inducible factor-1a (HIF1a) physiology in mouse skeletal muscle
STUDY_SUMMARY
Hypoxia-inducible factor (HIF)-1a is continuously synthesized and degraded in normoxia, whereas during hypoxia, HIF1a stabilization restricts cellular oxygen utilization. Less is known about HIF1a function(s) and sex-specific effects during normoxia in the basal state. Since skeletal muscle is the largest protein store in mammals and protein homeostasis has high energy demands, we determined HIF1a function at baseline during normoxia in skeletal muscle with the use of untargeted metabolomics. Skeletal muscle from mice with and without skeletal muscle HIF1a deletion were harvested and analyzed using untargeted metabolomics (hydrophilic interaction (HILIC) and reverse phase liquid chromatography (RPLC) methods). We identified differentially expressed metabolites in the glycolytic and TCA cycle pathways.
Role of Hypoxia-inducible factor-1a (HIF1a)in Skeletal Muscle Physiology (C2C12 myotube model)
STUDY_SUMMARY
Hypoxia-inducible factor (HIF)-1a is continuously synthesized and degraded in normoxia, whereas during hypoxia, HIF1a stabilization restricts cellular oxygen utilization. Less is known about HIF1a function(s) and sex-specific effects during normoxia in the basal state. Since skeletal muscle is the largest protein store in mammals and protein homeostasis has high energy demands, we determined HIF1a function at baseline during normoxia in C2C12 murine myotubes with the use of untargeted metabolomics. 114 samples of extracted metabolites from cells were analyzed using LCMS. We identified that metabolites, especially those in the glycolytic pathway and TCA cycle, were differentially expressed in cells with HIF1a KO compared to WT.
Analyzing Metabolic Alterations in the Gut, Blood, and Brain of Mice Induced by Running Exercise Through Gas Chromatography Mass Spectrometry.
STUDY_SUMMARY
Studying the metabolic impact of running exercise on the gut, blood, and specific brain regions like the hippocampus and brainstem is crucial for comprehending the broader health effects of physical activity. In our six-week study, we utilized a mouse model (C57BL/6 genotype) to investigate these metabolic changes. Employing gas chromatography coupled with mass spectrometry followed by metabolomics for a comprehensive analysis, our approach offers insights into how exercise influences metabolic processes, including brain function. Our findings hold the potential to shape more effective exercise strategies for enhancing overall health and cognitive function.
INSTITUTE
University of Puerto Rico, School of Medicine
DEPARTMENT
Biochemistry
LAST_NAME
Chorna
FIRST_NAME
Nataliya
ADDRESS
University of Puerto Rico, Medical Sciences Campus, San Juan, PR 00935
Analyzing Metabolic Alterations in the Gut, Hippocampus and Brainstem of Mice Induced by Running Exercise Through Liquid Chromatography Mass Spectrometry
STUDY_SUMMARY
Metabolic analysis of the impact of running exercise on the gut, hippocampus and brainstem is crucial as it provides insights into how exercise affects memory, mood, and vital physiological functions. In our six-week study, we utilized a mouse model (C57BL/6 genotype) to investigate these metabolic changes. Employing liquid chromatography coupled with mass spectrometry followed by metabolomics for a comprehensive analysis, our approach offers insights into how exercise influences metabolic processes, including brain function. Our findings hold the potential to shape more effective exercise strategies for enhancing overall health and cognitive function.
INSTITUTE
University of Puerto Rico, School of Medicine
DEPARTMENT
Biochemistry
LAST_NAME
Chorna
FIRST_NAME
Nataliya
ADDRESS
University of Puerto Rico, Medical Sciences Campus, San Juan, PR 00935
Coral endosymbiont growth is enhanced by metabolic interactions with bacteria
STUDY_SUMMARY
Bacteria are key contributors to microalgae resource acquisition, competitive performance, and functional diversity, but their potential metabolic interactions with coral microalgal endosymbionts (Symbiodiniaceae) have been largely overlooked. Here, we show that altering the bacterial composition of two widespread Symbiodiniaceae species, during their free-living stage, results in a significant shift in their cellular metabolism. Indeed, the abundance of monosaccharides and the key phytohormone indole-3-acetic acid (IAA) were correlated with the presence of specific bacteria, including members of the Labrenzia (Roseibium) and Marinobacter genera. Single-cell stable isotope tracking revealed that these two bacterial genera are involved in reciprocal exchanges of carbon and nitrogen with Symbiodiniaceae. We identified the provision of IAA by Labrenzia and Marinobacter, and this metabolite caused a significant growth enhancement of Symbiodiniaceae. By unravelling these interkingdom interactions, our work demonstrates how specific bacterial associates fundamentally govern Symbiodiniaceae fitness.
Metabolic profiling of newborn DBS samples associated with transit and false elevation of glutarylcarnitine (C5DC)
STUDY_SUMMARY
Background: Glutaric aciduria type-1 (GA-1) is a rare autosomal recessive metabolic disorder caused by a glutaryl coenzyme A dehydrogenase (GCDH) deficiency, affecting approximately 1 in 110,000 individuals globally. This enzymatic deficiency leads to abnormal elevations of glutaryl-CoA and its derivatives, specifically glutaric acid (GA), 3-hydroxyglutaric acid (3OHGA), and glutarylcarnitine (C5DC). Clinical manifestations encompass macrocephaly, developmental delays, and movement disorders. Early detection via genetic testing and newborn screening (NBS), utilizing GA-1 biomarkers in dried blood spot (DBS) samples, is vital for prompt intervention. Despite the NBS system, transit-elevated C5DC-containing DBS samples from falsely suspected GA-1 newborns sometimes yield normal results, posing diagnostic sensitivity and specificity challenges. Consequently, there is a growing need for alternative diagnostic tools. Comprehensive mass spectrometry-based untargeted metabolomics offers promise in identifying additional informative biomarkers for distinguishing falsely suspected GA-1 newborns from healthy counterparts. Methodology: In this prospective study, we obtained DBS samples with transit-elevated C5DC levels from falsely suspected GA-1 newborns (n=47) and matched control DBS samples from healthy newborns (n=47) through the NBS program. Metabolites were extracted and analyzed via liquid chromatography-high-resolution mass spectrometry (LC-HRMS). Subsequent multivariate and univariate statistical analyses and feature annotation enabled biomarker and pathway investigations for significantly altered metabolites. Results: Untargeted metabolomics analysis revealed alterations in 582 upregulated and 546 downregulated metabolites. The commonly used GA-1 biomarkers, including C5DC, exhibited no significant changes in the falsely suspected GA-1 DBS samples. Conversely, 155 endogenous metabolites displayed significant variations compared to the control group. Furthermore, our data identified novel altered metabolic biomarkers, such as N-Palmitoylcysteine, 3-hydroxylinoleoylcarnitine, Heptacarboxyporphyrin, and MG (0:0/20:1/0:0), along with perturbed metabolic pathways like sphingolipid and thiamine metabolism associated with the transient and falsely elevated C5DC levels in DBS samples. Conclusions: Our untargeted metabolomics investigation unveiled distinct metabolic pathways and biomarkers linked to the transient C5DC elevation in DBS samples from falsely suspected GA-1 newborns. These findings can enhance GA-1 diagnosis by serving as predictive indicators during NBS analysis. Validation studies are warranted to confirm the presence of these newly identified metabolic pathways and biomarkers in confirmed GA-1 cases.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
LAST_NAME
Al Mogren
FIRST_NAME
Maha
ADDRESS
Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia
Title: Myeloid cell-derived creatine in the hypoxic niche promotes glioblastoma growth
STUDY_TYPE
PBMC vs Tumor CD163+
STUDY_SUMMARY
Glioblastoma (GBM) is a malignancy dominated by the infiltration of tumor-associated myeloid cells (TAMCs). Examination of TAMC metabolic phenotypes in mouse models and human GBM patients identified the de-novo creatine metabolic pathway as a hallmark of TAMCs. Multi-omics analyses revealed that TAMCs surround the hypoxic peri-necrotic regions of GBM and express the creatine metabolic enzyme glycine amidinotransferase (GATM). Conversely, GBM cells located within these same regions are uniquely specific in expressing the creatine transporter (SLC6A8). Therefore, we hypothesized that TAMCs provide creatine to tumors, promoting GBM progression. Isotopic tracing demonstrated that TAMC-secreted creatine can be taken up by tumor cells. Creatine supplementation protected tumors from hypoxia-induced stress which was abrogated with genetic ablation or pharmacologic inhibition of SLC6A8. Lastly, inhibition of creatine transport using the clinically relevant compound, RGX-202-01, blunted tumor growth, and enhanced radiation therapy in-vivo. This work highlights that myeloid-to-tumor transfer of creatine promotes tumor growth in the hypoxic niche.
INSTITUTE
Northwestern University, Feinberg School of Medicine
O-GlcNAcase activity maintains stress granules for proximity-enhanced ATP production to ensure recovery from stress (Part 2)
STUDY_SUMMARY
Accurate disassembly of stress granules (SGs) after environmental stimuli release is essential for cells to maintain homeostasis , which requires ATP-consuming processes. However, the molecular mechanism whereby regulation of SGs programmatically disassembly and ATP restoration remain poorly understood in mammalian cells. Here we found that defect of OGA in cells leads to aggregates formation, severe autophagy and eventually apoptosis during stress recovery. OGA, which localized in SGs, had no effect on SGs formation but could protect SGs from rapid disassembly during stress recovery stage. Then the SGs localized glycolysis-related enzymes were reserved and concentrated in SGs during stress release for ATP production in a proximity manner, which was vital to guarantee cells resistant to stress and survival during recovery. Finally, supplementation of ATP to OGA knockdown cells during stress recovery significantly rescue cell from aggregates, autophagy and apoptosis. Together, these results describe a brand new mechanism on how OGA regulates the programmed disassembly of stress granules and restoration of ATP to safeguard cell viability in a very precisely programmed process, whose rate is rigorous regulated. This is a continuation of study ST002927 where a different set precursor/product ions were chosen for MS.
INSTITUTE
Zhejiang University
DEPARTMENT
Life Sciences Institute
LABORATORY
Shixian Lin
LAST_NAME
Chen
FIRST_NAME
Yulin
ADDRESS
Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang province, China
Deep Metabolic Phenotyping of Newborn Cord Blood Reveals Maternal-Fetal Interactions and Disease Risk
STUDY_SUMMARY
Metabolites are small molecules circulating in the mother, placental, and fetal blood that can have a profound effect on a developing fetus (1, 2). Many metabolites from pregnant mothers cross the placenta to provide energy, structural components, essential nutrients, and signals to the developing fetus (3, 4). Issues with proper transmission of metabolites to the fetus, whether through gestational diabetes, placental insufficiency, or other sources can permanently damage the fetus (5-7). However, quantification of many metabolites entering and exiting the fetus are unknown; associations between microbial metabolites in umbilical cords and disease have not been thoroughly investigated; and there remains a lack of quantifiable metabolic effects of some of the most common medications administered during pregnancy and parturition. Here we identified and quantified many metabolites with a gradient between arterial and venous cord blood; we demonstrated that exogenous metabolites in umbilical cords associate with many health outcomes; and we show that medications can profoundly alter the metabolic milieu of the fetus. We greatly expanded the number of metabolites that demonstrate a gradient between arterial and venous blood, indicating absorption by the fetus, including several essential fatty acids. The microbial metabolites 3-indolepropionic acid, hydroxyhippuric acid and others are associated with many newborn diseases. Lastly, we show that exogenous medications like bupivacaine and betamethasone can have a profound impact on newborn metabolic profile. This study is the most comprehensive study of umbilical cord metabolic and disease associations to date. It reveals important aspects of fetal biology, like the reliance on specific essential fatty acid and taurine. It suggests several interventions in pregnant mothers that may help newborn health, including new fatty acids. This study serves as a valuable reference for investigators wishing to better understand the impact of medications on the developing fetus and neonate.
Role of PI3K in Atrial Myopathy: Insights from Transgenic Mouse Models and Identification of a Dysregulated PI3K Lipid Profile in Individuals with Atrial Fibrillation - Part 1 of 2
STUDY_SUMMARY
Here, we assessed whether there was any evidence of dysregulation in IGF1R-PI3K signaling in veteran male athletes with/without AF by assessing their plasma lipid species. This was following evidence of dysregulation of the PI3K signaling cascade in the atria of mice that had undergone extreme exercise training, together with pathology in dnPI3K Tg(+/+) mice.
INSTITUTE
Baker Heart and Diabetes Institute
LABORATORY
Metabolomics
LAST_NAME
Tham
FIRST_NAME
Yow
ADDRESS
75 Commercial Rd, Melbourne, Victoria, 3004, Australia
Role of PI3K in Atrial Myopathy: Insights from Transgenic Mouse Models and Identification of a Dysregulated PI3K Lipid Profile in Individuals with Atrial Fibrillation - Part 2 of 2, Mus musculus
STUDY_SUMMARY
Lipidomic profiling was conducted on atrias and ventricles from 20 week old, male and female caPI3K and dnPI3K heterozygous and homozygous transgenic mice. These transgenic mice have cardiac specific overexpression of either a constitutively active PI3K transgene (to have increased PI3K activity in the heart, mimicking an athlete's heart) or dominate negative PI3K transgene (to have decreased PI3K activity in the heart, resulting in smaller hearts with increase susceptibility to a variety of cardiac insults).
Environmental Exposure in the New Hampshire Birth Cohort Study
STUDY_TYPE
Prospective Cohort Study
STUDY_SUMMARY
The New Hampshire Birth Cohort Study is a prospective cohort study of rural pregnant women and their children with the aim of investigating the effects of environmental exposures on birth outcomes and child health and development. Please contact Margaret Karagas at margaret.r.karagas@dartmouth.edu for questions related to the subject characteristics and outcomes. This research was supported by the Environmental influences on Child Health Outcomes (ECHO) Program, Office of The Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The New Hampshire Birth Cohort Study is an ECHO cohort project which is supported by the following ECHO Program Collaborators: ECHO Coordinating Center: Duke Clinical Research Institute, Durham, North Carolina: Smith PB, Newby KL, Benjamin DK; U2C OD023375 ECHO Data Analysis Center: Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland: Jacobson LP; Research Triangle Institute, Durham, North Carolina: Catellier D; U24 OD023382 RTI Children's Health Exposure Analysis Resource (CHEAR) Exposure Assessment Hub: Research Triangle Institute: Fennell T, UNC Untargeted Analysis Core, RTI CHEAR Hub: Sumner S; U2CES026544 Children's Health Exposure Analysis Resource Coordinating Center: Westat, Inc., Rockville, Maryland: OāBrien B; U24 ES026539
INSTITUTE
Dartmouth College
DEPARTMENT
Department of Epidemiology
LABORATORY
Karagas Lab
LAST_NAME
Karagas
FIRST_NAME
Margaret
ADDRESS
Geisel School of Medicine at Dartmouth, 1 Medical Center Drive, Williamson Translational Research Building, 7th Floor, Lebanon, NH 03756
Targeted analyses of microbial metabolites of xanthohumol
STUDY_SUMMARY
The Xanthohumol microbiome and signature (XMaS) study in healthy adults was a phase I, triple-masked, placebo-controlled clinical trial in healthy adults investigating effects of a natural product supplement of gut microbiome composition and metabolism. Xanthohumol, a flavonoid from the hops plant or placebo was administered to 27 healthy adults daily for eight weeks. Fecal, plasma, and 24-hr urine collections were obtained from participants at baseline and 2-week intervals. The aim of this study was to quantify major microbial metabolites of xanthohumol.
Targeted analysis of short chain fatty acids for the XMaS clinical trial.
STUDY_SUMMARY
The Xanthohumol microbiome and signature (XMaS) study in healthy adults was a phase I, triple-masked, placebo-controlled clinical trial in healthy adults investigating effects of a natural product supplement of gut microbiome composition and metabolism. Xanthohumol, a flavonoid from the hops plant or placebo was administered to 27 healthy adults daily for eight weeks. Fecal, plasma, and 24-hr urine collections were obtained from participants at baseline and 2-week intervals. The main objective of this study was to quantify short chain fatty acids from fecal samples of particiapnts in the XMaS trail
The Xanthohumol microbiome and signature (XMaS) study in healthy adults was a phase I, triple-masked, placebo-controlled clinical trial in healthy adults investigating effects of a natural product supplement of gut microbiome composition and metabolism. Xanthohumol, a flavonoid from the hops plant or placebo was administered to 27 healthy adults daily for eight weeks. Fecal, plasma, and 24-hr urine collections were obtained from participants at baseline and 2-week intervals. The main objective of this study was to quantify fecal bile acids from participants from the XMaS study.
Longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin
STUDY_SUMMARY
This study is part of a multi-part study, including a. longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin b. polar fecal metabolomics of a patient cohort c. fecal lipidomics of a patient cohort d. polar urinary metabolomics of a patient cohort e. polar metabolomics of in vitro digestions This specific part is part a. longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin
INSTITUTE
Ghent University
DEPARTMENT
Department of Translational Physiology, Infectiology and Public Health
This study is part of a multi-part study, including a. longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin b. polar fecal metabolomics of a patient cohort c. fecal lipidomics of a patient cohort d. polar urinary metabolomics of a patient cohort e. polar metabolomics of in vitro digestions This specific part is part b. polar fecal metabolomics of a patient cohort
INSTITUTE
Ghent University
DEPARTMENT
Department of Translational Physiology, Infectiology and Public Health
This study is part of a multi-part study, including a. longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin b. polar fecal metabolomics of a patient cohort c. fecal lipidomics of a patient cohort d. polar urinary metabolomics of a patient cohort e. polar metabolomics of in vitro digestions This specific part is part c. Fecal lipidomics of a patient cohort
INSTITUTE
Ghent University
DEPARTMENT
Department of Translational Physiology, Infectiology and Public Health
Hepatic lipid profiles of wild type and Cgref1-depleted mice of C57BL/6 strain obtained by an untargeted LC-MS/MS analysis
STUDY_SUMMARY
Among the range of experiments and observations recorded, we compared the lipid expression in the liver tissues between wild type and Cgref1-knockout mice of C57BL/6 strain using untargeted LC-MS/MS (n=3 per group). Particularly, the results revealed that Cgref1-knockout (KO) mice overall have lower levels of triglyceride and diglyceride species. Such finding provides supportive evidence that Cgref1 may promote de novo lipogenesis in the liver and increase the risk of fatty liver development.
Multi-omics Approach Reveals a Specific Profile in Severe Asthmatic Patients Treated with Mepolizumab or Omalizumab
STUDY_SUMMARY
The prevalence and severity of asthma continue to increase, and patients with uncontrolled severe asthma may require additional biological treatment. Given the pathophysiological heterogeneity underlying severe asthma, there is a lack of reliable biomarkers to monitor treatment response. In this context, this study aimed to identify potential biomarkers for monitoring the response to either Mepolizumab or Omalizumab in severe asthmatic patients using metabolomics and proteomics. Severe asthmatic patients who were prescribed either Mepolizumab (n=36) or Omalizumab (n=20) were followed during treatment, and serum samples were collected before treatment (T0), and both at 6 (T1) and 18 months (T2) after starting biological treatment. Treatment monitoring was performed by the analysis of a set of inflammatory metabolites and proteins using targeted metabolomic and proteomic approaches, along with the study of clinical variables such as the number of blood eosinophils, hospitalizations, and severe exacerbations. Mepolizumab and Omalizumab induced changes in the metabolomic and proteomic profiles of severe asthmatic patients. For both treatments, the greatest differences in metabolites were seen at T1 and maintained up to T2, whereas protein changes were seen throughout the treatment. Notable changes associated with anti-inflammatory effects included a decrease in several fatty acids, sphingosine-1-phosphate (S1P), L-arginine, and IL-10RA. Interestingly, S1P was the main feature that correlated significantly with eosinophilia in both treatments. These results unveil underlying molecular mechanisms that occur during Mepolizumab and Omalizumab treatments and provide potential biomarkers for monitoring treatment response in severe asthmatic patients.
INSTITUTE
CEMBIO
LAST_NAME
Contreras
FIRST_NAME
Nuria
ADDRESS
Urb. MonteprĆncipe s/n, Ctra. Boadilla del Monte km 5.3, Madrid 28668, Spain
Serum metabolomics reveals metabolic profile and potential biomarkers of ankylosing spondylitis
STUDY_SUMMARY
Ankylosing spondylitis (AS) is a chronic systemic inflammatory disease that significantly impairs physical function, quality of life, and work ability in young individuals. Nonetheless, the identification of early radiographic changes in AS is frequently delayed, and the diagnostic efficacy of biomarkers remains moderately effective, with unsatisfactory sensitivity and specificity. Hence, it is imperative to identify biomarkers that can facilitate early diagnosis, prognosis, and monitoring of AS. A total of 67 AS patients and 67 healthy controls were recruited to procure plasma samples for the purpose of screening potential biomarkers of AS via untargeted combined with targeted metabolomics approach utlizing UHPLC-QTOF-MS/MS and UHPLC-QQQ-MS/MS. Multivariate pattern recognition and univariate statistical analysis were employed to compare and elucidate the differential metabolites. The results indicated a notable divergence between the two groups, and a total of 170 different metabolites associated with the primary 6 metabolic pathways exhibiting a correlation with AS. Among those, 26 metabolites exhibited high sensitivity and specificity with area under curve (AUC) value were greater than 0.8. Subsequent targeted quantitative analysis discovered 3 metabolites, namely 3-amino-2-piperidone, hypoxanthine and octadecylamine, exhibiting excellent distinguishing ability based on the results of ROC curve and Random Forest model, thus qualifying as potential biomarkers for AS. Summarily, our non-targeted and targeted metabolomics investigations provide new insights into the metabolic profile and potential biomarker candidates of AS. These findings may provide additional diagnostic options for AS and enhance the understanding of the underlying pathophysiology of the condition.
INSTITUTE
Ningxia Medical University
LAST_NAME
Ma
FIRST_NAME
Xueqin
ADDRESS
1160 Shenli Street, Yinchuan, Ningxia, 750004, China
Investigate the impact of feeding time on the hexosamine biosynthetic pathway (HBP) in the mouse liver and heart using targeted metabolomics: biogenic amines
STUDY_SUMMARY
The overall goal of this project is to advance our understanding of post-translational mechanisms that mediate metabolic regulation of time-of-day-specific protein functions to orchestrate daily rhythms and maintain homeostasis in animals. Robust daily biological rhythms over the 24-hour (h) day-night cycles are key hallmarks of animal health span and are strongly regulated by circadian clocks. Circadian clocks are cell autonomous molecular timers present in the brain and in peripheral organs that enable animals to adapt to predictable daily changes in environment and regulate rhythmic processes such as sleep-wake cycles, feeding-fasting cycles, metabolism, hormonal signaling and neuronal excitability. Besides light, the dominant time cue for the brain clock, metabolic signals from clock-controlled feeding-fasting cycles represent the most potent time cue to entrain and synchronize peripheral clocks in key organs. Much effort has been dedicated to understanding the metabolic regulation of daily biological rhythms, but many important mechanisms are only just emerging. We recently established that metabolic signals from feeding-fasting cycles regulate daily biological rhythms in Drosophila through rhythmic O-linked-N-acetylglucosaminylation (O-GlcNAcylation). Protein O-GlcNAcylation is a nutrient sensitive posttranslational modification (PTM) that is tightly linked to metabolic status, as UDP-GlcNAc, the substrate of O-GlcNAcylation, is produced from hexosamine biosynthetic pathway (HBP), which integrates the metabolites from glucose, amino acid, lipid and nucleotide metabolism. We now propose to investigate whether feeding activity can regulate daily O-GlcNAcylation rhythm in mouse liver and heart and whether the levels of HBP metabolites in mouse liver and heart are affected by different feeding time within a day/night cycle. Here, we restricted the feeding time of C57BL/6 male mice to ZT12-24 (RF12-24. ZT, zeitgeber time; ZT0 indicates light on, while ZT12 indicates light off) v.s. ZT0-12 (RF0-12) for 3 weeks and collected liver and heart tissues every 4 hours over a 24-hour period. The liver and heart samples were subjected to targeted metabolomic analysis for HBP metabolites.
INSTITUTE
University of California, Davis
DEPARTMENT
Department of Entomology and Nematology
LABORATORY
Chiu lab
LAST_NAME
Chiu
FIRST_NAME
Joanna
ADDRESS
6352 Storer Hall, One Shields Avenue, Davis, CA 95616, USA
Investigate the impact of feeding time on the hexosamine biosynthetic pathway (HBP) in the mouse liver and heart using targeted metabolomics: primary metabolism
STUDY_SUMMARY
The overall goal of this project is to advance our understanding of post-translational mechanisms that mediate metabolic regulation of time-of-day-specific protein functions to orchestrate daily rhythms and maintain homeostasis in animals. Robust daily biological rhythms over the 24-hour (h) day-night cycles are key hallmarks of animal health span and are strongly regulated by circadian clocks. Circadian clocks are cell autonomous molecular timers present in the brain and in peripheral organs that enable animals to adapt to predictable daily changes in environment and regulate rhythmic processes such as sleep-wake cycles, feeding-fasting cycles, metabolism, hormonal signaling and neuronal excitability. Besides light, the dominant time cue for the brain clock, metabolic signals from clock-controlled feeding-fasting cycles represent the most potent time cue to entrain and synchronize peripheral clocks in key organs. Much effort has been dedicated to understanding the metabolic regulation of daily biological rhythms, but many important mechanisms are only just emerging. We recently established that metabolic signals from feeding-fasting cycles regulate daily biological rhythms in Drosophila through rhythmic O-linked-N-acetylglucosaminylation (O-GlcNAcylation). Protein O-GlcNAcylation is a nutrient sensitive posttranslational modification (PTM) that is tightly linked to metabolic status, as UDP-GlcNAc, the substrate of O-GlcNAcylation, is produced from hexosamine biosynthetic pathway (HBP), which integrates the metabolites from glucose, amino acid, lipid and nucleotide metabolism. We now propose to investigate whether feeding activity can regulate daily O-GlcNAcylation rhythm in mouse liver and heart and whether the levels of HBP metabolites in mouse liver and heart are affected by different feeding time within a day/night cycle. Here, we restricted the feeding time of C57BL/6 male mice to ZT12-24 (RF12-24. ZT, zeitgeber time; ZT0 indicates light on, while ZT12 indicates light off) v.s. ZT0-12 (RF0-12) for 3 weeks and collected liver and heart tissues every 4 hours over a 24-hour period. The liver and heart samples were subjected to targeted metabolomic analysis for HBP metabolites.
INSTITUTE
University of California, Davis
DEPARTMENT
Department of Entomology and Nematology
LABORATORY
Chiu lab
LAST_NAME
Chiu
FIRST_NAME
Joanna
ADDRESS
6352 Storer Hall, One Shields Avenue, Davis, CA 95616, USA
Comparisons of hepatic medium & long chain fatty acids expression between wild type and Cgref1-depleted mice of C57BL/6 strain by targeted GC-MS/MS
STUDY_SUMMARY
In this study, we performed a targeted analysis on the expression of hepatic medium & long chain fatty acids between wild type (n=3) and Cgref1-knockout mice (n=2) using targeted GC-MS/MS. The results, however, did not reveal significant differences between the two groups of mice. In the future, a larger sample size and optimization of normalization methods may be helpful to provide more meaningful insights.
Metabolite flux from temperature-acclimated diatom strains (drawdown experiment)
STUDY_SUMMARY
The temperature increase occurring in the surface ocean has fundamental implications for physiological rates and processes of marine microbes. Here we asked whether the temperature at which a marine diatom strain is acclimated affects carbon transfer to a co-cultured heterotrophic bacterium. Model systems were established in which the diatom Thalassiosira pseudonana was acclimated for three months at temperatures below (14Ā°C), equal to (20Ā°C), and above (28Ā°C) the temperature of optimal growth, and then inoculated with the heterotrophic bacterium Ruegeria pomeroyi. This deposition is for results obtained from a drawdown experiment of phytoplankton metabolites using R. pomeroyi conducted during this study.
Untargeted metabolomics analysis of plasma from viremic non-progressors (VNP), progressors, and healthy control (HC) patients using high-resolution, high-performance LC-MS/MS analysis.
STUDY_SUMMARY
Viremic Non-Progressors (VNPs) maintain normal CD4+ T-cell counts despite uncontrolled HIV-1 replication, but mechanisms leading to CD4+ T-cell preservation are incompletely characterized. We aim to generate a comprehensive understanding of this infrequent phenotype by comparing rates of cellular infection, immunophenotype, genomics, single-cell transcriptomics, metabolomics, and levels of plasma biomarkers in 16 VNPs and 29 HIV+ Progressors. During chronic infection VNPs show lower frequency of HIV-1-infected cells in periphery, which is associated with reduced CCR5 expression and higher prevalence of CCR5Ī32 heterozygosity. The CD8+ T-cell compartment displays a less cytotoxic and less activated phenotype in VNPs. Moreover, despite similar viremia, we found lower bystander CD4+ T-cell death in the VNPs, together with weaker IFN responses, reduced plasma levels of zonulin (a biomarker of intestinal permeability), altered tryptophan catabolism, and preserved LPS responsiveness in vitro. Overall, a complex multifactorial mechanism, suggesting gut-associated lymphoid tissue preservation, underlies resistance to HIV pathogenesis in VNPs.
INSTITUTE
IrsiCaixa
LAST_NAME
Martinez-Picado
FIRST_NAME
Javier
ADDRESS
Carretera de Canyet, s/n, 08916 Badalona, Barcelona
Automated preparation of plasma lipids, metabolites, and proteins for LC/MS-based analysis of a high-fat diet in mice
STUDY_TYPE
Method Development
STUDY_SUMMARY
We designed an automated liquid-liquid extraction method with minimal contamination and human intervention. This approach enables accurate and precise collection of metabolite, lipid fractions, and protein pellet from a small volume of mice plasma for multiomic analysis.
Metabolite flux from temperature-acclimated diatom strains (main experiment)
STUDY_SUMMARY
The temperature increase occurring in the surface ocean has fundamental implications for physiological rates and processes of marine microbes. Here we asked whether the temperature at which a marine diatom strain is acclimated affects carbon transfer to a co-cultured heterotrophic bacterium. Model systems were established in which the diatom Thalassiosira pseudonana was acclimated for three months at temperatures below (14Ā°C), equal to (20Ā°C), and above (28Ā°C) the temperature of optimal growth, and then inoculated with the heterotrophic bacterium Ruegeria pomeroyi. This deposition is for the results of diatom endometabolites obtained from the main experiment of this study.
Lipidomic analysis of demyelination and remyelination in the Plp1-iCKO-Myrf mouse model of demyelination
STUDY_SUMMARY
In this study, we obtained a longitudinal lipidomic profile of the brain, spinal cord, and serum using a genetic mouse model of demyelination, known as Plp1-iCKO-Myrf mice. This model has distinct phases of demyelination and remyelination over the course of 24 weeks, in which loss of motor function peaks during demyelination.
Transcriptional regulation of amino acid metabolism by KDM2B
STUDY_SUMMARY
Epigenetic and metabolic alterations in cancer cells are intertwined. The concentration of metabolites can influence the activity of chromatin modifiers, which in turn can act as metabolic sensors that translate changes in cellular metabolism to transcriptional reprogramming. In the present study, we investigated the role of histone demethylase KDM2B in the metabolic reprogramming of the triple-negative breast cancer (TNBC), in which KDM2B is selectively expressed at high levels. Knockdown of KDM2B in TNBC cell lines reduced their proliferation rate and tumor growth in vivo. Transcriptomic, proteomic, and metabolomic profiling demonstrated that the Serine-Glycine pathway and One Carbon metabolism (SGOC) and other amino acid biosynthetic and catabolic processes are downregulated by the knockdown of KDM2B. Additionally, we see reduction of metabolites produced via these pathways (purines, pyrimidines, formate, glutathione and NADPH). Importantly, the expression of the enzymes involved in the SGOC metabolic pathway (e.g. PHGDH, PSAT1, PSPH, SHMT2, MTHFD1L, MTHFD2 and DHFR) depends on c-MYC, NRF2, and ATF4 which our data show that they are under the positive regulatory control of KDM2B. The epistatic relationship between these factors, with the expression of the enzymes of the SGOC pathway and the effects of the KDM2B knockdown on chromatin occupancy and accessibility of the promoters of these factors is in progress and will be presented. Analysis of TCGA data showed positive and statistically significant correlations between KDM2B and the SGOC gene signature in TNBC patients. In addition, the metabolic pathway signature that distinguishes control and shKDM2B-transduced cells corresponds to the metabolic signature of a subset of TNBCs, which have been reported to carry poor prognosis. The present study highlights the role of the epigenetic factor KDM2B as an upstream regulator of the metabolic reprogramming of TNBC.
Analysis of Lipids Secreted from Fibroblast Young Cells
STUDY_SUMMARY
In this experimental study, we aimed to understand the potential factors within the secretions of young cells that could trigger the reverse aging of Mid-old cells. To investigate this phenomenon, we co-cultured young cells with Mid-old cells and observed a fascinating outcome: the Mid-old cells exhibited reverse aging and transformed into a more youthful state. To uncover the specific factors responsible for this reverse aging effect, we conducted a detailed analysis of the secreted factors from the young cells. Our analysis focused on a range of biomolecules, including lipids. However, despite our efforts, we did not identify any distinct factors that could be directly attributed to this remarkable reverse aging process.
INSTITUTE
Ajou University Medical Center
LAST_NAME
Kim
FIRST_NAME
Young Hwa
ADDRESS
206, World cup-ro, Yeongtong-gu, Suwon-si, Gyeonggi-do, Republic of Korea
Analysis of Metabolites Secreted from Fibroblast Young Cells
STUDY_SUMMARY
In this experimental study, we aimed to uncover the factors in young cell secretions that trigger the reverse aging of mid-old cells, co-culturing them and observing a striking transformation, although we could not identify the specific factors responsible for this rejuvenation
INSTITUTE
Ajou University Medical Center
LAST_NAME
Kim
FIRST_NAME
Young Hwa
ADDRESS
206, World cup-ro, Yeongtong-gu, Suwon-si, Gyeonggi-do, Republic of Korea
LC/MS detection for NADPH and NADP+ levels in KRAS-driven lung tumors with LKB1 or p53 deficiency, comparing cases with G6PD wild-type and G6PD knockout
STUDY_SUMMARY
In this study, LC/MS was employed to assess whether G6PD loss affects the NADPH and NADP+ pool in Lkb1-deficient KRAS-driven lung tumors with LKB1 or p53 deficiency.
INSTITUTE
Rutgers Cancer Institute of New Jersey
LAST_NAME
Guo
FIRST_NAME
Jessie
ADDRESS
Room 3020, 195 Little Albany Stree (Rutgers Cancer Institute of New Jersey)
LC/MS detection for GSH and GSSG levels in KRAS-driven lung tumors with LKB1 or p53 deficiency, comparing cases with G6PD wild-type and G6PD knockout
STUDY_SUMMARY
In this study, LC/MS was employed to assess whether G6PD loss affects the GSH and GSSG pool in Lkb1-deficient KRAS-driven lung tumors with LKB1 or p53 deficiency.
INSTITUTE
Rutgers Cancer Institute of New Jersey
LAST_NAME
Guo
FIRST_NAME
Jessie
ADDRESS
Room 3020, 195 Little Albany Stree (Rutgers Cancer Institute of New Jersey)
Metabolomics reveal the pathway of benzylisoquinoline alkaloids in Corydalis yanhusuo bulbs
STUDY_TYPE
spatial and temporal distribution
STUDY_SUMMARY
In general, bulbs of Corydalis yanhusuo can be divided into "mother-bulb (MB)" and "son-bulb (SB)" according to different parts. The mother bulbs are formed by the degeneration and re-expansion of their original stem and are used as medicinal material in production. Son bulbs emerge from axillary buds on horizontally elongated rhizomes, of which the larger bulb can also be used as medicine, while the smaller bulb is reserved as a seed stem for "seed". In this study, materials of C. yanhusuo bulbs were cultivated in the field, which was proposed and identified by Professor Da-xia Chen. Widely targeted metabolome sequencing of C. yanhusuo bulbs was performed by UPLC-ESI MS/MS system, and its metabolites were successfully identified and annotated in self-built database (the MetWare database). A total of 702 metabolites were identified in all samples, including 216 alkaloids, 120 lipids, 67 amino acids and their derivatives, 59 organic acids, 63 phenolic acids, 19 terpenoids, 28 flavonoids, 4 lignin and coumarins, 43 nucleotides and their derivatives, 1 tannin, 3 quinones and 79 other substances. The numbers of up-accumulated and down-accumulated metabolites in MB-A vs MB-C and SB-A vs SB-C were 135 and 148, 90 and 210, respectively. There were 184 kinds of DAMs between SB-A and MB-A (including 144 down-accumulated and 40 up-accumulated compounds in the MB-A samples) and 127 kinds of DAMs between SB-C and MB-C (including 57 down-accumulated and 40 up-accumulated compounds in the MB-C samples) .
INSTITUTE
Chongqing Academy of Chinese Materia Medica, Chongqing, China
LABORATORY
Department of Traditional Chinese Medicine
LAST_NAME
Zhao
FIRST_NAME
Xiao
ADDRESS
Nanshan stree, Chongqing, Nanan District, 400065, China
Lipidomics study of FASN inhibition in HT-29 and HCT 116 spheroids
STUDY_SUMMARY
Cancerous cells synthesize most of their lipids de novo to keep up with their rapid growth and proliferation. Fatty acid synthase (FAS) is a key enzyme in the lipogenesis pathway that is upregulated in many cancers and has gained popularity as a druggable target of interest for cancer treatment. The first FAS inhibitor discovered, cerulenin, initially showed promise for chemotherapeutic purposes until it was observed that it had adverse side effects in mice. TVB-2640 (Denifanstat), is part of the newer generation of inhibitors. With multiple generations of FAS inhibitors being developed, it is vital to understand their distinct molecular downstream effects to elucidate potential interactions in the clinic. Here, we profile the lipidome of two different colorectal cancer (CRC) spheroids treated with a generation 1 inhibitor (cerulenin) or a generation 2 inhibitor (TVB-2640). We observe that the cerulenin causes drastic changes to the spheroid morphology as well as alterations to the lipid droplets found within CRC spheroids. TVB-2640 causes higher abundances of polyunsaturated fatty acids (PUFAs) whereas cerulenin causes decreased abundance of PUFAs. The increase in PUFAs in TVB-2640 exposed spheroids indicates it is causing cells to die via a ferroptotic mechanism rather than a conventional apoptotic or necrotic mechanism.
Polar metabolites in cecal tissue of mice treated with or without ampicillin and tributyrin
STUDY_SUMMARY
The chromatin landscape integrates diverse cellular signals to regulate genome structure and subsequent biological functions, partly through posttranslational modifications (PTMs) on histone proteins. Many donor molecules for histone PTMs are metabolites and are therefore impacted by cellular metabolism and environmental cues. In this study, we aimed to investigate how chromatin and cellular metabolism are linked in the intestine. One class of metabolites in intestinal lumen is short chain fatty acids (SCFAs), which are generated by the commensal microbiota. We found that select histone PTMs (including acetylation, butyrylation, and propionylation) are located in intestinal epithelial cells and are dependent on the presence of microbes. Histone butyrylation is associated with active gene expression and regulated by the metabolite tributyrin, which increases metabolites related to butyrate metabolism and induces specific metabolic gene programs. Together, these studies demonstrate a physiological setting in which previously uncharacterized histone acylations are dynamically regulated through metabolites and associated with gene expression.
INSTITUTE
Case Western Reserve University
DEPARTMENT
Biochemistry
LABORATORY
Gates
LAST_NAME
Gates
FIRST_NAME
Leah
ADDRESS
Wood Building W456, 2109 Adelbert Road, Cleveland, Ohio 44106-4395
Examine the through-filter recovery of metabolites extracted from a complex bacterial medium
STUDY_SUMMARY
Based on this metabolomic protocol, the specific dataset submitted here addresses whether passing metabolite extracts through a 0.2 micron filter plate impacts the overall detection of metabolites. We recommend the use of filter plate to remove particulate, in turn, prolonging column and instrument life. Here we have tested the through-filter recovery of metabolites extracted from a rich, complex bacterial culture media (mega media) used to culture diverse gut bacterial species in our study. We select mega media as our biological matrix for this experiment, because it enables us to assess a diverse set of metabolites. Leveraging this dataset, we have observed that the ion-abundance a large number of molecular features detected in pre- vs. post-filtered samples closely correlate with each other. We have performed this experiment with two independent batches of mega media and observed consistent results. Collectively, our observations indicate a good retention of ion abundance of molecular features after passing them through the 0.2 micron membrane filter.
INSTITUTE
Duke University
DEPARTMENT
Biochemistry
LABORATORY
Han
LAST_NAME
Han
FIRST_NAME
Shuo
ADDRESS
307 Research Drive, Nanaline Duke Building, Room 159
Leishmania mexicana Promotes Pain-reducing Metabolomic Reprogramming In Cutaneous Lesions
STUDY_TYPE
Untargeted MS
STUDY_SUMMARY
Cutaneous leishmaniasis is characterized by extensive skin lesions, which are usually painless despite being associated with extensive inflammation. The molecular mechanisms responsible for this analgesia have not been identified. Through untargeted metabolomics, we found enriched anti-nociceptive metabolic pathways in L. mexicana-infected mice. Purines were elevated in infected macrophages and at the lesion site during chronic infection. These purines have anti-inflammatory and analgesic properties by acting through adenosine receptors, inhibiting TRPV1 channels, and promoting IL-10 production. We also found arachidonic acid metabolism enriched in the ear lesions compared to the non-infected controls. Arachidonic acid is a metabolite of anandamide (AEA) and 2-arachidonoylglycerol (2-AG). These endocannabinoids act on cannabinoid receptors 1 and 2 and TRPV1 channels to exert anti-inflammatory and analgesic effects. Our study provides evidence of metabolic pathways upregulated during L. mexicana infection that may mediate anti-nociceptive effects experienced by CL patients and identifies macrophages as a source of these metabolites.
INSTITUTE
The Ohio State University
DEPARTMENT
Pathology and Microbiology
LAST_NAME
Satoskar
FIRST_NAME
Abhay
ADDRESS
Evans Hall, 520 King Avenue, Columbus, OH, 43201, USA
Metabolomics Insights into Doxorubicin and 5-Fluorouracil Combination Therapy in Triple-Negative Breast Cancer: A Xenograft Model Study (Part 1)
STUDY_SUMMARY
Background: Breast cancer is one of the most prevalent malignancies and a leading cause of death among women worldwide. Among its subtypes, triple-negative breast cancer (TNBC), which poses significant clinical challenges due to its aggressive behavior and limited treatment options. Aim: This study explored the effects of doxorubicin (DOX) and 5-fluorouracil (5-FU) as monotherapies and in combination on MDA-MB-231 xenograft model. Employing advanced metabolomics analysis, the study was designed to investigate molecular alterations triggered by these treatments. Methods: State-of-the-art metabolomics analysis using Ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) was conducted including comprehensive plasma and tumor tissue sample profiling. Results: The study explored alterations induced by DOX, 5-FU, and their combination treatment. Each treatment group exhibited unique metabolic profiles in plasma and tumor analysis. Univariate and enrichment analyses identified alterations in metabolic pathways, including glycine and serine metabolism, spermidine and spermine biosynthesis, and purine and pyrimidine pathways. The combination of DOX and 5-FU significantly influenced plasma and tumor metabolites. The comprehensive metabolic profiling of both plasma and tumor samples shed light on the intricate changes within the tumor microenvironment and their systemic implications. Conclusion: The study findings offer insights into the metabolic vulnerabilities of TNBC in vivo induced by the studied chemotherapeutics. These findings highlight the involved metabolites and metabolic pathways in the response of MDA-MB-231 cells to DOX, 5-FU, and their combination which advance our understanding of TNBC treatment strategies, offering new possibilities for enhancing therapeutic outcomes.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Metabolomics Insights into Doxorubicin and 5-Fluorouracil Combination Therapy in Triple-Negative Breast Cancer: A Xenograft Model Study (Part 2)
STUDY_TYPE
LC/MS/MS
STUDY_SUMMARY
Background: Breast cancer is one of the most prevalent malignancies and a leading cause of death among women worldwide. Among its subtypes, triple-negative breast cancer (TNBC), which poses significant clinical challenges due to its aggressive behavior and limited treatment options. Aim: This study explored the effects of doxorubicin (DOX) and 5-fluorouracil (5-FU) as monotherapies and in combination on MDA-MB-231 xenograft model. Employing advanced metabolomics analysis, the study was designed to investigate molecular alterations triggered by these treatments. Methods: State-of-the-art metabolomics analysis using Ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) was conducted including comprehensive plasma and tumor tissue sample profiling. Results: The study explored alterations induced by DOX, 5-FU, and their combination treatment. Each treatment group exhibited unique metabolic profiles in plasma and tumor analysis. Univariate and enrichment analyses identified alterations in metabolic pathways, including glycine and serine metabolism, spermidine and spermine biosynthesis, and purine and pyrimidine pathways. The combination of DOX and 5-FU significantly influenced plasma and tumor metabolites. The comprehensive metabolic profiling of both plasma and tumor samples shed light on the intricate changes within the tumor microenvironment and their systemic implications. Conclusion: The study findings offer insights into the metabolic vulnerabilities of TNBC in vivo induced by the studied chemotherapeutics. These findings highlight the involved metabolites and metabolic pathways in the response of MDA-MB-231 cells to DOX, 5-FU, and their combination which advance our understanding of TNBC treatment strategies, offering new possibilities for enhancing therapeutic outcomes. This part of study involves comprehensive metabolomic profiling of the tumor tissue samples specifically and tumor growth assessment provide valuable insights into these treatments' efficacy and potential synergistic effects in TNBC.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Offline Two-dimensional Liquid Chromatography-Mass Spectrometry for Deep Annotation of the Fecal Metabolome following Fecal Microbiota Transplant
STUDY_SUMMARY
In this study, we describe a novel experimental strategy using multidimensional chromatography to facilitate compound identification in untargeted metabolomics. Pooled fecal metabolite extract samples were fractionated using an offline semi-preparative liquid chromatography. The resulting fractions were analyzed by an orthogonal LC-MS/MS method, and the data were searched against commercial, public and local spectral libraries. Multidimensional chromatography yielded more than a 3-fold improvement in identified compounds compared to the typical single-dimensional LC-MS/MS approach, and successfully identified several rare and novel compounds including atypical conjugated bile acid species. Most features identified by the new approach could be matched to features that were detectable, but not identifiable, in the original single-dimensional data. An evaluation of this approach in the context of patients with recurrent Clostridioides difficile infection receiving fecal microbiota transplants is also included. Overall, our approach represents a powerful strategy for deeper annotation of the metabolome that can be implemented with common commercially-available instrumentation, and should be applicable to any dataset requiring deeper annotation of the metabolome.
In this study, human microglia from induced pluripotent stem cells (iPSCs) derived from a TSC patient cohort were generated . With a comprehensively molecular and cellular characterization on TSC microglia, including transcriptomics, proteomics/phosphopreteomics, and lipidomics, patient-carrying TSC2 mutations lead to aberrant lipid metabolism were found, in particular, upregulated glycerophosphocholines and fatty acyls in TSC microglia, resulting in increased phagocytosis and inflammation. Strikingly, the dysregulated lipid metabolism in TSC microglia is driven by hyper-activation of mTOR-LPL pathway. Furthermore, cellular and electrophysiological assessments of neuron/microglia co-cultures revealed that TSC microglia directly affect neuronal development and excitability as well as neuronal network activity, which could be largely ameliorated by mTOR/LPL inhibition
Water soluble metabolomics in mice upon loss of SHMT
STUDY_SUMMARY
The enzyme SHMT interconverts the amino acids serine and glycine as part of the folate cycle. To explore the role of SHMT in amino acid homeostasis, Mice were treated with a small molecule inhibitor of SHMT (SHIN2) or had Shmt2 genetically knocked-out in a liver specific manner. Serum and liver samples were collected and underwent LC-MS metabolomics analysis.
Developmental Neurotoxicity of Deltamethrin Exposure on Hypothalamic Neurogenesis in Embryonic Zebrafish
STUDY_SUMMARY
In this study, to investigate DM effects at different stages of early life, zebrafish embryos were exposed to DM just before (10-16 hpf), at the onset of (16-24 hpf), at the peak of (24-36 hpf) hypothalamic neurogenesis and across 10-120 hpf with different dosage levels (0, 1, 100, and 250 nM).
INSTITUTE
College of Marine Food and Biological Engineering, Jimei University
LAST_NAME
Gao
FIRST_NAME
Longhua
ADDRESS
No. 185 Yinjiang Road, Jimei District, Xiamen City, Fujian Province, China
Metabolic characterization of the polar endometabolome of Triple-Negative Breast Cancer parental and cDDP-resistant cells (part 2)
STUDY_TYPE
NMR-based metabolomics of polar endometabolome of cultured cells
STUDY_SUMMARY
Platinum (Pt(II)) drugs, e.g. cisplatin (cDDP), are some of the most used chemotherapeutic agents, yet tumor acquired resistance and high toxicity are still current drawbacks. Metabolomics can measure the metabolic response of drug-exposed cells, unveiling insight into drug mechanisms and metabolic markers of drug efficacy, toxicity and resistance. The present 1H NMR metabolomics study aims to describe the effects of cDDP and Pd2Spm on the polar endometabolome of both MDA-MB-231 cDDP-sensitive and cDDP-resistant cell lines, aiming to describe metabolic markers of (i) resistance upon cDDP treatment, and (ii) the effect of Pd2Spm on the established cDDP-resistant cells. The former observations will give helpful insights about the metabolic features of cDDP-resistance during treatment, and enlighten on the potential role of Pd2Spm in metabolically affecting/tackling cDDP-resistance.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry and CICECO-Aveiro Institute of Materials
LAST_NAME
Carneiro
FIRST_NAME
Tatiana JoĆ£o
ADDRESS
Campus UniversitƔrio de Santiago, Aveiro, Aveiro, 3810-193, Portugal
Deciphering the metabolic heterogeneity of hematopoietic stem cells with single-cell resolution
STUDY_SUMMARY
Metabolic status is crucial for stem cell functions; however, the metabolic heterogeneity of endogenous stem cells has never been directly assessed. Here, we develop a platform for high-throughput single-cell metabolomics (hi-scMet) of hematopoietic stem cells (HSCs). By combining flow cytometric isolation and nanoparticle-enhanced laser desorption/ionization mass spectrometry, we routinely detected >100 features from single cells. We mapped the single-cell metabolomes of all hematopoietic cell populations, and HSC subpopulations with different division times, detecting 33 features whose levels exhibited trending changes during HSC proliferation. We found progressive activation of oxidative pentose phosphate pathway (OxiPPP) from dormant to active HSCs. Genetic or pharmacological interference with OxiPPP increased reactive oxygen species level in HSCs, reducing HSC self-renewal upon oxidative stress. Together, our work uncovers the metabolic dynamics during HSC proliferation, reveals a role of OxiPPP for HSC activation, and illustrates the utility of hi-scMet in dissecting metabolic heterogeneity of immunophenotypically defined cell populations.
INSTITUTE
Shanghai Jiao Tong University
LAST_NAME
CAO
FIRST_NAME
JING
ADDRESS
1954 Huashan Road, Shanghai, Shanghai, 200030, China
Amino acid catabolite markers for early prognostication of pneumonia in patients with COVID-19
STUDY_SUMMARY
Effective early-stage markers for predicting which patients are at risk of developing SARS-CoV-2 infection have not been fully investigated. Here, we performed comprehensive serum metabolome analysis of a total of 83 patients from two cohorts to determine that the acceleration of amino acid catabolism within 5 days from disease onset correlated with future disease severity. Increased levels of de-aminated amino acid catabolites involved in the de novo nucleotide synthesis pathway were identified as early prognostic markers that correlated with the initial viral load. We further employed mice models of SARS-CoV2-MA10 and influenza infection to demonstrate that such de-amination of amino acids and de novo synthesis of nucleotides were associated with the abnormal proliferation of airway and vascular tissue cells in the lungs during the early stages of infection. Consequently, it can be concluded that lung parenchymal tissue remodeling in the early stages of respiratory viral infections induces systemic metabolic remodeling and that the associated key amino acid catabolites are valid predictors for excessive inflammatory response in later disease stages.
INSTITUTE
Graduate School of Medicine, Kyoto University
DEPARTMENT
Center for Cancer Immunotherapy and Immunobiology
LABORATORY
Sugiura-lab
LAST_NAME
Sugiura
FIRST_NAME
Yuki
ADDRESS
Shogoin-Kawaramachi 53,, Sakyo-ku, Kyoto, Kyoto, 160-8582, Japan
Polar metabolomics of in vitro digestions of commercial cow's milk formula.
STUDY_TYPE
(un)targeted MS
STUDY_SUMMARY
This study is part of a multi-part study, including a. longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin b. polar fecal metabolomics of a patient cohort c. fecal lipidomics of a patient cohort d. polar urinary metabolomics of a patient cohort e. polar metabolomics of in vitro digestions. This specific part is part e. polar metabolomics of in vitro digestions
INSTITUTE
Ghent University
DEPARTMENT
Department of Translational Physiology, Infectiology and Public Health
Deciphering the metabolic heterogeneity of hematopoietic stem cells with single-cell resolution
STUDY_SUMMARY
Metabolic status is crucial for stem cell functions; however, the metabolic heterogeneity of endogenous stem cells has never been directly assessed. Here, we develop a platform for high-throughput single-cell metabolomics (hi-scMet) of hematopoietic stem cells (HSCs). By combining flow cytometric isolation and nanoparticle-enhanced laser desorption/ionization mass spectrometry, we routinely detected >100 features from single cells. We mapped the single-cell metabolomes of all hematopoietic cell populations, and HSC subpopulations with different division times, detecting 33 features whose levels exhibited trending changes during HSC proliferation. We found progressive activation of oxidative pentose phosphate pathway (OxiPPP) from dormant to active HSCs. Genetic or pharmacological interference with OxiPPP increased reactive oxygen species level in HSCs, reducing HSC self-renewal upon oxidative stress. Together, our work uncovers the metabolic dynamics during HSC proliferation, reveals a role of OxiPPP for HSC activation, and illustrates the utility of hi-scMet in dissecting metabolic heterogeneity of immunophenotypically defined cell populations.
INSTITUTE
Shanghai Jiao Tong University
LAST_NAME
CAO
FIRST_NAME
JING
ADDRESS
1954 Huashan Road, Shanghai, Shanghai, 200030, China
Activation induces shift in nutrient utilization that differentially impacts cell functions in human neutrophils
STUDY_SUMMARY
Neutrophils could utilize a variety of metabolic sources to support their vital functions as the first responders in innate immunity. Here, using in vivo and ex vivo isotopic tracing, we quantitatively examined the contributions of different nutrients under specific conditions, and found that human circulating neutrophils, in contrast to neutrophil cell line, rely on glycogen storage as a major metabolic source under resting state, but rapidly switch to primarily using extracellular glucose upon activation with various stimuli. This shift is driven by a substantial increase in glucose uptake, which is mechanistically mediated by the rapid phosphorylation and translocation of GLUT1, that dominates the simultaneous increase in gross glycogen cycling. Shift in nutrient utilization impacts neutrophil functions in a function-specific manner: Oxidative burst depends on glucose utilization; whereas NETosis and phagocytosis can be flexibly supported by either glucose or glycogen; and neutrophil migration and fungal control are enhanced by the shift from glycogen utilization to glucose utilization. This provides a quantitative and dynamic understanding of fundamental features in neutrophil metabolism, and elucidates how metabolic remodeling shapes neutrophil functions, which has broad health relevance.
This study is part of a multi-part study, including a. longitudinal polar fecal metabolomics of mice undergoing sensitization to beta-lactoglobulin b. polar fecal metabolomics of a patient cohort c. fecal lipidomics of a patient cohort d. polar urinary metabolomics of a patient cohort e. polar metabolomics of in vitro digestions This specific part is part d. polar urinary metabolomics of a patient cohort
INSTITUTE
Ghent University
DEPARTMENT
Department of Translational Physiology, Infectiology and Public Health
Reparative macrophages play a crucial role in limiting excessive fibrosis and promoting cardiac repair after myocardial infarction (MI), highlighting the significance of enhancing their reparative phenotype for wound healing. Metabolic adaptation orchestrates the phenotypic transition of macrophages; however, the precise mechanisms governing metabolic reprogramming of cardiac reparative macrophages remain poorly understood. In this study, we investigated the role of Nucleophosmin 1 (NPM1) in the metabolic and phenotypic shift of cardiac macrophages in the context of MI, and explored the effect of targeting NPM1 for ischemic tissue repair.
INSTITUTE
Renji Hospital, Shanghai Jiao Tong University School of Medicine
LAST_NAME
Zhan
FIRST_NAME
Zhenzhen
ADDRESS
160 Pujian Road, Shanghai, Shanghai, 200127, China
Folate depletion time-course in K562 cells with analysis for porphyrin metabolites
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 100 nM folic acid for 0, 1, 2, 4, 6, or 8 days followed my LC-MS targeting porphyrin metabolites. This is a reverse timecourse where all samples are harvested on the same day. Day 0 in 100 nM folic acid indicates 8 days culture in 2,000 nM folic acid.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Metabolomics studies on human colorectal cancer cell lines
STUDY_SUMMARY
Although targeting oxidative phosphorylation (OXPHOS) for cancer treatment is currently impeded due to dose-limiting toxicities, there remain opportunities through combinations that provide therapeutic benefits at doses attainable in patients. On the other hand, while glycolysis-deficient cancers are generally vulnerable to OXPHOS inhibition in preclinical models, the full extent of phenotypical and mechanistic consequences of inhibiting OXPHOS in cancers capable of glycolysis is not yet well understood. We aimed to clarify the response and underlying mechanisms of colorectal cancer (CRC) that commonly exhibit the glycolytic phenotype to OXPHOS inhibition and to identify potential approaches to render such cells more sensitive to OXPHOS inhibitors. The responses of glycolysis-competent CRC cells to targeting OXPHOS were tested using targeted meatbolomics.
Polar metabolite levels in K562 cells following C13-Serine and C13-Glycine tracing
STUDY_SUMMARY
Culture of K562 cells for 7 days in RPMI media containing 2000 nM folic acid or 100 nM folic acid. At day 8, media was changed to 2000 nM or 100 nM folic acid with unlabeled serine and glycine, or 2-C13-Serine and 2-C13-Glycine at RPMI levels. Amino acid tracing was performed for 24 hours. These samples were followed up by metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Identifying subgroups of childhood obesity by using multiplatform metabotyping
STUDY_SUMMARY
Obesity results from an interplay between genetic predisposition and environmental factors such as diet, physical activity, culture, and socioeconomic status. Personalized treatments for obesity would be optimal, thus necessitating the identification of individual characteristics to improve the effectiveness of therapies. For example, genetic impairment of the leptin-melanocortin pathway can result in rare cases of severe early-onset obesity. Metabolomics has the potential to distinguish between a healthy and obese status; however, differentiating subsets of individuals within the obesity spectrum remains challenging. Factor analysis can integrate patient features from diverse sources, allowing an accurate subclassification of individuals. This study presents a workflow to identify metabotypes, particularly when routine clinical studies fail in patient categorization. 110 children with obesity (BMI > +2 SDS) genotyped for nine genes involved in the leptin-melanocortin pathway (CPE, MC3R, MC4R, MRAP2, NCOA1, PCSK1, POMC, SH2B1, and SIM1) and two glutamate receptor genes (GRM7 and GRIK1) were studied; 55 harboring heterozygous rare sequence variants and 55 with no variants. Anthropometric and routine clinical laboratory data were collected, and serum samples processed for untargeted metabolomic analysis using GC-q-MS and CE-TOF-MS and reversed-phase U(H)PLC-QTOF-MS/MS in positive and negative ionization modes. Following signal processing and multialignment, multivariate and univariate statistical analyses were applied to evaluate the genetic trait association with metabolomics data and clinical and routine laboratory features. Neither the presence of a heterozygous rare sequence variant nor clinical/routine laboratory features determined subgroups in the metabolomics data. To identify metabolomic subtypes, we applied Factor Analysis, by constructing a composite matrix from the five analytical platforms. Six factors were discovered and three different metabotypes. Subtle but neat differences in the circulating lipids, as well as in insulin sensitivity could be established, which opens the possibility to personalize the treatment according to the patients categorization into such obesity subtypes. Metabotyping in clinical contexts poses challenges due to the influence of various uncontrolled variables on metabolic phenotypes. However, this strategy reveals the potential to identify subsets of patients with similar clinical diagnoses but different metabolic conditions. This approach underscores the broader applicability of Factor Analysis in metabotyping across diverse clinical scenarios.
INSTITUTE
CEU San Pablo University
LABORATORY
CEMBIO
LAST_NAME
Chamoso-SƔnchez
FIRST_NAME
David
ADDRESS
Urb. MonteprĆncipe. 28925 AlcorcĆ³n, Madrid (EspaƱa)
Integrating uterine microbiome and metabolome to advance the understanding of the uterine environment in dairy cows with metritis
STUDY_SUMMARY
Background: Metritis is a prevalent uterine disease that affects the welfare, fertility, and survival of dairy cows. The uterine microbiome from cows that develop metritis and those that remain healthy do not differ from calving until 2 days after calving, after which there is a dysbiosis of the uterine microbiome characterized by a shift towards opportunistic pathogens such as Fusobacteriota and Bacteroidota. Whether these opportunistic pathogens proliferate and overtake the uterine commensals could be determined by the type of substrates present in the uterus. The objective of this study was to integrate uterine metabolome and microbiome data to advance the understanding of metritis development in dairy cows. Holstein cows (n = 104) had uterine fluid collected at calving and at the day of metritis diagnosis. Cows with metritis (n = 52) were paired with cows without metritis (n = 52) based on days after calving. First, the uterine metabolome and microbiome were evaluated individually, and then integrated using network analyses. Results: The uterine metabolome differed both at calving and on the day of metritis diagnosis between cows with and without metritis. The uterine microbiome did not differ at calving but differed on the day of metritis diagnosis between cows with and without metritis. Omics integration was performed between 153 significant metabolites and 6 significant bacteria genera on the day of metritis diagnosis. A total of 49 metabolites were correlated with 3 bacteria genera (i.e. Fusobacteria, Porphyromonas and Bacteroides) on the day of metritis diagnosis. The main metabolites have been associated with attenuation of biofilm formation by commensal bacteria, pathogenic bacterial overgrowth, defense mechanisms against the immune system, tissue damage and inflammation, and immune dysregulation. Conclusions: The data integration presented herein helps advance the understanding of metritis development in dairy cows. The identified metabolites may be promising targets for future interventions aiming to reduce pathogenic bacterial growth in the uterus, and therefore, reducing the incidence of metritis.
Untargeted metabolomics of Quercus ilex seedlings under drought and Phytophthora cinnamomi inoculation stresses
STUDY_SUMMARY
Holm oak (Quercus ilex) is considered one of the major structural elements of the Mediterranean forests and the agrosilvopastoral Spanish ādehesaā, representing an outstanding example of ecological and socio-economic sustainability of forest ecosystems. The exotic pathogen Phytophthora cinnamomi is one of the most aggressive of woody species, and together drought is considered one of the main drivers of holm oak decline. The effect and responses of P. cinnamomi inoculation has been studied on the offspring of mother trees growing in declined and non-declined areas of two Andalusian populations (Cordoba and Huelva). Damage symptoms, mortality, and chlorophyll fluorescence have been evaluated in seedlings inoculated under humid and drought conditions. The effect and responses depended on the population, being more accused in Huelva than in Cordoba population. An integrative proteomic and metabolomic analysis revealed the involvement of different metabolic pathways in response to the pathogen in both populations, such as amino acid metabolism pathways in Huelva, and terpenoids and flavonoids biosynthesis in Cordoba. However, a differential response was not observed between seedlings inoculated under humid and drought conditions. A protective mechanism of the photosynthetic apparatus is launched in response to defective photosynthetic activity in inoculated plants, which seems to be more efficient in the Cordoba population. In addition, enzymes and metabolites of the phenylpropanoid and flavonoid biosynthesis pathways may confer higher resistance to Cordoba population. Some of these enzymes are proposed as markers of resilience, among which glyoxalase I, glutathione reductase, thioredoxin reductase, and cinnamyl alcohol dehydrogenase are candidates.
INSTITUTE
University of Cordoba
DEPARTMENT
Department of Biochemistry and Molecular Biology
LABORATORY
Agroforestry and Plant Biochemistry, Proteomics and Systems Biology
LAST_NAME
Tienda Parrilla
FIRST_NAME
Marta
ADDRESS
Campus de Rabanales, CĆ³rdoba, CĆ³rdoba, 14014, Spain
Tissue Lipidomic Profiling for Detection of Non-Small Cell Lung Cancer
STUDY_SUMMARY
UPLC-HRMS analysis was performed on AC and SCC patients. OPLSDA classfication was performed on tumor vs. ANT and DNT samples. Panels of discriminant features were identified. The biomarkers identified in discovery set samples for each binary classification were confirmed by using a set of validation samples, which were run separately. Additionally, paired analysis showed the abundance of discriminant compounds has significant altered in tumor tissues compared to corresponding DNT and ANT samples, reflecting the lipid characteristics related to tumor in situ and cancer cell signaling.
Metabolomic analysis reveal mechanisms of Shenzao dripping pill against myocardial ischemia
STUDY_SUMMARY
This study employed the advanced Ultra-high performance liquid chromatography-quadruple-Exactive Orbitrap mass spectrometry technology to detect biomarkers related to myocardial ischemia in rats. Additionally, it aimed to investigate the effect of the Shenzao dripping pill on rectifying metabolic disorders and preserving the metabolites in dynamic balance during myocardial ischemia, analyze potential metabolic pathways, and elucidate the mechanism of action of SZDP through the lens of metabolomics.
INSTITUTE
Guangdong Pharmaceutical University
DEPARTMENT
Engineering & Technology Research Center for Chinese Materia Medica Quality of the Universities of Guangdong Province
LABORATORY
Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State Administration of Traditional Chinese Medicine
LAST_NAME
Kuang
FIRST_NAME
Jiehui
ADDRESS
Waihuan East Road, 280, Guangzhou, Guangdong, 510006, China
EMAIL
kuang_jieh@163.com
STUDY_TYPE
Metabolomic analysis on serum samples of SZDP against MI
The role of gut microbiota in muscle mitochondria function, colon health, and sarcopenia: from clinical to bench
STUDY_SUMMARY
To further investigate if metabolites produced by probiotics could directly improve muscle growth, an in-vitro study was performed by utilizing C2C12 myoblasts and myotubes, as well as probiotic supernatant. The supernatant of probiotics was collected and LC-MS analysis was performed.
Metabolomics and glucose and glutamine labeled isotope tracing analysis in murine lung adenocarcinoma cells in the context of normal or active NRF2 pathway as well as HDAC and glutaminase inhibitor treatment.
STUDY_SUMMARY
Interplay between metabolism and chromatin signaling are implicated in cancer progression. However, whether and how metabolic reprogramming in tumors generates chromatin vulnerabilities remain unclear. Lung adenocarcinoma (LUAD) tumors frequently harbor aberrant activation of the NRF2 antioxidant pathway which drives aggressive and chemo-resistant disease. Using a chromatin-focused CRISPR screen we report that NRF2 activation sensitizes LUAD cells to genetic and chemical inhibition of class I histone deacetylases (HDAC). This association is observed across cultured cells, mouse models and patient-derived xenografts. Integrative epigenomic, transcriptomic and metabolomic analysis demonstrates that HDAC inhibition causes widespread redistribution of H4ac and its reader protein, which transcriptionally downregulates metabolic enzymes. This results in reduced flux into amino acid metabolism and de novo nucleotide synthesis pathways that are preferentially required for the survival of NRF2-active cancer cells. Together, our findings suggest NRF2 activation as a potential biomarker for effective repurposing of HDAC inhibitors to treat solid tumors. In this metabolomics experiment we characterize the changes in metabolic pathway flux in KP LUAD cells in response to HDAC and glutaminase inhibition. This dataset includes metabolomics of U-C13 glucose tracing (1h and 24h) and U-C13 glutamine (8h) of mouse LUAD cell lines with Kras overexpression and p53 knock-out (KP), carrying empty vector (EV) or overexpression of NRF2dNeh2 (NRF2) and treated with DMSO, Romidepsin or CB-839. 3 technical replicates were done per condition in 2 separate experiments, 1: glucose tracing and 2: glutamine tracing.
Effects of Microbiome Depletion on Radiation Biodosimetry Metabolomics
STUDY_SUMMARY
Development of novel biodosimetry assays and medical countermeasures is needed to obtain a level of radiation preparedness in the event of malicious or accidental mass exposures to ionizing radiation (IR). For biodosimetry, metabolic profiling with mass spectrometry (MS) platforms has identified several small molecules in easily accessible biofluids that are promising for dose reconstruction. As our microbiome has profound effects on biofluid metabolite composition, it is of interest how variation in the host microbiome may affect metabolomics based biodosimetry. Here, we chemically āknocked outā the microbiome of male and female C57BL/6 mice (Abx mice) and then irradiated (0, 3, or 8 Gy) them to determine the role of the host microbiome on biofluid radiation signatures (3 d serum).
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
The Track Dairy Cattle (trackDC) study is a longitudinal study in northern China that aims to track newborn calves to assess the development of gut microbiota during early life that contributes to cattle health and production. In this study, 36 newborn calves were randomly assigned to three groups and followed for two months after birth. The groups included a control group (CON), a rumen microbiota transplantation group (RMT), and a rumen fluid transplantation group (RFT) and intensive data has been collected. Blood samples were collected at 15, 35, and 56 days after birth.
The role of gut microbiota in muscle mitochondria function, colon health, and sarcopenia: from clinical to bench (2)
STUDY_SUMMARY
Gut microbes produce metabolites in the intestinal lumen, which can travel systemically to affect organs. Metagenomic analysis was performed on liquid chromatographyĆ¢ĀĀmass spectrometry (LC-MS) to detect differential metabolites between people with or without sarcopenia.
Gut microbiota and metabolites in estrus cycle and their changes in a menopausal transition rat model with typical neuroendocrine aging
STUDY_SUMMARY
Neuroendocrine alterations in the mid-life hypothalamus coupled with reproductive decline herald the initiation of menopausal transition. The certain feature and contribution of gut microflora and metabolites to neuroendocrine changes in the menopausal transition remain largely unknown. Fecal samples of rats experiencing different reproductive stages were collected and processed for 16S rRNA and liquid chromatography-mass spectrometry sequencing. The differences of gut microbiota and metabolites between young and middle-aged rats during proestrus and diestrus were analyzed and their relationships to neuroendocrine aging were then examined. This study documents specific gut microbial composition changes and concomitant shifting trends of metabolites during menopausal transition, which may initiate the gut-brain dysfunction in neuroendocrine aging.
We quantified metabolites of extracellular fluid samples from BAT and eWAT. Briefly, we collected the BAT_EF samples and eWAT_EF samples from 12 weeks chow diet C57BL/6J mice (n=5). We run the EF metabolomics using high ph HILIC method on Exploris 240.
Brown adipose tissue (BAT) metabolites in mice acclimated to cold (6ĖC) or thermoneutrality (30ĖC) for two weeks. N = 10 for mice housed at 6 ĖC and 9 for mice housed at 30 ĖC.
To determine the metabolic fate and nitrogen flux of BCAA in mouse brown adipocytes, we used 15N labeled BCAA tracing (Leu (NLM-142-1, CIL), Ile (NLM-292-0.25, CIL) and Val (NLM-316-0.5, CIL)) followed by LC-MS analysis.
13C BCAA tracing in differentiated brown adipocyte
STUDY_SUMMARY
To determine the metabolic fate and carbon flux of BCAA in mouse brown adipocytes, we used 13C labeled BCAA tracing (Leu (NLM-142-1, CIL), Ile (NLM-292-0.25, CIL) and Val (NLM-316-0.5, CIL)) followed by LC-MS analysis.
To determine the metabolic fate and nitrogen flux of BCAA in mouse brown adipocytes, we used 15N labeled BCAA tracing (Leu (NLM-142-1, CIL), Ile (NLM-292-0.25, CIL) and Val (NLM-316-0.5, CIL)) followed by LC-MS analysis.
Media_13C BCAA tracing in differentiated brown adipocyte
STUDY_SUMMARY
To determine the metabolic fate and carbon flux of BCAA in mouse brown adipocytes, we used 13C labeled BCAA tracing (Leu (NLM-142-1, CIL), Ile (NLM-292-0.25, CIL) and Val (NLM-316-0.5, CIL)) followed by LC-MS analysis.
Untargeted metabolomics analysis of murine colon content
STUDY_SUMMARY
we used the minimal consortium Oligo-Mouse Microbiota (OMM)12 to study the function of Coriobacteriia under defined conditions in gnotobiotic mice. OMM12 mice with or without addition of the dominant gut bacterium Eggerthella lenta were fed with diets varying in fat content or supplemented with primary bile acids.
A High-Fat Eucaloric Diet Induces Reprometabolic Syndrome of Obesity in Normal Weight Women - metabolomics
STUDY_SUMMARY
Objective: We examined the effects of one month of a eucaloric, high-fat (48% of calories) diet (HFD) on gonadotropin secretion in normal weight women to interrogate the role of free fatty acids and insulin in mediating the relative hypogonadotropic hypogonadism of obesity. Methods: Eighteen eumenorrheic women (BMI 18-25 kg/m2) were studied in the early follicular phase of the menstrual cycle before and after exposure to a HFD with frequent blood sampling for LH and FSH, followed by an assessment of pituitary sensitivity to GnRH. Mass spectrometrybased plasma metabolomic analysis was also performed. Paired testing and time series analysis were performed as appropriate. Results: Mean endogenous LH (unstimulated) was significantly decreased after the HFD (4.3 Ā±1.0 vs 3.8 Ā± 1.0, P<0.01); mean unstimulated FSH was not changed. Both LH (10.1 Ā± 1.0 vs 7.2 Ā± 1.0, P<0.01), and FSH (9.5 Ā± 1.0 vs 8.8 Ā± 1.0, P<0.01) response to 75 ng/kg of GnRH were reduced after the HFD. Mean LH pulse amplitude and LH interpulse interval were unaffected by the dietary exposure. Eucaloric HFD exposure did not cause weight change. Plasma metabolomics confirmed adherence with elevation of fasting free fatty acids (especially long-chain mono-, poly- and highly-unsaturated fatty acids) by the last day of the HFD. Conclusion: One-month exposure to a HFD successfully induced key reproductive and metabolic features of Reprometabolic Syndrome in normal weight women. Dietary factors may underly the gonadotrope compromise seen in obesity related subfertility and that therapeutic dietary interventions, independent of weight loss, may be possible.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
NMR- and MS-based omics reveal characteristic metabolome atlas and optimize biofluid earlydiagnostic biomarkers for esophageal squamous cell carcinoma (part-ā ”)
STUDY_SUMMARY
Metabolic changes precede malignant histology. However, it remains unclear whether detectable characteristic metabolome exists in esophageal squamous cell carcinoma (ESCC) tissues and biofluids for early diagnosis. We conducted NMR- and MS-based metabolomics on 1,153 matched ESCC tissues, normal mucosae, pre- and one-week post-operative sera and urines from 560 participants across three hospitals, with machine learning, logistic regression and WGCNA. Aberrations in 'alanine, aspartate and glutamate metabolism' proved to be prevalent throughout the ESCC evolution, and were reflected in 16 serum and 10 urine metabolic signatures that were consistently identified by NMR and MS in both discovery and validation sets. NMR-based simplified panels of any five serum or urine metabolites outperformed clinical serological tumor markers (AUC = 0.984 and 0.930, respectively), and were effective in distinguishing early-stage ESCC in test set (serum accuracy = 0.994, urine accuracy = 0.879). Collectively, NMR-based biofluid screening can reveal characteristic metabolic events of ESCC and be feasible for early detection (ChiCTR2300073613).
INSTITUTE
Radiology Department, Second Affiliated Hospital, Shantou University Medical College, Shantou
LAST_NAME
Lin
FIRST_NAME
Yan
ADDRESS
No. 69, Dongxia North Road, Shantou, Guangdong, China
NMR- and MS-based omics reveal characteristic metabolome atlas and optimize biofluid earlydiagnostic biomarkers for esophageal squamous cell carcinoma (part-ā ¢)
STUDY_SUMMARY
Metabolic changes precede malignant histology. However, it remains unclear whether detectable characteristic metabolome exists in esophageal squamous cell carcinoma (ESCC) tissues and biofluids for early diagnosis. We conducted NMR- and MS-based metabolomics on 1,153 matched ESCC tissues, normal mucosae, pre- and one-week post-operative sera and urines from 560 participants across three hospitals, with machine learning, logistic regression and WGCNA. Aberrations in 'alanine, aspartate and glutamate metabolism' proved to be prevalent throughout the ESCC evolution, and were reflected in 16 serum and 10 urine metabolic signatures that were consistently identified by NMR and MS in both discovery and validation sets. NMR-based simplified panels of any five serum or urine metabolites outperformed clinical serological tumor markers (AUC = 0.984 and 0.930, respectively), and were effective in distinguishing early-stage ESCC in test set (serum accuracy = 0.994, urine accuracy = 0.879). Collectively, NMR-based biofluid screening can reveal characteristic metabolic events of ESCC and be feasible for early detection (ChiCTR2300073613).
INSTITUTE
Radiology Department, Second Affiliated Hospital, Shantou University Medical College
LAST_NAME
Lin
FIRST_NAME
Yan
ADDRESS
No. 69, Dongxia North Road, Shantou, Guangdong, China, Shantou, Guangdong, China, 515041, China
NMR- and MS-based omics reveal characteristic metabolome atlas and optimize biofluid earlydiagnostic biomarkers for esophageal squamous cell carcinoma (part-ā )
STUDY_SUMMARY
Metabolic changes precede malignant histology. However, it remains unclear whether detectable characteristic metabolome exists in esophageal squamous cell carcinoma (ESCC) tissues and biofluids for early diagnosis. We conducted NMR- and MS-based metabolomics on 1,153 matched ESCC tissues, normal mucosae, pre- and one-week post-operative sera and urines from 560 participants across three hospitals, with machine learning, logistic regression and WGCNA. Aberrations in 'alanine, aspartate and glutamate metabolism' proved to be prevalent throughout the ESCC evolution, and were reflected in 16 serum and 10 urine metabolic signatures that were consistently identified by NMR and MS in both discovery and validation sets. NMR-based simplified panels of any five serum or urine metabolites outperformed clinical serological tumor markers (AUC = 0.984 and 0.930, respectively), and were effective in distinguishing early-stage ESCC in test set (serum accuracy = 0.994, urine accuracy = 0.879). Collectively, NMR-based biofluid screening can reveal characteristic metabolic events of ESCC and be feasible for early detection (ChiCTR2300073613).
INSTITUTE
Radiology Department, Second Affiliated Hospital, Shantou University Medical College, Shantou
LAST_NAME
Lin
FIRST_NAME
Yan
ADDRESS
No. 69, Dongxia North Road, Shantou, Guangdong, China
Metabolomics analysis of Cormus domestica (L.) fruits and the valorisation of an ethnobotanical heritage of culinary and medicinal uses in Italy
STUDY_TYPE
Untargeted MS-based metabolomics
STUDY_SUMMARY
Cormus domestica (L.) is a monophyletic wild fruit tree belonging to the Rosaceae family, with well-documented use in the Mediterranean region. Traditionally, these fruits are harvested and stored for at least 2 weeks before consumption. During this period, the fruit reaches its well-known and peculiar organolepticĀ and texture characteristics. However, the spread of more profitable fruit tree species, resulted in its progressive erosion. In this work weĀ performed proteomic and metabolomic fruit analyses at three times after harvesting to provide data on its chemical composition and nutritional and nutraceutical properties.
INSTITUTE
UniversitĆ degli Studi del Sannio (Benevento, Italy) and University of CĆ³rdoba (Spain)
DEPARTMENT
University of Sannio, Department of Science and Technology, via de Sanctis, BeneventoĀ 82100Ā Italy AND University of CĆ³rdoba, Department of Biochemistry and Molecular Biology, Spain
LABORATORY
Integrated Laboratory of Animal and Plant Biology for EnvironmentĀ andĀ Health (Italy) AND Agroforestry and Plant Biochemistry, Proteomics and Systems Biology (Spain)
LAST_NAME
Tienda
FIRST_NAME
Marta
ADDRESS
Campus de Rabanales, CĆ³rdoba, CĆ³rdoba, 14014, Spain
Lipidomics and plasma hormone reveal indicators of reproductive status in Florida manatees (Trichechus manatus latirostris)
STUDY_SUMMARY
Florida manatees (Trichechus manatus latirostris) are protected as a threatened species, and data are lacking regarding their reproductive physiology. This study aimed to (1) quantify plasma steroid hormones in Florida manatees from two field sites, Crystal River and Indian River Lagoon, at different gestational stages and to (2) determine the relationship between plasma progesterone concentrations and lipid biochemistry in relation to pregnancy status. Ultra-high performance liquid chromatography-tandem mass spectrometric analysis was used to measure plasma steroid hormones and lipids. Pregnant female manatees were morphometrically distinct from male and non-pregnant female manatees, characterized by larger body weight and maximal girth. Progesterone concentrations in manatees were also elevated during early gestation versus late gestation. Cholesterol, an important metabolic lipid and precursor for reproductive steroids, was not different between groups. Lipidomics quantified 949 lipids and plasma concentrations of a sphingolipid, ceramide non-hydroxy fatty acid-sphingosine and several glycerophospholipids, including lysophosphatidylcholine, phosphatidylethanolamines, plasmenyl-phosphatidylserines and monomethyl phosphatidylethanolamines, were associated with pregnancy status in the Florida manatee. This research contributes to improving knowledge of manatee reproductive physiology by providing data on plasma steroid hormones relative to reproductive status and by assessing how plasma lipids in healthy Florida manatees correspond to progesterone levels. This lipid panel has potential as a diagnostic approach to identify pregnant individuals in fresh and archived samples. These biochemical and morphometric indicators of reproductive status advance the understanding of manatee physiology.
Polar metabolite levels in K562 cells following PKC inhibition in 2,000 nM or 100 nM FA conditions
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 2000 nM or 100 nM folic acid for 6 days in the presence or absence of the PKC inhibitor, GF109203X. These samples were followed up by metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Metabolomic profiling of PMM2-CDG brain organoids by GC/MS
STUDY_SUMMARY
PMM2-CDG is a rare inborn error of metabolism caused by deficiency of the PMM enzyme, which leads to impaired protein glycosylation. While the disorder has primarily neurological presentation, there is limited knowledge about the specific brain-related changes that result from PMM deficiency. Utilizing 3D brain organoids derived from individuals with PMM2-CDG, we identified abnormal glucose metabolism in PMM2-CDG organoids, indicating disturbances in metabolic utilization. In this experiment, day 160 organoids were used.
Polar metabolite levels in MEL cells following PKC inhibition in 2,000 nM or 100 nM FA conditions
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 2000 nM or 100 nM folic acid for 6 days in the presence or absence of the PKC inhibitor, GF109203X. These samples were followed up by metabolomics targeting polar metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Porphyrin metabolite levels in K562 cells following PKC inhibition in 2,000 nM or 100 nM FA conditions
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 2000 nM or 100 nM folic acid for 6 days in the presence or absence of the PKC inhibitor, GF109203X. These samples were followed up by metabolomics targeting porphyrin metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Porphyrin metabolite levels in MEL cells following PKC inhibition in 2,000 nM or 100 nM FA conditions
STUDY_SUMMARY
Culture of MEL cells in RPMI media containing 2000 nM or 100 nM folic acid for 6 days in the presence or absence of the PKC inhibitor, GF109203X. These samples were followed up by metabolomics targeting porphyrin metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Porphyrin metabolite levels in K562 cells following 6 day culture in 2,000 nM or 100 nM folic acid
STUDY_SUMMARY
Culture of K562 cells in RPMI media containing 2000 nM or 100 nM folic acid for 6 days. These samples were followed up by metabolomics targeting porphyrin metabolites.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Identifying and mathematically modeling the time-course of extracellular metabolic markers associated with resistance to ceftolozane/tazobactam in Pseudomonas aeruginosa - Part 1
STUDY_SUMMARY
Extracellular bacterial metabolites have potential as markers of bacterial growth and resistance emergence, but have not been evaluated in dynamic in vitro studies. We investigated the dynamic metabolomic footprint of a multidrug-resistant hypermutable Pseudomonas aeruginosa isolate exposed to ceftolozane/tazobactam as continuous infusion (4.5g/day, 9g/day) in a hollow-fiber infection model over 7-9 days in biological replicates (n=5). Bacterial samples were collected at 0, 7, 23, 47, 71, 95, 143, 167, 191 and 215h, the supernatant quenched and extracellular metabolites extracted. Metabolites were analyzed via untargeted metabolomics, including hierarchical clustering and correlation with quantified total and resistant bacterial populations. The time-courses of five metabolites were mathematically modeled. These five (of 1921 detected) metabolites were from enriched pathways (arginine and central carbon metabolism). Absorbed L-arginine and secreted L-ornithine were highly correlated with the total bacterial population (r -0.79 and 0.82 respectively, p<0.0001). Ribose-5-phosphate, sedoheptulose-7-phosphate and trehalose-6-phosphate correlated with the resistant subpopulation (0.64, 0.64 and 0.67, respectively, p<0.0001), and were likely secreted due to resistant growth overcoming oxidative and osmotic stress induced by ceftolozane/tazobactam. Using PK/PD-based transduction models, these metabolites were successfully modeled based on the total or resistant bacterial populations. The models well described the abundance of each metabolite across the differing time-course profiles of biological replicates, based on bacterial killing and, importantly, resistant regrowth. These proof-of-concept studies suggest further exploration is warranted to determine the generalizability of these findings. The metabolites modeled in this work are not exclusive to bacterial cells. Future studies may use this approach to identify bacteria-specific metabolites correlating with resistance, which would ultimately be extremely useful for clinical translation.
INSTITUTE
Monash Institute of Pharmaceutical Sciences
DEPARTMENT
Monash Institute of Pharmaceutical Sciences
LABORATORY
Cornelia Landersdorfer
LAST_NAME
Landersdorfer
FIRST_NAME
Cornelia
ADDRESS
399 Royal Pd
EMAIL
dovile.anderson@monash.edu
NUM_GROUPS
6 groups with time points
TOTAL_SUBJECTS
NA
NUM_MALES
NA
NUM_FEMALES
NA
PUBLICATIONS
Identifying and mathematically modeling the time-course of extracellular metabolic markers associated with resistance to ceftolozane/tazobactam in Pseudomonas aeruginosa
NMR- and MS-based omics reveal characteristic metabolome atlas and optimize biofluid earlydiagnostic biomarkers for esophageal squamous cell carcinoma (part-ā ¤)
STUDY_SUMMARY
Metabolic changes precede malignant histology. However, it remains unclear whether detectable characteristic metabolome exists in esophageal squamous cell carcinoma (ESCC) tissues and biofluids for early diagnosis. We conducted NMR- and MS-based metabolomics on 1,153 matched ESCC tissues, normal mucosae, pre- and one-week post-operative sera and urines from 560 participants across three hospitals, with machine learning, logistic regression and WGCNA. Aberrations in 'alanine, aspartate and glutamate metabolism' proved to be prevalent throughout the ESCC evolution, and were reflected in 16 serum and 10 urine metabolic signatures that were consistently identified by NMR and MS in both discovery and validation sets. NMR-based simplified panels of any five serum or urine metabolites outperformed clinical serological tumor markers (AUC = 0.984 and 0.930, respectively), and were effective in distinguishing early-stage ESCC in test set (serum accuracy = 0.994, urine accuracy = 0.879). Collectively, NMR-based biofluid screening can reveal characteristic metabolic events of ESCC and be feasible for early detection (ChiCTR2300073613).
INSTITUTE
Radiology Department, Second Affiliated Hospital, Shantou University Medical College, Shantou
LAST_NAME
Lin
FIRST_NAME
Yan
ADDRESS
No. 69, Dongxia North Road, Shantou, Guangdong, China
Untargeted Metabolomics Reveals Unique Biomolecular Signatures in Overweight and Obesity Using UHPLC-ESI-QTOF-MS Analysis
STUDY_TYPE
LC/MS/MS
STUDY_SUMMARY
Aims: Obesity poses a multifaceted challenge to global public health, impacting individuals and society in various ways. Apart from the heightened susceptibility to chronic conditions such as diabetes, cardiovascular diseases, obesity significantly escalates healthcare costs. Effective public health strategies are essential for addressing issues related to early detection, diagnosis, and personalized treatment plans. This emphasizes the crucial need for a deep understanding of biochemical pathways, patient monitoring, and prognosis. In this context, metabolomics has become a valuable approach, focusing on the identification of metabolites in biofluids and tissues. Main Methods: In this study, an untargeted metabolomics-based method was employed to investigate metabolomic changes and their relationship to pathways in overweight and obese individuals. Plasma samples were collected from 29 healthy individuals with normal weight, 17 overweight individuals, and 28 obese individuals who met the inclusion criteria for the study. The plasma samples were analyzed using highly sensitive ultra-high-performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry. Results: Pantothenic acid and L-proline showed increased levels in the overweight group, whereas phenylacetaldehyde and glycerophosphocholine were notably decreased compared to the normal weight group. Conversely, the obese group exhibited elevated levels of specific metabolites, including L-leucine, L-tryptophan, phenylalanine, and tyrosine. On the contrary, the obese group demonstrated decreased levels of other metabolites such as 2,3-Diaminopropionic acid, and phenylacetaldehyde. Additionally, significant changes in metabolic pathways, such as pantothenate and CoA biosynthesis, and beta-alanine metabolism, were observed in the overweight group. In contrast, the obese group displayed significant alterations in phenylalanine and tyrosine metabolism, tryptophan metabolism, and beta oxidation of very long-chain fatty acids. Conclusion: The present investigation sheds light on the potential diagnostic significance of certain metabolites in obesity and the impact of their level changes on specific metabolic pathways. Additional studies are necessary to confirm the association of these metabolites in obesity and to confirm their diagnostic value.
INSTITUTE
Sharjah Institute for Medical Research
DEPARTMENT
Research institute of medical and health science
LABORATORY
Biomarker Discovery Group
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
NMR- and MS-based omics reveal characteristic metabolome atlas and optimize biofluid earlydiagnostic biomarkers for esophageal squamous cell carcinoma (part-ā £)
STUDY_SUMMARY
Metabolic changes precede malignant histology. However, it remains unclear whether detectable characteristic metabolome exists in esophageal squamous cell carcinoma (ESCC) tissues and biofluids for early diagnosis. We conducted NMR- and MS-based metabolomics on 1,153 matched ESCC tissues, normal mucosae, pre- and one-week post-operative sera and urines from 560 participants across three hospitals, with machine learning, logistic regression and WGCNA. Aberrations in 'alanine, aspartate and glutamate metabolism' proved to be prevalent throughout the ESCC evolution, and were reflected in 16 serum and 10 urine metabolic signatures that were consistently identified by NMR and MS in both discovery and validation sets. NMR-based simplified panels of any five serum or urine metabolites outperformed clinical serological tumor markers (AUC = 0.984 and 0.930, respectively), and were effective in distinguishing early-stage ESCC in test set (serum accuracy = 0.994, urine accuracy = 0.879). Collectively, NMR-based biofluid screening can reveal characteristic metabolic events of ESCC and be feasible for early detection (ChiCTR2300073613).
INSTITUTE
Radiology Department, Second Affiliated Hospital, Shantou University Medical College, Shantou
LAST_NAME
Lin
FIRST_NAME
Yan
ADDRESS
No. 69, Dongxia North Road, Shantou, Guangdong, China
Chronic stress dampens Lactobacillus johnsonii-mediated tumor suppression to enhance colorectal cancer progression
STUDY_SUMMARY
Colorectal cancer (CRC) development and outcome are impacted by modifiable risk factors, including psychological stress. The gut microbiota has also been shown to be linked to psychological factors. Here, we found a marked deteriorative effect of chronic stress in multiple CRC models, including chemically-induced (AOM/DSS), genetically engineered (APCmin/+), and xenograft tumor mouse models. RNA-seq data from colon tissues revealed that expression of stemness-related genes was upregulated in the stressed CRC group by activated Ī²-catenin signaling, which was further confirmed by results from ex vivo organoid analyses as well as in vitro and in vivo cell tumorigenicity assays. 16S rRNA sequencing of the gut microbiota showed that chronic stress disrupted gut microbes, and antibiotic treatment and fecal microbiota transplantation abolished the stimulatory effects of chronic stress on CRC progression. Stressed CRC mice displayed a significant decrease in Lactobacillus johnsonii (L. johnsonii) abundance, which was inversely correlated with tumor load. Moreover, protocatechuic acid (PCA) was identified as a beneficial metabolite produced by L. johnsonii based on metabolome sequencing and LCāMS/MS analysis. Replenishment of L. johnsonii or PCA blocked chronic stress-induced CRC progression by decreasing Ī²-catenin expression. Furthermore, PCA activated the cGMP pathway, and the cGMP agonist sildenafil abolished the effects of chronic stress on CRC. Altogether, these data identify that stress impacts the gut microbiome to support CRC progression.
INSTITUTE
China Pharmaceutical University
LAST_NAME
Cao
FIRST_NAME
Qiuhua
ADDRESS
No. 639 Longmian Avenue, Jiangning District, Nanjing, Jiangsu, 211198, China
Estrogen-mediated inhibition of purine metabolism and cell cycle arrest as a novel therapeutic approach in colorectal cancer cells
STUDY_SUMMARY
Purine metabolism is upregulated in various cancers including colorectal cancer (CRC). While previous research has elucidated the role of Estrogen (E2) in metabolism remodeling and ATP production, its effects on purine metabolism remained unexplored. This study investigates the impact of E2 signalling on purine metabolism in CRC cells. We demonstrate, for the first time, a protective effect of E2 on CRC cells by targeting the purine synthesis pathway through its receptor estrogen receptor Ī± (ERĪ±). A full metabolomic profiling, next generation sequencing (NGS) and integrated OMICS were conducted for HCT-116 cells treated with E2 with and without silencing ERĪ±. Our results revealed an enrichment of the purine metabolic pathway, with 27 genes in the de novo purine synthesis pathway downregulated in E2-treated CRC cells. Besides, E2-induced DNA damage, cell cycle arrest, and apoptosis are ERĪ±-dependent. Our findings suggest potential therapeutic avenues for CRC treatment through antimetabolites targeting purine synthesis, as E2 treatment reduces the expression of relevant metabolites.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
A small molecule macrophage migration inhibitory factor agonist ameliorates age-related myocardial intolerance to ischemia-reperfusion insults via metabolic regulation - Part 2
STUDY_SUMMARY
Macrophage migration inhibitory factor (MIF) is an innate cytokine that regulates both inflammatory and homeostatic responses. MIF is expressed by cardiomyocytes, where it exerts a protective action against ischemia-reperfusion (I/R) injury by activating AMP-activated protein kinase (AMPK). This effect is attenuated in the senescent heart due to an intrinsic, age-related reduction in MIF expression. We hypothesized that treating the aged heart with the small molecule MIF agonist (MIF20) can reinforce protective MIF signaling in cardiomyocytes, leading to a beneficial effect against I/R stress. The administration of MIF20 at the onset of reperfusion was found to not only decrease myocardial infarct size but also preserves systolic function in the aged heart. Protection from I/R injury was reduced in mice with cardiomyocyte-specific Mif deletion, consistent with the mechanism of action of MIF20 to allosterically increase MIF affinity for its cognate receptor CD74. We further found MIF20 to contribute to the maintenance of mitochondrial fitness and to preserve the contractile properties of aged cardiomyocytes under hypoxia/reoxygenation. MIF20 augments protective metabolic responses by reducing the NADH/NAD ratio, leading to a decrease in the accumulation of reactive oxygen species (ROS) in the aged myocardium under I/R stress. We also identify alterations in the expression levels of the downstream effectors PDK4 and LCAD, which participate in the remodeling of the cardiac metabolic profile. Data from this study demonstrates that pharmacologic augmentation of MIF signaling provides beneficial homeostatic actions on senescent myocardium under I/R stress.
Early time-restricted eating improves markers of cardiometabolic health but has no impact on nutrient absorption in healthy adults
STUDY_TYPE
Randomized Controlled Trial
STUDY_SUMMARY
Metabolomic analysis performed on 88 human plasma samples collected from 16 participants that received 2 treatments with 3 time points each. Samples were analyzed by UPLC-MS using a Waters Acquity UPLC and detected on a 4000 QTrap by multiple reaction monitoring (MRM) with negative mode electrospray ionization.
INSTITUTE
California Polytechnic State University, San Luis Obispo
LAST_NAME
La Frano
FIRST_NAME
Michael
ADDRESS
Cal Poly State University 1 Grand Avenue San Luis Obispo, CA 93407
Effects of Preanalytical Sample Collection and Handling on Comprehensive Metabolite Measurements in Human Urine Biospecimens
STUDY_TYPE
LCMS Investigation of Urine Metabolite Stability
STUDY_SUMMARY
This study evaluates the impact of sample handling conditions on urine metabolome profiles relative to a gold standard condition. The sample handling conditions included a combination of three variables, use of preservative (None, Borate, or Chlorhexidine), refrigeration storage time (0h or 24h), or freeze-thaws (either 1x or 3x freeze-thaws, thawed on ice, in a refrigerator, or at room temperature). Samples were taken from 13 participants, and then aliquoted and subjected to the various conditions. Comparisons between treatment effects were taken between pairs of samples within each participantās sample set, a within participant paired design.
INSTITUTE
National Center for Advancing Tranlastional Sciences
Short chain fatty acid (SCFA) analysis in a bioreactor model system exposed to PFAS and bisphenols.
STUDY_SUMMARY
An in vitro bioreactor model system based on the simplified human microbiome model (SIHUMIx) was used to investigate the direct effects of either perfluorooctanoic acid (PFOA), perfluorohexanoic acid (PFHxA) and perfluorobutanoic acid (PFBA) or bisphenol S (BPS) and bisphenol F (BPF) or a combined mixture on the microbiota. To determine any changes in the main fermentation products of the bacterial cultures three short chain fatty acids (SCFAs) were analyzed, namely acetate butyrate and propionate. Only the PFAS exposed cultures showed a significant increase in the concentrations of acetate (KW-test P= 0.0431) and butyrate (KW-test P= 0.0158) during exposure. In none of the exposure cultures nor in the control did the concentration of propionate change significantly.
Central Transcriptional Regulator Controls Growth and Carbon Storage under High Light Stress in Photosynthetic Microalgae Model Strains
STUDY_TYPE
Algae
STUDY_SUMMARY
Carbon capture efficiency and biochemical storage are some of the primary drivers of photosynthetic productivity and by extension crop yield. To elucidate the mechanisms governing yield phenotypes and carbon allocation regulatory elements, we selected two microalgae strains as simplified models of photosynthetic crops. The Picochlorum celeri TG2 isolate is one of the fastest growing algae and in this work is juxtaposed to a closely related, slower growing, isolate, TG1, of the same species with less than 2% genomic divergence. Through the application of a comprehensive systems biology light-stress response study, we observed a stark difference in carbon assimilation and storage rates, with the slower growing isolate accumulating almost three times the amount of starch compared to the fast-growing isolate. We characterized the carbon storage rates and allocation dynamics, with metabolic bottlenecks, and transport rates of intermediates underlying the variations in growth and composition in high light using instationary 13C-fluxomics experiments. High light stress analysis of transcriptomic dynamics during acclimation of the strains from low to high light identified a widespread response with up to 73% the annotated gene set significantly differentially expressed after only 1 hour. Broad transcriptional regulatory control was inferred by a rapid depletion of a global diel-responsive transcription factor closely related to a circadian-regulator in plants, as the single most distinct transcription factor. Transferring this factor to the slower variant increased yield, specific growth rate, and carbohydrate accumulation of the selected engineered strain, providing further evidence for a coordinating regulatory mechanism for this complex phenotype.
Identifying and mathematically modeling the time-course of extracellular metabolic markers associated with resistance to ceftolozane/tazobactam in Pseudomonas aeruginosa - Part 2
STUDY_SUMMARY
Extracellular bacterial metabolites have potential as markers of bacterial growth and resistance emergence, but have not been evaluated in dynamic in vitro studies. We investigated the dynamic metabolomic footprint of a multidrug-resistant hypermutable Pseudomonas aeruginosa isolate exposed to ceftolozane/tazobactam as continuous infusion (4.5g/day, 9g/day) in a hollow-fiber infection model over 7-9 days in biological replicates (n=5). Bacterial samples were collected at 0, 7, 23, 47, 71, 95, 143, 167, 191 and 215h, the supernatant quenched and extracellular metabolites extracted. Metabolites were analyzed via untargeted metabolomics, including hierarchical clustering and correlation with quantified total and resistant bacterial populations. The time-courses of five metabolites were mathematically modeled. These five (of 1921 detected) metabolites were from enriched pathways (arginine and central carbon metabolism). Absorbed L-arginine and secreted L-ornithine were highly correlated with the total bacterial population (r -0.79 and 0.82 respectively, p<0.0001). Ribose-5-phosphate, sedoheptulose-7-phosphate and trehalose-6-phosphate correlated with the resistant subpopulation (0.64, 0.64 and 0.67, respectively, p<0.0001), and were likely secreted due to resistant growth overcoming oxidative and osmotic stress induced by ceftolozane/tazobactam. Using PK/PD-based transduction models, these metabolites were successfully modeled based on the total or resistant bacterial populations. The models well described the abundance of each metabolite across the differing time-course profiles of biological replicates, based on bacterial killing and, importantly, resistant regrowth. These proof-of-concept studies suggest further exploration is warranted to determine the generalizability of these findings. The metabolites modeled in this work are not exclusive to bacterial cells. Future studies may use this approach to identify bacteria-specific metabolites correlating with resistance, which would ultimately be extremely useful for clinical translation.
INSTITUTE
Monash Institute of Pharmaceutical Sciences
DEPARTMENT
Drug Delivery, Disposition and Dynamics
LABORATORY
Cornelia Landersdorfer
LAST_NAME
Landersdorfer
FIRST_NAME
Cornelia
ADDRESS
399 Royal Pd
EMAIL
cornelia.landersdorfer@monash.edu
NUM_GROUPS
6 groups with time points
TOTAL_SUBJECTS
NA
NUM_MALES
NA
NUM_FEMALES
NA
PUBLICATIONS
Identifying and mathematically modeling the time-course of extracellular metabolic markers associated with resistance to ceftolozane/tazobactam in Pseudomonas aeruginosa
Untargeted lipidomics of WT and Cyp2c44(-/-) mice liver.
STUDY_TYPE
Untargeted Lipidomics
STUDY_SUMMARY
Cytochrome P450 epoxygenase Cyp2c44 and their metabolite epoxyeicosatrienoic acids or EETs promotes insulin sensitivity. Mice lacking Cyp2c44 exhibits hepatic insulin resistance. Insulin resistance is also intricately related to increased hepatic lipid accumulation and hyperlipidemia. Interestingly, Cyp2c44(-/-) mice in standard chow diet had significantly increased hepatic and plasma lipid levels compared to wild-type mice. To identify the nature of these lipids, with a focus on fatty acids, we performed lipidomic analysis of liver homogenates from SD-fed WT and Cyp2c44(-/-) mice. We identified 2425 lipids (1152 in negative mode and 1273 in positive mode) that passed both quality control filters set as 25% for QC RSD and 10% for QC/blank ratio. Principal component analysis revealed two distinct lipid clusters in livers of WT and Cyp2c44(-/-) mice. Heatmap analysis revealed a hierarchical clustering of significant differences occurring in lipid species between WT and Cyp2c44(-/-) livers. Volcano plot analysis of the 1152 lipids identified in the negative mode (which contain fatty acids) revealed 160 lipid species upregulated, 61 downregulated, and 931 not significantly changed in Cyp2c44(-/-) livers compared to WT livers. Analysis of fatty acids classes in negative mode identified a total of 146 fatty acids, with 49 upregulated, 3 downregulated and 94 unchanged in Cyp2c44(-/-) compared to WT livers. Among the fatty acids that are significantly upregulated in the livers of Cyp2c44(-/-), we detected the saturated fatty acids palmitic acid; the monosaturated oleic acid; and the polyunsaturated arachidonic, linoleic, eicosapentaenoic and docosahexaenoic acids. Importantly, arachidonic acid is the major substrate of Cyp2c epoxygenases, although linoleic, eicosapentaenoic and docosahexaenoic acids are also efficient alternative substrates. We thus hypothesized that Cyp2c44 also governs hepatic lipid metabolism.
INSTITUTE
Vanderbilt University Medical Center
LAST_NAME
Ghoshal
FIRST_NAME
Kakali
ADDRESS
B-3106 Medical Center North, 1161 21st Ave South, Nashville, TN 37232
A Non-Targeted Metabolomics Comparative Study on Plasma of Pfizer and Sinopharm COVID- 19 Vaccinated individuals, Assessed by (TIMS-QTOF) Mass Spectrometry.
STUDY_SUMMARY
COVID-19 is a contagious globally threatening infectious disease that accounted for an ongoing pandemic that manifested in multi-organs diseases and failures. The current study aimed to investigate the effectiveness of the Pfizer and Sinopharm vaccines in relation to metabolomic alterations and their association with immune pathways. The study employed a cross-sectional design and utilized an untargeted metabolomics-based approach. Plasma samples were collected from three groups: non- vaccinated participants, Sinopharm vaccinated participants, and Pfizer vaccinated participants. Comparative metabolomic analysis was performed using TIMS-QTOF, and a one-way ANOVA test was conducted using MetaboAnalyst Software. Out of the 105 detected metabolites, 72 showed statistically significant alterations (p<0.05) among the different groups. Several metabolites, including neopterin, pyridoxal, and syringic acid, were highly altered in individuals vaccinated with Pfizer. On the other hand, sphinganine, neopterin, and sphingosine were impacted in individuals vaccinated with Sinopharm. These metabolites could potentially serve as biomarkers for vaccine efficacy. Furthermore, both Pfizer and Sinopharm vaccinations were found to affect sphingolipid metabolism pathways and histidine metabolism pathways when compared to the control group. The Sinopharm group exhibited altered lysine degradation compared to the control group. When comparing the enriched pathways of the Pfizer and Sinopharm groups, purine metabolism was found to be affected. Additionally, perturbations in tryptophan metabolism and vitamin B6 metabolism were observed when comparing the Pfizer group with both the control and Sinopharm groups. These findings highlight the importance of metabolomics in assessing vaccine effectiveness and identifying potential biomarkers.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
MM.1S Myeloma tumor cells and tumors made by subcutaneously injecting 1M Luc+/GFP+ MM.1S cells treated with 50 Ī¼M BMS309403 and Vehicle (PBS with a matched concentration of 5% DMSO)
STUDY_SUMMARY
MM.1S Myeloma tumor cells were treated with 50 Ī¼M BMS309403 once over 24 hours in vitro or at 5mg/kg X3 per week over 32 days in vivo. In Vivo tumors were made by subcutaneously injecting 1M Luc+/GFP+ MM.1S cells mixed with Matrigel in a 1:1 ratio into the backs of 8-week old, female SCID-Beige. Vehicle (PBS with a matched concentration of 5% DMSO) was used as control for each condition.
INSTITUTE
Mainehealth
LAST_NAME
Reagan
FIRST_NAME
Michaela
ADDRESS
81 Research Drive, Scarborough, ME, Portland, ME, 04074, USA
Soil pH, developmental stages and geographical origin differently influence the root metabolomic diversity and root-related microbial diversity of Echium vulgare from native habitats
STUDY_SUMMARY
This study examines the impact of different factors (geographic location, soil pH, genetic cluster, developmental stage) on the metabolome and microbiome of wild Echium vulgare, as well as the microbiome-metabolome correlation contained therein. Samples analyzed are wild Echium vulgare plant roots collected at six locations in Austria; out of the six locations, four contain plant roots from two different developmental stages. Extraction of metabolites and subsequent UHPLC-HRMS analysis were performed to obtain data for metabolomics analysis.
INSTITUTE
Aristotle University of Thessaloniki
LAST_NAME
RodiÄ
FIRST_NAME
NebojŔa
ADDRESS
Stepe StepanoviÄa 5, Äonoplja, Vojvodina, Serbia
The metabolomic resetting effect of DY131 in cisplatin-induced AKI
STUDY_TYPE
MS
STUDY_SUMMARY
LC-MS/MS analyses were performed using renal tissues from cisplatin-induced AKI mice with or without DY131 treatment. The data revealed that DY131 alleviated cisplatin-induced mitochondrial dysfunction and energy metabolism disorder, as well as multiple metabolic disorders.
INSTITUTE
Children's Hospital of Nanjing Medical University
DEPARTMENT
Department of Nephrology, State Key Laboratory of Reproductive Medicine
LABORATORY
Nanjing Key Lab of Pediatrics, Jiangsu Key Laboratory of Pediatrics
LAST_NAME
Lu
FIRST_NAME
Lingling
ADDRESS
Guangzhou Road 72, Nanjing, Jiangsu, 210000, China
Retinoic acid receptor alpha activity in proximal tubules prevents kidney injury and fibrosis
STUDY_SUMMARY
Retinoid levels of all-trans-retinol, retinoic acid, and retinyl palmitate were measured in the kidney and serum of GCERRARaD (kidney proximal tubule RARalpha knockout mice) females 3 days or 3 months post-tamoxifen (n=5/group) and age-matched Wild Type females (n=4).
Proteomic and metabolomic signatures of rectal tumor discriminate patients with different responses to preoperative radiotherapy
STUDY_SUMMARY
Background: Neoadjuvant radiotherapy (neo-RT) is widely used in locally advanced rectal cancer (LARC) as a component of radical treatment. Despite the advantages of neo-RT, which typically improves outcomes in LARC patients, the lack of reliable biomarkers that predict response and monitor the efficacy of therapy, can result in the application of unnecessary aggressive therapy affecting patientsā quality of life. Hence, the search for molecular biomarkers for assessing the radio responsiveness of this cancer represents a relevant issue. Methods: Here, we combined proteomic and metabolomic approaches to identify molecular signatures, which could discriminate LARC tumors with good and poor responses to neo-RT. Results: The integration of data on differentially accumulated proteins and metabolites made it possible to identify disrupted metabolic pathways and signaling processes connected with response to irradiation, including ketone bodies synthesis and degradation, purine metabolism, energy metabolism, degradation of fatty acid, amino acid metabolism, and focal adhesion. Moreover, we proposed multi-component panels of proteins and metabolites which could serve as a solid base to develop biomarkers for monitoring and predicting the efficacy of preoperative RT in rectal cancer patients. Conclusions: We proved that an integrated multi-omic approach presents a valid look at the analysis of the global response to cancer treatment from the perspective of metabolomic reprogramming.
INSTITUTE
Institute of Bioorganic Chemistry Polish Academy of Sciences
Multi-Omics Plasma Signatures of Severe Injury with Influence of REBOA Intervention in a Swine Model.
STUDY_TYPE
Original Research
STUDY_SUMMARY
Swine experienced controlled severe injury involving combinations of trauma, hemorrhagic shock, and disseminated complex blast injury (DCBI). After a period of hemorrhagic shock, resuscitation was performed using Resuscitative Endovascular Balloon Occlusion of the Aorta (REBOA) and shed blood transfusion. Blood samples were collected from sham and experiment swine. Blood samples for experiment swine were collected at baseline and periodically through the monitored time course. The plasma fraction were then subjected to mass spectrometry based metabolomics. Results indicated intervention using REBOA following polytrauma in our swine model induced distinct multi-omics alterations as a function of placement location. Namely, aortic balloon placement in Zone-1 worsened ischemic damage and mitochondrial dysfunction, patterns that continued through the monitored time course.
Defective mitochondria remodelling in B cells leads to an aged immune response
STUDY_SUMMARY
The germinal centre (GC) reaction requires a unique bioenergetic supply. Although mitochondria are remodelled upon antigen stimulation, mitochondrial function in B cells is still poorly understood. To gain a better understanding of the role of mitochondria in B cell function, we generated mice that lack, specifically in B cells, Tfam, a transcription factor necessary for mitochondrial biogenesis. Tfam knock-out (KO) mice displayed a blockage of the GC reaction and established an immune response featured by the differentiation of activated B cells towards memory B cells and aged-related B cells, hallmarks of an aged immune response. Unexpectedly, GC blockage in Tfam KO mice did not cause defects in the bioenergetic supply, but this phenotype was associated with a defect in the remodelling of the lysosomal compartment in B cells. Therefore, these results may describe a new mitochondrial function for antigen presentation during the GC reaction, the abrogation of which may be the basis of an aged immune response.
Identification and validation of serum metabolite biomarkers for endometrial cancer diagnosis
STUDY_TYPE
Biomarker, Endometrial cancer, Machine learning, Mass spectrometry, Metabolite
STUDY_SUMMARY
Endometrial cancer (EC) stands as the most prevalent gynecological tumor in women worldwide. Notably, differentiation diagnosis of abnormity detected by ultrasound findings (e.g., thickened endometrium or mass in the uterine cavity) is essential and remains challenging in clinical practice. Herein, we identified a metabolic biomarker panel for differentiation diagnosis of EC using machine learning of high-performance serum metabolic fingerprints (SMFs) and validated the biological function. We first recorded the high-performance SMFs of 191 EC and 204 Non-EC subjects via particle-enhanced laser desorption/ionization mass spectrometry (PELDI-MS). Then, we achieved an area-under-the-curve (AUC) of 0.957-0.968 for EC diagnosis through machine learning of high-performance SMFs, outperforming the clinical biomarker of cancer antigen 125 (CA-125, AUC of 0.610-0.684, P < 0.05). Finally, we identified a metabolic biomarker panel of glutamine, glucose, and cholesterol linoleate with an AUC of 0.901-0.902 and validated the biological function in vitro. Therefore, our work would facilitate the development of novel diagnostic biomarkers for EC in clinics.
Plasma instead of serum avoids critical confounding of clinical metabolomics studies by platelets (Part 2/3 - Eicosadomics of isolated platelets)
STUDY_SUMMARY
Metabolomics is an emerging and powerful molecular profiling method supporting clinical investigations. Serum and plasma are commonly used without rational prioritization. Serum is collected after blood coagulation, a complex biochemical process involving active platelet metabolism. This may affect the metabolome and increase the variance as platelet counts and function may vary substantially in individuals. A multi-omics approach systematically investigating the suitability of serum and plasma for clinical studies demonstrated that metabolites correlated well (n=461, R2=0.991), whereas lipid mediators (n=104, R2=0.906) and proteins (n=322, R2=0.860) differed substantially between specimen. Independently, analysis of platelet releasates identified most biomolecules significantly enriched in serum when compared to plasma. A prospective, randomized, controlled parallel group metabolomics trial with acetylsalicylic acid administered for 7 days demonstrated that the apparent drug effects significantly differ depending on analyzed specimen. Only serum analyses of healthy individuals suggested a significant downregulation of TXB2 and 12-HETE, which were specifically formed during coagulation in vitro. Plasma analyses reliably identified acetylsalicylic acid effects on metabolites and lipids occurring in vivo such as a decrease in polyunsaturated fatty acids. The present data suggests that plasma should be preferred above serum for clinical metabolomics studies as the serum metabolome may be substantially confounded by platelets.
Plasma instead of serum avoids critical confounding of clinical metabolomics studies by platelets (Part 1/3 - Plasma and serum eicosadomics)
STUDY_SUMMARY
Metabolomics is an emerging and powerful molecular profiling method supporting clinical investigations. Serum and plasma are commonly used without rational prioritization. Serum is collected after blood coagulation, a complex biochemical process involving active platelet metabolism. This may affect the metabolome and increase the variance as platelet counts and function may vary substantially in individuals. A multi-omics approach systematically investigating the suitability of serum and plasma for clinical studies demonstrated that metabolites correlated well (n=461, R2=0.991), whereas lipid mediators (n=104, R2=0.906) and proteins (n=322, R2=0.860) differed substantially between specimen. Independently, analysis of platelet releasates identified most biomolecules significantly enriched in serum when compared to plasma. A prospective, randomized, controlled parallel group metabolomics trial with acetylsalicylic acid administered for 7 days demonstrated that the apparent drug effects significantly differ depending on analyzed specimen. Only serum analyses of healthy individuals suggested a significant downregulation of TXB2 and 12-HETE, which were specifically formed during coagulation in vitro. Plasma analyses reliably identified acetylsalicylic acid effects on metabolites and lipids occurring in vivo such as a decrease in polyunsaturated fatty acids. The present data suggests that plasma should be preferred above serum for clinical metabolomics studies as the serum metabolome may be substantially confounded by platelets.
Shotgun lipidomics of breast cancer endocrine therapy persisters
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Despite adjuvant endocrine therapy, the rate of ER+ breast cancer recurrence remains high. The metabolic state of cells that remain after endocrine therapy, specifically oxidative stress, is poorly understood. We demonstrate that endocrine-tolerant persister cells are oxidatively stressed after initiation of endocrine therapy. Furthermore, modulation of redox levels similarly modifies persister cell fitness. We find ferroptosis sensitivity and lipid peroxidation to be a dominant factor in the oxidative state, and persister cells are found to be sensitive to ferroptosis induction compared to parental cells. Persister cells exhibited an altered lipidome, with an increase in phospholipids having a predilection for peroxidation. We hypothesized that ferroptosis induction would be well-coordinated with endocrine therapy. Xenografts treated with a combination of fulvestrant and ferroptosis induction exhibited the greatest treatment effect, supporting the role of ferroptosis as a therapeutic strategy in ER+ breast cancer.
Synaptic vesicle lipidomics captures signatures of aging and synucleinopathy
STUDY_SUMMARY
Untargeted synaptic vesicle lipidomics analysis: Synaptic vesicles were isolated from six animals each of 1-month, 3-month, and 10-month old wild-type mice (JAX strain #: 000664), Ī±-synuclein knockout mice (JAX strain #: 023837), and Ī±-synuclein BAC mice (JAX strain #: 023837) using subcellular fractionation. Vesicles were concentrated using 3 kDa cutoff Amicon ultra centrifugal devices (Millipore). Pellets were resuspended in 100 Āµl water and homogenized. Protein concentration was determined using absorption at 280 nm. Samples were adjusted to the same concentration to a final volume of 200 Āµl in 50% methanol. All samples were mixed with a panel of 15 lipid internal standards (Splash LipidMix lot#330707), followed by lipid extraction as previously reported.1 Lipids were analyzed by LC-MS/MS on an Accucore C30 column (2.1 mm id x 150mm) under both positive and negative ion modes. Raw data files were searched and processed using LipidSearch 5.1 (Thermo Scientific) and normalized against the known amount of internal standard lipids for each sample, allowing for reliably quantitative measurements of all 15 categories of lipids.1 1 Bhawal, R., Fu, Q., Anderson, E.T., Gibson, G.E., and Zhang, S. (2021). Serum Metabolomic and Lipidomic Profiling Reveals Novel Biomarkers of Efficacy for Benfotiamine in Alzheimer's Disease. Int J Mol Sci 22.
Providing insight into the mechanism of action of Cationic Lipidated Oligomers (CLOs) using metabolomics
STUDY_TYPE
Biomedical research
STUDY_SUMMARY
The increasing resistance of clinically relevant microbes against current commercially available antimicrobials underpins the urgent need for alternative and novel treatment strategies. Cationic lipidated oligomers (CLOs) are innovative alternatives to antimicrobial peptides, and have reported antimicrobial potential. An understanding of their antimicrobial mechanism of action is required to rationally design future treatment strategies for CLOs, either in monotherapy or synergistic combinations. In the present study, metabolomics was used to investigate the potential metabolic pathways involved in the mechanisms of antibacterial activity of one CLO, C12-o-(BG-D)-10, which we have previously shown to be effective against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300. The metabolomes of MRSA ATCC 43300 at 1, 3 and 6 h following treatment with C12-o-(BG-D)-10 (48 Āµg/mL i.e., 3x MIC) were compared to those of the untreated controls. Our findings reveal that the studied CLO, C12-o-(BG-D)-10, disorganized the bacterial membrane as the first step towards its antimicrobial effect, as evidenced by marked perturbations in the bacterial membrane lipids and peptidoglycan biosynthesis observed at early time points i.e., 1, and 3 h. Central carbon metabolism, and biosynthesis of DNA, RNA, and arginine were also vigorously perturbed, mainly at early time points. Moreover, bacterial cells were under osmotic and oxidative stress across all time points, evident by perturbations of trehalose biosynthesis and pentose phosphate shunt. Overall, this metabolomics study has, for the first time, revealed that the antimicrobial action of C12-o-(BG-D)-10 may potentially stem from the dysregulation of multiple metabolic pathways.
INSTITUTE
Monash University
DEPARTMENT
Drug Delivery, Disposition and Dynamics
LABORATORY
Cornelia Landersdorfer
LAST_NAME
Hussein
FIRST_NAME
Maytham
ADDRESS
Monash Biomedicine Discovery Institute, Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia
Integrated metabolomics and proteomics of symptomatic and early pre-symptomatic states of colitis
STUDY_SUMMARY
Colitis has a multifactorial pathogenesis with a strong cross-talk among microbiota, hypoxia and tissue metabolism. Here, we aimed to characterize the molecular signature of the disease in symptomatic and pre-symptomatic stages of the inflammatory process at the tissue and fecal level. The study is based on two different murine models for colitis. High-resolution Magic Angle Spinning NMR on cryopreserved āintactā colon tissues and LC-MS/MS on colon tissue extracts were used to derive untargeted metabolomics and proteomics information, respectively. Solution NMR was used to derive metabolomic profiles of fecal extracts. By combining metabolomic and proteomic analyses of the tissues, we found increased anaerobic glycolysis, accompanied by altered citric acid cycle and oxidative phosphorylation in inflamed colons; these changes associate with inflammation-induced hypoxia taking place in colon tissues. Pre-symptomatic states can be discriminated from healthy samples before macroscopic inflammation is observed. Different colitis states are characterized by significantly different metabolomic profiles of fecal extracts, attributable to both the dysbiosis characteristic of colitis, as well as the dysregulated tissue metabolism. Strong and distinctive fecal metabolomic signatures can be detected before onset of symptoms. Therefore, untargeted metabolomics of tissues and fecal extracts provides a comprehensive picture of the changes accompanying the disease onset already at pre-clinical stages, highlighting the diagnostic potential of global metabolomics for inflammatory diseases.
INSTITUTE
University of Florence
LAST_NAME
Ghini
FIRST_NAME
Veronica
ADDRESS
via Luigi Sacconi, 6, Sesto Fiorentino, Firenze, 50019, Italy
Untargeted metabolomics revealed multiple metabolic perturbations in plasma of T2D patients in response to Liraglutide - Part 2
STUDY_SUMMARY
Despite the global efforts put into the clinical research and studies in order to protect against Type-2 diabetes mellitus (T2DM), the incidence of T2DM remains high causing a major health problem and impacting the health and care systems. Therefore, T2DM-related treatments and therapies are continuously invented for the clinical use, including Liraglutide. The last is a GLP-1 analogue and shows its beneficial health outcomes e.g., improved glycemic control, lower body weight, and reduced cardiovascular disease risks. The intrinsic mechanisms of these beneficial effects are not fully understood; however, our research group has previously published proteomics work demonstrating the involvement of certain important proteins in part in the beneficial health outcomes of Liraglutide. Since proteomics and metabolomics are complementary to each other in the context of the biological pathways, studying the metabolic impacts of Liraglutide on T2DM patients would add further information about the beneficial health outcomes of Liraglutide. Thus, herein, we performed an untargeted metabolomics approach for identifying metabolic pathways impacted by the treatment of Liraglutide in T2DM patients. Methods: Untargeted liquid chromatography coupled with mass spectrometry was used for metabolomics analysis of plasma samples collected from T2DM patients (n=20) before and after receiving Liraglutide treatment. Metabolic profiling and related pathway and network analyses were conducted. Results: The metabolic profiling analyses identified 93 endogenous metabolites were significantly affected by the Liraglutide treatments, which 49 metabolites up-regulated and 44 metabolites down-regulated. Moreover, the metabolic pathway analyses revealed that the most pronounced metabolite and metabolic pathways that were affected by the Liraglutide treatment was Pentose and glucuronate interconversion, suggesting the last may be a potential target of the Liraglutide treatment could be involved in part in the beneficial effects seen in T2DM patients, specially, we found that glucuronate interconversion pathway which is known by its role in eliminating toxic and undesirable substances from the human body, impacted in Liraglutide treated patients. The last findings ar consistence with our previous proteomics findings. Conclusion: These findings, taken together with our previous results, provide a deeper understanding of the underlying mechanisms involved in the beneficial effects of Liraglutide at the proteomic and metabolic levels in T2DM patients.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
LAST_NAME
Al Mogren
FIRST_NAME
Maha
ADDRESS
Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia
Spatiotemporal mapping of lipid disturbance in heart injury - Part 1
STUDY_SUMMARY
The project consists of 3 studies in total, in this study, mouse tissue were collected for LC-MS/MS based lipidomics. The heart tissue underwent I/R injury at different time points (0h, 3h, 24h, 3d, 7d) were separate into infarct, remote and border area for investigation.
Spatiotemporal mapping of lipid disturbance in heart injury - Part 2
STUDY_SUMMARY
The project consists of 3 studies in total, this is the second part, in this study, mouse plasma were collected for LC-MS/MS based lipidomics. The plasma sample from mouse underwent I/R injury at different time points (0h, 3h, 24h, 3d, 7d) were collected for investigation.
Spatiotemporal mapping of lipid disturbance in heart injury - Part 3
STUDY_SUMMARY
The project consists of 3 studies in total, this is the third part, in this study, pig plasma were collected for LC-MS/MS based lipidomics. The plasma sample from pig underwent I/R injury at different time points were collected for investigation.
Fluxomics of hormone deprivation in ER+ breast cancer cell lines
STUDY_TYPE
Fluxomics
STUDY_SUMMARY
Despite adjuvant treatment with endocrine therapies, estrogen receptor-positive (ER+) breast cancers recur in a significant proportion of patients. Recurrences are attributable to clinically undetectable endocrine-tolerant persister cancer cells that retain tumor-forming potential. We observed that persistence occurred stochastically without genetic predisposition. Genome-wide screening in persisters revealed a survival mechanism involving metabolic reprogramming with reliance upon mitochondrial respiration. Proteomic profiling showed reduced levels of glycolytic proteins in persisters. Metabolic tracing of glucose revealed an energy-depleted state in persisters where oxidative phosphorylation was required to generate ATP. Persisters exhibiting residual proliferation in human breast tumors following neoadjuvant endocrine therapy showed increased mitochondrial content. Pharmacological inhibition of oxidative phosphorylation suppressed the tumor-forming potential of persisters and synergized with the anti-estrogen fulvestrant to induce regression of patient-derived xenografts, supporting therapeutic targeting of mitochondrial metabolism to help eradicate residual disease.
Untargeted metabolomics analysis of three tea cultivars.
STUDY_TYPE
leaf tissue
STUDY_SUMMARY
To compare flavonoid metabolism in three tea cultivars, namely Camellia sinensis cv. "Jinfenghuang" (JFH), "Fuyun 6" (FY6) and "Bantianyao" (BTY), fresh tea leaves were plucked, rapidly ground to fine powders in liquid nitrogen and subjected to extraction by 70% methanol. The extracts were analyzed by UPLC-Quadrupole Time-of-Flight Mass Spectrometry (UPLC-QTOFMS) using an ACQUITY UPLC system in tandem to a SYNAPT G2-Si QTOF mass spectrometer (Waters, Milford, MA, USA), following the previously described method. Raw data were processed in Progenesis QI (v2.4, Nonlinear Dynamics) with default settings for peak picking and alignment. Metabolites were annotated by searching against our in-house MS/MS spectral library and public databases.
INSTITUTE
Fujian Agriculture and Forestry University
LAST_NAME
Yu
FIRST_NAME
Xiaomin
ADDRESS
Shangxia Dian Road No. 15, Fuzhou 350002, Fujian, China
Identification of Plasma Metabolites and Peptides as Diagnostic Biomarkers for Psoriasis Vulgaris through HILIC-HRMS-Based Metabolomics
STUDY_SUMMARY
In this work, we explored plasma metabolite and peptide biomarkers for PS through a three-step study using HILIC-LCHRMS based metabolomics. This involved a small-scale case-control study for discovery purposes, subsequent validation of significantly changed metabolic features using another larger group, and further refinement of the differential features using urticaria patient samples for psoriasis specificity.Finally,We identified 9 potential biomarkers specific to psoriasis, with 5 being structurally characterized. Receiver operating characteristic curve analysis revealed all these 9 biomarkers with AUC values above 0.80.
Targeted metabolomics in serum from high-fat diet fed control and MBCUCP1 KO mice. N = 6 for control and 7 for MBCUCP1 KO. Statistic: unpaired t-test. Data represented as z-score heatmap with each cell representing quantitated value for each mouse.
Metabolic responses of Amaranthus caudatus roots and leaves to zinc stress
STUDY_TYPE
GCMS-based untargeted and targeted analysis
STUDY_SUMMARY
During the last decades pollution with heavy metals became an important stress factor. Plants are characterized by significant biochemical plasticity and can adjust their metabolism to ensure survival under changing environmental conditions. In the most straightforward way these metabolic shifts can be addressed by the untargeted mass spectrometry-based metabolomics approach. However, so far this methodology was only minimally employed in studies of Zn-induced metabolic shifts in plants. Moreover, the genus Amaranthus is still not addressed in this respect. Therefore, here we propose, to the best of our knowledge, the first gas chromatography-mass spectrometry (GC-MS)-based metabolomics study of Zn2+-induced stress responses in Amaranthus caudatus plants. The GC-MS-based study was performed with root and leaf aqueous methanolic extracts after their lyophylization and sequential derivatization with methoxylamine hydrochloride and N-trimethylsilyl-N-methyl trifluoroacetamide. Thereby, 419 derivatives were detected, of which 144 could be putatively annotated. The metabolic shifts in seven-week old A.caudatus plants in response to a seven-day treatment with 300 Āµmol/L ZnSO4Ā·7H2O in nutrient solution were organ-specific and more pronounced in roots. The most of the responsive metabolites were up-regulated and dominated with sugars and sugar acids. These effects could be attributed to the involvement of these metabolites in osmoregulation, ROS scavenging and complexation of Zn2+ ions. Galactose was the most Zn2+-responsive root sugar that indicated its possible role in the binding of Zn2+ ions to the root cell walls. A 59-fold up-regulation of gluconic acid in roots clearly indicated its involvement in chelation of Zn2+. A high Zn2+āinduced up-regulation of salicylic acid in roots and shoots suggested a key role of this hormone in the activation of Zn2+ stress tolerance mechanisms. Thus, our study provides the first insight in the general trends in Zn-induced biochemical rearrangements and main adaptive metabolic shifts in A. caudatus plants.
INSTITUTE
K.A. Timiryazev Institute of Plant Physiology RAS, Moscow, Russia
LABORATORY
Laboratory of Analytical Biochemistry and Biotechnology
LAST_NAME
Frolov
FIRST_NAME
Andrej
ADDRESS
Botanicheskaya st. 35., Moskow, 127276, Russian Federation
Investigative needle core biopsies for multi-omics in Glioblastoma
STUDY_SUMMARY
Glioblastoma (GBM) is a primary brain cancer with an abysmal prognosis and few effective therapies. The ability to investigate the tumor microenvironment before and during treatment would greatly enhance both our understanding of disease response and progression, as well as the delivery and impact of therapeutics. Stereotactic biopsies are a routine surgical procedure performed primarily for diagnostic histopathologic purposes. The role of investigative biopsies ā tissue sampling for the purpose of understanding tumor microenvironmental responses to treatment using integrated multi-modal molecular analyses (āMulti-omicsā) has yet to be defined. Here we adapt stereotactic needle core biopsy tissue for highly resolved multi-omics analysis methods including single cell RNA sequencing, spatial-transcriptomics, metabolomics, proteomics, phosphoproteomics, T-cell clonotype analysis, and MHC Class I immunopeptidomics. Biopsy tissue was obtained from a single patient with recurrent GBM during one procedure. In a second patient, we analyzed multi-regional core biopsies to decipher spatial and genomic variance. Finally in a separate cohort of patients we investigated the utility of stereotactic biopsies as a method for generating patient derived xenograft models. Dataset integration across modalities showed good correspondence between spatial modalities and highlighted immune cell associated metabolic pathways and poor correlation between RNA expression and the tumor MHC Class I immunopeptidome. In conclusion, stereotactic needle biopsy cores are of sufficient quality for the purposes of investigative biopsy and can generate multi-omics data, providing data rich insight into a patientās disease process and tumor immune microenvironment and could be of potential value in evaluating treatment responses.
Heritability of RBC metabolites: baseline correlation of metabolites and markers of RBC health and stability
STUDY_SUMMARY
The goal of this study was to examine the genetic heritability of metabolites and to identify metabolites, present immediately after blood draw, that were highly correlated with red blood cells (RBCs) ex vivo survival. Extreme longevity in humans is known to be a heritable trait. In a well-established twin erythrocyte metabolomics and proteomics database, we identified the longevity factor spermidine and a cluster of correlated molecules with high heritability estimates. Erythrocyte spermidine is 82% heritable and significantly correlated with 59 metabolites and 22 proteins. Thirty-eight metabolites and 19 proteins were >20% heritable, with a mean heritability of 61% for metabolites and 49% for proteins. Correlated metabolites are concentrated in energy metabolism, redox homeostasis, and autophagy pathways. Erythrocyte mean cell volume (MCV)āan established heritable traitāwas consistently negatively correlated with the top 25 biomolecules most strongly correlated with spermidine, indicating that smaller MCVs are associated with higher concentrations of spermidine and correlated molecules.
Attenuation of Helicobacter pylori VacA toxin-induced cell death by modulation of intracellular taurine metabolism - Study #1
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Colonization of the human stomach with H. pylori strains producing active forms of a secreted toxin (VacA) is associated with an increased risk of peptic ulcer disease and gastric cancer, compared to colonization with strains producing hypoactive forms of VacA. Previous studies have shown that VacA causes cell vacuolation and mitochondrial dysfunction. In this study, we sought to define the cellular metabolic consequences of VacA intoxication. Untargeted metabolomic analyses revealed that several hundred metabolites were significantly altered in VacA-treated gastroduodenal cells (AGS and AZ-521) compared to control cells. Pathway analysis suggested that VacA caused alterations in taurine and hypotaurine metabolism. Treatment of cells with the purified active s1m1 form of VacA, but not hypoactive s2m1 or 6-27 VacA mutant proteins (defective in membrane channel formation), caused reductions in taurine and hypotaurine levels. Supplementation of the tissue culture medium with taurine or hypotaurine protected AZ-521 cells against VacA-induced cell death. Untargeted global metabolomics of AZ-521 cells or AGS cells intoxicated with VacA in the presence or absence of extracellular taurine showed that taurine was the main intracellular metabolite significantly altered by extracellular taurine supplementation. These results indicate that VacA causes alterations in cellular taurine metabolism and indicate that repletion of taurine is sufficient to attenuate VacA-induced cell death.
Attenuation of Helicobacter pylori VacA toxin-induced cell death by modulation of intracellular taurine metabolism _ Study #2
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Colonization of the human stomach with H. pylori strains producing active forms of a secreted toxin (VacA) is associated with an increased risk of peptic ulcer disease and gastric cancer, compared to colonization with strains producing hypoactive forms of VacA. Previous studies have shown that VacA causes cell vacuolation and mitochondrial dysfunction. In this study, we sought to define the cellular metabolic consequences of VacA intoxication. Untargeted metabolomic analyses revealed that several hundred metabolites were significantly altered in VacA-treated gastroduodenal cells (AGS and AZ-521) compared to control cells. Pathway analysis suggested that VacA caused alterations in taurine and hypotaurine metabolism. Treatment of cells with the purified active s1m1 form of VacA, but not hypoactive s2m1 or 6-27 VacA mutant proteins (defective in membrane channel formation), caused reductions in taurine and hypotaurine levels. Supplementation of the tissue culture medium with taurine or hypotaurine protected AZ-521 cells against VacA-induced cell death. Untargeted global metabolomics of AZ-521 cells or AGS cells intoxicated with VacA in the presence or absence of extracellular taurine showed that taurine was the main intracellular metabolite significantly altered by extracellular taurine supplementation. These results indicate that VacA causes alterations in cellular taurine metabolism and indicate that repletion of taurine is sufficient to attenuate VacA-induced cell death.
Plasma instead of serum avoids critical confounding of clinical metabolomics studies by platelets (Part 3/3 - Plasma and serum metabolomics)
STUDY_SUMMARY
Metabolomics is an emerging and powerful molecular profiling method supporting clinical investigations. Serum and plasma are commonly used without rational prioritization. Serum is collected after blood coagulation, a complex biochemical process involving active platelet metabolism. This may affect the metabolome and increase the variance as platelet counts and function may vary substantially in individuals. A multi-omics approach systematically investigating the suitability of serum and plasma for clinical studies demonstrated that metabolites correlated well (n=461, R2=0.991), whereas lipid mediators (n=104, R2=0.906) and proteins (n=322, R2=0.860) differed substantially between specimen. Independently, analysis of platelet releasates identified most biomolecules significantly enriched in serum when compared to plasma. A prospective, randomized, controlled parallel group metabolomics trial with acetylsalicylic acid administered for 7 days demonstrated that the apparent drug effects significantly differ depending on analyzed specimen. Only serum analyses of healthy individuals suggested a significant downregulation of TXB2 and 12-HETE, which were specifically formed during coagulation in vitro. Plasma analyses reliably identified acetylsalicylic acid effects on metabolites and lipids occurring in vivo such as a decrease in polyunsaturated fatty acids. The present data suggests that plasma should be preferred above serum for clinical metabolomics studies as the serum metabolome may be substantially confounded by platelets.
Attenuation of Helicobacter pylori VacA toxin-induced cell death by modulation of intracellular taurine metabolism - Study #3
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Colonization of the human stomach with H. pylori strains producing active forms of a secreted toxin (VacA) is associated with an increased risk of peptic ulcer disease and gastric cancer, compared to colonization with strains producing hypoactive forms of VacA. Previous studies have shown that VacA causes cell vacuolation and mitochondrial dysfunction. In this study, we sought to define the cellular metabolic consequences of VacA intoxication. Untargeted metabolomic analyses revealed that several hundred metabolites were significantly altered in VacA-treated gastroduodenal cells (AGS and AZ-521) compared to control cells. Pathway analysis suggested that VacA caused alterations in taurine and hypotaurine metabolism. Treatment of cells with the purified active s1m1 form of VacA, but not hypoactive s2m1 or 6-27 VacA mutant proteins (defective in membrane channel formation), caused reductions in taurine and hypotaurine levels. Supplementation of the tissue culture medium with taurine or hypotaurine protected AZ-521 cells against VacA-induced cell death. Untargeted global metabolomics of AZ-521 cells or AGS cells intoxicated with VacA in the presence or absence of extracellular taurine showed that taurine was the main intracellular metabolite significantly altered by extracellular taurine supplementation. These results indicate that VacA causes alterations in cellular taurine metabolism and indicate that repletion of taurine is sufficient to attenuate VacA-induced cell death.
Attenuation of Helicobacter pylori VacA toxin-induced cell death by modulation of intracellular taurine metabolism - Study #4
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Colonization of the human stomach with H. pylori strains producing active forms of a secreted toxin (VacA) is associated with an increased risk of peptic ulcer disease and gastric cancer, compared to colonization with strains producing hypoactive forms of VacA. Previous studies have shown that VacA causes cell vacuolation and mitochondrial dysfunction. In this study, we sought to define the cellular metabolic consequences of VacA intoxication. Untargeted metabolomic analyses revealed that several hundred metabolites were significantly altered in VacA-treated gastroduodenal cells (AGS and AZ-521) compared to control cells. Pathway analysis suggested that VacA caused alterations in taurine and hypotaurine metabolism. Treatment of cells with the purified active s1m1 form of VacA, but not hypoactive s2m1 or 6-27 VacA mutant proteins (defective in membrane channel formation), caused reductions in taurine and hypotaurine levels. Supplementation of the tissue culture medium with taurine or hypotaurine protected AZ-521 cells against VacA-induced cell death. Untargeted global metabolomics of AZ-521 cells or AGS cells intoxicated with VacA in the presence or absence of extracellular taurine showed that taurine was the main intracellular metabolite significantly altered by extracellular taurine supplementation. These results indicate that VacA causes alterations in cellular taurine metabolism and indicate that repletion of taurine is sufficient to attenuate VacA-induced cell death.
Investigation of polar metabolites by targeted LC-MS analysis from mouse adult or embryonic CSF and from adult serum.
STUDY_SUMMARY
To maximize discovery from samples with limited amounts, we optimized a method to detect the thyroid hormones ( 3,5,3ā-L-triiodothyronine (T3) and 3,5,3ā5ā-L-tetraiodothyronine (T4)) as well as polar metabolites from central carbon metabolism from mouse adult or embryonic CSF and from adult mouse serum. Samples were ran on reverse phase and HILIC chromatography respectively. For polar metabolite analysis, we employed targeted metabolomics profiling of a panel of 200 compounds. We interrogated relative changes between fresh and frozen adult (male or female) CSF compared to fresh and frozen embryonic CSF. This data presents results from polar HILIC chromatography
INSTITUTE
Boston Childrens Hospital
DEPARTMENT
Pathology
LABORATORY
Kanarek Lab
LAST_NAME
Petrova
FIRST_NAME
Boryana
ADDRESS
300 Longwood Av
EMAIL
boryana.petrova@childrens.harvard.edu
PHONE
617
NUM_GROUPS
7
TOTAL_SUBJECTS
19
NUM_MALES
8
NUM_FEMALES
6
STUDY_COMMENTS
embryos also investigated. samples split for fresh and frozen conditions
Investigation of 3,5,3ā-L-triiodothyronine (T3) and 3,5,3ā5ā-L-tetraiodothyronine (T4) by targeted LC-MS analysis from mouse adult or embryonic CSF and from adult serum.
STUDY_SUMMARY
To maximize discovery from samples with limited amounts, we optimized a method to detect the thyroid hormones ( 3,5,3ā-L-triiodothyronine (T3) and 3,5,3ā5ā-L-tetraiodothyronine (T4)) as well as polar metabolites from central carbon metabolism from mouse adult or embryonic CSF and from adult mouse serum. Samples were ran on reverse phase and HILIC chromatography respectively. For T3 and T4 analysis we utilized targeted analysis using information from previously ran synthetic standards. We interrogated relative changes between fresh and frozen adult (male or female) CSF compared to fresh and frozen embryonic CSF. This data presents the levels of T3 and T4 as interrogated by reverse phase chromatography
INSTITUTE
Boston Childrens Hospital
DEPARTMENT
Pathology
LABORATORY
Kanarek Lab
LAST_NAME
Petrova
FIRST_NAME
Boryana
ADDRESS
300 Longwood Av
EMAIL
boryana.petrova@childrens.harvard.edu
PHONE
6173557433
NUM_GROUPS
7
TOTAL_SUBJECTS
19
NUM_MALES
8
NUM_FEMALES
6
STUDY_COMMENTS
embryos also investigated. samples split for fresh and frozen conditions
Spectral information for 3,5,3ā-L-triiodothyronine (T3) and 3,5,3ā5ā-L-tetraiodothyronine (T4) from synthetic standards
STUDY_SUMMARY
We utilized synthetic standards for 3,5,3ā-L-triiodothyronine (T3) and 3,5,3ā5ā-L-tetraiodothyronine (T4) to obtain high-quality MS2-level information that would aid compound identification from biological matrixes
In this study, a subset of plasma samples were processed using an in-house two phase extraction protocol based on the protocol originally proposed by Matayash et al. The resulting non-polar layer was then analyzed on a Thermo ScientificTM TranscendTM Duo LX-2 UHPLC system interfaced with high resolution Thermo ScientificTM Orbitrap ID-XTM TribidTM mass spectrometer with a HESI ionization source. This dataset was collected as a single batch and was used for the pipeline's ability to detect failed injections, which in this study, was simulated through the substitution of an empty vial for a missing study sample.
Data-dependent and -independent acquisition lipidomics analysis reveals the tissue-dependent effect of metformin on lipid metabolism
STUDY_SUMMARY
Free fatty acids and polar metabolite analysis were conducted on six mouse tissues and plasma using HILIC chromatography coupled to a Q Exactive Plus mass spectrometer. This enabled a comprehensive assessment of fatty acid and metabolic changes induced by metformin across various tissues.
Data-dependent and -independent acquisition lipidomics analysis reveals the tissue-dependent effect of metformin on lipid metabolism (Adipose tissue measurements)
STUDY_SUMMARY
Lipidomics analysis of 6 mouse tissues and plasma using MS/MS combining BRI-DIA and DDA allowed a systemic evaluation of lipidomic changes induced by metformin in different tissues. We observed that 1) the degrees of lipidomic changes induced by metformin treatment overly correlated with tissue concentrations of metformin; 2) the impact on lysophosphorylcholine and cardiolipins was positively correlated with tissue concentrations of metformin, while neutral lipids such as triglycerides did not correlate with the corresponding tissue metformin concentrations.
Dynamic exometabolomics reveals metabolic adaptations of Staphylococcus epidermidis to pH-mimicking skin and bloodstream
STUDY_SUMMARY
A common human skin coloniser, Staphylococcus epidermidis (SE), is often the cause of infections associated with medical devices. Strains that retain this pathogenic and commensal potential coexist in human skin, belonging to clonal lineages A/C and B, respectively. The pH is an abiotic factor that changes during infection when SE is transferred from host skin to blood. However, pathogenicity mechanisms are poorly understood in this organism and recognizing how it deals with increments of pH, is relevant to design effective prevention and treatment strategies against SE infections. To investigate the metabolic adaptations of A/C and B representative strains to an increase in pH, we mimicked the pH conditions of skin and blood (5.5 and 7.4). Biomass formation, growth media pH and 1H-NMR exometabolomic data were measured until the stationary phase was reached, at the two selected pH values. Higher biomass was reached for both strains when grown at pH 7.4. For all experimental conditions, media pH significantly changed during growth, revealing several pH adaptation mechanisms. SE metabolism relies on saccharides and amino acids uptake, citric acid cycle, mixed acid and alcoholic fermentations at a higher extent in blood than at skin pH. This study highlighted accumulation of extracellular formate by the pathogenic strain at blood pH, consistent with the absence of a formate dehydrogenase gene in this strain. This, most likely constitutes a virulence factor that, together with higher media acidification, impacts on host invasion. This work depicts the relevance of specific metabolic processes in determining SE commensalism and pathogenicity, thus providing helpful information towards new design strategies against SE infections.
Targeted MS analysis of maternal liver and serum and embryonic CSF and liver, 3h post saline or PolyI:C administrated to mother. Metabolome changes in embryonic CSF (E12.5), embryonic liver (E12.5), maternal liver or maternal serum 3h hours following PolyI:C or saline injection in mother. Older experiment run included of embryonic CSF at 48 hour collection point, mice at E14.5
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Targeted MS analysis of embryonic CSF. Metabolome changes in embryonic CSF and serum and maternal serum at E14.5, 48h post Saline or PolyI:C injected into mother.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Untargeted MS analysis of embryonic and maternal CSF. Metabolome changes in embryonic and maternal CSF and serum at E14.5, Saline VS PolyI:C injected into mother, collection at 48 hours.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Untargeted MS analysis of maternal liver, CSF, serum, and embryonic CSF and liver, with saline and PolyI:C administrated to mother. Metabolome changes in embryonic CSF 3 hours prior to PolyI:C injection in mother. Metabolome changes in embryonic CSF, maternal and embryonic liver 48 hours following PolyI:C injection in mother.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Untargeted MS analysis of embryonic and maternal CSF. Generating a reference metabolome database for embryonic CSF from CD-1 mice. Samples include wild type embryonic CSF and mock treated empty tubes. Samples were extensively analyzed and curated using untargeted metabolomics analysis.
INSTITUTE
Boston Children's Hospital, Harvard Medical School
Analysis of lipid profiles of N2A-Pz1-KO cells expressing tdTOMATO-vector (mock), TMEM120A, or TMEM120B
STUDY_TYPE
Lipidomic
STUDY_SUMMARY
N2A-Pz1-KO cells expressing tdTOMATO-vector (mock), TMEM120A, or TMEM120B were used to detect differences in levels of lipids. TMEM120A is a protein without a clear function that we previously identified to inhibit mechanically activated PIEZO2 channels. TMEM120A has structural similarity a lipid modifying enzyme (ELOVL7), and has been proposed to be necessary for adipocyte maturation & triglyceride production. We performed LC-MS/MS to determine whether TMEM120A can modify the lipid content of cells where it is expressed and utilize that data to assess whether those lipids can inhibit PIEZO2 channels. This study revealed that TMEM120A expressing cells do have larger levels of phosphatidic acid and lysophosphatidic acid, and subsequent functional studies revealed that these lipids do inhibit PIEZO2.
INSTITUTE
Rutgers University
DEPARTMENT
Pharmacology, Physiology and Neuroscience; Metabolomics Shared Resource
T cell Immune Escape in Blast Crisis Transformation of Chronic Myeloid Leukemia
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
We report that leukemia secreted cytokines reduce miR-142 levels in T cells, causing T cell loss and exhaustion and therefore promoting CML BC transformation. Our homemade synthetic miR-142 increased T cell antileukemic activity and significantly prolonged the survival of CML BC murine and PDX models.
Prediction of metabolites associated with somatic mutations in cancers by using genome-scale metabolic models and mutation data
STUDY_SUMMARY
Background Oncometabolites, often generated as a result of a gene mutation, show pro-oncogenic function when abnormally accumulated in cancer cells. Identification of such mutation-associated metabolites will facilitate developing treatment strategies for cancers, but is challenging due to the large number of metabolites in a cell and the presence of multiple genes associated with cancer development. Results Here we report the development of a computational workflow that predicts metabolite-gene-pathway sets. Metabolite-gene-pathway sets present metabolites and metabolic pathways significantly associated with specific somatic mutations in cancers. The computational workflow uses both cancer patient-specific genome-scale metabolic models (GEMs) and mutation data to generate metabolite-gene-pathway sets. A GEM is a computational model that predicts reaction fluxes at a genome scale, and can be constructed in a cell-specific manner by using omics data. The computational workflow is first validated by comparing the resulting metabolite-gene pairs with multi-omics data (i.e., mutation data, RNA-seq data, and metabolome data) from acute myeloid leukemia and renal cell carcinoma samples collected in this study. The computational workflow is further validated by evaluating the metabolite-gene-pathway sets predicted for 18 cancer types, by using RNA-seq data publicly available, in comparison with the reported studies. Therapeutic potential of the resulting metabolite-gene-pathway sets is also discussed. Conclusions Validation of the metabolite-gene-pathway set-predicting computational workflow indicates that a decent number of metabolites and metabolic pathways appear to be significantly associated with specific somatic mutations. The computational workflow and the resulting metabolite-gene-pathway sets will help identify novel oncometabolites, and also suggest cancer treatment strategies.
INSTITUTE
Korea Advanced Institute of Science and Technology (KAIST)
LAST_NAME
Lee
FIRST_NAME
Sang Mi
ADDRESS
291 Daehak-ro, Yuseong-gu, Daejeon, 34141, South Korea
Integrative Analysis of Cytokine and Lipidomics Datasets Following Mild Traumatic Brain Injury in the Rat
STUDY_SUMMARY
Traumatic brain injury (TBI) is a significant source of disability in the United States and around the world and may lead to long-lasting cognitive deficits and decreased quality of life for patients across injury severities. Following the primary injury phase, TBI is characterized by com-plex secondary cascades that involve altered homeostasis and metabolism, faulty signaling, neu-roinflammation, and lipid dysfunction. The objectives of the present study were to (1) assess po-tential correlations between lipidome and cytokine changes after closed-head mild TBI (mTBI), and (2) examine reproducibility of our acute lipidomic profiles following TBI. Cortices from 54 Sprague Dawley male and female rats were analyzed by ultra-high-performance liquid chromatography mass spectrometry (LC-MS) in both positive and negative ionization modes and multiplex cytokine analysis after single (smTBI) or repetitive (rmTBI) closed-head impacts, or sham conditions. Tissue age was a variable, given that two cohorts (n= 26 and n=28) were initially run a year-and-a-half apart, creating inter-batch variations. We annotated the lipidome datasets using an in-house data dictionary based on exact masses of precursor and fragment ions and removed features with statis-tically significant differences between sham control batches. Our results indicate that lipids with high fold change between injury groups moderately correlate with the cytokines eotaxin, IP-10, and TNF-a. Additionally, we show a significant decrease of the pro-inflammatory markers, IL-1b and IP-10, TNF-a, and RANTES in the rmTBI samples relative to sham control. We discuss the major challenges in correlating high dimensional lipidomic data with functional cytokine profiles and the implications for understanding the biological significance of two related but disparate analysis modes in the study of TBI, an inherently heterogeneous neurological disorder.
INSTITUTE
Georgia Institute of Technology
DEPARTMENT
The Wallace H. Coulter Department of Biomedical Engineering
Lactiplantibacillus plantarum intervention on serum untargeted metabolome of gestational fecal microbiome transplant germ-free mice
STUDY_SUMMARY
The study aims to investigate whether administration of probiotic L. plantarum could influence profiling of serum untargeted metabolome of germ-free mice. Results showed that there were 23 metabolites significantly different in intervention group.
Lactiplantibacillus plantarum intervention on cecum bile acids of gestational fecal microbiome transplant germ-free mice
STUDY_SUMMARY
The study aims to investigate whether administration of probiotic L. plantarum could influence profiling of cecum bile acids of germ-free mice. L. plantarum during gestation of humanized GF mice showed that allolithocholic acid was significantly increased. Other bile acids showed no significant difference.
Lactiplantibacillus plantarum intervention on cecum tryptophans of gestational fecal microbiome transplant germ-free mice
STUDY_SUMMARY
The study aims to investigate whether administration of probiotic L. plantarum could influence profiling of cecum tryptophans of germ-free mice. L. plantarum intervention during gestation of humanized GF mice showed no significant difference on tryptophansās content.
Lactiplantibacillus plantarum intervention on fecal bile acids of gestational fecal microbiome transplant germ-free mice
STUDY_SUMMARY
The study aims to investigate whether administration of probiotic L. plantarum could influence profiling of fecal bile acids of germ-free mice. L. plantarum during gestation of humanized GF mice showed that Ī±-MCA, UCA, ACA, Ī²-UDCA, and isoLCA were significantly increased. Other bile acids showed no significant difference.
Parallel pheromonal, metabolite, and lipid analyses reveal patterns associated with early life transitions and ovary activation in honey bee (Apis mellifera) queens
STUDY_SUMMARY
We used a novel pheromone detection method to quantify retinue pheromone (QRP) concurrently with shotgun metabolomics and lipidomics analysis to determine what changes in pheromones and small molecules may underpin differences in age, laying status, and acceptance by workers in honey bee queens.
INSTITUTE
University of British Columbia
DEPARTMENT
Life Sciences Institute
LAST_NAME
Alcazar Magana
FIRST_NAME
Armando
ADDRESS
2350 Health Sciences Mall, Vancouver, BC, V6T1Z3, Canada
Cellular adaptation to cancer therapy along a resistance continuum
STUDY_SUMMARY
Recent research has shed light on the role of non-genetic plasticity in transient drug tolerance and the acquisition of stable resistance. However, the dynamics of cell state transitions occurring in the adaptation to cancer therapies remain elusive and require a systems-level longitudinal framework. Here we demonstrate that resistance develops through trajectories of cell state transitions accompanied by a progressive increase in cell fitness, which we denote the āresistance continuumā. This cellular adaptation involves a step-wise assembly of gene expression programs and epigenetically reinforced cell states underpinned by phenotypic plasticity stress adaptation and metabolic reprogramming. Through systematic genetic perturbations, we identify an acquisition of progressive metabolic dependencies, exposing a spectrum of vulnerabilities that can be potentially exploited therapeutically. The concept of the resistance continuum highlights the dynamic nature of cellular adaptation and calls for complementary therapies directed at the mechanisms underlying adaptive cell state transitions.
Reinforcing the Evidence of Mitochondrial Dysfunction in Long COVID Patients using a Multiplatform Mass Spectrometry-based Metabolomics Approach
STUDY_SUMMARY
Despite the recent and increasing knowledge surrounding COVID-19 infection, the underlying mechanisms of the persistence of symptoms long time after the acute infection are still not completely understood. Here, a multiplatform mass spectrometry-based approach was used for metabolomic and lipidomic profiling of human plasma samples from Long COVID patients (n=40) to reveal mitochondrial dysfunction when compared with individuals fully recovered from acute mild COVID-19 (n=40). Untargeted metabolomic analysis using CE-ESI(+/ā)-TOF-MS and GC-Q-MS was performed. Additionally, a lipidomic analysis using LC-ESI(+/ā)-QTOF-MS based on an in-house library revealed 471 lipid species identified with high confidence annotation level. The integration of complementary analytical platforms has allowed a comprehensive metabolic and lipidomic characterization of plasma alterations in Long COVID disease that found 46 relevant metabolites which allowed to discriminate between Long COVID and fully recovered patients. We report specific metabolites altered in Long COVID, mainly related to a decrease in the amino acid metabolism and ceramide plasma levels, and an increase in the tricarboxylic acid (TCA) cycle, reinforcing the evidence of an impaired mitochondrial function. The most relevant alterations shown in this study will help to better understand the insights of Long COVID syndrome by providing a deeper knowledge of the metabolomic basis of the pathology.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
Chemistry and Biochemistry
LABORATORY
CEMBIO
LAST_NAME
Martinez
FIRST_NAME
Sara
ADDRESS
UrbanizaciĆ³n MonteprĆncipe, 28660, Boadilla del Monte, Madrid, Spain
Targeted metabolomics was performed to measure polar metabolites in both positive and negative ionization mode on left ventricular tissue acquired from pre-mortem healthy donor hearts as classified by formal pathological examination and stored at the Sydney Heart Bank. The minimum number of observations for young (age ā¤ 25 years) and old (age ā„ 50 years) cohorts using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Activation of GFRAL+ Neurons Induces Hypothermia and Glucoregulatory Responses Associated with Nausea and Torpor
STUDY_SUMMARY
GFRAL-expressing neurons actuate aversion and nausea, are targets for obesity treatment and may mediate metformin effects by long-term GDF15-GFRAL agonism. If GFRAL+ neurons acutely regulate glucose and energy homeostasis is however underexplored. Here, we report that cell-specific activation of GFRAL+ neurons using a variety of techniques causes a torpor-like state, including hypothermia, the release of stress hormones, a shift from glucose to lipid oxidation, and impaired insulin sensitivity, glucose tolerance and skeletal muscle glucose uptake but augmented glucose uptake in visceral fat. Metabolomic analysis of blood and transcriptomics of muscle and fat indicate alterations in ketogenesis, insulin signaling, adipose tissue differentiation and mitogenesis, and energy fluxes. Our findings reveal that acute GFRAL+ neuron activation induces endocrine and gluco- and thermoregulatory responses associated with nausea and torpor. While chronic activation of GFRAL signaling promotes weight loss in obesity, these results show that acute activation of GFRAL+ neurons causes hypothermia and hyperglycemia.
13C-palmitate labeling experiment in ICC13-7 treated with DMSO or Infigratinib
STUDY_SUMMARY
Genomic alterations that activate FGFR2 are common in intrahepatic cholangiocarcinoma (ICC) and confer sensitivity to treatment with FGFR inhibitors. However, the depth and duration of responses are often limited. Here, we conducted integrative transcriptomic and metabolomic analysis of patient-derived models to define the pathways that fuel tumor growth downstream of oncogenic FGFR2 signaling in ICC and to uncover compensatory mechanisms associated with pathway inhibition. We find FGFR2-mediated activation of NF-ļ«B maintains a highly glycolytic phenotype. Conversely, FGFR inhibition blocks glucose uptake and glycolysis while inciting a series of adaptive changes, including switching fuel source utilization to favor fatty acid oxidation and increasing mitochondrial fusion and autophagy. Accordingly, FGFR inhibitor efficacy is potentiated by combined mitochondrial targeting, an effect enhanced in xenograft models by intermittent fasting. Thus, we show that oncogenic FGFR2 signaling drives NF-kB-dependent glycolysis in ICC and that metabolic reprogramming in response to FGFR inhibition confers new targetable vulnerabilities.
Characterization of metabolic changes upon FGFR inhibition in ICC13-7 Xenograft
STUDY_SUMMARY
Genomic alterations that activate FGFR2 are common in intrahepatic cholangiocarcinoma (ICC) and confer sensitivity to treatment with FGFR inhibitors. However, the depth and duration of responses are often limited. Here, we conducted integrative transcriptomic and metabolomic analysis of patient-derived models to define the pathways that fuel tumor growth downstream of oncogenic FGFR2 signaling in ICC and to uncover compensatory mechanisms associated with pathway inhibition. We find FGFR2-mediated activation of NF-ļ«B maintains a highly glycolytic phenotype. Conversely, FGFR inhibition blocks glucose uptake and glycolysis while inciting a series of adaptive changes, including switching fuel source utilization to favor fatty acid oxidation and increasing mitochondrial fusion and autophagy. Accordingly, FGFR inhibitor efficacy is potentiated by combined mitochondrial targeting, an effect enhanced in xenograft models by intermittent fasting. Thus, we show that oncogenic FGFR2 signaling drives NF-ļ«B-dependent glycolysis in ICC and that metabolic reprogramming in response to FGFR inhibition confers new targetable vulnerabilities.
Complete absence of GLUT1 does not impair human terminal erythroid differentiation
STUDY_SUMMARY
The Glucose transporter 1 (GLUT1) is one of the most abundant proteins within the erythrocyte membrane and is required for glucose and dehydroascorbic acid (Vitamin C precursor) transport. It is widely recognized as a key protein for red cell structure, function, and metabolism. Previous reports highlighted the importance of GLUT1 activity within these uniquely glycolysis-dependent cells, in particular for increasing antioxidant capacity needed to avoid irreversible damage from oxidative stress in humans. However, studies of glucose transporter roles in erythroid cells are complicated by species-specific differences between humans and mice. Here, using CRISPRmediated gene editing of immortalized erythroblasts and adult CD34+ hematopoietic progenitor cells, we generate committed human erythroid cells completely deficient in expression of GLUT1. We show that absence of GLUT1 does not impede human erythroblast proliferation, differentiation, or enucleation. This work demonstrates for the first-time generation of enucleated human reticulocytes lacking GLUT1. The GLUT1-deficient reticulocytes possess no tangible alterations to membrane composition or deformability in reticulocytes. Metabolomic analyses of GLUT1-deficient reticulocytes reveal hallmarks of reduced glucose import, downregulated metabolic processes and upregulated AMPK-signalling, alongside alterations in antioxidant metabolism, resulting in increased osmotic fragility and metabolic shifts indicative of higher oxidant stress. Despite detectable metabolic changes in GLUT1 deficient reticulocytes, the absence of developmental phenotype, detectable proteomic compensation or impaired deformability comprehensively alters our understanding of the role of GLUT1 in red blood cell structure, function and metabolism. It also provides cell biological evidence supporting clinical consensus that reduced GLUT1 expression does not cause anaemia in GLUT1 deficiency syndrome.
INSTITUTE
University of Colorado
LAST_NAME
Stephenson
FIRST_NAME
Daniel
ADDRESS
Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA
In this study, plasma samples were processed using an in-house one phase extraction protocol as previously published (Siddiqa, A., et al., A pilot metabolomic study of drug interaction with the immune response to seasonal influenza vaccination. npj Vaccines, 2023. 8(1): p. 92. ). The resulting extract was then analyzed on a Thermo ScientificTM TranscendTM Duo LX-2 UHPLC system interfaced with high resolution Thermo ScientificTM Orbitrap ID-XTM TribidTM mass spectrometer with a HESI ionization source. This dataset was collected over 17 batches and was used to evaluate the QA/QC metrics produced by the pipeline and resulting MS2 annotations generated from a complimentary AcquireX (deep scan) MS2 dataset were compared to compound discoverer.
INSTITUTE
Jackson Laboratory for Genomic Medicine
LABORATORY
Shuzhao Li Laboratory
LAST_NAME
Joshua
FIRST_NAME
Mitchell
ADDRESS
10 Discovery Dr, Farmington CT 06032
EMAIL
joshua.mitchell@jax.org
PUBLICATIONS
Common data models to streamline metabolomics processing and annotation, and implementation in a Python pipeline Joshua Mitchell, Yuanye Chi, Maheshwor Thapa, Zhiqiang Pang, Jianguo Xia, Shuzhao Li doi: https://doi.org/10.1101/2024.02.13.580048
Influence of Lifestyle Factors on Epigenetic and Proteomic Biomarkers: A Cross-Sectional Comparison
STUDY_TYPE
Cross-sectional Study
STUDY_SUMMARY
This cross-sectional study investigated differences in the plasma metabolome in two groups of adults that were of similar age but varied markedly in body composition and dietary and physical activity patterns. Study participants included 52 adults in the lifestyle group (LIFE) (28 males, 24 females) and 52 in the control group (CON) (27 males, 25 females). The results using an extensive untargeted UPLC-HRMS analysis with 10,535 metabolite peaks identified 83 metabolites and 16 metabolic pathways that differentiated LIFE and CON groups. A novel metabolite signature of positive lifestyle habits emerged from this analysis highlighted by lower plasma levels of numerous bile acids, an amino acid profile characterized by higher histidine and lower glutamic acid, glutamine, Ī²-alanine, phenylalanine, tyrosine, and proline, an elevated vitamin D status, higher levels of beneficial fatty acids and gut microbiome catabolism metabolites from plant substrates, and reduced levels of N-glycan degradation metabolites and environmental contaminants. This study established that the plasma metabolome is strongly associated with body composition and lifestyle habits. The robust lifestyle metabolite signature identified in this study is consistent with an improved life expectancy and a reduced risk for chronic disease.
INSTITUTE
Appalachian State University
DEPARTMENT
Department of Biology
LABORATORY
Human Performance Laboratory
LAST_NAME
Nieman
FIRST_NAME
David
ADDRESS
North Carolina Research Campus, 600 Laureate Way, Kannapolis, NC 28081
Inhibition of Asparagine Synthetase Effectively Retards Polycystic Kidney Disease Progression, investigated with targeted metabolomics in Tam-Cre;Pkd1ĪC/flox mouse model kidneys.
STUDY_SUMMARY
Polycystic Kidney Disease (PKD) is a genetic disorder characterized by bilateral cyst formation. We showed that PKD cells and kidneys display metabolic alterations, including the Warburg effect and glutaminolysis, sustained in vitro by the enzyme asparagine synthetase (ASNS). Here, we used antisense oligonucleotides (ASO) against Asns in orthologous and slowly progressive PKD murine models and show that treatment leads to a drastic reduction of total kidney volume (measured by MRI) and a prominent rescue of renal function in the mouse. Mechanistically, the upregulation of an ATF4-ASNS axis in PKD is driven by the amino acid response (AAR) branch of the integrated stress response (ISR). Metabolic profiling of PKD or control kidneys treated with Asns-ASO or Scr-ASO revealed major changes in the mutants, several of which are rescued by Asns silencing in vivo. Indeed, ASNS drives glutamine-dependent de novo pyrimidine synthesis and proliferation in cystic epithelia. Notably, while several metabolic pathways were completely corrected by Asns-ASO, glycolysis was only partially restored. Accordingly, combining the glycolytic inhibitor 2DG with Asns-ASO further improved efficacy. Our studies identify a new therapeutic target and novel metabolic vulnerabilities in PKD. Altogether, targeted metabolomics analysis performed in Tam-Cre;Pkd1ĪC/flox mouse model kidneys corroborates the central role of ASNS in the metabolic rewiring occurring in PKD, highlighting the therapeutic potential of its inhibition.
Glucose Hypometabolism Prompts RAN Translation and Exacerbates C9orf72-related ALS/FTD Phenotypes
STUDY_SUMMARY
The most prevalent genetic cause of both amyotrophic lateral sclerosis and frontotemporal dementia is a (GGGGCC)n nucleotide repeat expansion (NRE) occurring in the first intron of the C9orf72 gene (C9). Brain glucose hypometabolism is consistently observed in C9-NRE carriers, even at pre-symptomatic stages, although its potential role in disease pathogenesis is unknown. Here, we identified alterations in glucose metabolic pathways and ATP levels in the brains of asymptomatic C9-BAC mice. We found that, through activation of the GCN2 kinase, glucose hypometabolism drives the production of dipeptide repeat proteins (DPRs), impairs the survival of C9 patient-derived neurons, and triggers motor dysfunction in C9-BAC mice. We also found that one of the arginine-rich DPRs (PR) can directly contribute to glucose metabolism and metabolic stress. These findings provide a potential mechanistic link between energy imbalances and C9-ALS/FTD pathogenesis and suggest a feedforward loop model that opens several opportunities for therapeutic intervention.
Inhibition of Asparagine Synthetase Effectively Retards Polycystic Kidney Disease Progression, investigated with targeted tracing metabolomics analysis in MEF cells using 15N2-glutamine.
STUDY_SUMMARY
Polycystic Kidney Disease (PKD) is a genetic disorder characterized by bilateral cyst formation. We showed that PKD cells and kidneys display metabolic alterations, including the Warburg effect and glutaminolysis, sustained in vitro by the enzyme asparagine synthetase (ASNS). Here, we used antisense oligonucleotides (ASO) against Asns in orthologous and slowly progressive PKD murine models and show that treatment leads to a drastic reduction of total kidney volume (measured by MRI) and a prominent rescue of renal function in the mouse. Mechanistically, the upregulation of an ATF4-ASNS axis in PKD is driven by the amino acid response (AAR) branch of the integrated stress response (ISR). Metabolic profiling of PKD or control kidneys treated with Asns-ASO or Scr-ASO revealed major changes in the mutants, several of which are rescued by Asns silencing in vivo. Indeed, ASNS drives glutamine-dependent de novo pyrimidine synthesis and proliferation in cystic epithelia. Notably, while several metabolic pathways were completely corrected by Asns-ASO, glycolysis was only partially restored. Accordingly, combining the glycolytic inhibitor 2DG with Asns-ASO further improved efficacy. Our studies identify a new therapeutic target and novel metabolic vulnerabilities in PKD. Of interest, in these tracing studies we could confirm that the pyrimidine biosynthesis pathway is increased and rescued by silencing of Asns.
Lipidomics analyses in model membranes, isolated mitochondria and cellular systems to study how the local lipid environment affects BAX and BAK function during apoptosis.
STUDY_SUMMARY
To investigate how the local lipid environment affects BAX and BAK function during apoptosis, we performed quantitative analyses of different lipid classes (glycerophospholipids, fatty acids, ceramides and sphingomyelins) in cultured cells, isolated mitochondria and lipid nanodics formed by Styrene-Malic Acid (SMA) co-polymers. Ceramides, sphingomyelins, fatty acids and cardiolipins were analyzed by Liquid Chromatography coupled to Tandem Mass Spectrometry (LC-MS/MS). For glycerophospholipids (PC, PE, PI, PS, PG, PA) we applied direct infusion MS approaches (Shotgun Lipidomics).
INSTITUTE
University of Cologne
DEPARTMENT
Faculty of Medicine and University Hospital of Cologne, Cluster of Excellence Cellular Stress Responses in Aging-associated Diseases (CECAD)
Comprehensive Untargeted LC-MS Metabolomics Analysis of HMGCS2 Knockout iPSCs(Cell Pellet)
STUDY_SUMMARY
Human induced pluripotent stem cells (hIPSCs) are a key tool for biomedical research. In this study, human induced pluripotent stem cells of the KOLF2.2J lineage were generated with null allele mutation for HMGCS2. These cell lines were cultured on several types of media with and without various additives and were differentiated into primitive syncytium. Biochemical phenotyping, i.e., untargeted metabolomics and lipidomics profiling, was performed on both differentiated and undifferentiated cells using a Thermo Scientific Q Exactive HF-X Mass Spectrometer coupled to a Thermo Scientific Transcend LX-2 Duo UHPLC system, equipped with an HES-II ionization source, using both positive and negative ionization modes. This dataset, combined with the other -omics level data generated through MorPhiC on these cell lines, provides a molecular foundation for understanding the implications of specific gene deletions in these cell lines and their derivative cell types (e.g., primitive syncytium). This study includes data generated from the cell pellets of the above cell lines. This study was funded, in part, through UM1HG012651 which established the JAX MorPhiC Center, a MorPhiC Phase 1 Data Production Research and Development Center at the Jackson Laboratory for Genomic Medicine.
Comprehensive Untargeted LC-MS Metabolomics Analysis of HMGCS2 Knockout iPSCs(Supernatant/Media)
STUDY_SUMMARY
Human induced pluripotent stem cells (hIPSCs) are a key tool for biomedical research. In this study, human induced pluripotent stem cells of the KOLF2.2J lineage were generated with null allele mutation for HMGCS2. These cell lines were cultured on several types of media with and without various additives and were differentiated into primitive syncytium. Biochemical phenotyping, i.e., untargeted metabolomics and lipidomics profiling, was performed on both differentiated and undifferentiated cells using a Thermo Scientific Q Exactive HF-X Mass Spectrometer coupled to a Thermo Scientific Transcend LX-2 Duo UHPLC system, equipped with an HES-II ionization source, using both positive and negative ionization modes. This dataset, combined with the other -omics level data generated through MorPhiC on these cell lines, provides a molecular foundation for understanding the implications of specific gene deletions in these cell lines and their derivative cell types (e.g., primitive syncytium). This study includes data generated from the cell pellets of the above cell lines. This study was funded, in part, through UM1HG012651 which established the JAX MorPhiC Center, a MorPhiC Phase 1 Data Production Research and Development Center at the Jackson Laboratory for Genomic Medicine.
Metabolomics of patients with Plasmodium vivax malaria
STUDY_SUMMARY
Background: Malaria is the leading parasitic disease worldwide, with P. vivax being a major challenge for its control. Several studies have indicated metabolomics as a promising tool for combating the disease. Objective: The study evaluated plasma metabolomic profiles of patients with recurrent and non-recurrent P. vivax malaria in the Brazilian Amazon. Methods: Metabolites extracted from the plasma of P. vivax-infected patients were subjected to LC-MS analysis. Untargeted metabolomics was applied to investigate the metabolic profile of the plasma in the two groups. Results: Overall, 51 recurrent and 59 non-recurrent patients were included in the study. Longitudinal metabolomic analysis revealed 52 and 37 significant metabolite features from the recurrent and non-recurrent participants, respectively. Recurrence was associated with disturbances in eicosanoid metabolism. Comparison between groups suggest alterations in vitamin B6 (pyridoxine) metabolism, tyrosine metabolism, 3-oxo-10-octadecatrienoate Ī²-oxidation, and alkaloid biosynthesis II. Integrative network analysis revealed enrichment of other metabolic pathways for the recurrent phenotype, including the butanoate metabolism, aspartate and asparagine metabolism, and N-glycan biosynthesis. Conclusion: The metabolites and metabolic pathways predicted in our study suggest potential biomarkers of recurrence and provide insights into targets for antimalarial development against P. vivax.
INSTITUTE
University of Sao Paulo
LAST_NAME
Gardinassi
FIRST_NAME
Luiz Gustavo
ADDRESS
Av. dos Bandeirantes, 3900, Campus UniversitĆ”rio, RibeirĆ£o Preto, SP, Brazil
Glucose Hypometabolism Prompts RAN Translation and Exacerbates C9orf72-related ALS/FTD Phenotypes - Study 2
STUDY_SUMMARY
The most prevalent genetic cause of both amyotrophic lateral sclerosis and frontotemporal dementia is a (GGGGCC)n nucleotide repeat expansion (NRE) occurring in the first intron of the C9orf72 gene (C9). Brain glucose hypometabolism is consistently observed in C9-NRE carriers, even at pre-symptomatic stages, although its potential role in disease pathogenesis is unknown. Here, we identified alterations in glucose metabolic pathways and ATP levels in the brains of asymptomatic C9-BAC mice. We found that, through activation of the GCN2 kinase, glucose hypometabolism drives the production of dipeptide repeat proteins (DPRs), impairs the survival of C9 patient-derived neurons, and triggers motor dysfunction in C9-BAC mice. We also found that one of the arginine-rich DPRs (PR) can directly contribute to glucose metabolism and metabolic stress. These findings provide a potential mechanistic link between energy imbalances and C9-ALS/FTD pathogenesis and suggest a feedforward loop model that opens several opportunities for therapeutic intervention.
Mannose is crucial for mesoderm specification and symmetry breaking in gastruloids.
STUDY_SUMMARY
Patterning and growth are fundamental features of embryonic development that must be tightly coordinated. To understand how metabolism impacts early mesoderm development, we used mouse embryonic stem cell-derived gastruloids, that co-expressed glucose transporters with the mesodermal marker T/Bra. While the glucose mimic, 2-deoxy-D-glucose (2-DG), blocked T/Bra expression and abolished axial elongation in gastruloids, removal of glucose did not phenocopy 2-DG treatment despite a decline in glycolytic intermediates occurring under both conditions. As 2-DG could also act as a competitive inhibitor of mannose in protein glycosylation, we added mannose together with 2-DG and found that it could rescue the mesoderm specification both in vivo and in vitro. We further showed that blocking production and intracellular recycling of mannose abrogated mesoderm specification. Proteomics analysis revealed that mannose reversed glycosylation of the Wnt pathway regulator, Secreted Frizzled Receptor, Frzb. Our study showed how mannose is crucial for mesoderm specification in gastruloids.
INSTITUTE
Dept of Genetics, University of Cambridge
LAST_NAME
Dingare
FIRST_NAME
Chaitanya
ADDRESS
Downing Site, Cambridge, Cambridgeshire, CB2 3EH, United Kingdom
Metabolomic profiling of ALG13-CDG brain organoids
STUDY_SUMMARY
ALG13-CDG is a rare inborn error of metabolism caused by deficiency of the ALG13 enzyme, which is believed to lead to impaired protein glycosylation. While the disorder has primarily neurological presentation, there is limited knowledge about the specific brain-related changes that result from ALG13 deficiency. We used 3D brain organoids derived from fibroblasts of the individuals with ALG13-CDG to probe the brain-specific metabolic changes in ALG13-CDG.
Serum metabolites in inherited retinal degenerations
STUDY_SUMMARY
The diagnosis of inherited retinal degeneration (IRD) is challenging owing to its phenotypic and genotypic complexity. Clinical information is important before a genetic diagnosis is made. Metabolomics studies the entire picture of bioproducts, which are determined using genetic codes and biological reactions. We demonstrated that the common diagnoses of IRD, including retinitis pigmentosa (RP), cone-rod dystrophy (CRD), Stargardt disease (STGD), and Biettiās crystalline dystrophy (BCD), could be differentiated based on their metabolite heatmaps. Hundreds of metabolites were identified in the volcano plot compared with that of the control group in every IRD except BCD, considered as potential diagnosing markers. The phenotypes of CRD and STGD overlapped but could be differentiated by their metabolomic features with the assistance of a machine learning model with 100% accuracy. Moreover, EYS-, USH2A-associated, and other RP, sharing considerable similar characteristics in clinical findings, could also be diagnosed using the machine learning model with 85.7% accuracy. Further study would be needed to validate the results in the external dataset. By incorporating mass spectrometry and machine learning, a metabolomics-based diagnostic workflow for the clinical and molecular diagnoses of IRD was proposed in our study.
Pulmonary maternal immune activation does not extend through the placenta but leads to fetal metabolic adaptation - Fetal liver
STUDY_SUMMARY
Maternal immune system activation (MIA) during pregnancy can disrupt the fetal environment, causing postnatal susceptibility to disorders. How the placenta and the fetus respond to acute MIA over time is unknown. Here, we characterized the response to acute maternal pulmonary inflammation across time in maternal and fetal organs using multi-omics. Unlike maternal organs which mounted strong innate immune responses, the placenta upregulated tissue-integrity genes, likely to prevent fetal exposure to infections, and downregulated growth-associated genes. Subsequently, the placenta upregulated biosynthesis and endoplasmic reticulum stress genes in order to return to homeostasis. These responses likely protected the fetus, since we observed no immune response in fetal liver. Instead, likely due to nutrient depletion, the fetal liver displayed metabolic adaptations, including increases in lipids containing docosahexaenoic acid, crucial for fetal brain development. Our study shows, for the first time, the integrated temporal response to pulmonary MIA across maternal and fetal organs.
INSTITUTE
University of Copenhagen
DEPARTMENT
Department of Biology
LABORATORY
Section for Computational and RNA Biology
LAST_NAME
Sandelin
FIRST_NAME
Albin
ADDRESS
Copenhagen University Ole Maaloes Vej 5, DK2200, Copenhagen N, Denmark
Effect of high fat diet on heart metabolome of CHCHD10 mutant mice
STUDY_SUMMARY
Mutations in CHCHD10, a mitochondrial protein with undefined functions, are associated with autosomal dominant mitochondrial diseases. Chchd10 knock-in mice harboring a heterozygous S55L mutation (equivalent to human pathogenic S59L) develop a fatal mitochondrial cardiomyopathy caused by CHCHD10 aggregation and proteotoxic mitochondrial integrated stress response (mtISR). In mutant hearts, mtISR is accompanied by a metabolic rewiring characterized by increased reliance on glycolysis rather than fatty acid oxidation. To counteract this metabolic rewiring, heterozygous S55L mice were subjected to chronic high fat diet (HFD) to decrease insulin sensitivity and glucose uptake and enhance fatty acid utilization in the heart. HFD ameliorated the ventricular dysfunction of mutant hearts and significantly extended the survival of mutant female mice affected by severe pregnancy-induced cardiomyopathy. Gene expression profiles confirmed that HFD increased fatty acid utilization and ameliorated cardiomyopathy markers. Importantly, HFD also decreased accumulation of aggregated CHCHD10 in the S55L heart, suggesting activation of quality control mechanisms. Overall, our findings indicate that metabolic therapy can be effective in mitochondrial cardiomyopathies associated with proteotoxic stress.
Effect of high fat diet on heart lipidome in CHCHD10 mutant mice
STUDY_SUMMARY
Mutations in CHCHD10, a mitochondrial protein with undefined functions, are associated with autosomal dominant mitochondrial diseases. Chchd10 knock-in mice harboring a heterozygous S55L mutation (equivalent to human pathogenic S59L) develop a fatal mitochondrial cardiomyopathy caused by CHCHD10 aggregation and proteotoxic mitochondrial integrated stress response (mtISR). In mutant hearts, mtISR is accompanied by a metabolic rewiring characterized by increased reliance on glycolysis rather than fatty acid oxidation. To counteract this metabolic rewiring, heterozygous S55L mice were subjected to chronic high fat diet (HFD) to decrease insulin sensitivity and glucose uptake and enhance fatty acid utilization in the heart. HFD ameliorated the ventricular dysfunction of mutant hearts and significantly extended the survival of mutant female mice affected by severe pregnancy-induced cardiomyopathy. Gene expression profiles confirmed that HFD increased fatty acid utilization and ameliorated cardiomyopathy markers. Importantly, HFD also decreased accumulation of aggregated CHCHD10 in the S55L heart, suggesting activation of quality control mechanisms. Overall, our findings indicate that metabolic therapy can be effective in mitochondrial cardiomyopathies associated with proteotoxic stress.
Effect of high fat diet on serum fatty acids, lipidome and metabolome in CHCHD10 mutant mice
STUDY_SUMMARY
Mutations in CHCHD10, a mitochondrial protein with undefined functions, are associated with autosomal dominant mitochondrial diseases. Chchd10 knock-in mice harboring a heterozygous S55L mutation (equivalent to human pathogenic S59L) develop a fatal mitochondrial cardiomyopathy caused by CHCHD10 aggregation and proteotoxic mitochondrial integrated stress response (mtISR). In mutant hearts, mtISR is accompanied by a metabolic rewiring characterized by increased reliance on glycolysis rather than fatty acid oxidation. To counteract this metabolic rewiring, heterozygous S55L mice were subjected to chronic high fat diet (HFD) to decrease insulin sensitivity and glucose uptake and enhance fatty acid utilization in the heart. HFD ameliorated the ventricular dysfunction of mutant hearts and significantly extended the survival of mutant female mice affected by severe pregnancy-induced cardiomyopathy. Gene expression profiles confirmed that HFD increased fatty acid utilization and ameliorated cardiomyopathy markers. Importantly, HFD also decreased accumulation of aggregated CHCHD10 in the S55L heart, suggesting activation of quality control mechanisms. Overall, our findings indicate that metabolic therapy can be effective in mitochondrial cardiomyopathies associated with proteotoxic stress.
Pulmonary maternal immune activation does not extend through the placenta but leads to fetal metabolic adaptation - Maternal liver
STUDY_SUMMARY
Maternal immune system activation (MIA) during pregnancy can disrupt the fetal environment, causing postnatal susceptibility to disorders. How the placenta and the fetus respond to acute MIA over time is unknown. Here, we characterized the response to acute maternal pulmonary inflammation across time in maternal and fetal organs using multi-omics. Unlike maternal organs which mounted strong innate immune responses, the placenta upregulated tissue-integrity genes, likely to prevent fetal exposure to infections, and downregulated growth-associated genes. Subsequently, the placenta upregulated biosynthesis and endoplasmic reticulum stress genes in order to return to homeostasis. These responses likely protected the fetus, since we observed no immune response in fetal liver. Instead, likely due to nutrient depletion, the fetal liver displayed metabolic adaptations, including increases in lipids containing docosahexaenoic acid, crucial for fetal brain development. Our study shows, for the first time, the integrated temporal response to pulmonary MIA across maternal and fetal organs.
INSTITUTE
University of Copenhagen
DEPARTMENT
Department of Biology
LABORATORY
Section for Computational and RNA Biology
LAST_NAME
Sandelin
FIRST_NAME
Albin
ADDRESS
Copenhagen University Ole Maaloes Vej 5, DK2200, Copenhagen N, Denmark
Pulmonary maternal immune activation does not extend through the placenta but leads to fetal metabolic adaptation - Maternal blood
STUDY_SUMMARY
Maternal immune system activation (MIA) during pregnancy can disrupt the fetal environment, causing postnatal susceptibility to disorders. How the placenta and the fetus respond to acute MIA over time is unknown. Here, we characterized the response to acute maternal pulmonary inflammation across time in maternal and fetal organs using multi-omics. Unlike maternal organs which mounted strong innate immune responses, the placenta upregulated tissue-integrity genes, likely to prevent fetal exposure to infections, and downregulated growth-associated genes. Subsequently, the placenta upregulated biosynthesis and endoplasmic reticulum stress genes in order to return to homeostasis. These responses likely protected the fetus, since we observed no immune response in fetal liver. Instead, likely due to nutrient depletion, the fetal liver displayed metabolic adaptations, including increases in lipids containing docosahexaenoic acid, crucial for fetal brain development. Our study shows, for the first time, the integrated temporal response to pulmonary MIA across maternal and fetal organs.
INSTITUTE
University of Copenhagen
DEPARTMENT
Department of Biology
LABORATORY
Section for Computational and RNA Biology
LAST_NAME
Sandelin
FIRST_NAME
Albin
ADDRESS
Copenhagen University Ole Maaloes Vej 5, DK2200, Copenhagen N, Denmark
While there is strong evidence for interactions between the microbiota-gut-brain axis and host physiology in the context of chronic stress, limited research has investigated the role of the microbiome in host response to acute stress. Determining the underlying mechanisms by which stress-induced microbiota changes may provoke functional changes in the gut and brain is critical for developing future therapeutics to alleviate the adverse consequences of traumatic stress. Here, we aimed to identify a biological signature of gut metabolites that are significantly altered following exposure to acute restraint stress. Adult male C57Bl/6 conventional, germ-free and colonized germ-free mice underwent a 15-minute restraint stress exposure. Caecal contents were collected from naĆÆve mice and stressed mice, either immediately or 45 minutes following stress. Caecal contents underwent untargeted metabolomics analysis.
While there is strong evidence for interactions between the microbiota-gut-brain axis and host physiology in the context of chronic stress, limited research has investigated the role of the microbiome in host response to acute stress. Determining the underlying mechanisms by which stress-induced microbiota changes may provoke functional changes in the gut and brain is critical for developing future therapeutics to alleviate the adverse consequences of traumatic stress. Here, we aimed to identify a biological signature of gut metabolites that are significantly altered following exposure to acute restraint stress. Adult male C57Bl/6 conventional, germ-free and colonized germ-free mice underwent a 15-minute restraint stress exposure. Caecal contents were collected from naĆÆve mice and stressed mice, either immediately or 45 minutes following stress. Colonic mucosa underwent untargeted metabolomics analysis.
Unraveling Salivary Metabolome in Children with Eosinophilic Esophagitis
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
In this pilot study, we performed global untargeted salivary metabolomics. We focused on salivary metabolomics as oral mucosa is the initial interface between the triggering antigens and the host mucosal immune system,9 saliva is rich yet understudied biofluid, and saliva can be collected safely, rapidly, and non-invasively (compared to esophageal and plasma samples) making it uniquely suited for testing pediatric populations.10 Our primary aim was to gain novel insights into the upstream metabolomic alterations in pediatric EoE by profiling salivary metabolome in children with EoE, and our secondary aim was to assess the translational potential of salivary metabolites.
Targeting SOX13 inhibits the assembly of respiratory chain supercomplexes to overcome ferroptosis-resistance in gastric cancer
STUDY_SUMMARY
Therapeutic resistance represents a bottleneck to treatment in advanced gastric cancer (GC). Ferroptosis is an iron-dependent form of non-apoptotic cell death and is associated with anti-cancer therapeutic efficacy. Further investigations are required to clarify the underlying mechanisms. Ferroptosis-resistant GC cell lines are constructed. Dysregulated mRNAs between ferroptosis-resistant and parental cell lines are identified. The expression of SOX13/SCAF1 is manipulated in GC cell lines where relevant biological and molecular analyses are performed. Molecular docking and computational screening are performed to screen potential inhibitors of SOX13. We show that SOX13 boosts protein remodeling of electron transport chain (ETC) complexes by directly transactivating SCAF1. This leads to increased supercomplexes (SCs) assembly, mitochondrial respiration, mitochondrial energetics and chemo- and immune-resistance. Zanamivir, reverts the ferroptosis-resistant phenotype via directly targeting SOX13 and promoting TRIM25-mediated ubiquitination and degradation of SOX13. Here we show, SOX13/SCAF1 are important in ferroptosis-resistance, and targeting SOX13 with zanamivir has therapeutic potential. We conducted untargeted metabolomic analysis of Erastin-resis SNU-668 cells transfected with shRNA-SOX13 or shRNA-NC.
Metabolomic study of T effector and T regulatory cells in severe allergic patients
STUDY_SUMMARY
Metabolism has a profound impact on T cell fate and function. Uncovering the metabolome of circulating human CD4+ T effector memory (Teff) and T regulatory (Treg) cells would enable better understanding of Th2-driven diseases, such as allergy or asthma. Here, we demonstrated that in healthy humans, energy metabolism and functions of memory CD4+ Teff cells mainly relied on amino acids, whereas Treg cells predominantly used fatty acids. Arginine and phenylalanine increased T cell receptorinduced glycolysis and oxidative phosphorylation in total and memory CD4+ T cells, but high levels of phenylalanine limited CD4+ T cell proliferation via disrupting mitochondrial respiration and activation of L-phenylalanine oxidase, IL4I1. Accordingly, lowest levels of phenylalanine were linked with the pathogenic Th2a cells, and impaired Treg cells in patients with the most severe forms of allergies. It all suggests that phenylalanine is a metabolic checkpoint of pathogenic Th2 cells development.
INSTITUTE
Universidad San Pablo-CEU, CEU Universities
DEPARTMENT
QuĆmica y BioquĆmica
LAST_NAME
VillaseƱor
FIRST_NAME
Alma
ADDRESS
UrbanizaciĆ³n MonteprĆncipe, n/s, Boadilla del Monte, Madrid, 28003, Spain
Circulating N-lactoyl-amino acids and N-formyl-methionine reflect mitochondrial dysfunction and predict mortality in septic shock
STUDY_SUMMARY
he aim of this study was to identify circulating markers that correlated with severity and mortality in sepsis. We performed metabolomic profiling on 4 cohorts: 1) patients with septic shock, 2) patients with bacteremia without sepsis, 3) cardiogenic shock, and 4) healthy ambulatory controls. We found a set of metabolites that out outperformed lactate in their ability to distinguish survivors and non-survivors of sepsis.
INSTITUTE
Massachusetts General Hospital
LAST_NAME
Sharma
FIRST_NAME
Rohit
ADDRESS
185 Cambridge Street, 6th Floor, Boston, MA, 02114, USA
Endothelial-Dependent Vascular Reactivity After Cardiopulmonary Bypass is Associated with Unique Metabolomic Signatures
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Cardiopulmonary bypass (CPB), an extracorporeal method necessary for the surgical correction of complex congenital heart defects, incites significant inflammatory and vascular changes. Along with these changes are alterations in cellular metabolism that promote energy production to deal with this stress. Utilizing laser-doppler perfusion monitoring coupled with iontophoresis (LDPMI) in patients undergoing corrective heart surgery, we hypothesized that temporal, untargeted metabolomics could be performed to assess the link between metabolism and vascular function. Globally, we found 2404 unique metabolites in the plasma of patients undergoing CPB. Metabolites related to arginine biosynthesis were the most altered in the CPB period. When examining metabolic profiles in correlation with endothelial-dependent (acetylcholine, ACh) or endothelial-independent (sodium nitroprusside, SNP) vascular reactivity, purine metabolism was most consistently associated with either vascular response. With ACh-mediated responses, L-acetylcarnitine levels were most strongly associated, while L-glutamine levels were associated with both ACh and SNP responsiveness. These data give insight into the metabolic landscape of children undergoing CPB for corrective heart surgery and provide detail into how these metabolites relate to physiological aberrations in the vasculature.
Untargeted metabolomic analysis of lumenal contents from AMX-treated and untreated mice
STUDY_SUMMARY
Antibiotics cause collateral damage to resident microbes that is associated with various health risks. To-date, studies have largely focused on impacts of antibiotics on large intestinal and fecal microbiota. Here, we employ a GI-wide integrated multiomic approach to reveal that amoxicillin (AMX) treatment reduces overall bacterial abundance, bile salt hydrolase activity and unconjugated bile acids in the small intestine (SI). An accompanying loss of fatty acids and increase in acyl-carnitines in the large intestine corresponded with spatially-distinct expansions of proteobacteria. Parasutterella excrementihominis utilized fatty acid biosynthesis, becoming dominant in the SI while multiple Klebsiella species employed fatty acid oxidation during expansion in the large intestine. Depletion of bile acids and lipids may contribute to AMX-induced dysbiosis in the lower GI. To test this, we demonstrate that restoration of unconjugated bile acids can mitigate losses of commensals in the large intestine while also inhibiting the expansion of Proteobacteria during AMX treatment.
Effect of liver Acox1 knockout on serum lipidome in mice
STUDY_SUMMARY
The liver gene expression of the peroxisomal Ī²-oxidation enzyme acyl-coenzyme A oxidase 1 (ACOX1), which catabolizes very long chain fatty acids (VLCFA), increases in the context of obesity. To check if liver peroxisomal fatty acids beta-oxidation deficiency will affect whole body metabolic homeostasis through circulating lipids. We analyzed serum samples from 5 WT and 5 Acox1-LKO mice.
Mitochondrial complex I promotes kidney cancer metastasis
STUDY_SUMMARY
Most kidney cancers display metabolic dysfunction but how this relates to cancer progression in humans is unknown. We infused 13C-labeled nutrients during surgical tumour resection in over 80 patients with kidney cancer. Labeling from [U-13C]glucose varies across subtypes, indicating that the kidney environment alone cannot account for all metabolic reprogramming in these tumours. Compared to the adjacent kidney, clear cell renal cell carcinomas (ccRCC) display suppressed labelling of tricarboxylic acid (TCA) cycle intermediates in vivo and in organotypic cultures ex vivo, indicating that suppressed labeling is tissue intrinsic. Infusions of [1,2-13C]acetate and [U-13C]glutamine in patients, coupled with measurements of respiration in mitochondria isolated from kidneys and tumours, reveal electron transport chain (ETC) defects in ccRCC. However, ccRCC metastases unexpectedly have enhanced TCA cycle labeling compared to primary ccRCCs, indicating a divergent metabolic program during metastasis in patients. In mice, stimulating respiration or NADH recycling in kidney cancer cells is sufficient to promote metastasis, while inhibiting ETC complex I decreases metastasis. These findings indicate that metabolic properties and liabilities evolve during kidney cancer progression in humans, and that mitochondrial function is limiting for metastasis but not for growth at the original site.
INSTITUTE
University of Texas Southwestern Medical Center at Dallas
To combat the global burden of malaria, development of new drugs to replace or complement current therapies are urgently required. Here we show that the compound MMV1557817 is a selective, nanomolar inhibitor of both Plasmodium falciparum and Plasmodium vivax aminopeptidases M1 and M17, leading to inhibition of end stage haemoglobin digestion in asexual parasites. MMV1557817 can kill sexual stage P. falciparum, is active against murine malaria and did not show any shift in activity against a panel of parasites resistant to other antimalarials. MMV1557817-resistant P. falciparum exhibited a slow growth rate that was quickly outcompeted by wild type parasites and were sensitised to the current clinical drug, artemisinin. Overall, these results confirm MMV1557817 as a lead compound for further drug development and highlight the potential of dual inhibition of M1 and M17 as an effective multi-species drug targeting strategy.
INSTITUTE
Monash University
LAST_NAME
Siddiqui
FIRST_NAME
Ghizal
ADDRESS
381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia
Muscle-type specific stress responses explain the onset of external ophthalmoplegia in mitochondrial disease
STUDY_SUMMARY
Mitochondrial dysfunctions elicit progressive tissue-specific stress responses that can be protective or deleterious. Here, we show that even different muscles of an individual react differently to mitochondrial disease. In mitochondrial myopathy (MM), the extraocular muscles (EOMs) are affected first, followed by exercise intolerance in the large lower limb muscles. Both muscle types show clear signs of respiratory chain deficiency. However, the limbs upregulate the mitochondrial integrated stress response (ISRmt) that drives glucose carbons to anabolic repair pathways, while EOMs present no signs of ISRmt. In contrast, in EOMs, Pdk4 activation inhibits pyruvate metabolism, while beta-oxidation of fatty acids is induced - despite the reliance of beta-oxidation on a functional respiratory chain for ATP production. The data suggest that the inability to upregulate ISRmt and consequent deleterious fuel choices sensitize EOMs to early weakness and atrophy, explaining ophthalmoplegia, the most common MM sign. The distinct responses to disease even in muscles of a single individual predict different responses to treatment, which is essential knowledge when designing interventions.
Exploring The Impact of Two Novel DNA Minor Groove Binders on HCT-116 Cells: A Comprehensive Multi-Omics Analysis Using Mass Spectrometry
STUDY_TYPE
LC/MS/MS
STUDY_SUMMARY
Colorectal cancer (CRC) poses a significant global health challenge, necessitating innovative therapeutic approaches. Despite advancements, current treatments encounter obstacles such as chemotherapy resistance and adverse effects due to non-selective targeting. DNA Minor Groove Binders (MGBs) present promising alternatives, targeting DNA structure without causing permanent damage. In this study, two novel MGB compounds were synthesized, MGB30 and MGB32, resembling distamycin, a natural DNA-binding agent. These compounds bind reversibly to the DNA minor groove, influencing DNA structure and inhibiting cancer growth-related enzymes. Our study aims to explore the unique effects of MGB30 and MGB32 on the metabolomic profiles of treated HCT-116 cells using TIMS-QTOF-UHPLC-MS. Objectives include comprehensive analysis, comparison of effects, identification of altered pathways, and insights into MGB compound mechanisms. Additionally, we established four biological replicates for each treatment condition. Advanced statistical analyses, including the two-tailed independent Student's t-test and one-way analysis of variance (ANOVA), were utilized to minimize false discoveries. Our analysis generated a comprehensive dataset from 12 samples, identifying 75 distinct metabolites. The significance of this study lies in elucidating the molecular mechanisms of action of MGB30 and MGB32, crucial for their development as CRC drug candidates.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
BT-474 cells fed with [13C2] acetate and treated with 1 uM Fasnall or 1 uM GSK2194069 for 24 h
STUDY_TYPE
Intracellular metabolomics, [13C2] acetate
STUDY_SUMMARY
BT-474 breast cancer cells fed with [13C2] acetate and treated with 1 uM Fasnall or 1 uM GSK2194069 for 24 h. Cells were grown in RPMI-1640 with 10% dialyzed FBS.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Impact of early-life exposure to a potent aryl hydrocarbon receptor ligand on gut microbiota and host glucose homeostasis in C57BL/6J male mice (Part I)
STUDY_SUMMARY
This study aimed to explore the association between early-life exposure to a potent aryl hydrocarbon receptor (AHR) agonist and persistent disruptions in the microbiota, leading to impaired metabolic homeostasis later in life. This study utilized metagenomics, NMR- and mass spectrometry-based metabolomics, and biochemical assays to analyze the gut microbiome composition and function, as well as the physiological and metabolic effects of early-life exposure to 2,3,7,8-tetrachlorodibenzofuran (TCDF) in conventional, germ-free (GF), and Ahr-null mice. The impact of TCDF on Akkermansia muciniphila (A. muciniphila) in vitro was assessed using optical density (OD 600), flow cytometry, transcriptomics, and mass spectrometry-based metabolomics. TCDF-exposed mice exhibited disruption in the gut microbiome community structure and function, characterized by lower abundances of A. muciniphila, lower levels of cecal short chain fatty acids (SCFAs) and indole-3-lactic acid (ILA), and a reduction in gut hormones GLP-1 and PYY. Importantly, microbial and metabolic phenotypes associated with early-life POP exposure were transferable to GF recipients in the absence of POP carry-over. In addition, AHR-independent interactions between POPs and the microbiota were observed, significantly affected the growth, physiology, gene expression, and metabolic activity of A. muciniphila, resulting in suppressed activity along the ILA pathway.
Impact of early-life exposure to a potent aryl hydrocarbon receptor ligand on gut microbiota and host glucose homeostasis in C57BL/6J male mice (Part II)
STUDY_SUMMARY
This study aimed to explore the association between early-life exposure to a potent aryl hydrocarbon receptor (AHR) agonist and persistent disruptions in the microbiota, leading to impaired metabolic homeostasis later in life. This study utilized metagenomics, NMR- and mass spectrometry-based metabolomics, and biochemical assays to analyze the gut microbiome composition and function, as well as the physiological and metabolic effects of early-life exposure to 2,3,7,8-tetrachlorodibenzofuran (TCDF) in conventional, germ-free (GF), and Ahr-null mice. The impact of TCDF on Akkermansia muciniphila (A. muciniphila) in vitro was assessed using optical density (OD 600), flow cytometry, transcriptomics, and mass spectrometry-based metabolomics. TCDF-exposed mice exhibited disruption in the gut microbiome community structure and function, characterized by lower abundances of A. muciniphila, lower levels of cecal short chain fatty acids (SCFAs) and indole-3-lactic acid (ILA), and a reduction in gut hormones GLP-1 and PYY. Importantly, microbial and metabolic phenotypes associated with early-life POP exposure were transferable to GF recipients in the absence of POP carry-over. In addition, AHR-independent interactions between POPs and the microbiota were observed, significantly affected the growth, physiology, gene expression, and metabolic activity of A. muciniphila, resulting in suppressed activity along the ILA pathway.
BT474 breast cancer cell line grown in 20% D2O-containing RPMI-1640 medium treated with Fasnall and GSK2194069
STUDY_TYPE
Free fatty acid analysis, D2O tracing
STUDY_SUMMARY
BT474 breast cancer cell line grown in 20% D2O-containing medium treated with Fasnall and GSK2194069. Cells were grown for 24 h in RPMI-1640 with 10% dialyzed FBS.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Metabolomics analysis of murine xenograft tumors derived from human breast cancer cell line MCF7 1 h after isotopic glucose bolus. The NSG female mice were ~55 days after grafting tumors. [U-13C6] D-Glucose bolus was delivered intraperitoneally with a 10 mg/kg dose of Fasnall.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Nucleotide metabolism of tumor interstitial fluid in one murine model of pancreatic cancer
STUDY_SUMMARY
The experiment focus on the nucleotide metabolism of tumor interstitial fluid in one murine model of pancreatic cancer. Briefly, KPC FC1245 cells were genetically engineered using doxycycline inducible CRISPR/Cas9 system platform to target specifically cytidine deaminase (sgCda). sgCda and the control sgNT were then orthotopically injected in the head of the pancreas of 8-10 weeks old C57BL/6J female mice. When the tumor weight reached approximately 0.2-0.4g, mice were sacrificed and tumor interstitial fluid was collected as reported in the Collection section of this study. Standard curves for glutamine, cytidine, uridine, glucose, UTP and UDP were prepared and extracted along with the interstitial fluid samples. The concentrations of glutamine, cytidine, uridine, UDP and UTP were measured by LC-MS, and glucose was measured by GC-MS. A decrease in uridine content, and accordingly in UDP and UTP, and a concomitant accumulation of cytidine was observed in sgCda tumors. On the other hand, no differences were observed in glucose and glutamine abundance.
NMR-based metabolomics combined with metabolic pathway analysis reveals metabolic heterogeneity of colorectal cancer tissue at different anatomical locations and stages
STUDY_SUMMARY
Colorectal cancer (CRC) still remains the leading cause of cancer death worldwide. This study aimed to profile the metabolic differences of colorectal cancer tissues (CCT) at different stages and sites, as compared with their adjacent noncancerous tissues (ANT), to investigate the temporal and spatial heterogeneities of metabolic characterization. Our NMR-based metabolomics fingerprinting revealed that many of the metabolite levels were significantly altered in CCT compared to ANT and esophageal cancer tissues, indicating deregulations of glucose metabolism, one-carbon metabolism, glutamine metabolism, amino acid metabolism, fatty acid metabolism, TCA cycle, choline metabolism, etc. A total of five biomarker metabolites, including glucose, glutamate, alanine, valine and histidine, were identified to distinguish between early and advanced stages of CCT. Metabolites that distinguish the different anatomical sites of CCT include glucose, glycerol, glutamine, inositol, succinate, and citrate. Those significant metabolic differences in CRC tissues at different pathological stages and sites suggested temporal and spatial heterogeneities of metabolic characterization in CCT, providing a metabolic foundation for further study on biofluid metabolism in CRC early detection.
INSTITUTE
Radiology Department, Second Affiliated Hospital, Shantou University Medical College, Shantou
LAST_NAME
Lin
FIRST_NAME
Yan
ADDRESS
No. 69, Dongxia North Road, Shantou, Guangdong, China, Shantou, Guangdong, China, 515041, China
Diet modulates the protective effects of dimethyl fumarate mediated by the immunometabolic neutrophil receptor HCA2 - Metabolomics
STUDY_SUMMARY
Monomethyl fumarate (MMF) and its prodrug dimethyl fumarate (DMF) are currently the most widely used agents for the treatment of multiple sclerosis (MS). However, not all patients benefit from DMF. We hypothesized that the variable response of patients may be due to their diet. In support of this hypothesis, mice subjected to experimental autoimmune encephalomyelitis (EAE), a model of MS, did not benefit from DMF treatment when fed a lauric acid-rich (LA) diet, in contrast to normal chow (NC) or, more so, a high-fiber (HFb) diet. DMF lacked efficacy in the LA diet fed group despite similar resorption and preserved effects on plasma lipids.
Diet modulates the protective effects of dimethyl fumarate mediated by the immunometabolic neutrophil receptor HCA2 - Lipidomics
STUDY_SUMMARY
Monomethyl fumarate (MMF) and its prodrug dimethyl fumarate (DMF) are currently the most widely used agents for the treatment of multiple sclerosis (MS). However, not all patients benefit from DMF. We hypothesized that the variable response of patients may be due to their diet. In support of this hypothesis, mice subjected to experimental autoimmune encephalomyelitis (EAE), a model of MS, did not benefit from DMF treatment when fed a lauric acid-rich (LA) diet, in contrast to normal chow (NC) or, more so, a high-fiber (HFb) diet. DMF lacked efficacy in the LA diet fed group despite similar resorption and preserved effects on plasma lipids.
Untargeted serum metabolomics in the Parkinson's Environment and Genes (PEG) Study
STUDY_SUMMARY
This project aims to evaluate the serum metabolome of Parkinsonās disease (PD) patients relative to unaffected controls in the Parkinsonās Environment and Genes (PEG) Study. Background: Untargeted high-resolution metabolomic profiling provides simultaneous measurement of thousands of metabolites. Metabolic networks based on these data can help uncover disease-related perturbations across interconnected pathways. Objective: Identify metabolic disturbances associated with PD in the PEG population-based study using untargeted metabolomics. Methods: We provide serum-based untargeted metabolomics data derived from liquid chromatography with high-resolution mass spectrometry (LC-HRMS). LC-HRMS detected 4,762 metabolites for analysis (HILIC: 2716 metabolites; C18: 2046 metabolites).
New class of heterospirocyclic compounds present strong and rapid activity against artemisinin- and multidrug-resistant P. falciparum parasites
STUDY_SUMMARY
Malaria remains a significant health burden and a leading contributor to global mortality rates. Increasing drug resistance creates an urgent demand for novel treatment options. We have synthesised a new class of heterospirocyclic compounds with novel chemical connectivities. Compounds 25 and 26 display antimalarial activity within 24 h and have similar potency against a panel of drug-resistant strains of Plasmodium falciparum, the most virulent of human malaria parasites, including parasites resistant to the frontline artemisinin antimalarials. C25 and C26 do not induce major toxicity in kidney- and hepatic-derived human cell lines, highlighting their specificity. Untargeted metabolomics analysis of P. falciparum infected red blood cells revealed that the mechanism of action of C25 involves disruption of the pyrimidine biosynthesis pathway and haemoglobin catabolism. These heterospirocyclic compounds represent a promising opportunity for antimalarial drug development and could prove relevant against drug resistant malaria.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Diet-omics in the Study of Urban and Rural Crohn disease Evolution (SOURCE) cohort
STUDY_SUMMARY
Crohn disease (CD) burden has increased with globalization/urbanization, and the rapid rise is attributed to environmental changes rather than genetic drift. The Study Of Urban and Rural CD Evolution (SOURCE, n=380) has considered diet-omics domains simultaneously to detect complex interactions and identify potential beneficial and pathogenic factors linked with rural-urban transition and CD. We characterize exposures, diet, ileal transcriptomics, metabolomics, and microbiome in newly diagnosed CD patients and controls in rural and urban China and Israel. We show that time spent by rural residents in urban environments is linked with changes in gut microbial composition and metabolomics, which mirror those seen in CD. Ileal transcriptomics highlights personal metabolic and immune gene expression modules, that are directly linked to potential protective dietary exposures (coffee, manganese, vitamin D), fecal metabolites, and the microbiome. Bacteria-associated metabolites are primarily linked with host immune modules, whereas diet-linked metabolites are associated with host epithelial metabolic functions.
INSTITUTE
Sheba hospital
LAST_NAME
Braun
FIRST_NAME
Tzipi
ADDRESS
Sheba hospital, Ramat Gan, Ramat Gan, 52621, Israel
Loss of dihydroceramide desaturase drives neurodegeneration by disrupting endoplasmic reticulum and lipid droplet homeostasis in glial cells
STUDY_TYPE
Untargeted Metabolomics & Lipidomics
STUDY_SUMMARY
Dihydroceramide desaturases convert dihydroceramides to ceramides, the precursors of all complex sphingolipids. Reduction of DEGS1 dihydroceramide desaturase function causes pediatric neurodegenerative disorder hypomyelinating leukodystrophy-18 (HLD-18). We discovered that infertile crescent (ifc), the Drosophila DEGS1 homolog, is expressed primarily in glial cells to promote CNS development by guarding against neurodegeneration. Loss of ifc causes massive dihydroceramide accumulation and severe morphological defects in cortex glia, including endoplasmic reticulum (ER) expansion, failure of neuronal ensheathment, and lipid droplet depletion. RNAi knockdown of the upstream ceramide synthase schlank in glia of ifc mutants rescues ER expansion, suggesting dihydroceramide accumulation in the ER drives this phenotype. RNAi knockdown of ifc in glia but not neurons drives neuronal cell death, suggesting that ifc function in glia promotes neuronal survival. Our work identifies glia as the primary site of disease progression in HLD-18 and may inform on juvenile forms of ALS, which also feature elevated dihydroceramide levels.
INSTITUTE
Washington University in St. Louis
DEPARTMENT
Genetics, Medicine, Chemistry
LABORATORY
Skeath and Patti Laboratories
LAST_NAME
Cho
FIRST_NAME
Kevin
ADDRESS
1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA
Traumatic brain injury (TBI) is a global public health problem with 50-60 million incidents per year, most of which are considered mild (mTBI) and many of these repetitive (rmTBI). Despite their massive implications, the pathologies of mTBI and rmTBI are not fully understood, with a paucity of information on brain lipid dysregulation following mild injury event(s). To gain more insight on mTBI and rmTBI pathology, a non-targeted spatial lipidomics workflow utilizing ultrahigh resolution mass spectrometry imaging was developed to map brain region-specific lipid alterations in rats following injury. Discriminant multivariate models were created for regions of interest including the hippocampus, cortex, and corpus callosum to pinpoint lipid species that differentiated between injured and sham animals. A multivariate model focused on the hippocampus region differentiated injured brain tissues with an area under the curve of 0.994 using only four lipid species. Lipid classes that were consistently discriminant included polyunsaturated fatty acid-containing phosphatidylcholines (PC), lysophosphatidylcholines (LPC), LPC-plasmalogens (LPC-P) and PC potassium adducts. Many of the polyunsaturated fatty acid-containing PC and LPC-P selected have never been previously reported as altered in mTBI. The observed lipid alterations indicate that neuroinflammation and , oxidative stress and disrupted sodium-potassium pumps are important pathologies that could serve to explain cognitive deficits associated with rmTBI. Therapeutics which target or attenuate these pathologies may be beneficial to limit persistent damage following a mild brain injury event.
Metabolomics of Murine WT or MCJ KO CD19-BBz CD8 CAR-T cells
STUDY_SUMMARY
MCJ/DnaJC15 is an endogenous negative regulator of Complex I and mitochondrial respiration. The goal of these experiments are to characterize the metabolic profile of murine WT or MCJ KO CD8 CD19-BBz CAR-T cells after 3 expansions (6 days) with IL-2 (60IU/ml).
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
Metabolism of chimeric antigen receptor (CAR) T cells is emerging as an important area to improve CAR-T cell therapy in cancer treatment. Mitochondrial respiration is essential for survival and function of CAR-T cells, but developing strategies to specifically enhance mitochondrial respiration has been challenging. Here we identify MCJ/DnaJC15, an endogenous negative regulator of mitochondrial Complex I, as a metabolic target to enhance mitochondrial respiration in CD8 CAR-T cells. Loss of MCJ in CD8 CAR-T cells increases their in vitro and in vivo efficacy against mouse B cell leukemias. MCJ deficiency in TCR- specific CD8 cells also increases their efficacy against solid tumors in vivo. Furthermore, we reveal that human CD8 cells express MCJ and that silencing MCJ expression increases mitochondrial metabolism and anti-tumor activity of human CAR-T cells. Thus, targeting MCJ to enhance mitochondrial metabolism is a promising therapeutic strategy to improve the efficacy of adoptive T cell therapies.
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
Lipidomic analysis of cryopreserved human cardiac tissue from young and ageing adults
STUDY_SUMMARY
Untargeted lipidomic analysis was performed to measure lipids in left ventricular heart tissue from pre-mortem healthy donor hearts, as classified by formal pathological examination. Hearts were stored at the Sydney Heart Bank. Samples were divided into young (age ā¤ 25 years) and old (age ā„ 50 years) cohorts. Lipidomic analysis used liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a high resolution Q-Exactive HF-X Quadrupole-Orbitrap mass spectrometer, operated in both positive and negative ionisation mode.
INSTITUTE
University of Sydney
DEPARTMENT
Medicine and Health
LABORATORY
Lipid Metabolism and Neurochemistry
LAST_NAME
Don
FIRST_NAME
Anthony
ADDRESS
The Hub, Charles Perkins Centre, D17, The University of Sydney, NSW, 2006
Metabolism of chimeric antigen receptor (CAR) T cells is emerging as an important area to improve CAR-T cell therapy in cancer treatment. Mitochondrial respiration is essential for survival and function of CAR-T cells, but developing strategies to specifically enhance mitochondrial respiration has been challenging. Here we identify MCJ/DnaJC15, an endogenous negative regulator of mitochondrial Complex I, as a metabolic target to enhance mitochondrial respiration in CD8 CAR-T cells. Loss of MCJ in CD8 CAR-T cells increases their in vitro and in vivo efficacy against mouse B cell leukemias. MCJ deficiency in TCR- specific CD8 cells also increases their efficacy against solid tumors in vivo. Furthermore, we reveal that human CD8 cells express MCJ and that silencing MCJ expression increases mitochondrial metabolism and anti-tumor activity of human CAR-T cells. Thus, targeting MCJ to enhance mitochondrial metabolism is a promising therapeutic strategy to improve the efficacy of adoptive T cell therapies.
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
Metabolism of chimeric antigen receptor (CAR) T cells is emerging as an important area to improve CAR-T cell therapy in cancer treatment. Mitochondrial respiration is essential for survival and function of CAR-T cells, but developing strategies to specifically enhance mitochondrial respiration has been challenging. Here we identify MCJ/DnaJC15, an endogenous negative regulator of mitochondrial Complex I, as a metabolic target to enhance mitochondrial respiration in CD8 CAR-T cells. Loss of MCJ in CD8 CAR-T cells increases their in vitro and in vivo efficacy against mouse B cell leukemias. MCJ deficiency in TCR- specific CD8 cells also increases their efficacy against solid tumors in vivo. Furthermore, we reveal that human CD8 cells express MCJ and that silencing MCJ expression increases mitochondrial metabolism and anti-tumor activity of human CAR-T cells. Thus, targeting MCJ to enhance mitochondrial metabolism is a promising therapeutic strategy to improve the efficacy of adoptive T cell therapies.
INSTITUTE
University of Colorado School of Medicine
LABORATORY
Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon
LAST_NAME
Cendali
FIRST_NAME
Francesca
ADDRESS
13199 East Montview Boulevard, Aurora, CO, 80045, USA
Untargeted Metabolomics for Exploring Metabolomic Profile of Maple Syrup Urine Disease Sick Patients
STUDY_TYPE
Untargeted LCMS
STUDY_SUMMARY
Abstract non-newborn: Background: A malfunction in the activity of the branched-chain alpha-ketoacid dehydrogenase (BCKAD) complex results in maple syrup urine disease (MSUD), a genetically inherited illness. Three amino acidsāleucine, isoleucine, and valineāare typically broken down by this complex. Abnormal activity in this process, therefore, can affect vital body systems and result in metabolic dysregulation associated with the consequences of the disease. The therapy and follow-up of ill MSUD patients are greatly aided by many researched endogenous metabolites as well as dysregulated biomarkers and pathways. Objectives: Our goal is to add to the increasing knowledge of information about sick MSUD with relation to MSUD newborns and the pathways that are involved in improving patient outcomes by utilizing untargeted metabolomics to examine the unique profile of MSUD in sick MSUD patients. Methods: This study evaluated the metabolic changes in the dry blood spot (DBS) of 14 sick MSUD patients and 14 healthy controls utilizing untargeted metabolomics studies performed with liquid chromatographyāmass spectrometry. Findings: Based on metabolomics analysis,7754 metabolites were found to be highly dysregulated.Out of them,3716 were up-regulated and 4038 were down-regulated.1557 of the annotated metabolites were found to be endogenous metabolites. The research found possible biomarkers for MSUD, including Glutathioselenol and dUDP, which were elevated in sick MSUD relative to healthy controls and LysoPI downregulated in sick MSUD. Moreover, the Sphingolipid metabolism, selenocompound metabolism and porphyrin metabolism pathways were the most impacted in MSUD newborns.This study shows 92 endogenous metabolites between newborn MSUD and sick MSUD. In summary, our findings shows that metabolomics is a noninvasive approach to understanding the pathophysiology of the medical condition and a potentially useful technique for assessing novel biomarkers in the early detection of sick MSUD.Further research is required regarding the relationship of these dysregulated metabolites to compromised pathways.
Untargeted Metabolomic Profile Of Chili Pepper (Capsicum Chinensed) Developmental Cycle
STUDY_SUMMARY
To explore the temporal dynamics of metabolites in chili peppers at various developmental stages, we employed a non-targeted metabolomics method based on liquid chromatography-mass spectrometry (LC-MS). The metabolome data collection began on the day of flowering (0 days post-anthesis), and continued for 7, 16, 30, 50, 55, and 60 DPA. This comprehensive dataset paved the way for future studies on metabolite changes and biological processes in chili peppers throughout their life cycle.
Assessment and partial characterization of candidate genes in dihydrochalcone and arbutin biosynthesis in an apple-pear hybrid by de novo transcriptome assembly
STUDY_SUMMARY
The goal of the study was to determine the phenolic profile of young and old leaves, as well as fruit of apple (Malus x domestica), pear (Pyrus communis) and an intergeneric apple-pear hybrid. Three independent replicates were obtained for each genotype from the germplasm collection at Fondazione Edmund Mach (Italy) and analyzed by a targeted phenolic LC/MS-MS method. In addition, candidate genes from apple, pear and apple-pear hybrid retrieved from a de novo transcriptome assembly were expressed in E. coli and recombinant proteins were tested (in triplicate) to determine the conversion of hydroquinone to arbutin. Combining RNA-Seq, in silico functional annotation prediction, targeted gene expression analysis and expression ā metabolite correlations with the data submitted to Metabolomics Workbench, we identified candidate genes for functional characterisation, resulting in the identification of active arbutin synthases in the hybrid and parental genotypes. We found that the putative arbutin synthases of pear (PcAS) and apple-pear hybrid (HybAS) were able to convert hydroquinone into arbutin. Interestingly, also one out of two putative arbutin synthases isolated from apple (MdAS1) could produce arbutin in vitro. However, the metabolomic profiling of phenolic compounds showed that apple lacks of arbutin and was found to accumulate the precursor hydroquinone in traces in young and old leaves of apple. Although quercetin was accumulated in similar amounts in the same tissues, a luminiscence-based assay showed that quercetin was converted only 25% compared to activity towards hydroquinone in the tested conditions. In summary, the metabolomic profiling submitted to Metabolomics workbench also shows that: 1) arbutin is accumulated mainly in young leaves of pear, followed by the apple-pear hybrid and was found in traces in apple fruit; 2) rutin was found mainly in pear and apple-pear hybrid tissues; 3) phenolic profile of apple is dominated by phloridzin and undetectable in all pear tissues analyzed, with young leaves being the tissue showing highest accumulation.
INSTITUTE
Fondazione Edmund Mach
LAST_NAME
Miranda Chavez
FIRST_NAME
Simon David
ADDRESS
Via Mach, 1, San Michele all'Adige, Trento, 38098, Italy
O-GlcNAcase activity maintains stress granules for proximity-enhanced ATP production to ensure recovery from stress (Part 3)
STUDY_SUMMARY
Accurate disassembly of stress granules (SGs) after environmental stimuli release is essential for cells to maintain homeostasis , which requires ATP-consuming processes. However, the molecular mechanism whereby regulation of SGs programmatically disassembly and ATP restoration remain poorly understood in mammalian cells. Here we found that defect of OGA in cells leads to aggregates formation, severe autophagy and eventually apoptosis during stress recovery. OGA, which localized in SGs, had no effect on SGs formation but could protect SGs from rapid disassembly during stress recovery stage. Then the SGs localized glycolysis-related enzymes were reserved and concentrated in SGs during stress release for ATP production in a proximity manner, which was vital to guarantee cells resistant to stress and survival during recovery. Finally, supplementation of ATP to OGA knockdown cells during stress recovery significantly rescue cell from aggregates, autophagy and apoptosis. Together, these results describe a brand new mechanism on how OGA regulates the programmed disassembly of stress granules and restoration of ATP to safeguard cell viability in a very precisely programmed process, whose rate is rigorous regulated. This is a continuation of study ST002927 and ST002936 where an in vitro reaction was performed with 13C-glucose on purified stress granules, to validate the conclusion.
INSTITUTE
Zhejiang University
DEPARTMENT
Life Sciences Institute
LABORATORY
Shixian Lin
LAST_NAME
Chen
FIRST_NAME
Yulin
ADDRESS
Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang province, China
Inactivation of mitochondrial MUL1 E3 ubiquitin ligase inhibits lipogenesis and prevents diet-induced obesity in mice
STUDY_TYPE
Basic Research
STUDY_SUMMARY
Characterize the role of mitochondrial MUL1 E3-ubiquitin ligase on energy metabolism and lipogenesis using Mul1 deficient, Mul1(-/-), mice. MUL1 protein is involved in mitochondrial dynamics, and innate immune response but its primary function might be the regulation of lipogenesis under conditions of nutritional overload. Using metabolic cages, we monitored whole body energy expenditure, metabolism, and thermoregulation of the Mul1(-/-) mice under standard diet (ND) or high fat diet (HFD). We examined the effect of Mul1 inactivation on body weight, HFD-induced adiposity, fatty liver, glucose intolerance, and insulin resistance. We performed global metabolomics, lipidomic, and genome-wide mRNA sequencing using liver from Mul1(+/+) and Mul1(-/-) animals on HFD. The expression level of key proteins involved in lipogenesis and their regulation in the absence of MUL1 was monitored by SDS-PAGE and Western blot analysis. We found that Mul1(-/-) animals have a metabolic phenotype that confers robust resistance to HFD-induced obesity. Several metabolic and lipidomic pathways are perturbed in the liver of Mul1(-/-) animals on HFD, particularly the one driven by Stearoyl-CoA Desaturase 1 (SCD1), a key regulator of lipid metabolism and obesity. In addition, key enzymes involved in lipogenesis and fatty acid oxidation such as ACC1, FASN, AMPK, and CTP1 were also modulated. The concerted deregulation of these enzymes, in the absence of MUL1, causes reduced fat storage and increased fatty acid oxidation. We identified a new function of mitochondrial MUL1 E3 ubiquitin ligase in the regulation of lipogenesis and adiposity, particularly during conditions of HFD. Inactivation of MUL1 provides resistance to HFD-induced obesity and can be a therapeutic target for the treatment of obesity, type 2 diabetes, and nonalcoholic fatty liver disease (NAFLD).
Chronic manganese exposure disrupts glutamatergic signaling and alters behavior in APP/PSEN1 mice.
STUDY_TYPE
untargeted metabolomics analysis
STUDY_SUMMARY
Pathological changes in AlzheimerĆ¢ĀĀs disease (AD) begin decades before cognitive symptoms appear. Chronic exposure to environmental neurotoxins like manganese (Mn) from sources such as air pollution and water can impact disease development across decades. Mn toxicity can lead to motor and neuropsychiatric symptoms resembling Parkinson's disease, attributed to its accumulation in globus pallidus. In contrast, the impact of chronic lower-level exposures on normal and neuropathological brain function is unknown. We tested whether 10 months of a high Mn diet (2400 ppm) could alter aspects of glutamate signaling in APP/PSEN1 mice to further exacerbate cognitive impairments compared to control fed animals (70 ppm). This study supports the potential role for chronic environmental exposures in the development of AlzheimerĆ¢ĀĀs disease associated neuropathology.
A Longitudinal Study in Rheumatoid Arthritis Unveils Metabolomic Biomarkers Preceding Clinical Onset, Assessing Disease Severity, and Anticipating Treatment Response to csDMARDs
STUDY_SUMMARY
Rheumatoid arthritis (RA) is a bundle of systemic inflammatory diseases mainly affecting the joints, complicating the identification of biomarkers for early diagnosis, predicting disease progress and therapeutic outcomes. This study scrutinizes a longitudinal cohort of RA, inclusive of follow-ups, alongside OA, UA and ACPA/RF-RA and healthy controls, aiming to discover plasma metabolic markers that can precede RA onset, assess disease activity, and forecast treatment efficacy. Our investigation revealed substantial metabolic alterations at both the pathway and individual metabolite levels across RA, at-risk or RA and healthy control. The drug response predictive models constructed on critical differential metaboites showed optimal performance. Additionally, our longitudinal data sheds light on the molecular impacts on metabolism of csDMARDs in RA.
INSTITUTE
West China Hospital of Sichuan University
LAST_NAME
Zhu
FIRST_NAME
Chenxi
ADDRESS
West China Hospital, Sichuan University, 37# Guoxue Xiang, Chengdu, Sichuan, 610041, China.
In 2016, the National Institutes of Health (NIH) launched an initiative to study ME/CFS. The NIH Division of Intramural Research developed an exploratory clinical research program to perform deep phenotyping on a cohort of PI-ME/CFS volunteers and healthy volunteers (HV) as controls. Prior to the SARS-CoV-2 pandemic, this study recruited a cohort of well-characterized PI-ME/CFS patients and applied modern broad and deep scientific measures to describe their biophenotype compared to HVs. The aim was to identify relevant group differences that could generate new hypotheses about the pathogenesis of PI-ME/CFS and provide direction for future research. Over 75 scientists and clinicians across 15 of the 27 institutes that comprise the NIH contributed to this multi-disciplinary work. Importantly, we developed rigorous inclusion criteria which comprised detailed medical and psychological evaluations to minimize diagnostic misattribution. A relatively homogenous population was recruited in whom symptoms were initiated after infection. This study aimed to investigate the underlying pathophysiological mechanisms. The volunteers underwent a multi-dimensional evaluation that included a wide range of physiological measures, physical and cognitive performance testing, and biochemical, microbiological, and immunological assays of blood, cerebrospinal fluid, muscle, and stool. Novel measurement techniques were developed to query issues such as physical capacity, effort preference, and deconditioning that may confound the results. Multi-omic measurements of gene expression, proteins, metabolites, and lipids were performed in parallel on collected samples.
Property and Activity Refinement of Dihydroquinazolinone-3-carboxamides as Orally Efficacious Antimalarials that Target PfATP4
STUDY_SUMMARY
The development of new antimalarial classes is pertinent because of resistance against the current antimalarial treatments. To contribute to the global effort to create new antimalarial therapies, we previously disclosed initial findings on the optimization of the dihydroquinazolinone-3-carboxamide class that targets PfATP4. The preliminary optimization generated analogs, such as 4, that exhibited potent in vitro asexual stage activity but only showed modest oral efficacy in a P. berghei mouse model attributed to its low aqueous solubility and modest metabolic stability. Here we report on correcting these parameters to improve in vivo efficacy. We show that the incorporation of heterocycle systems in the 8-position of the scaffold markedly improved aqueous solubility without a significant loss of asexual parasite activity. Certain configurations of pyrazoles in the 8-position were found to provide the greatest attainable balance between parasite activity, aqueous solubility, and metabolic stability. We were able to show that modifications made to the optimized analogs, such as WJM992 did not perturb the sensitivity to PfATP4 drug-resistant parasites or alter on-target activity in a PfATP4-associated parasite cytosolic Na+ flux assay and gave a distinct metabolic signature indicative of other PfATP4 inhibitors. The optimized analogs showed an appreciable efficacy in malaria mouse models and blocked sexual stage gamete development preventing transmission to mosquitoes.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Depression symptoms modifies differently plasma metabolites in pre- and post-menopausal women
STUDY_SUMMARY
Depression will be the disease with the highest incidence worldwide by 2030. Data indicate that postmenopausal women have a higher incidence of mood disorders, and this high vulnerability seems to be related to hormonal changes and weight gain. Although research evaluating the profile of metabolites in mood disorders is advancing, further research, maintaining consistent methodology, is necessary to reach a consensus. Therefore, the objective of the present study was to carry out an exploratory analysis of the plasma polar metabolites and lipids of pre- and postmenopausal women to explore whether the profile is affected by depression. The study was performed in accordance with the principles of the Declaration of Helsinki and was approved by the Human Research Ethics Committee of the Universidade Federal de SĆ£o Paulo (nĀŗ 0624/2019) and all participants signed the informed consent. 42 premenopausal and 67 postmenopausal women had the depression symptoms assessed by the Beck Depression's Inventory (BDI). Lipids and polar metabolites were extracted from plasma and analyzed in an ultra-performance liquid chromatography system (UHPLC) coupled to a spectrometer with a triple-quadrupole analyzer operating with an electrospray ionization source (ESI) in positive and negative mode. Lipids and polar metabolites analyses was performed using MetaboAnalyst 5.0. Ten metabolites were significantly affected by depression symptoms in postmenopause, including Adenosine, Guanosine, Proline, Citrulline, Lysine, and Carnitine, which were down-regulated, and Dimethylglycine, Glutathione, Creatine, and Methionine that were up-regulated. In premenopausal women with depression, Oxidized Glutathione was down-regulated, and Dimethylglycine and 4-hydroxyproline were up-regulated. Seven lipids were were significantly affected by depression symptoms in pre-menopausal women PC(36:1)/PC(18:1(9Z)/18:0), PC(19:0/19:0), and LTB4 were up-regulated and PC(18:0/14:0)/GPCho(18:0/14:0), SM(d18:1/16:0), LysoPC(18:1(9z))/LPC 18:1, and Azelaoyl-PAF were down-regulated. In post-menopausal women PC(16:1(9z)/16:1(9z)) and 14,15-DHET were down-regulated and acetylcarnitine was up-regulated. Though there seems to be a relationship in the occurrence of obesity and depression in women the hormonal status influenced the effect of depression in women and that this status may be taken into account when searching for a marker for depression.
INSTITUTE
Federal University of SĆ£o Paulo
LAST_NAME
Boldarine
FIRST_NAME
Valter
ADDRESS
Rua Botucatu, 862, 2Āŗ floor, SĆ£o Paulo, SĆ£o Paulo, 04026-000, Brazil
METTL3-mediated chromatin contacts promote stress granule phase separation through metabolic reprogramming during senescence
STUDY_SUMMARY
METTL3 is the catalytic subunit of the methyltransferase complex, which mediates m6A modification to regulate gene expression. In addition, METTL3 regulates transcription in an enzymatic activity-independent manner by driving changes in high-order chromatin structure. However, how these functions of MTC are coordinated remains unknown. Here we show that the methyltransferase complex coordinates its enzymatic activity-dependent and independent functions to regulate cellular senescence, a state of stable cell growth arrest. Specifically, METTL3-mediated chromatin loops induce Hexokinase 2 expression through the three-dimensional chromatin organization during senescence. Elevated Hexokinase 2 expression subsequently promotes liquid-liquid phase separation, manifesting as stress granule phase separation, by driving metabolic reprogramming. This correlates with an impairment of translation of cell-cycle related mRNAs harboring polymethylated m6A sites. In summary, our results report a coordination of m6A-dependent and -independent function of the methyltransferase complex in regulating senescence through phase separation driven by metabolic reprogramming.
Campylobacter infection of young children in Colombia and its impact on the gastrointestinal metabolome
STUDY_SUMMARY
Campylobacter infections are a leading cause of bacterial-derived gastroenteritis worldwide with particularly profound impacts on pediatric patients in low-and-middle income countries. It remains unclear how Campylobacter impacts these hosts, though it is becoming increasingly evident that it is a multifactorial process that depends on the host immune response, the gastrointestinal microbiota, various bacterial factors, and host nutritional status. Since these factors likely vary between adult and pediatric patients in different regions of the world, it is important that studies define these attributes in well characterized clinical cohorts in diverse settings. In this study, we analyzed the metabolomic profiles of asymptomatic and symptomatic pediatric patients in Colombia that were either infected or uninfected with Campylobacter during a case-controlled study on acute diarrheal disease. Using computational models, we identified fecal metabolites that were associated with Campylobacter infection and found that glucose-6-phosphate and homovanillic acid were the strongest predictors of infection in these pediatric patients, which suggest that colonocyte metabolism are impacted during infection.
A multimodal drug-diet-immunotherapy combination restrains melanoma progression and metastasis - tumor lipidomics
STUDY_SUMMARY
Lipidomics profiling of immunocompetent B16F10 model of melanoma to examine lipid levels in tumors following reduced oleic acid content in diet combined with the stearyl CoA desaturase inhibitor CAY10566.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
A multimodal drug-diet-immunotherapy combination restrains melanoma progression and metastasis - plasma lipidomics
STUDY_SUMMARY
Lipidomics profiling of immunocompetent B16F10 model of melanoma to examine lipid levels in mouse plasma following reduced oleic acid content in diet combined with the stearyl CoA desaturase inhibitor CAY10566.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Untargeted plasma metabolomics on bacterial culture supernatants
STUDY_SUMMARY
As part of our pipeline to identify microbial metabolites in bloodstream infections, we performed untargeted metabolomics on bacterial cell culture supernatants. We focused on metabolites that were significantly altered in both our human cohort (PR001807) and mouse model of bloodstream infection
Untargeted Metabolomic Profile of Septic Shock in the Emergency Department
STUDY_SUMMARY
Septic shock is a life-threatening condition that arises from a dysregulated host response to infection, resulting in the failure of multiple organ systems and the rapid decline of the patientās health status. As a medical emergency, early recognition and timely interventions are crucial for improving patient outcomes, particularly in the emergency department setting where septic patients are initially treated. Despite the importance of early identification, current diagnostic tools remain challenges in predicting septic shock at the time of admission. To address these limitations, this study aimed to develop a highly sensitive and specific platform for predicting septic shock in the emergency department. By implementing a supervised algorithm to the metabolic profile, a model based on a 22-metabolite panel was developed to accurately predict septic shock. This approach has the potential to optimize the clinical management of septic shock in the emergency department.
INSTITUTE
National Taiwan University
DEPARTMENT
Department of Chemistry
LABORATORY
Cheng-Chih Hsu's lab
LAST_NAME
Hong
FIRST_NAME
Yu
ADDRESS
No. 1, Sec. 4, Roosevelt Rd., Taipei, Taipei, 106, Taiwan
A clade of receptor-like cytoplasmic kinases and 14-3-3 proteins coordinate the inositol hexaphosphate accumulation
STUDY_SUMMARY
Inositol hexaphosphate (InsP6) is the major storage form of phosphorus in seeds. Reducing seed InsP6 content is a breeding objective in agriculture, as InsP6 negatively impacts animal nutrition and the environment. Nevertheless, how InsP6 accumulation is regulated remains largely unknown. Here, we identify a clade of receptor-like cytoplasmic kinases (RLCKs), named Inositol Polyphosphate-related Cytoplasmic Kinases 1-6 (IPCK1-IPCK6), deeply involved in InsP6 accumulation. The InsP6 concentration is dramatically reduced in seeds of ipck quadruple (T-4m/C-4m) and quintuple (C-5m) mutants, accompanied with the obviously increase of phosphate (Pi) concentration. The plasma membrane-localized IPCKs recruit IPK1 involved in InsP6 synthesis, and facilitate its binding and activity via phosphorylation of GRF 14-3-3 proteins. IPCKs also recruit IPK2s and PI-PLCs required for InsP4/InsP5 and InsP3 biosynthesis respectively, to form a potential IPCK-GRF-PLC-IPK2-IPK1 complex. Our findings therefore uncover a previously uncharacterized regulatory mechanism of InsP6 accumulation governed by IPCKs, shedding new light on the mechanisms of InsP biosynthesis in eukaryotes.
Fructose 1,6-bisphosphate quantification in HepG2 liver cancer cell line.
STUDY_TYPE
LC/MS Quantitative Analysis
STUDY_SUMMARY
Quantification of specific fructose 1,6-bisphosphate (FBP) concentrations in HepG2 cells. A minimum of 1 million cells per replicate and condition will be grown and treated. Glucose-starved cells will be either treated with 10 mM glucose, 2.5 Ī¼M Oligomycin, 50 mM 2-deoxy-D-glucose (2-DG), or not treated. Cells will be lysed and processed for FBP quantification using an isotope FBP standard by LC/MS.
Aspartate tracing in Wildtype and UQCRC2 Knockout 143B Cells
STUDY_SUMMARY
Ubiquinone (UQ), the only known electron carrier in the mammalian electron transport chain (ETC), delivers electrons to both oxygen (O2) and fumarate as terminal electron acceptors. As fumarate has a lower reduction potential than UQ, fumarate reduction is only thermodynamically favorable when ubiquinol, the reduced form of UQ, accumulates. Paradoxically, some tissues reduce fumarate without ubiquinol buildup, suggesting another mechanism enables fumarate reduction in mammals. Here, we identify rhodoquinone (RQ), a novel mammalian electron carrier that directs electrons to fumarate, instead of O2, as the favored terminal electron acceptor. RQ, which is undetectable in cultured mammalian cells, is enriched in tissues that catalyze fumarate reduction. RQ and UQ-directed ETC circuits support distinct programs of mitochondrial function. Through expression of a bacterial enzyme that converts UQ into RQ and development a novel RQ analog, we demonstrate that reprogramming the mammalian ETC from the UQ to RQ circuit renders cells highly resistant to hypoxia exposure. Thus, we establish RQ as a fundamental component of the mammalian ETC and unveil reprogramming the ETC to the RQ-circuit as a tractable strategy to treat hypoxia-related diseases. Wildtype 143B and UQCRC2 knockout cells were seeded and treated with media containing 13C4-Aspartate. The cells were extracted for metabolomics and ran on the LC/MS on the pHILIC method.
Metabolomics study on frozen tissue derived from the tumor and adjacent non-malignant liver tissue (NML) of patient afflicted with fibrolamellar carinoma (FLC)
STUDY_TYPE
LC-MS quantitative analysis
STUDY_SUMMARY
Fibrolamellar carcinoma (FLC) is a rare, early-onset liver cancer. The five-year survival rate is low, and there is a critical need for new therapeutics. The primary driver in FLC is the fusion oncoprotein, DNAJ-PKAc, which remains challenging to target therapeutically. It is critical, therefore, to expand understanding of the FLC molecular landscape to identify druggable pathways/targets. To date, only one study has attempted to characterize the FLC proteome and metabolome, but with modest sample size (proteomicsān = 16 patient samples; metabolomicsān = 10 patient samples) and protein detection (n = 4620 proteins). We have performed the most comprehensive characterization of FLC in both proteomics (n = 23 patient samples; n = 8485 proteins) and metabolomics (n = 26 patient samples; n = 135 metabolites). Targeted metabolomics on central carbon metabolism (polar metabolite extraction) was performed followed by extensive quantitative and qualitative assessment of its relationship with the proteome of FLC to gain insight on how the metabolic network is constructed in this cancer. Frozen patient tissue was derived from both primary and metastatic tumors as well as adjacent non-malignant liver tissue (NML). Primary and metastatic tumors served as our FLC cohort while NMLs served as our control cohort.
Multi-Omics Analysis Revealed Significant Metabolic Changes in Brain Cancer Cells Treated with Paclitaxel and/or Topotecan
STUDY_SUMMARY
Glioblastoma (GB) is the most common primary malignant brain tumor, representing approximately 57% of all gliomas and 48% of all primary malignant central nervous system (CNS) tumors. Despite the best standard therapies, glioblastoma survivors have a brief survival time, about 24 months on average. The treatment is troublesome because the cancer cells may not respond well to specific therapies as they grow within an extensive network of blood vessels. A multi-omics approach to provide information on biomolecules from multiple layers appears promising for systematically and holistically understanding complex biology. However, till now, only few studies have utilized omics analysis to investigate the impact of anticancer drugs on GBM. Our study aimed to evaluate the impacts of the anticancer medications (paclitaxel 5.3 Ī¼g/mL and topotecan 0.26 Ī¼M) solely and in pairwise combination on the metabolic and proteomic signatures of the U87 cell line while utilizing the accurate ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) analytical technology. The studied cancer cells wear treated with DMSO (control group), paclitaxel 5.3 ĀµM, topotecan 0.26 ĀµM, and a combination of paclitaxel 5.3 ĀµM and topotecan 0.26 ĀµM. Using One-way ANOVA, we observed 14 significantly altered metabolites compared to those cells treated with DMSO. For combination treatment (paclitaxel and topotecan), 10 metabolites were significantly dysregulated. The sparse partial least squares-discriminant analysis (sPLS-DA) showed minimal overlapping, indicating a difference between the four groups. While for proteomics, a total of 79 proteins were significantly dysregulated among the groups. These findings can aid in identifying new biomarkers associated with the utilized drugs and creating a map for targeted therapy. EIF3F, GNB2L1, HINT2, and RPA3 were shown to be significantly upregulated in the combination group when compared to the control. Moreover, ribosome, apoptosis, HIF-1 signaling, arginine and proline, glutathione, purine metabolism, apelin signaling pathway, and glycolysis were significantly altered in the combination group.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Unveiling cellular changes in leukaemia cell line K-562 after cannabidiol treatment through lipidomics
STUDY_SUMMARY
The present study was aimed at revealing the metabolic changes that occurred in the cellular lipid pattern of acute and chronic myeloid leukaemia cells following treatment with cannabidiol (CBD). CBD is a non-psychoactive compound present in Cannabis sativa L., which has shown an antiproliferative action in these type of cancer cells, to determine significant alterations of the cell metabolism attributable to the induction of apoptosis, previously observed from in vitro studies. Control and treated cells of chronic myeloid leukaemia (K562) were studied through an untargeted lipidomics approach. Treatment was carried out with CBD at a concentration of 23 ĀµM CBD for 48 h. After the extraction of the lipid content from cell lysates, the samples were analysed by UHPLC-QTOF-MS/MS both in the positive and the negative ionization modes.
Unveiling cellular changes in leukaemia cell line HL-60 after cannabidiol treatment through lipidomics
STUDY_SUMMARY
The present study was aimed at revealing the metabolic changes that occurred in the cellular lipid pattern of acute and chronic myeloid leukaemia cells following treatment with cannabidiol (CBD). CBD is a non-psychoactive compound present in Cannabis sativa L., which has shown an antiproliferative action in these type of cancer cells, to determine significant alterations of the cell metabolism attributable to the induction of apoptosis, previously observed from in vitro studies. Control and treated cells of acute myeloid leukaemia (HL-60) were studied through an untargeted lipidomics approach. Treatment was carried out with CBD at a concentration of 10 Ī¼M for 48 h. After the extraction of the lipid content from cell lysates, the samples were analysed by UHPLC-QTOF-MS/MS both in the positive and the negative ionization modes.
Metabolite profiling in the liver from fasted eIF4ES209A (S209A) mice compared to fasted wild type (WT) mice
STUDY_SUMMARY
Fasting is associated with a range of health benefits, including increased longevity, enhanced brain function, and improved metabolism. How fasting signals elicit changes in the proteome to establish metabolic programs that underlie lipid catabolism and the production of ketone bodies, an essential alternative fuel of energy, remain poorly understood. Here we show that paradoxically, while global translation is downregulated during fasting, hepatocytes selectively remodel the translatome to sustain lipid metabolism and ketogenesis. We discovered that phosphorylation of the major cap binding protein, eukaryotic translation initiation factor (P-eIF4E), is induced during fasting. By employing genome-wide unbiased polysome sequencing, we show that P-eIF4E is responsible for controlling the translation of the entire ketogenesis pathway, including the master regulator of fatty acid oxidation in the liver, peroxisome proliferator-activated receptor alpha (PPAR-alpha). Importantly, P-eIF4E regulates those mRNAs through a specific translation regulatory element within their 5ā untranslated regions. Genetic inhibition of P-eIF4E interrupts ketogenesis and fatty acid oxidation upon fasting. In addition, our findings reveal a new signaling property of fatty acids (FAs) derived from adipose tissue lipolysis, which are elevated during fasting. We uncovered that FAs bind and induce AMPK kinase activity that in turn enhances the phosphorylation of the kinase that phosphorylates eIF4E, the mitogen-activated protein kinase-interacting kinase (MNK). The AMPK-MNK axis controls ketogenesis revealing a new lipid-mediated kinase signaling pathway that links ketogenesis to translation control. We further show that genetically inhibiting P-eIF4E also impairs ketogenesis in response to a ketogenic diet. Certain types of cancers, such as pancreatic cancers use ketone bodies as an energy source which may rely on P-eIF4E suggesting a novel point of vulnerability. Our findings reveal that upon a ketogenic diet, treatment with eFT508 (a clinic P-eIF4E inhibitor) restrains pancreatic tumor growth in vivo. Importantly, restoring circulating Ī²-hydroxybutyrate (BHB) or overexpression of PPAR-alpha in tumor cells ablate the effect of eFT508, demonstrating a systemic and tumor intrinsic role of P-eIF4E during tumorigenesis in response to a ketogenic diet. Thus, our findings unveil a novel fatty acid-induced signaling pathway that activates selective translation, which underlies the rapid cellular response to fasting and ketogenesis and provides a tailored diet intervention therapy for cancer.
A study of 9 common breath VOCs in 504 healthy subjects using PTR-TOF-MS
STUDY_SUMMARY
This study utilized Proton-Transfer-Reaction Mass Spectrometry (PTR-MS) to analyze the exhaled breath profiles of 504 healthy adults, focusing on nine common breath volatile organic compounds (VOCs): acetone, acetaldehyde, acetonitrile, ethanol, isoprene, methanol, propanol, phenol, and toluene. The concentrations of these VOCs exhibited log normal distributions, with distinct differences observed based on different gender, age, and smoking history groups. Male group showed higher average VOC concentrations of acetaldehyde, acetonitrile, isoprene, and toluene. Age-related effects were evident, particularly in propanol and methanol, whose concentrations increased significantly with age. Smoking history also influenced VOC concentrations, with significant differences among non-smokers, current smokers, and ex-smokers. Ex-smokers demonstrated lower VOC levels than current smokers, indicating a normalization post-smoking cessation. Positive correlations were identified between breath VOC concentrations and years of smoking. The study enhances understanding of demographic influences on exhaled VOC profiles, contributing valuable insights for potential applications in health monitoring and clinical diagnosis. The use of PTR-MS was crucial for real-time VOC measurement, offering advantages over conventional methods. The research lays a foundation for future developments in non-invasive disease detection. The results highlight the potential of breath analysis as a valuable tool for health assessment.
INSTITUTE
Breathonix Pte Ltd
LAST_NAME
Zihao
FIRST_NAME
Xing
ADDRESS
83 Science Park, #02-01A, The Curie, Singapore, Singapore, Singapore, 118258, Singapore
Molecular signatures of xenograft colon cancer models treated with topotecan: A Mass Spectrometry-Based Study
STUDY_SUMMARY
Colorectal cancer (CRC) is one of the most common cancers worldwide. Despite improvement in standardized screening methods and the development of promising therapies, the 5-year survival rates are as low as 10% in the metastatic setting. Metabolomics, the study of metabolites on a large scale, has provided new insight into disease diagnosis and biomarkers identification. We chose topotecan as an anti-cancer drug in this context because, as far as we know, there are no studies examining the effect of this anti-cancer drug on metabolic alterations in CRC. In this study, untargeted metabolomic analysis study was performed to compare between four animal groups; HCT-116 xenograft models treated with topotecan, untreated HCT-116 xenograft models (vehicle controls), positive controls, and negative controls, using UHPLC-ESIQTOF-MS platform. One way ANOVA analysis discovered 53 statistically significant metabolites among all four groups (p <0.05). T-test revealed that 15 metabolites were statistically significant among vehicle controls and negative controls. Also, 20 metabolites were statistically significant among the potential respondersto topotecan and the vehicle controls. In addition, only 1 metabolite was statistically significant among the positive and negative control. Ultimately, by analyzing metabolomic profiles, our findings can create a map that can be utilized to assess the anticancer activity of topotecan in CRC. More studies with larger number of models are needed to verify the implication of the significantly altered metabolites and metabolic pathways in diagnosing and treating CRC.
INSTITUTE
Sharjah Institute for Medical Research
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Integrated multi-omics unveil the impact of the phosphinic compounds, desmethylphosphinothricin and its keto-analogue, on Escherichia coli metabolism
STUDY_SUMMARY
Desmethylphosphinothricin (L-Glu-Ī³-PH) is the phosphinic analogue of glutamate with a carbon-phosphorus (C-P) bond. In L-Glu-Ī³-PH the phosphinic group acts as a bioisostere of glutamate Ī³-carboxyl group allowing the molecule to be a substrate of Escherichia coli glutamate decarboxylase, a pyridoxal 5ā-phosphate (PLP)-dependent Ī±-decarboxylase. In addition, the L-Glu-Ī³-PH decarboxylation product, GABA-PH, is further metabolized by bacterial GABA-transaminase, another PLP-dependent enzyme, and succinic semialdehyde dehydrogenease, a NADP+ -dependent enzyme. The product of these consecutive reactions, the so-called GABA shunt, lead to the formation of succinate-PH, the phosphinic analogue of succinate, a TCA cycle intermediate. Notably, L-Glu-Ī³-PH displays an antibacterial activity of the same order of well-established antibiotics in E. coli. The dipeptide L-Leu-Glu-Ī³-PH was shown to display a higher efficacy, likely as a consequence of an improved penetration into the bacteria. Herein, with the aim of further understanding the intracellular effects of L-Glu-Ī³-PH, 1H NMR-based metabolomics and LC-MS-based shotgun proteomics were used. This study included also the keto-analogue of L-Glu-Ī³-PH, Ī±-ketoglutarate-Ī³-PH (Ī±-KG-Ī³-PH), which also exhibits antimicrobial activity. L-Glu-Ī³-PH and Ī±-KG-Ī³-PH were found to similarly impact the bacterial metabolism, though the overall effect of Ī±-KG-Ī³-PH is more pervasive, and not exclusively because of its intracellular conversion into L-Glu-Ī³-PH. Notably, both molecules impact the pathways where aspartate, glutamate and glutamine are used as precursors for the biosynthesis of related metabolites, activate the acid stress response and deprive cells of nitrogen. This work highlights the multi-target drug potential of L-Glu-Ī³-PH and Ī±-KG-Ī³-PH and paves the way for their exploitation as antimicrobials.
INSTITUTE
ITQB NOVA
LAST_NAME
GonƧalves
FIRST_NAME
LuĆs
ADDRESS
Avenida Republica, Oeiras, Not USCanada, 2780-157 Oeiras, Portugal
Metabolomic investigation of tear fluid biomarkers obtained from participants enrolled in the BiZiFED nutrition study
STUDY_SUMMARY
Zinc is an essential micronutrient, and its deficiency is particularly prevalent amongst the female population of Pakistan, leading to a substantial health burden on women of reproductive age and their children. Biofortification of wheat with zinc may offer a cost-effective strategy to attempt to improve nutritional status on a national scale. Biofortification with Zinc and Iron for Eliminating Deficiency (BiZiFED) is research program that aims to evaluate the impact of consuming zinc biofortified wheat flour on the zinc status and associated health outcomes of vulnerable communities northwest Pakistan. Of the health outcomes measured, tear biofluid was collected as a non-invasive method of determining whether zinc supplementation altered tear metabolomic biomarkers. In a crossover trial, 25 women of reproductive age received standard, non-biofortified wheat flour, and another 25 women received zinc-biofortified wheat flour for 8 weeks. At the end of this period, the nutritional intervention was switched for a further 8 weeks, whereby those individuals receiving the zinc-biofortified flour subsequently received the standard flour, whilst those receiving standard flour subsequently received the zinc-biofortified flour. The tear biofluid was collected using Schirmer strips after a 2-week baseline period and after 8 weeks and 16 weeks of the study. Metabolomic analysis was conducted using the MxP Quant 500 kit (Biocrates). Following multiple comparisons, four metabolites were significantly correlated with plasma zinc concentration, two demonstrating a positive correlation and two demonstrating a negative correlation. Five metabolites were significantly altered in response to zinc supplementation with biofortified wheat flour. These results highlight the potential of using tear biofluids as an alternative source for metabolomic biomarkers for the assessment of zinc status of individuals enrolled in nutritional studies.
INSTITUTE
Institute for Global Food Security
LAST_NAME
Pan
FIRST_NAME
Xiaobei
ADDRESS
19 Chlorine Gardens, Belfast, Antrim, BT9 5DL, United Kingdom
Metabolic profile evolution in relapsed/refractory B-NHL patients treated with CD19.CAR-T cell therapy and implications in clinical outcome
STUDY_SUMMARY
Plasma metabolomics analysis was performed on 44 patients with relapsed/refractory B-cell non Hodgkin lymphoma (r/r/B-NHL) infused with approved CD19.CAR-T cell products at the time of pre-lymphodepletion (PLD) and at day +1, +7, and +30 after CAR-T cell infusion. At the PLD time point, a metabolic profile characterized by high lipoproteins and lactate and low glucose contributed to poor outcome prediction in association with high lactate dehydrogenase levels. At day+1, higher plasma levels of lipid metabolism products and lower glucose and glycoproteins levels were observed in tisa-cel- compared to axi-cel-treated patients. At day+30, a higher content of lactate, as well as phenylalanine/tryptophan amino acids which represent the precursors of the key players of myeloid derived suppressor cells were found in patients who relapsed over time compared to patients who maintained complete remission until 1-year. Our data show complex metabolomic changes that track the evolution of the disease and drug activity in the first 30 days of CAR-T cell therapy. Notably, specific metabolic signatures at PLD and day+30, together with a day+30 metabolic profile related to myeloid immunosuppression, are associated with poor outcome.
Dynamics of Gut Metabolome and Microbiome Maturation during Early Life - untargeted GC, LECO tof MS
STUDY_SUMMARY
Early-life gut microbiome-metabolome crosstalk has a pivotal role in the maintenance of host physiology. However, our understanding on early-life gut microbiome-metabolome maturation trajectories in humans remains limited. This study aims to explore the longitudinal patterns of gut metabolites during early life, and how they are related to gut microbiota composition in birth cohort samples of n = 670 children collected at 2.5 (n=272), 6 (n=232), 14 (n=289), and 30 months (n=157) of age. Factor analysis showed that breastfeeding has an effect on several metabolites including secondary bile acids. We found that the prevalent gut microbial abundances were associated with metabolite levels, especially in the 2.5 months-olds. We also demonstrated that the prevalent early colonizers Bacteroides, Escherichia and Bifidobacterium abundances associated with microbial metabolites bile acids especially in the breastfed infants. Taken together, our results suggests that as the microbiome matures during the early-life there is an association with the metabolome composition in an analogous fashion to how the genome information mature during early life.
Dynamics of Gut Metabolome and Microbiome Maturation during Early Life - Short chain fatty acid (SCFA) data (targeted GC)
STUDY_SUMMARY
Early-life gut microbiome-metabolome crosstalk has a pivotal role in the maintenance of host physiology. However, our understanding on early-life gut microbiome-metabolome maturation trajectories in humans remains limited. This study aims to explore the longitudinal patterns of gut metabolites during early life, and how they are related to gut microbiota composition in birth cohort samples of n = 670 children collected at 2.5 (n=272), 6 (n=232), 14 (n=289), and 30 months (n=157) of age. Factor analysis showed that breastfeeding has an effect on several metabolites including secondary bile acids. We found that the prevalent gut microbial abundances were associated with metabolite levels, especially in the 2.5 months-olds. We also demonstrated that the prevalent early colonizers Bacteroides, Escherichia and Bifidobacterium abundances associated with microbial metabolites bile acids especially in the breastfed infants. Taken together, our results suggests that as the microbiome matures during the early-life there is an association with the metabolome composition in an analogous fashion to how the genome information mature during early life.
Dynamics of Gut Metabolome and Microbiome Maturation during Early Life - Bile acid data (targeted LCMS)
STUDY_SUMMARY
Early-life gut microbiome-metabolome crosstalk has a pivotal role in the maintenance of host physiology. However, our understanding on early-life gut microbiome-metabolome maturation trajectories in humans remains limited. This study aims to explore the longitudinal patterns of gut metabolites during early life, and how they are related to gut microbiota composition in birth cohort samples of n = 670 children collected at 2.5 (n=272), 6 (n=232), 14 (n=289), and 30 months (n=157) of age. Factor analysis showed that breastfeeding has an effect on several metabolites including secondary bile acids. We found that the prevalent gut microbial abundances were associated with metabolite levels, especially in the 2.5 months-olds. We also demonstrated that the prevalent early colonizers Bacteroides, Escherichia and Bifidobacterium abundances associated with microbial metabolites bile acids especially in the breastfed infants. Taken together, our results suggests that as the microbiome matures during the early-life there is an association with the metabolome composition in an analogous fashion to how the genome information mature during early life.
Untargeted metabolomics analysis on single cells and cell clusters of TNBC cell lines using mass spectrometry
STUDY_SUMMARY
Triple-negative breast cancer (TNBC) is more aggressive than other subtypes of breast cancerļ¼ and 40% of TNBC patients undergo recurrence and metastasis after treatments. In the metastasis process, clustered cancer cells (cell clusters) exhibited a higher ability of invasion and metastasis. On the other hand, metabolic reprogramming is closely related to the process of cancer metastasis. To explore the metabolic heterogeneity between single cells and cell clusters, mass spectrometry-based untargeted metabolomic analysis was performed on single cells and cell clusters constructed from two TNBC cell lines (MDA-MB-231 and MDA-MB-453). As a result, metabolic profiling between single cells and cell clusters were explored and metabolites contributing to cell clustering were identified.
INSTITUTE
Nanjing Medical University
LAST_NAME
Wang
FIRST_NAME
Zhongcheng
ADDRESS
101Longmian Avenue, Jiangning District, Nanjing 211166, P.R. China
FetalĀ growth delay caused byĀ loss ofĀ non-canonical imprintingĀ isĀ resolvedĀ late inĀ pregnancy and culminates in offspring overgrowth
STUDY_SUMMARY
Germline epigenetic programming, including genomic imprinting, substantially influences offspring development. Polycomb Repressive Complex 2 (PRC2) plays an important role in Histone 3 Lysine 27 trimethylation (H3K27me3)-dependent imprinting, loss of which leads to growth and developmental changes in mouse offspring. In this study, we show that offspring from mouse oocytes lacking the PRC2 protein Embryonic Ectoderm Development (EED) were initially developmentally delayed, characterised by low blastocyst cell counts and substantial growth delay in mid-gestation embryos. This initial developmental delay was resolved as offspring underwent accelerated fetal development and growth in late gestation resulting in offspring that were similar stage and weight to controls at birth. The accelerated development and growth in offspring from Eed-null oocytes was associated with remodelling of the placenta, which involved an increase in fetal and maternal tissue size, conspicuous expansion of the glycogen enriched cell population and delayed parturition. Despite placental remodelling and accelerated offspring fetal growth and development, placental efficiency and fetal blood glucose levels were low, and the fetal blood metabolome was unchanged. Moreover, while expression of the H3K27me3-imprinted gene and amino acid transporter Slc38a4 was increased, fetal blood levels of individual amino acids were similar to controls, indicating that placental amino acid transport was not enhanced. Genome-wide analyses identified extensive transcriptional dysregulation and DNA methylation changes in affected placentas, including a range of imprinted and non-imprinted genes. Together, while deletion of Eed in growing oocytes resulted in fetal growth and developmental delay and placental hyperplasia, our data indicate a remarkable capacity for offspring fetal growth to be normalised despite inefficient placental function and the loss of H3K27me3-dependent genomic imprinting.
Using Untargeted Metabolomics as a New Approach for Understanding Honeybee Toxicity
STUDY_SUMMARY
The impact of crop protection products on pollinating insects, particularly honeybees, is a critical consideration for ecological safety. Beyond acute toxicity, sublethal effects can potentially disrupt honeybee behavior, compromising pollination efficiency. Traditional assessment methods, reliant on visual observations, may not be optimal for capturing sublethal effects. In this study, we delved into molecular mechanisms underlying sublethal effects, employing three distinct platforms: whole-tissue transcriptomics, spatial transcriptomics, and metabolomics. Our findings reveal valuable insights into honeybee toxicity, with certain approaches demonstrating superiority in elucidating tissue-specific and treatment-specific responses, especially untargeted metabolomics tend to be a valuable tool for new bee-friendly agrochemical development. This research will aid in screening new molecules for pollinator safety and contribute to developing strategies that enhance crop yield while maintaining ecological sustainability.
INSTITUTE
Corteva Agriscience
LAST_NAME
Yang
FIRST_NAME
Kundi
ADDRESS
9330 Zionsville Rd, Indianapolis, IN 46268, Indianapolis, Indiana, 46077, USA
The central role of creatine and polyamines in fetal growth restriction
STUDY_SUMMARY
Placental insufficiency is often associated with fetal growth restriction (FGR), a condition associated with both short- and long-term complications for newborns. In this study, we performed a comprehensive transcriptomic and metabolomic characterization of the placenta of newborns born small for gestational age (SGA) and appropriate for gestational age (AGA). Transcriptome and metabolome data obtained from villous tissue biopsies and verified in 3D trophoblast organoids revealed that the defective placentas associated with FGR are characterized by metabolic reprogramming consisting of adaptation to hypoxia and alteration of arginine metabolism, particularly at the level of synthesis of polyamines and creatine phosphate. Arginine was utilized in defective SGA placentas to synthesize phosphocreatine, an energy-rich compound that is critical for ATP production and represents a positive metabolic adaptation that supports trophoblast fitness. In addition, SGA placentas were characterized by a deficit in polyamines due to overexpression of SAT1. SAT1 catalyzes the acetylation of spermine and spermidine and promotes their excretion from the trophoblast, resulting in a deficit of polyamines that cannot be compensated by arginine or polyamine supplementation unless SAT1 expression is disrupted. In conclusion, our study improves the understanding of metabolic adaptations associated with placental dysfunction and provides valuable insights for future therapeutic interventions
Glutamine Tracing Assay in RquA AAV Injected Mice with a Standard Curve
STUDY_SUMMARY
The extraction of RquA AAV injected livers from C57BL/6 mice. Metabolites from the liver were used to measure fumarate reduction and succinate oxidation. Glutamine tracing was used to determine forward and reverse direction of the TCA cycle. Standards were used to create a standard curve by diluting down a stock of fumarate and succinate standard mixture. The standard curve was used to determine picomoles of metabolites in the TCA cycle.
INSTITUTE
UMass Chan Medical School
LAST_NAME
UMass Chan
FIRST_NAME
Spinelli Lab
ADDRESS
55 Lake Avenue North, Worcester, Massachusetts, 01605, USA
Protein restriction slows the development and progression of Alzheimer's disease in mice
STUDY_SUMMARY
Dietary protein is a critical regulator of metabolic health and aging. Low protein diets are associated with healthy aging in humans, and many independent groups of researchers have shown that dietary protein restriction (PR) extends the lifespan and healthspan of mice. Here, we examined the effect of PR on metabolic health and the development and progression of Alzheimerās disease (AD) in the 3xTg mouse model of AD. We found that PR has metabolic benefits for 3xTg mice and non-transgenic controls of both sexes, promoting leanness and glycemic control in 3xTg mice and rescuing the glucose intolerance of 3xTg females. We found that PR induces sex-specific alterations in circulating metabolites and in the brain lipidome, downregulating sphingolipid subclasses including ceramides, glucosylceramides, and sphingomyelins in 3xTg females. Consumption of a PR diet starting at 6 months of age reduced AD pathology in conjunction with reduced mTORC1 activity, increased autophagy, and had cognitive benefits for 3xTg mice. Finally, PR improved the survival of 3xTg mice. Our results demonstrate that PR slows the progression of AD at molecular and pathological levels, preserves cognition in this mouse model of AD, and suggests that PR or pharmaceutical interventions that mimic the effects of this diet may hold promise as a treatment for AD.
Measuring UQ/UQH2 in WT and RquA 143B cell in Hypoxia
STUDY_SUMMARY
Cultured cells were used to measure the ratio of UQ to UQH2 in WT and RquA 143B cells. Cells were exposed to hypoxia to see if there was a difference in UQ:UQH2 ratio.
INSTITUTE
UMass Chan Medical School
LAST_NAME
UMass Chan
FIRST_NAME
Spinelli Lab
ADDRESS
55 Lake Avenue North, Worcester, Massachusetts, 01605, USA
Glutamine Tracing Assay in WT and SDHB KO 143B cells Treated with HKJS003
STUDY_SUMMARY
WT and succinate dehydrogenase iron sulfur subunit B(SDHB) KO 143B cells were treated with a small molecule analog of Rhodoquinone, HKJS003. Glutamine tracing was used to determine the fumarate reduction and succinate oxidation of the treated cells. Other metabolites such as ATP,ADP,AMP,Glutathione, Oxidized glutathione, UTP, lactic acid, and pyruvic acid were also measured.
INSTITUTE
UMass Chan Medical School
LAST_NAME
UMass Chan
FIRST_NAME
Spinelli Lab
ADDRESS
55 Lake Avenue North, Worcester, Massachusetts, 01605, USA
EVALUATION OF THE INFLUENCE OF ENVIRONMENTAL CONDITIONS ON THE METABOLOME OF Psychotria viridis LEAVES Ruiz & Pav.: Metabolomics approach
STUDY_SUMMARY
Psychotria viridis Ruiz & Pav. has gained particular attention due to its use in the ayahuasca drink. This work aims to deepen the understanding of the composition of specialized metabolites in the leaves of Psychotria viridis Ruiz & Pav. and investigate the influence of the growing environment as well as seasonality on the composition. The specimens were grown in the open field and others in the shaded environment of rubber tree (Hevea brasiliensis L.) cultivation. These specimens make up a clonal population of the mother-plant. The collection was made in the four seasons. After a three-phase extraction method on the leaves, the aqueous phase was analyzed on an ultra-high performance liquid chromatography coupled to electrospray ionization and Orbitrap mass spectrometry (UHPLC-ESI-Orbitrap-MS) system. Acquired data were processed using MS-DIAL 4.9 and MetaboAnalyst 5.0 for multivariate and pathway activity analysis. Chemical variations were investigated employing principal component analysis (PCA), hierarchical cluster analysis (HCA) and partial least squares discriminant analysis (PLS-DA). The most important identified compounds for differentiation according to the seasonality were flavonoids. The pathways with significant variation in response to the seasonality were related to energy generation through biosynthesis and consumption of carbohydrates: Ascorbate and aldarate metabolism, Pentose and glucuronate interconversions and Citrate cycle. On the other hand, Biosynthesis of flavonoid, flavones and flavonols were the biochemical pathways associated with the influence of the cultivation location in full sun or shade in an intercrop being a plant response to oxidative stress. Keywords: Psychotria viridis, seasonality, abiotic stress, liquid chromatography-mass spectrometry; metabolomics.
Obesity, sex, and depot drive distinct lipid profiles in murine white adipose tissue
STUDY_SUMMARY
To investigate how the lipid composition of the adipose tissue changes with obesity, sex, and depot, we performed comprehensive untargeted lipidomics on the whole adipose tissue of the inguinal and perigonadal adipose depot from lean and obese, male and female mice. The screen allowed us to identify lipid signatures sufficient to differentiate different adipose tissue samples. Such that, increased levels of OxTG and CL, increase in DG and Cer, and increase in HexCer were all able to differentiate adipose tissue based on obesity, sex, or depot, respectively.
Untargeted NMR-lipidomic study in R7, R7sgRON, R7shDEK cells
STUDY_SUMMARY
Recurrent and metastatic breast cancer is frequently treatment resistant. A wealth of evidence suggests that reprogrammed lipid metabolism supports cancer recurrence. Overexpression of the RON and DEK proteins in breast cancer is associated with poor outcome. Both proteins promote cancer metastasis in laboratory models, but effects on lipid metabolite levels remain unknown. To measure RON- and DEK-dependent steady-state lipid metabolite levels, an Nuclear Magnetic Resonance (NMR)-based approach was utilized. The observed differences were then used to identify a lipid metabolism-related gene expression signature that is prognostic of overall survival (OS), distant metastasis free survival (DMFS), post-progression survival (PPS), and recurrence free survival (RFS) in patients with breast cancer. RON loss led to decreased cholesterol and sphingomyelin levels, while DEK loss increased total fatty acid levels and decreased free glycerol levels. Lipid-related genes were then queried to define a signature that predicts breast, ovarian, and lung cancer patient survival. Taken together, RON and DEK differentially regulate lipid metabolism in a manner that predicts and may promote breast cancer metastasis and recurrence.
A small molecule macrophage migration inhibitory factor agonist ameliorates age-related myocardial intolerance to ischemia-reperfusion insults via metabolic regulation - Part 1
STUDY_SUMMARY
Microphage migration inhibitory factor (MIF) is an innate cytokine that regulates both inflammatory and homeostatic responses. MIF is expressed by cardiomyocytes, where it exerts a protective action against ischemia-reperfusion (I/R) injury by activating AMP-activated protein kinase (AMPK). This effect is attenuated in the senescent heart due to an intrinsic, age-related reduction in MIF expression. We hypothesized that treating the aged heart with the small molecule MIF agonist (MIF20) can reinforce protective MIF signaling in cardiomyocytes, leading to a beneficial effect against I/R stress. The administration of MIF20 at the onset of reperfusion was found to not only decrease myocardial infarct size but also preserves systolic function in the aged heart. Protection from I/R injury was reduced in mice with cardiomyocyte-specific Mif deletion, consistent with the mechanism of action of MIF20 to allosterically increase MIF affinity for its cognate receptor CD74. We further found MIF20 to contribute to the maintenance of mitochondrial fitness and to preserve the contractile properties of aged cardiomyocytes under hypoxia/reoxygenation. MIF20 augments protective metabolic responses by reducing the NADH/NAD ratio, leading to a decrease in the accumulation of reactive oxygen species (ROS) in the aged myocardium under I/R stress. We also identify alterations in the expression levels of the downstream effectors PDK4 and LCAD, which participate in the remodeling of the cardiac metabolic profile. Data from this study demonstrates that pharmacologic augmentation of MIF signaling provides beneficial homeostatic actions on senescent myocardium under I/R stress. This experimental is part of case 4535 showing the Lipidomics experimental data.
EVALUATION OF THE INFLUENCE OF ENVIRONMENTAL CONDITIONS ON THE METABOLOME OF Psychotria viridis LEAVES Ruiz & Pav.: Lipidomics approach
STUDY_SUMMARY
Psychotria viridis Ruiz & Pav. has gained particular attention due to its use in the ayahuasca drink. This work aims to deepen the understanding of the composition of specialized metabolites in the leaves of Psychotria viridis Ruiz & Pav. and investigate the influence of the seasonality on the composition. The specimens were grown in the open field and others in the shaded environment of rubber tree (Hevea brasiliensis L.) cultivation. These specimens make up a clonal population of the mother-plant. The collection was made in the four seasons. After a three-phase extraction method on the leaves, the organic phase was analyzed on an ultra-high performance liquid chromatography coupled to electrospray ionization and Orbitrap mass spectrometry (UHPLC-ESI-Orbitrap-MS) system. Acquired data were processed using MS-DIAL 4.9 and MetaboAnalyst 5.0 for multivariate and pathway activity analysis. Chemical variations were investigated employing principal component analysis (PCA), hierarchical cluster analysis (HCA) and partial least squares discriminant analysis (PLS-DA). Through PCA was shown that samples tend to differentiate just according seasonality what was not observed according cultivation area. The most important identified compounds for differentiation according to the seasonality were polyunsaturated lipids with a carbon chain longer than 21 carbons. The pathway with significant variation in response to the seasonality was related to oxidative stress: alpha-linolenic acid metabolism. In this context, the change in the alpha-linolenic acid pathway could be associated with the adaptation of leaves to the drop in temperature that naturally occurs in winter compared to other seasons of the year under analysis. Keywords: Psychotria viridis, seasonality, abiotic stress, liquid chromatography-mass spectrometry; lipidomics.
The Ataxia-Telangiectasia Mutated Kinase Inhibitor AZD0156 is a Potent Inhibitor of Plasmodium Phosphatidylinositol 4-Kinase and is an Attractive Candidate for Repositioning Against Malaria
STUDY_SUMMARY
Here we interrogated the in vitro metabolic effects of AZD0156 on the Plasmodium falciparum 3D7 parasite strain using ultra-high performance liquid chromatography mass-spectrometry (UHPLC-MS). The resulting metabolic fingerprints provide information on the parasite biochemical pathways affected by pharmacologic intervention. The metabolic effects were compared to their parent strain. The metabolic fingerprints provided show that certain biochemical pathways are affected by the drug inhibitory effect.
INSTITUTE
Pennsylvania State University
LAST_NAME
Llinas
FIRST_NAME
Manuel
ADDRESS
W126 Millenium Science Complex, University Park, PA 16802
Lipidomic analysis of Axon Regeneration in Xenopus laevis Chiasm
STUDY_SUMMARY
CNS injuries of the anuran amphibian, Xenopus laevis, are uniquely befitted for studying the molecular compositions of neuronal regeneration of retinal ganglion cells (RGC) due to a functional recovery of optic axons disparate to adult mammalian analogues. RGCs and their optic nerve axons undergo irreversible neurodegeneration in glaucoma and associated optic neuropathies, resulting in blindness in mammals. Conversely, Xenopus demonstrates RGC lifetime-spanning regenerative capabilities after optic nerve crush, inciting opportunities to compare de novo regeneration and develop efficient pharmaceutical approaches for vision restoration. Studies revealing lipidome alterations during optic nerve regeneration are sparse and could serve as a solid foundation for these underlying molecular changes. We profile the lipid changes in a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that were either had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 12 and 27 days post optic nerve crush. Following euthanasia, the optic nerves were collected for lipidomic analysis. A modified Bligh and Dyer method was used for lipid extraction, followed by untargeted mass spectrometry lipid profiling with a Q-Exactive Orbitrap Liquid Chromatography-Mass Spectrometer (LC MS-MS) coupled with Vanquish Horizon Binary UHPLC LC-MS system.
Lipidomic analysis of Axon Regeneration in Xenopus laevis Retina
STUDY_SUMMARY
CNS injuries of the anuran amphibian, Xenopus laevis, are uniquely befitted for studying the molecular compositions of neuronal regeneration of retinal ganglion cells (RGC) due to a functional recovery of optic axons disparate to adult mammalian analogues. RGCs and their optic nerve axons undergo irreversible neurodegeneration in glaucoma and associated optic neuropathies, resulting in blindness in mammals. Conversely, Xenopus demonstrates RGC lifetime-spanning regenerative capabilities after optic nerve crush, inciting opportunities to compare de novo regeneration and develop efficient pharmaceutical approaches for vision restoration. Studies revealing lipidome alterations during optic nerve regeneration are sparse and could serve as a solid foundation for these underlying molecular changes. We profile the lipid changes in a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that were either had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 12 and 27 days post optic nerve crush. Following euthanasia, the optic nerves were collected for lipidomic analysis. A modified Bligh and Dyer method was used for lipid extraction, followed by untargeted mass spectrometry lipid profiling with a Q-Exactive Orbitrap Liquid Chromatography-Mass Spectrometer (LC MS-MS) coupled with Vanquish Horizon Binary UHPLC LC-MS system.
Lipidomic analysis of serum from WT, liver-specific Gclc KO, liver-specific Nrf2 KO, and liver-specific Gclc-Nrf2 DKO mice.
STUDY_SUMMARY
The rate-limiting enzyme in glutathione (GSH) synthesis is Gclc. GSH synthesis is suggested to be implicated in lipogenesis. Loss of GSH synthesis is suggested to increase the activity of NRF2. To examine the impact of GSH and NRF2 on lipid abundance in serum, we performed lipidomic analysis of serum from WT, liver-specific Gclc KO, liver-specific Nrf2 KO, and liver-specific Gclc-Nrf2 DKO mice.
INSTITUTE
University of Rochester Medical Center
LAST_NAME
Harris
FIRST_NAME
Isaac
ADDRESS
601 Elmwood Ave, Rochester, New York, 14642-0001, USA
Suppression of prostaglandin I2ātype I interferon axis induces extramedullary hematopoiesis to promote cardiac repair after myocardial infarction
STUDY_SUMMARY
Background: Immune cells are closely associated with all processes of cardiac repair following myocardial infarction (MI), including the initiation, development, and resolution of inflammation. Spleen extramedullary hematopoiesis (EMH) serves as a critical source of emergency mature blood cells that are generated through the self-renewal and differentiation of hematopoietic stem/progenitor cells (HSPCs). However, how EMH responds to MI and the role of EMH in cardiac repair post-MI remains unclear. Methods: To assess the role of spleen EMH in MI, a Tcf21CreERScfflox/flox MI mouse model with inhibited EMH was constructed. GFP+ HSCs sorted from eGFP mouse spleen by flow cytometry and injected into Tcf21CreERScfflox/flox mice to test the sources of local inflammatory cells during MI. Using highly specific liquid chromatography-tandem mass spectrometry and single-cell RNA sequencing, we analyzed the lipidomic profile of arachidonic acid metabolites and the transcriptomes of HSPCs in the spleen after MI. Results: We found that MI enhanced EMH, as reflected by the increase in spleen weight and volume and the number of HSPCs in the spleen. The lack of EMH in Scf-deficient mice exacerbated tissue injury post-MI. Analyzing the transcriptome of spleen HSPCs post-MI, we found the type I interferon (IFN) pathway significantly inhibited in HSC/multipotent progenitor subclusters and the absence of type I IFN signaling enhanced the MI-induced spleen EMH. Lipidomics analysis revealed that prostaglandin I2 (PGI2) was markedly reduced in the spleen. Mechanistically, PGI2 suppressed MI-induced EMH through a PGI2 receptor (IP)-cAMP-453p-SP1 cascade in spleen HSPCs. Finally, hematopoietic cell-specific IP-deficient mice exhibited enhanced EMH and improved cardiac recovery post-MI, which mitigated the adverse secondary outcomes of treatment with cicaprost, a PGI2 analog and anti-inflammatory agent. Conclusions: Together, our findings revealed that a PGI2āIFN axis was involved in spleen EMH after MI, providing new mechanistic insights into spleen EMH post-MI and offering a new therapeutic target for treating ischemic cardiac injury.
INSTITUTE
Tianjin Medical University
LAST_NAME
Lv
FIRST_NAME
Huizhen
ADDRESS
Qixiangtai Road 22th, Tianjin, Tianjin, 300070, China
Exploration of Zeb1-dependent changes in the lipidome of MDA-MB-231 cells
STUDY_SUMMARY
Human breast cancer MDA-MB-231 wildtype (WT) cells and the stably transduced MDA-MB-231 shZeb1 (stable Zeb1 knockdown) and shCtrl cell lines (control cell line for the stable Zeb1 knockdown) (Spaderna et al. 2008, DOI: 10.1158/0008-5472.CAN-07-5682) were analyzed for their phospholipid profile by UPLC-MS/MS.
Effects of acute mitoregulin loss on cardiac mitochondrial lipids in mice
STUDY_SUMMARY
Cardiac mitochondrial lipidome analysis in 7-week-old male aMHC-Cas9 transgenic mice, harvested at 4 weeks post-injection with either AAV:guMtln (sgRNAs to delete Mtln) or AAV:GFP control.
Mitochondrial respiration impairment in microglia dampens response to demyelinating injury but is not sufficient to induce an aging phenotype
STUDY_SUMMARY
Microglia are necessary for CNS function during development and play roles in aging, Alzheimerās Disease (AD) and the response to demyelinating injury1ā5. Mitochondrial respiratory chain (RC) controls macrophage-dependent immune responses6ā9. However, whether mitochondrial RC is essential to microglia function is not known. We conditionally deleted the mitochondrial complex III subunit Rieske Iron-Sulfur Protein (RISP) in the microglia of adult mice to assess the requirement of microglial RC for survival, proliferation, and adult CNS function in vivo. Surprisingly, mitochondrial RC function was not required for survival or proliferation of microglia in vivo. RNA-seq analysis showed that loss of RC function in microglia caused changes in gene expression distinct from aged or disease-associated microglia (DAM). Microglia-specific loss of mitochondrial RC function did not affect cognitive decline during aging or in the 5xFAD model of Alzheimerās disease (AD). However, Abeta plaque coverage decreased and microglial interaction with Abeta plaques increased in the hippocampus of 5xFAD mice with mitochondrial RC-deficient microglia. Microglia-specific loss of mitochondrial RC function did impair remyelination following an acute, reversible demyelinating event. Thus, mitochondrial respiration in microglia is dispensable for maintenance of normal cognitive function but is essential to maintain a proper response to CNS demyelinating injury.
Lipidomic analysis of Axon Regeneration in Xenopus laevis Tectum
STUDY_SUMMARY
CNS injuries of the anuran amphibian, Xenopus laevis, are uniquely befitted for studying the molecular compositions of neuronal regeneration of retinal ganglion cells (RGC) due to a functional recovery of optic axons disparate to adult mammalian analogues. RGCs and their optic nerve axons undergo irreversible neurodegeneration in glaucoma and associated optic neuropathies, resulting in blindness in mammals. Conversely, Xenopus demonstrates RGC lifetime-spanning regenerative capabilities after optic nerve crush, inciting opportunities to compare de novo regeneration and develop efficient pharmaceutical approaches for vision restoration. Studies revealing lipidome alterations during optic nerve regeneration are sparse and could serve as a solid foundation for these underlying molecular changes. We profile the lipid changes in a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that were either had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 12 and 27 days post optic nerve crush. Following euthanasia, the optic nerves were collected for lipidomic analysis. A modified Bligh and Dyer method was used for lipid extraction, followed by untargeted mass spectrometry lipid profiling with a Q-Exactive Orbitrap Liquid Chromatography-Mass Spectrometer (LC MS-MS) coupled with Vanquish Horizon Binary UHPLC LC-MS system.
Temperature change elicits lipidome adaptation in the simple organisms Mycoplasma mycoides and JCVI-syn3B
STUDY_SUMMARY
Cell membranes mediate interactions between life and its environment, with lipids determining their properties. Understanding how cells adjust their lipidomes to tune membrane properties is crucial yet poorly defined due to the complexity of most organisms. We used quantitative shotgun lipidomics to study temperature adaptation in the simple organism Mycoplasma mycoides and the minimal cell, JCVI-syn3B. We show that lipid abundances follow a universal logarithmic distribution across eukaryotes and bacteria, with comparable degrees of lipid remodeling for adaptation regardless of lipidomic or organismal complexity. Lipid features analysis demonstrates head group-specific acyl chain remodeling as characteristic of lipidome adaptation; its deficiency in Syn3B is associated with impaired homeoviscous adaptation. Temporal analysis reveals a two-stage cold adaptation process: swift cholesterol and cardiolipin shifts followed by gradual acyl chain modifications. This work provides an in-depth analysis of lipidome adaptation in minimal cells, laying a foundation to probe the design principles of living membranes.
Metabolomics studies on mouse cardiac samples on a Western diet
STUDY_SUMMARY
Targeted metabolomics was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure polar metabolites in both positive and negative ionization mode on cardiac mice tissue acquired after a 30 week dietary intervention.
Metabolomics studies on mouse liver samples on a Western diet
STUDY_SUMMARY
Targeted metabolomics was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS).to measure polar metabolites in both positive and negative ionization mode on cardiac mice tissue acquired after a 30 week dietary intervention.
The impact of grass- and grain-finishing on metabolomic profiles of North American Black Angus Beef cattle.
STUDY_SUMMARY
The goal of this study was to compare meat metabolomes (pectoralis profundus) of Black Angus cattle from two commercial US beef finishing systems (pasture-finished on Western U.S. rangeland; n=18 and grain-finished in a Midwest U.S. feedlot; n=18).
Exploration of Zeb1-dependent changes in the redox-lipidome of MDA-MB-231 cells
STUDY_SUMMARY
Human breast cancer MDA-MB-231 wildtype (WT) cells and the stably transduced MDA-MB-231 shZeb1 (stable Zeb1 knockdown) and shCtrl cell lines (control cell line for the stable Zeb1 knockdown) (Spaderna et al. 2008, DOI: 10.1158/0008-5472.CAN-07-5682) were treated with DMSO or RSL3 (1 or 10 ĀµM) for 2 h, 4 h, 6 h or 24 h. The cell pellets were collected and analyzed for their oxidized phospholipid profile by UPLC-MS/MS. Please note that one sample set was measured three times with the same sample-ID, but with different methods (Ox-PE, Ox-PC, Ox-PI), therefore each sub-class has their own raw-data file marked by their corresponding abbreviation (Ox-PE, Ox-PC, Ox-PI; e.g. "210514_MDA_ZEB1_oxPE_dil_UD_std_1ul_JZ_oxPE_MRM_003.wiff", "210514_MDA_ZEB1_oxPC_dil_UD_std_1ul_JZ_oxPC_MRM_002.wiff" or "210514_MDA_ZEB1_oxPI_dil_UD_std_1ul_JZ_oxPI_MRM_001.wiff").
Exploration of EMT-dependent changes in the lipidome of KPC cells
STUDY_SUMMARY
KPC cell lines that are mesenchymal (lines KPC550 and KPC701), mesenchymal/epithelial mixed (lines KPC524 and KPC438), epithelial (lines KPC661 and KPC792) and KPC cell lines with Zeb1 knockout (lines KPCZ519, KPCZ436, KPCZ426) were analyzed for their phospholipid profile by UPLC-MS/MS.
Rev-erb's Role in Muscle Metabolism: Mass Spectrometry Analysis in WT and Rev-erb skeletal muscle specific knockout Mice
STUDY_SUMMARY
We used WT and Rev-erb skeletal muscle specific knockout mouse models to explore the changes in muscle metabolites at different time points, aiming to clarify the regulatory mechanism of Rev-erb on metabolism. The study design involved taking 3 mice each from the WT and muscle-specific knockout models at ZT10 and ZT22 (ZT, Zeitgeber Time), respectively, and performing mass spectrometry to detect muscle metabolites (Glycolysis, TCA, Amino acids, and Carnitine).Metabolomics profiling of muscles revealed robust changes in amino acids, glycolytic intermediates, and carnitines between KO and WT muscles, particularly at ZT22.
Exploration of EMT-dependent changes of phosphatidylethanolamine profiles in KPC allografts
STUDY_SUMMARY
Cryo-conserved tumors from subcutaneous allografts as described (Krebs et al. 2017, DOI: 10.1038/ncb3513) were retrieved. Three tumors derived from mesenchymal KPC cell lines (lines KPC550 and KPC701) and epithelial KPC cell lines (lines KPC438 and KPC661) were analyzed for their phosphatidylethanolamine profile by UPLC-MS/MS.
Exploration of RSL3-induced and Chlorido[N,Nā-disalicylidene-1,2-phenylenediamine]iron(III) complex-induced changes in the lipidome of MDA-MB-231 breast cancer cells
STUDY_SUMMARY
Chlorido[N,Nā-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs) exhibit potent anti-cancer properties through incompletely understood molecular mechanisms. Here, we treated human MDA-MB-231 triple-negative breast cancer cells with the glutathione peroxidase (GPX)4 inhibitor RSL3 or chlorido[N,Nā-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs) and analyzed their phospholipid profile by targeted lipidomics. SCs decreased the cellular proportion of polyunsaturated fatty acids (PUFAs) in phospholipids, which barely changed upon short-term treatment with RSL3.
Exploration of RSL3 or Chlorido[N,Nā-disalicylidene-1,2-phenylenediamine]iron(III) complex-induced changes in the phospholipid oxidation of MDA-MB-231 breast cancer cells
STUDY_SUMMARY
Chlorido[N,Nā-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs) exhibit potent anti-cancer properties through incompletely understood molecular mechanisms. Here, we treated human MDA-MB-231 triple-negative breast cancer cells with the glutathione peroxidase (GPX)4 inhibitor RSL3 or chlorido[N,Nā-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs) and analyzed their oxidized phospholipid profile by targeted lipidomics. SCs induce extensive (hydroper)oxidation of arachidonic acid and adrenic acid in membrane phospholipids, particularly phosphatidylethanolamines (PE) and phosphatidylinositols (PC). In this process, SCs have demonstrated superior efficacy compared to the GPX4 inhibitor RSL3, an established ferroptosis inducer. Please note that one sample set was measured three times with the same sample-ID, but with different methods (oxPE, oxPC, oxPI), therefore each sub-class has their own raw-data file marked by their corresponding abbreviation (oxPE, oxPC, oxPI; e.g. "210309_MDA_Timecourse_RSL3_oxPE_dil_UD_Std_1ul_SFT.wiff", "210324_Rescue_Gust_compounds_oxPE_dil_UD_std_1ul_SFT.wiff", "210309_MDA_Timecourse_RSL3_oxPC_dil_UD_Std_1ul_SFT.wiff", "210324_Rescue_Gust_compounds_oxPC_dil_UD_std_1ul_SFT.wiff" or "210324_Rescue_Gust_compounds_oxPI_dil_UD_std_1ul_SFT.wiff", ).
A Covalent Creatine Kinase Inhibitor Ablates Glioblastoma Migration and Sensitizes Tumors to Oxidative Stress.
STUDY_TYPE
Cki treatment on glioblastoma
STUDY_SUMMARY
Glioblastoma is a Grade 4 primary brain tumor defined by therapy resistance, diffuse infiltration, and near-uniform lethality. The underlying mechanisms are unknown, and no treatment has been curative. Using a recently developed kinase inhibitor (CKi), we explored the role of this inhibitor on GBM biology in vitro. While CKi minimally impacted GBM cell proliferation and viability, it significantly affected migration. In established GBM cell lines and patient-derived xenografts, CKi ablated both the migration and invasion of GBM cells. CKi also hindered radiation-induced migration. RNA-seq revealed a decrease in invasion-related genes, with an unexpected increase in glutathione metabolism and ferroptosis protection genes post-CKi treatment. The effects of CKi could be reversed by the addition of cell-permeable glutathione. Carbon-13 metabolite tracing indicated heightened glutathione biosynthesis post-CKi treatment. Combinatorial CKi blockade and glutathione inhibition or ferroptosis activation abrogated cell survival. Our data demonstrated that CKi perturbs promigratory and anti-ferroptotic roles in GBM, identifying the creatine kinase axis as a druggable target for GBM treatment.
INSTITUTE
Northwestern University, Feinberg School of Medicine
SLC25A48 controls mitochondrial choline import and metabolism
STUDY_SUMMARY
Choline is an essential nutrient for cellular metabolism, including the biosynthesis of phospholipids, neurotransmitters, and one-carbon metabolism. A critical step in choline catabolism is the mitochondrial import and synthesis of choline-derived methyl donors, such as betaine. However, the underlying mechanisms and the biological significance of mitochondrial choline metabolism remain poorly understood. Here, we report that a previously uncharacterized mitochondrial inner-membrane protein, SLC25A48, controls mitochondrial choline transport and catabolism in vivo. We show that SLC25A48 is highly expressed in brown adipose tissue and is required for whole-body cold tolerance, thermogenesis, mitochondrial respiration, and mitochondrial membrane integrity. Choline uptake into the mitochondria via SLC25A48 facilitates the synthesis of betaine and purine nucleotides, whereas loss of SLC25A48 resulted in increased production of reactive oxygen species and imbalanced mitochondrial lipids. Notably, human cells carrying a single nucleotide polymorphism on the SLC25A48 gene and cancer cells lacking SLC25A48 exhibited elevated oxidative stress and impaired cell proliferation. Together, the present study identified SLC25A48 as a mitochondrial carrier that mediates choline catabolism and plays a critical role in mitochondrial function and cell survival.
Malic Enzyme 2 maintains metabolic state and anti-tumor immunity of CD8+ T cells
STUDY_SUMMARY
For metabolite analysis, naive ME2+/+ and ME2-/- CD8+ T cells were isolated and activated with anti-CD3 (4Ī¼g/ml) and anti-CD28 (1Ī¼g/ml). Cells were washed twice with ice-cold PBS and metabolites were extracted with ice-cold 80% methanol. The extracts were analyzed by LC-MS/MS.
Metabolomics analysis of human spermatozoa reveals impaired metabolic pathways in asthenozoospermia (MS data)
STUDY_SUMMARY
Background: Infertility is a major health issue, affecting 15% of reproductive-age couples with male factors contributing to 50% of cases. Asthenozoospermia, or low sperm motility, is a common cause of male infertility with complex etiology, involving genetic and metabolic alterations, inflammation, and oxidative stress. However, the molecular mechanisms behind low motility are unclear. In this study, we used a metabolomics approach to identify metabolic biomarkers and pathways involved in sperm motility. Methods: We compared the metabolome and lipidome of spermatozoa of men with normozoospermia (n = 44) and asthenozoospermia (n = 22) using untargeted LC-MS and the metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal fluid redox status to assess the oxidative stress in the ejaculate. Results: We identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of the spermatozoaās metabolomics and lipidomics data showed a clear separation between groups. Spermatozoa of asthenozoospermic men presented lower levels of several amino acids, and increased levels of energetic substrates and lysophospholipids. However, the metabolome and redox status of the seminal fluid was not altered in asthenozoospermia. Conclusions: Our results indicate impaired metabolic pathways associated with redox homeostasis and amino acid, energy, and lipid metabolism in asthenozoospermia. Taken together, these findings suggest that the metabolome and lipidome of human spermatozoa are key factors influencing their motility and that oxidative stress exposure during spermatogenesis or sperm maturation may be in the etiology of decreased motility in asthenozoospermia.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry
LAST_NAME
Guerra-Carvalho
FIRST_NAME
BƔrbara
ADDRESS
Campus UniversitƔrio de Santiago, 3810-193 Aveiro, Portugal
FDX2-KO induces global down-regulation of iron-sulfur cluster-containing proteins and senescence-like growth arrest or death in ovarian cancer cells
STUDY_SUMMARY
Recent studies report molecular mechanisms underlying iron-sulfur cluster (Fe-S) biosynthesis and suggest its importance in health and disease. However, a role for Fe-S biosynthesis in cancer contexts remains unclear. Here we report that FDX2, an Fe-S assembly factor, is indispensable for maintenance of cellular Fe-S-containing proteins (Fe-S protein(s)) and proliferation of ovarian cancer (OVC) cells. CRISPR-screening of all metabolism-related genes in OVC cells identified several Fe-S assembly genes as essential for OVC growth. Using an inducible FDX2-KO OVC line, we found that FDX2 loss promotes either senescence-like growth arrest or cell death, depending on TP53 status. Mechanistically, FDX2-loss caused global but differential post transcriptional down-regulation of Fe-S proteins, in turn perturbing respiration, iron-regulation and redox homeostasis, all associated with DNA damage. These results demonstrate significant roles for Fe-S biosynthesis in OVC proliferation and survival and provide information about how the cellular Fe-S-protein network responds to disruptions in Fe-S assembly.
INSTITUTE
Tohoku University
DEPARTMENT
Graduate School of Medicine
LABORATORY
Department of Biochemical Oncology
LAST_NAME
Tanuma
FIRST_NAME
Nobu-hiro
ADDRESS
2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan
We analyzed metabolites in the retina, choroid/RPE, and plasma from WT and PHGDH heterozygous mice that were fed either a control or serine/glycine-deprived diet.
We analyzed metabolites in the retina, choroid/RPE, and plasma from WT and PHGDH heterozygous mice that were fed either a control or serine/glycine-deprived diet.
WT and PHGDH(+/-) mice fed control or -SG diet: Back of eye
STUDY_SUMMARY
We analyzed metabolites in the retina, choroid/RPE, and plasma from WT and PHGDH heterozygous mice that were fed either a control or serine/glycine-deprived diet. This study contains back of eye (choroid/RPE) samples.
Impact of serine supplementation following treatment with serine/glycine-depleted diet
STUDY_SUMMARY
We analyzed metabolites in the retina, choroid/RPE, plasma, and paw skin from mice that were previously on control or serine/glycine-depleted diet and then switched over to a control or serine-supplemented diet.
Impact of serine supplementation following treatment with serine/glycine-depleted diet
STUDY_SUMMARY
We analyzed metabolites in the retina, choroid/RPE, plasma, and paw skin from mice that were previously on control or serine/glycine-depleted diet and then switched over to a control or serine-supplemented diet.
Impact of serine supplementation following treatment with serine/glycine-depleted diet
STUDY_SUMMARY
We analyzed metabolites in the retina, choroid/RPE, plasma, and paw skin from mice that were previously on control or serine/glycine-depleted diet and then switched over to a control or serine-supplemented diet.
Metabolomics analysis within healthy donors and end-stage heart failure with a focus on ischaemic cardiomyopathy with diabetes
STUDY_TYPE
Human heart failure left ventricular myocardial metabolomics study across multiple disease phenotypes.
STUDY_SUMMARY
This study involves the analysis of water soluble metabolites within healthy and human heart failure conditions with a focus on ischaemic cardiomyopathy with diabetes and is part of a project which also integrates lipidomics, proteomics, RNA-seq, and histological analyses to identify molecular mechanisms influencing disease phenotypes. Ischaemic cardiomyopathy (ICM) is the most common cause of heart failure (HF) and often coexists with diabetes mellitus (DM). Yet, their combined effects are seldom investigated and are poorly understood. Herein, we performed multi-omic analyses of end-stage ICM with DM (ICM-DM) against ICM-No DM, non-ischaemic (dilated) cardiomyopathy with DM (NICM-DM), NICM-No DM, and healthy age-matched donors (AMD). Tissue was sourced from pre-mortem human left ventricular myocardium. Though fatty acid oxidation (FAO) proteins were down-regulated in ICM-DM relative to AMD and other HF, the unique ICM-DM down-regulation of acylcarnitines, perilipin, and ketone body, amino acid, and glucose metabolising proteins indicated FAO may not be entirely impaired. Oxidative phosphorylation appeared reduced in HF but exacerbated in ICM-DM, consistent with purportedly increased oxidative stress. Extracellular matrix proteins including collagens were up-regulated principally in ICM-DM despite the absence of macroscopic scar tissue. These findings were supported histologically and in metabolomic and RNA sequencing analyses.
Lipidomics analysis within healthy donors and end-stage heart failure with a focus on ischaemic cardiomyopathy with diabetes
STUDY_TYPE
Human heart failure left ventricular myocardial metabolomics study across multiple disease phenotypes.
STUDY_SUMMARY
This study involves the analysis of lipids within healthy and human heart failure conditions with a focus on ischaemic cardiomyopathy with diabetes and is part of a project which also integrates metabolomics, proteomics, RNA-seq, and histological analyses to identify molecular mechanisms influencing disease phenotypes. Ischaemic cardiomyopathy (ICM) is the most common cause of heart failure (HF) and often coexists with diabetes mellitus (DM). Yet, their combined effects are seldom investigated and are poorly understood. Herein, we performed multi-omic analyses of end-stage ICM with DM (ICM-DM) against ICM-No DM, non-ischaemic (dilated) cardiomyopathy with DM (NICM-DM), NICM-No DM, and healthy age-matched donors (AMD). Tissue was sourced from pre-mortem human left ventricular myocardium. Though fatty acid oxidation (FAO) proteins were down-regulated in ICM-DM relative to AMD and other HF, the unique ICM-DM down-regulation of acylcarnitines, perilipin, and ketone body, amino acid, and glucose metabolising proteins indicated FAO may not be entirely impaired. Oxidative phosphorylation appeared reduced in HF but exacerbated in ICM-DM, consistent with purportedly increased oxidative stress. Extracellular matrix proteins including collagens were up-regulated principally in ICM-DM despite the absence of macroscopic scar tissue. These findings were supported histologically and in metabolomic and RNA sequencing analyses.
Metabolomics analysis of human spermatozoa reveals impaired metabolic pathways in asthenozoospermia
STUDY_SUMMARY
Background: Infertility is a major health issue, affecting 15% of reproductive-age couples with male factors contributing to 50% of cases. Asthenozoospermia, or low sperm motility, is a common cause of male infertility with complex etiology, involving genetic and metabolic alterations, inflammation, and oxidative stress. However, the molecular mechanisms behind low motility are unclear. In this study, we used a metabolomics approach to identify metabolic biomarkers and pathways involved in sperm motility. Methods: We compared the metabolome and lipidome of spermatozoa of men with normozoospermia (n = 44) and asthenozoospermia (n = 22) using untargeted LC-MS and the metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal fluid redox status to assess the oxidative stress in the ejaculate. Results: We identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of the spermatozoaās metabolomics and lipidomics data showed a clear separation between groups. Spermatozoa of asthenozoospermic men presented lower levels of several amino acids, and increased levels of energetic substrates and lysophospholipids. However, the metabolome and redox status of the seminal fluid was not altered in asthenozoospermia. Conclusions: Our results indicate impaired metabolic pathways associated with redox homeostasis and amino acid, energy, and lipid metabolism in asthenozoospermia. Taken together, these findings suggest that the metabolome and lipidome of human spermatozoa are key factors influencing their motility and that oxidative stress exposure during spermatogenesis or sperm maturation may be in the etiology of decreased motility in asthenozoospermia.
INSTITUTE
University of Aveiro
DEPARTMENT
Department of Chemistry
LAST_NAME
Guerra-Carvalho
FIRST_NAME
BƔrbara
ADDRESS
Campus UniversitƔrio de Santiago, 3810-193 Aveiro, Portugal
Metabolomic analysis of Axon Regeneration in Xenopus laevis Retina
STUDY_SUMMARY
We profile the metabolite changes in the retina of a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that either had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 12 and 27 days post optic nerve crush. Following euthanasia, the tissues were collected for metabolomic analysis. Samples were pooled for each category (crush, sham, and control) at n =3 to obtain sufficient metabolite concentrations for analysis. Metabolites were extracted using a Precellys Homogenizer and a serial extraction method: (1) 1:1 Methanol/Water and (2) 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. Metabolites were identified and quantified using Compound Discoverer 3.3 and isotopic internal metabolites standards.
Metabolomic analysis of Axon Regeneration in Xenopus laevis Tectum
STUDY_SUMMARY
We profile the metabolite changes in the tectum of a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that either had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 12 and 27 days post optic nerve crush. Following euthanasia, the tissues were collected for metabolomic analysis. Samples were pooled for each category (crush, sham, and control) at n =3 to obtain sufficient metabolite concentrations for analysis. Metabolites were extracted using a Precellys Homogenizer and a serial extraction method: (1) 1:1 Methanol/Water and (2) 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. Metabolites were identified and quantified using Compound Discoverer 3.3 and isotopic internal metabolites standards.
Metabolomic analysis of Axon Regeneration in Xenopus laevis Optic Nerve
STUDY_SUMMARY
We profile the metabolite changes in the optic nerve of a transgenic line of 1 year old Xenopus laevis Tg(islet2b:gfp) frogs that either had a monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Tg(islet2b:gfp) frogs were allowed to recover for 12 and 27 days post optic nerve crush. Following euthanasia, the tissues were collected for metabolomic analysis. Samples were pooled for each category (crush, sham, and control) at n =3 to obtain sufficient metabolite concentrations for analysis. Metabolites were extracted using a Precellys Homogenizer and a serial extraction method: (1) 1:1 Methanol/Water and (2) 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) profiling using a Q-Exactive Orbitrap instrument coupled with Vanquish Horizon Binary UHPLC LC-MS system. Metabolites were identified and quantified using Compound Discoverer 3.3 and isotopic internal metabolites standards.
Phosphate availability conditions caspofungin tolerance, capsule attachment and titan cell formation in Cryptococcus neoformans
STUDY_SUMMARY
There is a pressing need for new antifungal drugs to treat invasive fungal diseases. Unfortunately, the echinocandin drugs that are fungicidal against other important fungal pathogens are ineffective against Cryptococcus neoformans, the causative agent of life-threatening meningoencephalitis in immunocompromised people. Contributing mechanisms for echinocandin tolerance are emerging with connections to calcineurin signaling, the cell wall, and membrane composition. In this context, we discovered that a defect in phosphate uptake impairs the tolerance of C. neoformans to the echinocandin caspofungin.
96-plex metabolomics studies on nutrient-deprived endothelial cells
STUDY_SUMMARY
A NeuCode tag which allows for cost efficient incorporation of isotopes is used to enable a 96-plex assay for small molecule metabolomics. A 96-well plate of cell lysates is analyzed simultaneously, providing absolute quantitation of 38 metabolites across 96 samples in a single LC-MS injection.
INSTITUTE
Saint Louis University
LAST_NAME
Armbruster
FIRST_NAME
Michael
ADDRESS
4448 Swiss Stone Ln E Apt 3C, Ypsilanti, Michigan, 48197, USA
Systematic evaluation of the hepatic de novo lipogenic substrate supply network reveals a hierarchical mitochondrial-cytosolic control structure
STUDY_SUMMARY
Hepatic de novo lipogenesis (DNL) is a fundamental physiologic process that becomes pathogenically elevated during metabolic disease. Both cytosolic citrate and acetate supply DNL, but their upstream inputs and pathways to DNL remain unclear. We traced 13C-labeled lactate/pyruvate and acetate into fatty acids in mitochondrial citrate carrier (CiC) and pyruvate carrier (MPC), ATP-citrate lyase (ACLY), acetyl-CoA synthetase 2 (ACSS2) liver-specific knockout (LivKO), and selective double LivKO (DLivKO) mice. We show that MPC and ACLY gate the major mitochondrial-cytosolic acetyl-CoA production pathway that operates in parallel to a cytosolic ACSS2 pathway. Given surprisingly persistent DNL after CiC + ACSS2 DLivKO, we considered a ketone-routed cytosolic acetyl-CoA production pathway as an alternative pathway feeding DNL. CiC LivKO markedly increased acetoacetate- and the ketogenic amino acid leucine-supplied DNL, consistent with ketones functioning as mitochondrial-citrate reciprocal DNL substrates. By delineating a structure for the DNL substrate supply network, these basic findings may inform strategies to therapeutically modulate DNL.
Quantification of bile acids in amoxicillin treated mice
STUDY_SUMMARY
Conventionally housed mice were orally treated with amoxicillin for 24 hours with and without cholic acid (48h) and small intestinal contents were collected for targeted MS analysis. Samples were flash frozen and stored at -80C prior to analysis. We found that amoxicillin drastically reduced unconjugated and secondary bile acids in the small intestine, while oral cholic acid supplementation produced only a modest recovery in intestinal cholic acid while also reducing the ratio of conjugated to unconjugated bile acids in the small intestine.
Interaction between NSCLC cells, CD8+ T cells and immune checkpoint inhibitors potentiates coagulation and promotes metabolic remodeling- new cues on CAT-VTE
STUDY_SUMMARY
Cancer-related thrombosis (CAT) and venous thromboembolism (VTE) are common cancer-related sequelae linked with high mortality, therefore finding predictive indicators is critical. Immune checkpoint inhibitors (ICI) are utilized in cancer immunotherapy to activate T lymphocytes against cancer cells. Recent retrospective investigations found that some individuals experienced increased VTE after receiving ICI. We postulated that ICI (anti-CTLA4 and anti-PD1) effect on immunological and cancer cells, as well as their interaction, contributes for an elevated risk of CAT-VTE. Our objective is to investigate this molecular interaction in order to uncover potential prothrombotic indicators. We pharmacologically modulated non-small cell lung cancer (NSCLC) cell lines in co-culture with CD8+ T lymphocytes (T CD8+) obtained from healthy blood donors. Nuclear Magnetic Resonance (NMR) metabolic remodeling analysis revealed differences in extracellular metabolite concentrations after T CD8+ and ICI treatment.
Metabolic profiling and synergistic therapeutic strategies unveil the cytotoxic potential of selenium-chrysin (SeChry) in NSCLC cells
STUDY_SUMMARY
Lung cancer ranks as the predominant cause of cancer-related mortalities on a global scale. Despite progress in therapeutic interventions, encompassing surgical procedures, radiation, chemotherapy, targeted therapies and immunotherapy, the overall prognosis remains unfavorable. Imbalances in redox equilibrium and disrupted redox signaling, common traits in tumors, play crucial roles in malignant progression and treatment resistance. Cancer cells, often characterized by persistent high levels of ROS resulting from genetic, metabolic, and microenvironmental alterations, counterbalance this by enhancing their antioxidant capacity. Cysteine availability emerges as a critical factor in chemoresistance, shaping the survival dynamics of non-small cell lung cancer (NSCLC) cells. Selenium-chrysin (SeChry) was disclosed as a modulator of cysteine intracellular availability. This study comprehensively characterizes the metabolism of SeChry in NSCLC. SeChry treatment induces notable metabolic shifts, particularly in selenocompounds metabolism, impacting crucial pathways such as glycolysis, gluconeogenesis, the tricarboxylic acid (TCA) cycle, and amino acid metabolism. Additionally, SeChry affects the levels of key metabolites such as acetate, lactate, glucose, and amino acids, contributing to disruptions in redox homeostasis and cellular biosynthesis.
Quantification of amoxicillin in mouse intestinal contents
STUDY_SUMMARY
Conventionally housed mice were orally treated with amoxicillin for 24 hours with and without cholic acid (48h) and small intestinal and cecal contents were collected for targeted MS analysis. Samples were flash frozen and stored at -80Ā°C prior to analysis. We found that amoxicillin levels in the small intestine were 0-40 pmol/mg of contents and no significant difference in amoxicillin concentration was detected between small intestine and cecum or between mice receiving cholic acid vs amoxicillin alone.
Integrative LC-MS and GC-MS Metabolic Profiling Unveils Dynamic Changes during Barley Malting
STUDY_TYPE
Untargeted LC-MS and GC-MS analysis
STUDY_SUMMARY
Malting, a crucial process for beer production, involves complex biochemical transformations affecting sensory attributes and product quality. Limited knowledge of metabolic alterations during malting hinders the ability to enhance malt quality. This study uses untargeted GC-MS and LC-MS metabolite profiling to characterize metabolic dynamics through the malting process. After data processing, a total of 4980 known metabolites were identified across six stages: dry seed, post-steeping, germination (DOG1, DOG3, DOG5), and kilned, about 82% of these showed significant changes during malting. Statistical analysis revealed stage-dependent shifts in metabolite profiles, highlighting the importance of the first 3 days of germination and kilning in determining the final metabolite content of finished malt. Dynamic changes in chemical classes and metabolic pathways provided insights into processes critical for malt quality and beer production. Additionally, metabolites associated with antimicrobial properties and stress responses were identified, underscoring the interplay between barley and microbial metabolic processes during malting. This comprehensive profiling advances our understanding of malting and suggests potential markers for process monitoring and quality control, ultimately enhancing malt quality and beer production.
High expression of oleoyl-ACP-hydrolase underpins life-threatening respiratory viral diseases
STUDY_SUMMARY
Although respiratory viral infections cause significant morbidity and mortality, it is unclear why some individuals succumb to severe disease. In patients hospitalized with avian A(H7N9) influenza, we investigated early drivers underpinning hypercytokinemia and fatal disease. Our transcriptomics studies identified differential expression of 10 early host genes, defined by 16 probe sets, between patients who recovered and died. Seven probe sets were specific for the same host gene encoding for a key enzyme mediating free fatty acid production, oleoyl-ACP-hydrolase (OLAH). High OLAH levels in fatal A(H7N9) patients were detected early after hospital admission and persisted until patients died. Conversely, patients who recovered had minimal OLAH expression throughout their hospital stay. High OLAH levels were also detected in patients hospitalized for severe infections with seasonal influenza virus, SARS-CoV-2, respiratory syncytial virus (RSV) and for multisystem inflammatory syndrome in children (MIS-C), while the main catalytic product of OLAH, oleic acid, was increased in hospitalized compared to ambulatory patients. Among healthy individuals and those with mild infections, however, OLAH was minimally detected. To understand how OLAH drives disease severity, we generated olah deficient mice. In contrast to wild-type mice, lethal influenza infection of olah-/- mice led to survival, milder disease, and markedly reduced lung viral loads, tissue damage, infection-driven pulmonary innate cell infiltration and inflammatory milieu. This phenotype was associated with differential lipid droplet dynamics, and reduced viral infection and inflammatory responses in macrophages. Supplementation of oleic acid, the main product of OLAH, increased influenza infection of macrophages and their inflammatory potential. Increased infectivity in the presence of olah was dependent on lipid droplet usage. Our findings define how expression of the key host enzyme, OLAH, drives life-threatening inflammation associated with respiratory viruses and propose OLAH as a potential early target for diagnosis and treatment of patients with severe disease.Ā
INSTITUTE
Peter Doherty Institute for Infection and Immunity
Atopic dermatitis and STAT3 double null mutant skin biopsies during healing.
STUDY_SUMMARY
To do this we developed a matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) workflow for high throughput lipid analysis of cutaneous biopsies as well as a cell culture model of wound healing. A total of 51 samples taken from normal and healing tissue in healthy and AD participants were analyzed using this workflow. The goal of our study was to (a) elucidating a broader understanding of the metabolic pathways initiated during wound healing, with a specific interest in lipid signatures, and (b) identify potential therapeutic targets for patients with barrier deficiencies or impaired healing.
Season-dependent variation in the contents of glucosinolates and S-methyl-L-cysteine sulfoxide and their hydrolysis in Brassica oleracea
STUDY_SUMMARY
Brassica vegetables are rich in phytochemicals, such as glucosinolates and S methyl L cysteine sulfoxide, which are sulfur-containing secondary metabolites. Upon cell damage, glucosinolates and S methyl L cysteine sulfoxide are hydrolyzed by endogenous enzymes to form bioactive breakdown products. Glucosinolate hydrolysis can result in the formation of isothiocyanates, or in presence of specifier proteins, nitriles, or epithionitriles can be formed. Isothiocyanates as well as some S methyl L cysteine sulfoxide derived volatile organosulfur compounds, such as S methyl methanethiosulfinate, show beneficial effects on human health. To optimize the quality of B. oleracea vegetables towards increased potential to release isothiocyanates from glucosinolates as well as S methyl L cysteine sulfoxide levels and hydrolysis, these compounds were monitored over the harvest seasons of different B. oleracea cultivars in three consecutive years. Strong seasonal shifts in glucosinolate hydrolysis outcome were observed in red cabbages. Summer red cabbage formed up to 40-fold more health-promoting isothiocyanates compared to red cabbage harvested in autumn. Decreasing temperatures and shorter days with lower light intensities in autumn enhanced nitrile and epithionitrile formation.This correlated with an increased abundance of epithiospecifier proteins in autumn red cabbage. On the opposite, higher volatile organosulfur compound levels were found in autumn red cabbage. The demonstrated seasonal shifts in glucosinolate hydrolysis behavior seem to be cultivar-specific since only little, or no change in isothiocyanate formation was observed in white cabbage and broccoli. By elucidating of the impact of abiotic growth factors on the regulatory mechanisms of glucosinolate degradation, cultivation conditions of B. oleracea vegetables can be optimised, and thus, their health value can be improved.
INSTITUTE
Leibniz Institute of Vegetable and Ornamental Crops (IGZ) e.V.
High Fat Feeding Alters Circulating Triglyceride Profile by Decreasing SCD Activity and Depleting omega-3 Fatty Acids
STUDY_SUMMARY
The study was to examine, in control and high-fat-fed rats, the effects of acute feeding (a single meal) on circulating lipid profiles. Acute feeding is expected to impact circulating lipid profiles due to lipids in chylomicron derived from the diet, and the acute feeding response may be altered with a HFD to account for some of the chronic effects of a HFD. The study was to examine the effects of a high fat diet (HFD) on circulating TG profiles in rats and test the hypothesis that a HFD reduces TGs composed of long-chain PUFAs, as previously seen in rats maintained on a HNaLK diet. Thus, we performed lipidomics analysis on plasma samples collected both in the pre- and postprandial states in the control and HFD groups. Lipidomics analysis was combined with general metabolomics analysis, which provides measurements of saturated FFAs and other metabolic indicators such as glycerol (as lipolysis indicator) or 3-hydroxybutyrate (ketosis indicator), not measured in our LC-MS-based lipidomics assay. The results provided novel insights into the differential behaviors of individual TGs and identified specific changes (i.e., decreased stearoyl-CoA desaturase [SCD] activity and depletion of omega-3 FFAs) associated with short-term high-fat feeding.
INSTITUTE
University of Southern California
DEPARTMENT
Keck School of Medicine USC
LAST_NAME
Youn
FIRST_NAME
Jang-Hyun
ADDRESS
CSC 214, 2250 Alcazar Street, Health Sciences Campus, Los Angeles CA 90033
Metabolomic profiling of cultured TRAMP-C2 cells in the presence or absence of PD-L1.
STUDY_SUMMARY
Recent evidence suggests that PD-L1, well-known as the ligand of the immune inhibitory receptor PD-1, can have cell-intrinsic effects in cancer and immune cells. One such cell-intrinsic effect is modulation of cellular metabolism, including regulation of mTOR activity and glycolysis. Here, we analyzed the metabolome of cultured mouse prostate cancer cells (TRAMP-C2) expressing PD-L1 or with PD-L1 deleted via CRISPR/Cas9.
Natural rubber (NR) is an important material with excellent physical properties. Unlike synthetic rubber from petroleum, NR contains non-rubber components such as proteins, lipids, and metal ions. The non-rubber components are known to affect the properties of NR. In this study, latex samples of Hevea brasiliensis were collected for nine months and their metabolites were comprehensively analyzed by mass spectrometry. NR was made from the same latex samples used for the mass spectrometry, and their vulcanization, tensile and thermal-aging properties were assessed. These results suggest that the metabolite composition in the latex and the NR properties changed seasonally. Correlation analysis between the metabolites and the properties of NR indicated that different metabolites affected different properties. A regression model of NR properties using metabolites as the explanatory variables suggests that about five metabolites need to be considered when examining the relationship between properties and metabolites. This method, which combines comprehensive analysis and characterization of NR, contributes to studies aimed at elucidating how the superior properties of NR are brought about.
INSTITUTE
Sumitomo Riko Co., Ltd.
DEPARTMENT
Fundamental Material Development Laboratory
LAST_NAME
Nobuyuki
FIRST_NAME
Hiraoka
ADDRESS
1, Higashi 3-chome, Komaki-shi, Aichi 485-8550, Japan
Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue
STUDY_SUMMARY
The central nervous system has been implicated in the age-induced reduction in adipose tissue lipolysis. SLC7A14 is a lysosomal membrane protein highly expressed in the brain. Herein, we investigated the possible role of hypothalamic SLC7A14 in the age-induced lipolysis reduction. In this study, we demonstrated the expression of SLC7A14 was reduced in proopiomelanocortin (POMC) neurons of aged mice. Overexpression of SLC7A14 in POMC neurons alleviated the age-induced reduction in white adipose tissue (WAT) lipolysis, whereas SLC7A14 deletion mimicked the age-induced lipolysis impairment. Moreover, POMC SLC7A14 regulated WAT lipolysis independently of sympathetic nerves in WAT. Metabolomics analysis revealed that POMC SLC7A14 increased the primary bile acid taurochenodeoxycholic acid (TCDCA) content, which mediated the SLC7A14 knockout- or age-induced WAT lipolysis impairment. Furthermore, SLC7A14-increased TCDCA content is dependent on intestinal apical sodium-dependent bile acid transporter (ASBT), which is regulated by intestinal sympathetic afferent nerves. Finally, SLC7A14 regulated the intestinal sympathetic afferent nerves by inhibiting mTORC1 signaling through inhibiting TSC1 phosphorylation. Collectively, our study suggests the function for central SLC7A14 and an upstream mechanism for the mTORC1 signaling pathway. Moreover, our data provides insights into the braināgutāadipose tissue crosstalk in age-induced lipolysis impairment.
INSTITUTE
Shanghai Institutes for Biological Sciences (SIBS) Chinese Academy of Sciences (CAS)
Short chain fatty acid (SCFA) analysis of human stool samples under different sample logistics
STUDY_SUMMARY
We sought to understand how sample logistics, i.e. sample storage directly upon sampling and during transportation to the processing laboratory, impacts on the microbiota in terms of the SCFA concentration in 6 different donors.Effects of oxygen exposure were negligible. At ambient temperature, SCFA were stable for 24 h and 48 h, respectively, while at 4 Ā°C, SCFA profiles were maintained for 48 h.
Untargeted Dialysate Metabolomics Identification and Detection of Novel Neurochemicals via Benzoyl Chloride Derivatization
STUDY_SUMMARY
This study combines these strategies to obtain in-depth untargeted chemical identification of dialysate. We target the rat dorsal and ventral striatum, given its importance in motivation, movement, and reward processing and identify 489 compounds. Despite the depth of analysis, many well-known neurochemicals, such as all neurotransmitters, were not identified in the untargeted method. These compounds could be detected using BzCl derivation in a targeted method though. By tracking isotopic BzCl feature pair detection and filtering of MS/MS spectra by known benzoyl fragment ions, we found 872 unique features in dialysate suggesting many unknown compounds remain to be identified in dialysate.
Metabolic Alterations in Aneurysmal Subarachnoid Hemorrhage
STUDY_SUMMARY
Aneurysmal subarachnoid hemorrhage (aSAH) is a severe type of stroke that is associated with poor outcome. A subset of patients with aSAH will develop secondary complications, most notably delayed cerebral ischemia (DCI), which potentiates neurological injury. In this study, we investigate the relationship between cerebrospinal fluid (CSF) iron accumulation, brain metabolism, and neuronal injury in aSAH patients with or without DCI. We collected longitudinal CSF samples of patients immediately after hospitalization and 5-8 days after onset of ictus. CSF was analyzed with metabolomics to determine metabolic alterations associated with aSAH and DCI. Metabolomic profiling of the CSF samples uncovered significant dysregulation of metabolic pathways associated with energy generation and amino acid utilization, consistent with mitochondrial dysfunction. Using machine learning, we identified a set of metabolites that predicted ICU length of stay (LOS). aSAH alters the CSF metabolome involved in mitochondrial function and a subset of these metabolites are predictive of ICU stay. These results identify potential biomarkers for mitochondrial pathology and provide insight into alterations in brain iron metabolism triggered by aSAH.
PyINETA: Open-source platform for INADEQUATE-JRES integration in NMR metabolomics
STUDY_SUMMARY
This study developed a new data analysis tool for metabolomics to integrate information of NMR experiments INADEQUATE and JRES. This deposition is for samples used for the validation of this new tool. Three mice were fed with a 13C-labeled diet and the distribution of 13C-labeled metabolites was examined using INADEQUATE and JRES.
Comparative analysis of breast cancer metabolomes highlights fascin's central role in regulating key pathways related to disease progression
STUDY_SUMMARY
Omics technologies provide useful tools for the identification of novel biomarkers in many diseases, including breast cancer, which is the most diagnosed cancer in women worldwide. We and others have reported a central role for the actin-bundling protein (fascin) in regulating breast cancer disease progression at different levels. However, whether fascin expression promotes metabolic molecules that could predict disease progression has not been fully elucidated. Here, fascin expression was manipulated via knockdown (fascinKD+NORF) and rescue (fascinKD+FORF) in the naturally fascin-positive (fascinpos+NORF) MDA-MB-231 breast cancer cells. Whether fascin dysregulates metabolic profiles that are associated with disease progression was assessed using untargeted metabolomics analyses via liquid chromatography-mass spectrometry. An overall of 12,226 metabolites were detected in the tested cell pellets. Fascinpos+NORF cell pellets showed 2,510 and 3,804 significantly dysregulated metabolites compared to their fascinKD+NORF counterparts. Fascin rescue (fascinKD+FORF) revealed 2,710 significantly dysregulated cellular metabolites compared to fascinKD+NORF counterparts. 101 overlapped cellular metabolites between fascinKD+FORF and fascinpos+NORF were significantly dysregulated in the fascinKD+NORF cells. Analysis of the significantly dysregulated metabolites by fascin expression revealed their involvement in the metabolism of sphingolipid, phenylalanine, tyrosine and tryptophan biosynthesis, and pantothenate and CoA biosynthesis, which are critical pathways for breast cancer progression. Our findings of fascin-mediated alteration of metabolic pathways could be used as putative poor prognostic biomarkers and highlight other underlying mechanisms of fascin contribution to breast cancer progression.
INSTITUTE
King Faisal Specialist Hospital and Research Centre (KFSHRC)
ASCT2 is a major contributor to serine uptake in cancer cells
STUDY_SUMMARY
The non-essential amino acid serine is a critical nutrient for cancer cells due to its diverse biosynthetic functions. While some tumors can synthesize serine de novo, others are auxotrophic and therefore reliant on serine uptake. Importantly, despite several transporters being known to be capable of transporting serine, the transporter(s) that mediate serine uptake in cancer cells are not known. Here, we characterize the amino acid transporter ASCT2 (SLC1A5) as a major contributor to serine uptake in cancer cells. ASCT2 is well-known as a glutamine transporter in cancer, and our work demonstrates that serine and glutamine compete for uptake through ASCT2. We further show that ASCT2-mediated serine uptake is essential for purine nucleotide biosynthesis and that ERĪ± promotes serine uptake by directly activating SLC1A5 transcription. Together, our work defines an additional important role for ASCT2 as a serine transporter in cancer and evaluates ASCT2 as a potential therapeutic target.
Untargeted metabolomics of rhizosphere soil from 4-years Panax ginseng that was treated with endo-borneol under field condition
STUDY_SUMMARY
To study the rhizosphere soil matabolism of 4-years Panax ginseng that were leaf-sprayed with endo-borneol at different concentrations (0, 0.1 mg/L, and 100 mg/L) under field condition, liquid chromarography tandem mass spectrometry was used for examination. Each treatment is set with 6 replicates for assessing nontargeted metabolomics. The LC-MS/MS analysis was performed using a Thermo UHPLC-Q Exactive HF-X system equipped with an ACQUITY HSS T3 column. As a result, a total of 296 and 569 metabolites were identified in negative ion mode and positive mode, respectively.
INSTITUTE
Yunnan University
DEPARTMENT
School of Agriculture
LABORATORY
Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China
LAST_NAME
Xing-Yu
FIRST_NAME
Ji
ADDRESS
Road Fengyuan, District Panlong, Kunming, Yunnan, China
An untargeted metabolic profiling strategy for the dissection of the oat (Avena sativa L.) plant innate immune response to various Pseudomonas syringae pathovars
STUDY_TYPE
Plant Metabolomics
STUDY_SUMMARY
One of the most important characteristics that plants utilise to successfully defend themselves is the ability to rapidly identify potential threats in the surrounding environment. Plants rely on the perception of microbe-derived molecular pattern chemicals for this recognition, which initiates a number of induced defence reactions that ultimately increase plant resistance. The metabolome acts as a metabolic fingerprint of the biochemical activities that take place in a biological system under particular conditions and therefore provides a functional readout of the cellular mechanisms involved in a biological system. In this study, an untargeted metabolomics approach was applied to decipher the biochemical processes involved in oat plant defence responses to inoculation with various pathovars of Pseudomonas syringae (pathogenic and non-pathogenic on oat) such as P. syringae pv. coronafaciens (Ps-c), -pv. tabaci (Ps-t), -pv. tomato DC3000 (DC3000) and -pv. tomato DC3000 hrcC mutant (hrcCā) and thereby identify signatory markers that are involved in host or nonhost defence responses. At the seedling growth stage, metabolic alterations in the Dunnart oat cultivar (tolerant to Ps-c) in response to inoculation with the respective Pseudomonas syringae pathovars were examined. Following inoculation, plants were monitored for symptom development and harvested at 2-, 4- and 6 d.p.i.. Methanolic metabolite extracts were prepared, and ultra-high-performance liquid chromatography (UHPLC) connected to a qTOF high-definition mass spectrometer was used to analyse the extracts. Chemometric modelling and multivariate statistical analysis revealed host- and time-related metabolic alterations that point to host and nonhost interactions in response to bacterial inoculation/infection. Metabolic profiles from further multivariate data analyses revealed a range of metabolite classes involved in the respective defence responses, including phenolic amides, saponins, phenolic acids, flavonoids, fatty acids, amino acids and alkaloids. The findings in this study allowed the elucidation of metabolic changes involved in oat defence responses to a range of pathovars of Pseudomonas syringae and ultimately contributed to a more comprehensive view of the oat plant metabolism under biotic stress during host vs nonhost interactions.
INSTITUTE
University of Johannesburg
DEPARTMENT
Biochemistry
LABORATORY
Plant Metabolomics
LAST_NAME
Pretorius
FIRST_NAME
Chanel
ADDRESS
Corner Kingsway and University Road, Auckland Park, Johannesburg, 2092
Needle leachates of Pinus koraiensis from May to September
STUDY_SUMMARY
To study the allelopathy in agroforestry system (Pinus koraiensis and Panax ginseng in this context), we identified the compounds of the needle leachates during the growing season of Panax ginseng (May~September). Three replicates were set for each month, leachates were extracted with ethyl acetate and concentrated for GC-MS analysis. As a result, a total of 795 metabolites were identified in all the samples.
INSTITUTE
Yunnan University
DEPARTMENT
School of Agriculture
LAST_NAME
Xing-Yu
FIRST_NAME
Ji
ADDRESS
Road Fengyuan, District Panlong, Kunming, Yunnan, China
Determination the concentration of endo-borneol from needle leachates of Pinus koraiensis flushed by precipitation via indoor simulated leaching)
STUDY_SUMMARY
The precipitation from April to October (2016-2021) was provided by local weather burea in Benxi. The monthly precipitation and duration in Benxi is classified into high, medium, and low level. Under each precipitation, the concentration of endo-borneol is quantified by indoor simulated leaching. The simulated leachates were extracted by ethyl acetate and then analyzed by GC/MSD. As a result, the peak area of endo-borneol ranges from 2006.257 to 27106.75 under tested precipitation.
INSTITUTE
Yunnan University
DEPARTMENT
School of Agriculture
LABORATORY
Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China
LAST_NAME
Xing-Yu
FIRST_NAME
Ji
ADDRESS
Road Fengyuan, District Panlong, Kunming, Yunnan, China
Complementary Omics Analyses of The Brain Of A Kainic Acid Epilepsy Model Reveal Alterations In Purine And Energy Metabolism With Metabolic Implications for miRNA Cargos Within Milk-Exosomes
STUDY_SUMMARY
Augmentations in energy and glucose metabolism have been widely implicated in epilepsy, a neurological disorder that manifests as seizures. Evaluating the metabolic underpinnings of epilepsy using systems approaches to identify potential treatment targets will especially benefit ~30% of patients that struggle with drug-resistant epilepsy. Recently, bovine milk-derived exosomes and their associated miRNA cargos were demonstrated to be bioavailable in the brain, improve spatial learning, and decrease seizure severity. Here, we assessed the metabolic changes associated with epilepsy and this exosome diet on brain tissue isolated from a kainic acid mouse model of epilepsy via metabolomics and proteomics. Mice fed either an RNA-sufficient exosome diet (ERS) or an RNA-insufficient exosome diet (ERD). After dietary interventions, mice were injected with kainic acid (KA+) or vehicle solution (KA-) (n=24; n=6 in each category). We show that central carbon, purine, and redox metabolism pathways were primarily affected by seizures in addition to relatively understudied pathways such as hexosamine and nicotinamide salvage. We also show that the interaction effects of dietary exosomes and their associated miRNAs were most prominent in the presence of seizure. Proteomics data revealed a correlation of proteins and metabolites related to glycolytic and energy production pathways, implicating flux.
INSTITUTE
University of Nebraska-Lincoln
LAST_NAME
Alicia
FIRST_NAME
Johnson
ADDRESS
433 Bolivar Street New Orleans, LA 70112
EMAIL
jadameclab@gmail.com
PHONE
(504) 568-1610
NUM_GROUPS
4
TOTAL_SUBJECTS
24
STUDY_COMMENTS
Raw metabolite values (with no processing) have been given.
Integrative analysis of serum and fecal metabolome and the microbiome that herald Crohn Disease flare - serum
STUDY_SUMMARY
We investigated the relationship between the host and gut microbiota in the context of Crohn Disease (CD), which is a relapsing-remitting condition. We analyzed paired omics of 80 CD patients and 43 controls, including 202 serum and 294 fecal metabolomics, and 258 microbiome samples. Our findings suggest that CD patients have an inverse shift in energy utilization from sugars and fat between serum and feces. In the serum, we noted a decrease in metabolites related to starch, sucrose, and tricarboxylic acid (TCA) cycle, as well as an increase in metabolites linked to fat in contrast to the feces, where we noted higher sugars and TCA cycle metabolites, and lower fat metabolites. Interestingly, fecal sugars were specifically linked with oral bacteria mislocated to the CD gut, while unsaturated fat derivatives of arachidonic acid were linked with R. gnavus and Fusobacteria. We identified consistent metabolite alterations in CD patients, which were also present in clinical/biomarkers active CD, and pre-flare samples of patients who experienced flare. Using pre-flare samples, we developed models that predicted a subsequent flare using metabolic serum and fecal signatures. We validated the fecal metabolomics predictions in another, similarly designed, independent CD cohort. Finally, we developed a clinical lab-based index [UA/Cr ratio+log2(CRP)] based on the serum metabolomics model, which was also predictive in the validation cohort. Here are the serum metabolomics samples.
INSTITUTE
Sheba hospital
LAST_NAME
Braun
FIRST_NAME
Tzipi
ADDRESS
Sheba hospital, Ramat Gan, Ramat Gan, 52621, Israel
Integrative analysis of serum and fecal metabolome and the microbiome that herald Crohn Disease flare - feces
STUDY_SUMMARY
We investigated the relationship between the host and gut microbiota in the context of Crohn Disease (CD), which is a relapsing-remitting condition. We analyzed paired omics of 80 CD patients and 43 controls, including 202 serum and 294 fecal metabolomics, and 258 microbiome samples. Our findings suggest that CD patients have an inverse shift in energy utilization from sugars and fat between serum and feces. In the serum, we noted a decrease in metabolites related to starch, sucrose, and tricarboxylic acid (TCA) cycle, as well as an increase in metabolites linked to fat in contrast to the feces, where we noted higher sugars and TCA cycle metabolites, and lower fat metabolites. Interestingly, fecal sugars were specifically linked with oral bacteria mislocated to the CD gut, while unsaturated fat derivatives of arachidonic acid were linked with R. gnavus and Fusobacteria. We identified consistent metabolite alterations in CD patients, which were also present in clinical/biomarkers active CD, and pre-flare samples of patients who experienced flare. Using pre-flare samples, we developed models that predicted a subsequent flare using metabolic serum and fecal signatures. We validated the fecal metabolomics predictions in another, similarly designed, independent CD cohort. Finally, we developed a clinical lab-based index [UA/Cr ratio+log2(CRP)] based on the serum metabolomics model, which was also predictive in the validation cohort. Here are the fecal metabolomics samples.
INSTITUTE
Sheba hospital
LAST_NAME
Braun
FIRST_NAME
Tzipi
ADDRESS
Sheba hospital, Ramat Gan, Ramat Gan, 52621, Israel
Randomized phase II trial of pre-operative fulvestrant with or without enzalutamide for ER+/Her2- primary breast cancer: effects on tumor immune microenvironment and clinical outcomes.
STUDY_SUMMARY
This randomized phase II trial of 4 months of neoadjuvant fulvestrant (Fulv) alone or with enzalutamide (Combo) assessed whether the addition of AR blockade to Fulv would limit residual tumor at time of surgery, as measured by modified preoperative endocrine predictive index (PEPI) score. Eligible patients were women with ER+/HER2- primary BC cT2 or greater. Lithium-heparin (LiHep) plasma samples were obtained every 4 weeks until surgery and one month after surgery then analyzed by MS-based metabolomics.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Effects of LDAH overexpression on the lipidome of oxLDL-treated mouse peritoneal macrophages
STUDY_SUMMARY
Mouse peritoneal macrophages from LDAH-transgenic mice and wild-type littermate controls were harvested 5 days after aged 3% thioglycolate injection. 4ā5 Ć 106 cells were plated in 60mm culture dishes, and cultured overnight in DMEM supplemented with 10% heat-inactivated fetal bovine serum (FBS). The media was replaced by DMEM-1% FBS supplemented with Hi-TBAR oxLDL (50 Ī¼g/ml, Alfa Aesar J65261) for 48h.
Effects of LDAH deficiency on the lipidome of oxLDL-treated mouse peritoneal macrophages
STUDY_SUMMARY
Lipid droplet-associated hydrolase (LDAH) has a lipase structure and high affinity for lipid droplets of macrophages/foam cells. However, LDAH's functions and lipid substrates remain poorly understood. Mouse peritoneal macrophages from LDAH-knockout mice and their wild-type control littermates were harvested 5 days after aged 3% thioglycolate injection. 4ā5 million cells were plated in 60mm culture dishes, and cultured overnight in DMEM supplemented with 10% heat-inactivated fetal bovine serum (FBS). The media was replaced by DMEM-1% FBS supplemented with Hi-TBAR oxLDL (50 Ī¼g/ml, Alfa Aesar J65261) for 48h.
Kinetic changes in the phospholipid composition of fibroblasts induced by cytotoxic stress
STUDY_SUMMARY
Time-dependent analysis (0, 0.17, 1, 6, 24, 48 h) of thephosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and phosphatidylglycerol fatty acid composition of NIH-3T3 mouse fibroblasts treated with cytotoxic stressors (staurosporine, cycloheximide, etoposide, thapsigargin, valinomycin, myrtucommulone) or depleted from serum. Please note that parts of the data were previously uploaded to project PR001114 [doi: 10.21228/M8Q39V] and reanalyzed for the current project (PI).
Changes in the phosphatidylcholine fatty acid composition in subcellular fractions of fibroblasts induced by valinomycin
STUDY_SUMMARY
Determination of the impact of valinomycin on the subcellular phosphatidylcholine fatty acid composition of NIH-3T3 fibroblasts challenged with valinomycin for 48 h and disrupted by repeated passage through a syringe. Nuclear, mitochondrial, membrane, and cytosolic fractions were obtained by differential centrifugation.
Time-dependent changes in the phosphatidylcholine fatty acid composition of fibroblasts treated with acetyl-CoA carboxylase inhibitors
STUDY_SUMMARY
Inhibition of acetyl-CoA carboxylase in NIH-3T3 fibroblasts by soraphen A or 5-(tetradecyloxy)-2-furoic acid (TOFA) and time-dependent analysis (0.17 h, 1 h, 6 h, 24 h, 48 h) of the phosphatidylcholine fatty acid composition.
Effect of ACC inhibition on the proportion of PUFAs in phosphatidylcholines in stressed fibroblasts
STUDY_SUMMARY
Consequences of acetyl-CoA carboxylase inhibition by 5-(tetradecyloxy)-2-furoic acid (TOFA) on the phosphatidylcholine fatty acid composition of NIH-3T3 fibroblasts treated with cytotoxic stressors (staurosporine, cycloheximide, etoposide, thapsigargin, valinomycin, myrtucommulone), exposed to tumor necrosis factor Ī±, or deprived of serum for 48 h.
EBC metabolomics in non-small cell lung cancer patients and control individuals
STUDY_SUMMARY
A total of 18 EBC differential metabolites (FDR < 0.05) were identified, which included phenylalanine and tryptophan, dipeptides related to BCAAs, unsaturated, oxidized and hydroxyl fatty acids, and other compounds. Among them, 8 differential metabolites had AUC > 0.8, exhibiting distinct levels in the NSCLC and control EBC samples. Two OPLS-DA models were established from the differential metabolites with FDR < 0.05 and FDR < 0.01, with a 5-metabolite panel and a 4-metabolite panel, respectively. The OPLS-DA model achieved 86.2% sensitivity, 83.3% specificity and 84.9% accuracy for the 5-metabolite panel, and 79.3% sensitivity, 83.3% specificity and 81.1% accuracy for the 4-metabolite panel. The EBC differential metabolites between the NSCLC and control groups showed a potential to distinguish NSCLC from controls, which may provide assistance for noninvasive diagnosis of lung cancer, once they are validated in large cohort studies.
Global Metabolomics and U-13C6-Glucose Tracing in Bone Marrow Derived Macrophages (BMDM)
STUDY_SUMMARY
We evaluated alterations in cellular metabolism via comprehensive metabolic profiling of BMDMs derived from control and trained HSCLT. Thus, we performed ultra-high-performance mass spectrometry (UHPLC-MS) analysis of control and trained donor HSCLT-derived BMDMs following 6hr IC stimulation. We identified 206 metabolites, of which 88 were differentially abundant in control vs. trained BMDMs. Notably, we observed reductions in global metabolite abundance across multiple pathways in trained HSCLT-derived BMDMs including multiple amino acid, nucleotide, glutathione, and fatty acid synthesis pathways among others. In agreement with our Seahorse data, BMDMs from trained HSCLT exhibited a reduction in the abundance of metabolites in and downstream of the glycolysis pathway following IC stimulation, compared to BMDMs from control donor HSCLT. This phenotype was particularly evident in pathways constituting central carbon metabolism, including glycolysis itself (green), the pentose phosphate pathway (red), the tricarboxylic acid cycle (TCA, blue), as well as alternative carbon metabolites (grey). Furthermore, we observed significant reductions in products of central carbon metabolism, including levels of ATP, AMP (gold), and amino acids (grey) as well as fatty acids, pentose phosphates and lactate. Along these lines, even in the absence of IC stimulation there were significant reductions in metabolites associated with several pathways, primarily amino acid metabolism, nucleotide metabolism, and fatty acid metabolism (42 metabolites total). Given the significant reduction in glycolytic metabolites and their downstream products in IC-stimulated BMDMs derived from trained HSCLT, we also measured changes in glucose fate by culturing the BMDMs for 24h in media containing U-13C-glucose. 6hr prior to harvest, we stimulated them with IC. This approach facilitates analysis of IC-induced glycolytic activity levels and details changes in glucose fate based on isotopic labeling of downstream metabolites with 13C, while also providing information on global (13C labeled + unlabeled) metabolite levels. Consistent with our global metabolic analysis and Seahorse assays above, we noted reduced global metabolite levels as well as broad reductions in overall 13C peak areas in multiple metabolic pathways downstream of glucose, including lower glycolysis (lactate production), the pentose phosphate pathway, TCA cycle and TCA cycle-derived amino acids, based on measurement of the peak area of 13C labeled isotope species (M+#). To further evaluate potential shifts in glucose utilization we also expressed the data as a proportion of M+x# 13C labeled plus M+0 unlabeled metabolite isotopologues. Of note, in trained HSCLT donor-derived BMDMs we identified a slight though significant increase in the proportion of M+6 13C labeling in upper glycolysis (hexose phosphate), which culminated in preferential entry into the pentose phosphate pathway (based on increased though non-significant proportions of M+6 13C labeled phospho-gluconate, M+7 13C labeled sedoheptulose 1/7-p) and M+5 labeled pentose phosphates, likely indicative of 13C label cycling through the non-oxidative PPP. On the other hand, BMDMs from trained donor HSCLT exhibited a trending reduction in the proportion of 13C labeling of lactate and a significant reduction in the proportion of 13C labeling in the TCA cycle indicative of reduced glucose oxidation through central carbon metabolic pathways downstream of pyruvate. Furthermore, we observed reduced 13C labeling in key amino acids derived from central carbon metabolism including alanine and glutamate (purple), reflecting reduced contribution of glucose to amino acid synthesis and consistent with reduced global levels of these metabolites (Fig. 6D). As these data could be consistent with increased glucose entry into fatty acid synthesis pathways, we measured 13C label accumulation into palmitate but did not identify significant changes in 13C labeling. Taken together, these data are consistent with broad metabolic remodeling of IC-stimulated BMDMs derived from trained HSCLT, characterized by smaller metabolite pools and preferential routing of glucose into the PPP, with concomitant reduction of glucose entry into the TCA cycle.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Bevers
FIRST_NAME
Shaun
ADDRESS
12801 E 17th Ave Bldg. RC-1S Rm L2400 Aurora CO 80045
Lipidome profiling in non-alcoholic steatohepatitis identifies phosphatidylserine synthase 1 as a regulator of hepatic lipoprotein metabolism
STUDY_SUMMARY
Non-alcoholic fatty liver disease and more progressive non-alcoholic steatohepatitis (NASH) are characterized by defective lipid metabolism, which causes hepatic steatosis and disease progression. However, the changes in lipid metabolism in NASH are incompletely understood. Using lipidome profiling in livers of eight mouse strains, that differ substantially in susceptibility to NASH and liver fibrosis, as well as in patients with NASH, we show that phosphatidylserine (PS) accumulation and preservation of PS synthase 1 (PSS1) expression is associated with resistance to NASH. Mechanistically, PSS1 overexpression in the liver reduces hepatic steatosis through remodeling of the hepatic and liver-derived VLDL lipidome in mice with NASH. Specifically, we show an increase in VLDL ceramide content that suppresses the expression and activity of lipoprotein lipase (LPL) in skeletal muscle, thereby reducing VLDL-triglyceride clearance, fatty acid uptake and lipid accumulation in skeletal muscle. In addition, remodelling of lipoprotein composition inhibits the LDL receptor in the liver, likely contributing to the reduction in hepatic steatosis. Together, this study provides a unique resource describing lipidome changes in NASH, and identifies PSS1 as a novel regulator of hepatic lipoprotein metabolism.
Control Of Major Bleeding After Trauma (COMBAT) Clinical Research Trial Metabolomics
STUDY_SUMMARY
Denver Health has conducted the Control of Major Bleeding After Trauma (COMBAT) clinical research study, which aimed to improve trauma care for all patients (https://www.denverhealth.org/services/trauma/for-professionals-2/research-publications/combattrial). The study began in April 2014 and concluded in March of 2017. In collaboration with the University of Colorado Anschutz Medical Campus, longitudinal plasma samples collected from the COMBAT trial were analyzed by MS-based metabolomics and proteomics (data available on ProteomeXchange).
Increased Cholesterol Synthesis Drives Neurotoxicity in Patient Stem Cell-Derived Model of Multiple Sclerosis - cellular lipidomics
STUDY_SUMMARY
Lipidomics analysis was performed on directly induced neural stem/progenitor cell (iNSC) lines derived from fibroblasts of patients with progressive multiple sclerosis (PMS) versus age matched controls (AMC) treated or untreated with cholesterol synthesis inhibitor simvastatin.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Increased Cholesterol Synthesis Drives Neurotoxicity in Patient Stem Cell-Derived Model of Multiple Sclerosis - media lipidomics
STUDY_SUMMARY
Lipidomics analysis was performed on conditioned media from cultured iNSCs derived from fibroblasts of progressive multiple sclerosis patients versus age-matched controls.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Increased Cholesterol Synthesis Drives Neurotoxicity in Patient Stem Cell-Derived Model of Multiple Sclerosis - cellular metabolomics
STUDY_SUMMARY
Fibroblasts from skin biopsies from patients with progressive multiple sclerosis (PMS) and age-matched controls (AMC) were derived and reprogrammed to yield induced neural stem cells (iNSCs). The resulting iNSCs were cultured in the presence of 13C6 glucose for 24h to assess changes in the intracellular metabolome, in particular changes in the flux of glucose-derived carbon into the TCA cycle along with steady state levels of compounds in other areas of the metabolome.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Increased Cholesterol Synthesis Drives Neurotoxicity in Patient Stem Cell-Derived Model of Multiple Sclerosis - media metabolomics
STUDY_SUMMARY
Fibroblasts from skin biopsies from patients with progressive multiple sclerosis (PMS) and age-matched controls (AMC) were derived and reprogrammed to yield induced neural stem cells (iNSCs). The resulting iNSCs were cultured in the presence of 13C6 glucose for 24h to assess changes in the intracellular metabolome, in particular changes in the flux of glucose-derived carbon into the TCA cycle along with steady state levels of compounds in other areas of the metabolome. The data in this study are from the assessment of the conditioned culture media at the end of the 24h incubation period.
INSTITUTE
University of Colorado Denver
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Urine Metabolite Profiling in Indian Male Population at High Altitude using Liquid Chromatography ā Mass Spectrometry: A Longitudinal Pilot Study
STUDY_SUMMARY
People who visit high altitude (HA) for work, pilgrimage, recreation purposes or deployment are subjected to environmental stresses such as low temperature, atmospheric pressure leading to hypoxia, high radiations, dry air, non-availability of fresh food and vegetables, etc. These environmental stressors pose significant physiological effects on the human body. Among these challenges, hypobaric hypoxia at high altitude affects the aerobic metabolism and thereby reducing the supply of metabolic energy. These alterations could further lead to extreme environment-related maladaptation as evidenced by changes in metabolites and metabolic pathways. In order to investigate the variation in metabolite prfile, urine samples were collected from 16 individual at base line (BL) and at High altitude (HA, 4420m). Untargeted urine metabolic profiling was performed using liquid chromatographyāmass spectrometry (LC-MS) in conjunction with statistical analysis. Univariate and Multivariate statistical analysis revealed 33 metabolites based on fold change, (FC >2 and < 0.5), VIP score (>1) and p value (<0.05). These 33 metabolites were primarily associated with pathways related to Phenylalanine, tyrosine and tryptophan biosynthesis, Phenylalanine metabolism, Biotin metabolism, Cysteine and methionine metabolism, Tyrosine metabolism, Alanine, aspartate and glutamate metabolism, Pentose and glucuronate interconversions, Vitamin B6 metabolism, Citrate cycle (TCA cycle) and Porphyrin metabolism. Further, ROC analysis detected 5 metabolites i.e., 2-Tetrahydrothiopheneacetic acid (AUC: 0.97266), 1-Benzyl-7,8-dimethoxy-3-phenyl-3H-pyrazolo[3,4-c]isoquinoline (AUC: 0.92969), Abietin (AUC:0.917), 4,4'-Thiobis-2-butanone (AUC: 0.917), and Hydroxyisovaleroyl carnitine (AUC: 0.910) with a high range of sensitivity and specificity. In summary, this present longitudinal study demonstrated metabolic changes in human exposed to high altitude, via utilising the potential application of LC-MS metabolomics. These findings will shed light on the impact of hypoxic exposure on metabolic adaptation and provide a better understanding about the pathophysiological mechanism of HA related illnesses correlated to tissue hypoxia.
INSTITUTE
Defence Institute of Physiology and Allied Sceinces
Impact of partial body shielding from very high dose rates on untargeted metabolomics in biodosimetry
STUDY_SUMMARY
A realistic exposure to ionizing radiation (IR) from an improvised nuclear device will likely include individuals that are partially shielded from the initial blast delivered at a very high-dose rate (VHDR). As different tissues have varying levels of radiosensitivity, e.g. hematopoietic vs. gastrointestinal tissues, the effects of shielding on radiation biomarkers needs to be addressed. Here, we explore how biofluid (urine and serum) metabolite signatures from male and female C57BL/6 mice exposed to VHDR (5 ā 10 Gy/sec) total body irradiation (TBI, 0, 4, and 8 Gy) compare to individuals exposed to partial body irradiation (PBI) (lower body irradiated [LBI] or upper body irradiated [UBI] at an 8 Gy dose) using a data-independent acquisition untargeted metabolomics approach. Although sex differences were observed in the spatial groupings of urine signatures from TBI and PBI mice, a metabolite signature (N6,N6,N6-trimethyllysine, carnitine, propionylcarnitine, hexosamine-valine-isoleucine, taurine, and creatine) previously developed from variable dose rate experiments was able to identify individuals with high sensitivity and specificity irrespective of radiation shielding. A panel of serum metabolites composed from previous untargeted studies on nonhuman primates had excellent performance for separating irradiated cohorts; however, a multi-omic approach to complement the metabolome could increase dose estimation confidence intervals. Overall, these results support the inclusion of small molecule markers in biodosimetry assays without substantial interference from upper or lower body shielding.
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Biological stress metabolomics in Duchenne muscular dystrophy rodent model
STUDY_TYPE
Biological Stress Discovery Targeted Metabolomics
STUDY_SUMMARY
Duchenne muscular dystrophy is a severe neuromuscular wasting disease that is caused by a primary defect in dystrophin protein and involves organism-wide comorbidities such as cardiomyopathy, metabolic and mitochondrial dysfunction, and non-progressive cognitive impairments. Physiological stress exposure in the mdx mouse model of Duchenne muscular dystrophy results in phenotypic abnormalities that include locomotor inactivity, hypotension, and increased morbidity. Severe and lethal stress susceptibility in mdx mice corresponds to metabolic dysfunction in several coordinated metabolic pathways within dystrophin-deficient skeletal muscle, as well as prolonged elevation in mdx plasma corticosterone levels that extends beyond the WT stress response. Here we performed a targeted mass spectrometry-based plasma metabolomics screen focused on biological stress pathways in healthy and dystrophin-deficient mdx mice exposed to mild scruff stress. One-third of the stress-relevant metabolites interrogated displayed significant elevation or depletion in mdx plasma after scruff stress and were restored to WT by skeletal muscle-specific dystrophin expression. The metabolic pathways of mdx mice altered by scruff stress are associated with regulation of the hypothalamic-pituitary-adrenal axis, locomotor tone, neurocognitive function, redox metabolism, cellular bioenergetics, and protein catabolism. Our data suggest that a mild stress triggers an exaggerated, multi-system metabolic response in mdx mice.
Differentiation between cytotoxic stress-induced PUFA biosynthesis and PUFA incorporation into phospholipids by metabolic flux studies in fibroblasts
STUDY_SUMMARY
Metabolic flux studies addressing the incorporation of i) deuterium labeled polyunsaturated fatty acids (20:4, 22:4, 22:5, 22:6) having one or two acetate-d3 incorporated or ii) one arachidonic acid-d8 or its elongation product adrenic acid-d8 into cellular phosphatidylcholines of NIH-3T3 fibroblasts that were treated with TOFA, VAL or a combination of both
Analysis of oxidized arachidonoyl-phosphatidylethanolamine in valinomycin-treated fibroblasts
STUDY_SUMMARY
Treatment of NIH-3T3 fibroblasts with valinomycin for 48 h and analysis of isomeric arachidonoyl phosphatidylethanolamines with two oxygens incorporated.
We aimed to characterize the rewarming ischemia phase during LuTx by measuring organ temperature and comparing metabolome profiles in tissue obtained at the end versus the start of implantation.
DINCH and MINCH reach adipose and liver tissues and levels are partially retained during recovery period
STUDY_SUMMARY
This study focused on determining the concentrations of DINCH, MINCH, and secondary metabolites in adipose tissue depots following treatment and recovery, using targeted metabolomics. Additionally, these concentrations were measured in the liver, the primary organ for biotransformation. Analysis of DINCH, MINCH, and secondary metabolite concentrations revealed significant variability both between and within individual tissues in males and females. Post-treatment, DINCH was present in all examined tissues, with the highest concentrations found compared to its metabolites. Among the metabolites, MINCH and OH-MINCH were predominant in the liver, while MINCH was the sole metabolite detected in adipose tissue. Following the recovery period, all DINCH metabolites were reduced to trace levels. Notably, there was a significant reduction of DINCH concentrations in the liver and brown adipose tissue (BAT), but no reduction in visceral adipose tissue (VIS AT) and subcutaneous adipose tissue (SC AT).
Effect of feeding and the mTORC1 activity on metabolism in Caenorhabditis elegans
STUDY_SUMMARY
Cellular growth and organismal development are remarkably complex processes that require the nutrient-responsive kinase mTORC1. Anticipating that important mTORC1 functions remained to be identified, we employed genetic and bioinformatic screening in C. elegans to uncover mechanisms of mTORC1 action. Here we show that during larval growth nutrients induce an extensive reprogramming of gene expression and alternative mRNA splicing by acting through mTORC1. mTORC1 regulates mRNA splicing and production of protein-coding mRNA isoforms largely independently of its target p70S6K, by increasing the activity of the serine/arginine-rich (SR) protein RSP-6 (SRSF3/7) and other splicing factors. mTORC1-mediated mRNA splicing regulation is critical for growth, mediates nutrient control of mechanisms that include energy, nucleotide, amino acid, and other metabolic pathways, and may be conserved in humans. Although mTORC1 inhibition delays aging, mTORC1-induced mRNA splicing promotes longevity, suggesting that when mTORC1 is inhibited enhancement of this splicing might provide additional anti-aging benefits.
INSTITUTE
Hiroshima University
LAST_NAME
Ogawa
FIRST_NAME
Takafumi
ADDRESS
1-3-1 Kagamiyama, Higashihiroshima, Hiroshima, 739-8530, Japan
Untargeted Lipidomic Profiling of Canine Cancer Cell Lines
STUDY_SUMMARY
Cell sensitivity to ferroptosis is closely associated with cellular lipid composition and unsaturation. To further characterize the 31 canine cell lines tested for ferroptosis sensitivity, we collected untargeted lipidomic data for each line in duplicate. Omics characterizations of cancer cell lines and integration with viability data in efforts such as the Cancer Cell Line Encyclopedia (CCLE) and the Cancer Dependency Map (www.depmap.org) have yielded powerful resources to explore causal and predictive biomarkers of response to genetic and small molecule perturbations. Our data facilitate a similar approach for the canine lines included in this study. Lipidomics data covered over 750 individual species falling into 18 main lipid classes. Total concentrations varied widely between individual cell lines, as did relative lipid composition. Likewise, the fraction of cellular polyunsaturated fatty acids (PUFAs), previously reported as a contributor to ferroptosis sensitivity, varied from 20% to almost 70%. Thus, the panel of tested canine cell lines represented a wide spectrum of cellular lipid composition.
Glucosinolate profile and specifier protein activity determine the glucosinolate hydrolysis product formation in kohlrabi (Brassica oleracea var. gongylodes) in a tissue-specific way
STUDY_SUMMARY
Glucosinolates, commonly found in Brassica vegetables, are hydrolyzed by myrosinase to form bioactive isothiocyanates unless specifier proteins redirect the degradation to less bioactive nitriles and epithionitriles. Here, the tissue-specific impact of specifier proteins on the outcome of glucosinolate hydrolysis in nine kohlrabi tissues was investigated. Glucosinolate and their hydrolysis product profiles, epithiospecifier protein and myrosinase activity and protein abundance patterns of key glucosinolate biosynthesis, transport and hydrolysis proteins were determined and correlated to the metabolites in the kohlrabi tissues. Leaf tissues were rich in glucoraphanin, while bulb tissues contained more glucoerucin. Moreover, a higher proportion of isothiocyanates was formed in leaf stalk, bulb peel, stem and root, tissues with relatively higher ESM1 abundance and a higher ratio of myrosinase activity to ESP activity. This study shows that tissue-specific glucosinolate hydrolysis is mediated by glucosinolate biosynthesis and profiles, myrosinase activity and specifier protein as well as modifier protein abundance.
INSTITUTE
Leibniz Institute of Vegetable and Ornamental Crops (IGZ) e.V.
Proteomic and metabolomic profiling of methicillin resistant versus methicillin sensitive Staphylococcus aureus using a simultaneous extraction protocol
STUDY_SUMMARY
Background: Understanding the biology of methicillin resistant Staphylococcus aureus (MRSA) is crucial to unlocking insights for new targets in our fight against this antimicrobial resistant priority pathogen. Although proteomics and metabolomic profiling offer the potential to elucidating such biological markers, reports of meth-odological approaches for carrying this out in S. aureus isolates remain limited. We describe the use of a dual-functionality methanol extraction method for the concur-rent extraction of protein and metabolites from S. aureus and report on the com-parative analysis of the proteomic and metabolomic profiles of MRSA versus methi-cillin sensitive S. aureus (MSSA). Methods: Bacterial reference strains MRSA ATCC43300 and MSSA ATCC25923 were used . The conventional urea methodology was used for protein extraction and a methanol based method was used for concurrent proteins and metabolites extraction. Proteomic and metabolomic profiling was carried out using TimsTOF mass spectrometry. Data processing was carried out using the MaxQuant version 2.1.4.0 Results: This study represents the first report on the utilization of the methanol ex-traction method for concurrent protein and metabolite extraction in Gram positive bacteria. Our findings demonstrate good performance of the method for the dual extraction of proteins and metabolites from S. aureus with demonstration of repro-ducibility.Comparison of MRSA and MSSA strains revealed 407 proteins with significantly different expression levels. Enrichment analysis of those proteins re-vealed distinct pathways involved in fatty acid degradation, metabolism and beta-lactam resistance. Penicillin-binding protein PBP2a, the key determinant of MRSA resistance, exhibited distinct expression patterns in MRSA isolates. Metabolomic analysis identified 146 metabolites with only one exclusive to the MRSA. The enriched pathways identified were related to arginine metabolism and biosynthesis. Conclusion: Our findings demonstrate the effectiveness of the methanol-based dual-extraction method, providing simultaneous insights into the proteomic and metabolomic landscapes of S. aureus strains. These findings demonstrate the utility of proteomic and metabolomic profiling for elucidating the biological basis of antimicrobial resistance.
INSTITUTE
Mohammed Bin Rashid University of Medicine and Health Science
LAST_NAME
Boucherabine
FIRST_NAME
Syrine
ADDRESS
Dubai Healthcare city building 14, Dubai, Dubai, 00000000, United Arab Emirates
The adaptation of tumor cells to metabolic stress is crucial for tumor development. We demonstrate that the translation of NRF2, a master regulator of antioxidant response, is controlled by the OGDH-METTL3 axis in melanoma for glucose starvation adaptation. To elucidate the regulatory mechanism of the OGDH-METTL3 axis during glucose starvation, we conducted co-immunoprecipitation and mass spectrometry analysis to identify the binding partner of OGDH that governs both the OGDH-METTL3 axis and tumor cell adaptation to glucose deprivation by detecting succinyl-CoA in total cells or cytoplasm.
INSTITUTE
Xiamen University
LAST_NAME
Xu
FIRST_NAME
Kejun
ADDRESS
Xiangshan Street 4221, Xiangan District, Xiamen, Fujian, 361100, China
ADSL deficiency drives mitochondrial dysfunction and ERK2 dysregulation in a linear genotype to phenotype correlation
STUDY_SUMMARY
Adenylosuccinate lyase (ADSL) deficiency (ADSLd) is a rare autosomal recessive defect of purine metabolism associated with a wide range of clinical manifestations. Despite ADSL implication in purine synthesis, no additional molecular alterations have been identified as a cause of ADSLd besides the accumulation of toxic substrates. Here we uncover a novel association between ADSLd and mitochondrial dysfunction, which is characterized by an increase in fragmentation and reduction in respiration and ATP production. The extent of this mitochondrial dysfunction is directly proportional to the pathological manifestations of ADSLd, which are predominantly observed in tissues that rely heavily on mitochondria. Our analysis also unveils a striking defect in mitochondrial dynamics and transport, which are associated with the suppression of ERK2 and AKT function. Remarkably, the mitochondrial phenotype can be rescued in part by overexpression of a constitutive form of ERK2 or through the administration of purine intermediates. This scenario provides an alternative explanation of ADSLd onset, reorienting research towards developing innovative therapeutic strategies based on the restoration of mitochondrial metabolism.
An integrated LC-MS analysis of the biometric characteristics of different time cohorts of race walkers - untargeted
STUDY_SUMMARY
To obtain metabolic changes during endurance exercise, we conducted a metabolomics study in which 19 race walkers were recruited to collect blood at four time points: before, during, after, and after rest until day two using both non-targeted and targeted methods. A total of 859 metabolites and 411 lipids were identified by non-targeted methods, and 465 metabolites were quantitatively identified by targeted methods. This extensive data set provides a valuable resource for understanding systemic metabolic changes caused by race walking and helps researchers identify biomarkers of exercise performance.
INSTITUTE
The First Affiliated Hospital of Dalian Medical University
LAST_NAME
Zhang
FIRST_NAME
yunshu
ADDRESS
No.222 Zhongshan Road, Xigang District, Dalian City, Liaoning Province
To obtain the metabolic alterations during endurance exercise, we performed a metabolomics study that enrolled 19 race-walking athletes using untargeted and targeted approaches. In total, 859 metabolites and 411 lipids were identified in the untargeted methods, and 465 metabolites were quantified from targeted methods. This extensive dataset offers a valuable resource for understanding the systemic metabolic changes induced by race walking and assists researchers in identifying biomarkers of athletic performance.
INSTITUTE
The First Affiliated Hospital of Dalian Medical University
LAST_NAME
Zhang
FIRST_NAME
yunshu
ADDRESS
No.222 Zhongshan Road, Xigang District, Dalian City, Liaoning Province
Dissecting the Genetic Basis of UV-B Responsive Metabolites in Rice
STUDY_SUMMARY
UV-B, an important environmental factor, has been shown to affect the yield and quality of rice (Oryza sativa) worldwide. However, the biochemical and genetic basis of the metabolome underlying the response to UV-B stress remain elusive in rice. In this study, we performed comprehensive metabolic profiling of leaves from 160 diverse rice accessions under UV-B and normal-light conditions using a widely targeted metabolomics approach. Our results revealed substantial differences in the accumulation of metabolites between the two major rice subspecies indica and japonica, especially after UV-B treatment, shedding light on the possible role and mechanism of metabolome changes in subspecies differentiation and the stress response.
INSTITUTE
Industrial Crops Institute of Hubei Academy of Agricultural Sciences
Untargeted lipidomics profiling of LPL-overexpression and LPL-knockdown microglia
STUDY_SUMMARY
In this study, human microglia from induced pluripotent stem cells (iPSCs) derived from a TSC patient cohort were generated. Upregulated glycerophosphocholines and fatty acyls were found in TSC microglia, resulting in increased phagocytosis and inflammation. Strikingly, the dysregulated lipid metabolism in TSC microglia is driven by hyper-activation of mTOR-lipoprotein lipase (LPL) pathway. Furthermore, cellular and electrophysiological assessments of neuron/microglia co-cultures revealed that TSC microglia directly affect neuronal development and excitability as well as neuronal network activity, which could be largely ameliorated by mTOR/LPL inhibition. Through integrated multi-omics analysis, levels of glycerophosphocholines were found increased in the microglia of humans with tuberous sclerosis complex. To determine whether glycerophosphocholines are regulated by the LPL gene, we performed experiments with LPL overexpression in microglia and LPL knockdown in microglia.
INSTITUTE
Stjude children research hospital
LAST_NAME
Chu
FIRST_NAME
Mengqi
ADDRESS
262 Danny Tomas Place, Memphis, Tennessee, 38104, USA
Incorporation of oleate-d9 and elaidate-d17 in long-chain base of sphingolipids in Huh7 cells.
STUDY_SUMMARY
We analyzed hydrolyzed long-chain bases (LCBs) in Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17 in the presence or absence of myriocin, a pharmacological inhibitor of SPT, the initial rate-limiting enzyme of sphingolipid biosynthesis.
Incorporation of co-treated oleate-d9 and elaidate-d17 in long-chain base of sphingolipids in Huh7 cells.
STUDY_SUMMARY
We analyzed hydrolyzed long-chain bases (LCBs) in Huh7 cells treated with a combination of BSA-oleate-d9 and BSA-elaidate-d17. We aimed to confirm the incorporation of oleate and elaidate in the LCB of sphingolipids using their deuterated versions and put them both in direct competition for SPT.
Noninvasive multiomic measurement of cell type repertoires in human urine
STUDY_SUMMARY
Background: Early detection of the cell type changes underlying several genitourinary tract diseases largely remains an unmet clinical need, whereas existing assays, if available, lack the cellular resolution afforded by an invasive biopsy. While messenger RNA in urine could reflect dynamic signal that facilitates early detection, current measurements primarily detect single genes and thus do not capture the full spectrum of cell type specific contributions. Methods: We isolated and sequenced the cellular and cell-free RNA from urine samples (n = 6 healthy controls and n = 12 kidney stones) alongside the metabolome. We analyzed the resulting urine transcriptomes and metabolomes by comparing the bulk gene expression, deconvolving the noninvasively measurable cell type contributions, and comparing to the plasma cell-free transcriptome. Results: We primarily observed signal originating from genitourinary tract cell types in addition to cell types from high-turnover solid tissues beyond the genitourinary tract. Integration of urinary transcriptomic and metabolomic measurements identified various metabolic pathways involved in amino acid metabolism overlap with metabolic subsystems associated with proximal tubule function. Conclusions: Noninvasive whole transcriptome measurements of human urine reflect signal from hard-to-biopsy tissues exhibiting low representation in the blood at cell type resolution.
INSTITUTE
CZ Biohub
LAST_NAME
DeFelice
FIRST_NAME
Brian
ADDRESS
1291 Welch Rd., Rm. G0821 (SIM1), Stanford CA, California, 94305, USA
Secretion of sphingomyelin from Huh7 cells treated with various fatty acids including of oleate-d9 and elaidate-d17.
STUDY_SUMMARY
We analyzed sphingomyelin (SM) in fresh and spent media from Huh7 cells treated with BSA-EtOH, BSA-palmitate, BSA-stearate, BSA-oleate-d9 or BSA-elaidate-d17. We aimed to determine the secretory flux of SM induced by various fatty acid treatments.
Secretion of phosphatidylcholine from Huh7 cells treated with oleate-d9 or elaidate-d17
STUDY_SUMMARY
We analyzed PC in the fresh and spent media of Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17 to determine their incorporation into secreted phospholipids. We aimed to determine if SM secretory flux in response to fatty acid treatments correlated to PC secretory flux.
Secretion of sphingomyelin from Huh7 cells treated with oleate-d9 or elaidate-d17 while modulating SPT flux.
STUDY_SUMMARY
We analyzed SM in the fresh and spent media from Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17. We validated their reduction in abundance through pharmacological inhibition of SPT with myriocin.
Bile acidāinduced gut mitochondrial dysfunction promotes Enterobacteriaceae colonization and associates with dysbiosis in Crohn's disease
STUDY_SUMMARY
Here, we investigated whether bile acids influence gut colonization by Enterobacteriaceae. In an external human dataset, we found association between mucosal Enterobacteriaceae and the expression of mitochondrial damageārelated genes. In a murine model, increased intestinal BA availability enhanced colonization by E. coli in an oxygen-dependent manner. Overall, this study establishes BAs as a potential bridge that facilitates interactions between commensals and the host epithelium, laying the essential foundation for future investigations into various intestinal diseases.
Metabolomics analysis of Glioblastoma (GBM) cell line U251 labeled by 13C-glutamine after treatment with pimozide
STUDY_SUMMARY
Glioblastoma (GBM) cell line U251 was treated with antipsychotic drug pimozide (3 uM) for 24 hr, and then labeled with 13C-glutamine (2 mM) for 1 hr. Cells were collected and extracted for metabolites and analyzed by LC-MS. Our data showed that pimozide treatment significantly increased 13C-labeled glutamine uptake and subsequent consumption, including anaplerosis of metabolites for tricarboxylic acid (TCA) cycle and de novo fatty acid synthesis derived from glutamine-mediated reductive carboxylation process.
INSTITUTE
The Ohio State University
LAST_NAME
Guo
FIRST_NAME
Deliang
ADDRESS
Department of Radiation Oncology, Ohio State Comprehensive Cancer Center, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, and College of Medicine at The Ohio State University, Columbus, OH 43210, USA.
Human milk microbiome across lactation stages and free glutamate concentrations in healthy Ecuadorian women
STUDY_SUMMARY
Project Summary: To examine the differences in microbiome and glutamate concentrations in human breast milk among different lactation stages (colostrum-, transition-, and mature-milk), a longitudinal cohort study for healthy Ecuadorian women was conducted. This deposition is for the glutamate analysis.
Deep lipidomic profiling reveals sex dimorphism of lipid metabolism in fibro-calcific aortic valve disease
STUDY_SUMMARY
In-depth lipidomics profiling was performed on human tricuspid and bicuspid aortic valve tissue at different pathological stages, including mildly diseased, fibrotic and calcific tissue sections. Untargeted and targeted analysis were employed to enable high identification coverage and accurate quantification of lipids. Chromatographic separation modes (C18 RPC, C30 RPC) was combined with MS analysis using various acquisition modes (full MS, DDA, SRM) to allow for the identification of 1084 lipid molecular species across 28 lipid subclasses, of which 480 lipids were semi-absolutely quantified.
Malate dehydrogenase (MDH) inhibition assay: Fasnall does not affect MDH activity in vitro
STUDY_TYPE
Intracellular metabolomics, medium metabolomics
STUDY_SUMMARY
The enzymatic activity of porcine malate dehydrogenase (MDH) was reconstructed using NADH and oxaloacetate as substrates. LC-MS/MS was used to monitor the reaction by measuring the accumulation of malic acid and NAD+, as well as the consumption of oxaloacetate and NADH. Fasnall does not affect MDH activity in vitro.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Non-Invasive Diagnosis of Moyamoya Disease Using Serum Metabolic Fingerprints and Machine Learning
STUDY_SUMMARY
Moyamoya disease (MMD) is a progressive cerebrovascular condition that elevates the risk of intracranial ischemia and hemorrhage. Timely diagnosis and intervention can considerably lower the chances of new stroke occurrences in MMD patients. However, existing diagnostic techniques are both invasive and costly, with few reports on non-invasive diagnosis using MMD biomarkers. To tackle this challenge, we conducted non-targeted metabolomics analysis using LDI-MS on serum from 288 samples (validation cohort: MMD/HC: 115/115; discovery cohort: MMD/HC: 29/29) to identify patients with MMD. We then created a diagnostic model leveraging deep learning algorithms, which demonstrated remarkable accuracy in distinguishing the MMD group from the HC group (AUC = 0.977, 95% CI of 0.945 to 1.000). This method represents a promising new approach for MMD diagnosis. Additionally, our findings may have wider implications for the diagnosis of other neurological disorders.
INSTITUTE
Shanghai Jiao Tong University affiliated Renji Hospital
LAST_NAME
Xu
FIRST_NAME
Yudian
ADDRESS
Shanghai, 200127, P. R. China, Shanghai, Shanghai, 200127, China
An untargeted metabolomic analysis of acute AFB1 treatment in liver, breast, and lung cells
STUDY_SUMMARY
Aflatoxin B1 (AFB1) is a class 1 carcinogen and mycotoxin known to contribute to the development of hepatocellular carcinoma (HCC), growth impairment, altered immune system modulation, and malnutrition. AFB1 is synthesized by Aspergillus flavus and is known to widely contaminate foodstuffs, particularly maize, wheat, and groundnuts. The mechanism in which AFB1 causes genetic mutations has been well studied, however its metabolomic effects remained largely unknown. A better understanding of how AFB1 disrupts metabolism would provide insight into how this mycotoxin leads to carcinogenesis, growth impairment, and/or immunomodulation, and may reveal potential targets for pharmacological or nutritional interventions to protect against these effects. The current study evaluated the metabolomic effects of various doses (2.5 ĀµM, 5 ĀµM, 10uM) of AFB1 treatment to HepG2 (liver), MDA-MB-231 (breast), and A549 (lung) cells. Treated and control cellsā metabolomic profiles were evaluated via ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Univariate and multivariate analyses revealed significant alterations in metabolite concentrations from each dose of AFB1 treatment in each cell type. Pathway analysis was then used to understand broader biochemical functions affected by AFB1 treatment in each cell type. HepG2 cell pathway analyses revealed significant pathway perturbations in lipid metabolism, carnitine synthesis, catecholamine biosynthesis, purine metabolism, and spermidine and spermine biosynthesis. Analysis of A549 cells found a greater emphasis of perturbations on various amino acids along with lipid synthesis-related pathways, and catecholamine biosynthesis. Finally, analysis of treated MDA-MB-231 cells found spermidine and spermine biosynthesis, carnitine synthesis, plasma membrane-related pathways (phosphatidylcholine synthesis and alpha linolenic acid and linoleic acid metabolism), and various amino acid metabolism pathways to be most affected. These highlighted pathways should be targeted in future investigations to evaluate their potential in mitigating or preventing the development of negative health effects associated with AFB1 exposure.
Incorporation of oleate-d9 and elaidate-d17 in broader lipidome of Huh7 cells.
STUDY_SUMMARY
We analyzed several lipid classes including LPC, LPE, PC, PE, DG, and TG in Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17 to determine their incorporation into the broader lipidome.
Incorporation of oleate-d9 and elaidate-d17 in broader lipidome of Huh7 cells.
STUDY_SUMMARY
We analyzed several lipid classes including LPC, LPE, PC, PE, DG, and TG in Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17 to determine their incorporation into the broader lipidome.
Incorporation of oleate-d9 and elaidate-d17 in sphingolipids in Huh7 cells while modulating SPT flux.
STUDY_SUMMARY
We analyzed sphingolipids in Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17 to determine their incorporation into the long-chain base (LCB) of intact sphingolipids. We validated their reduction in abundance through pharmacological inhibition of SPT with myriocin.
Incorporation of oleate-d9 and elaidate-d17 in sphingolipids in Huh7 cells while modulating SPT flux.
STUDY_SUMMARY
We analyzed sphingolipids in Huh7 cells treated with BSA-oleate-d9 or BSA-elaidate-d17 to determine their incorporation into the long-chain base (LCB) of intact sphingolipids. We validated their reduction in abundance through pharmacological inhibition of SPT with myriocin.
Structural and systems characterization of phosphorylation on metabolic enzymes identifies sex-specific metabolic reprogramming in obesity
STUDY_TYPE
Polar metabolomics analysis of diet induced obesity
STUDY_SUMMARY
C57BL/6J mice were obtained from The Jackson Laboratories and housed in a specific pathogen-free facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care using laminar flow cages at 21Ā°C. Mouse health is monitored and maintained by veterinary staff under the supervision of a veterinarian. Male and female mice fed ad libitum either a standard chow diet, a high fat diet (HFD, S3282, Bio-Serv), or a HFD supplemented with 1.5% 2(3)ātertābutylā4 hydroxyanisole (BHA, S7958, Bio-Serv) for 16 weeks beginning at age 8 weeks. All mice are euthanized at age 24 weeks following an overnight fast by inhalation of isoflurane followed by cervical dislocation, and the livers are snap frozen in liquid nitrogen upon removal.
Bempedoic acid improves diet-induced steatosis independent of hepatic ACLY
STUDY_SUMMARY
To test the effect of BPA on hepatic steatosis, we placed WT and LAKO mice on WD for 6 weeks, with 10 mg/kg BPA daily oral gavage over the last 3 weeks, a dose previously shown to be effective in reducing steatosis. Quantification of lipids by mass spectrometry demonstrated that ACLY deficiency increased abundance of TAGs and DAGs, while BPA treatment reduced these lipids in both genotypes. Moreover, ACLY deficiency reduced phospholipid (PC and PE) abundance.
INSTITUTE
Salk Institute for Biological Studies
LAST_NAME
KUNA
FIRST_NAME
RAMYA
ADDRESS
10010 N Torrey Pines Rd, La Jolla, California, 92037, USA
Sphingolipid secretory flux from Huh7 SPTLC3 KO cells
STUDY_SUMMARY
We analyzed 13C labeling on sphingolipids in fresh and spent media from Huh7 control and SPTLC3 KO cells treated with BSA-palmitate and the stable isotope tracers [13C]serine and [13C]glycine to measure sphingolipid secretory flux.
Obesity and fatty liver diseases-metabolic dysfunction-associated steatotic liver disease (MASLD and MASH) affect over a third of the global population and are exacerbated in individuals with reduced functional aldehyde dehydrogenase 2 (ALDH2), observed in approximately 560 million people. Current treatment to prevent disease progression to cancer remains inadequate, requiring innovative approaches. We observe that Aldh2-/- and Aldh2-/-Sptbn1+/- mice develop phenotypes of human Metabolic Syndrome (MetS) and MASH with a striking accumulation of endogenous aldehydes such as 4-hydroxynonenal (4-HNE). While phospholipids are often modified by reactive aldehydes that accumulate in the absence of ALDH2, to understand the mechanisms for the differences in liver metabolism in ASKO mice, we analyzed liver metabolomics and lipidomics from mice models. Briefly, C57BL/6 mice (n=15) were from 3 groups (WT, Aldh2-/-(ko), Aldh2-/-Sptbn1+/-(double), n=5 per group) and fed normal chow diet for 10 months. For quality control, 6 QC samples were also included in the analysis (total 21 samples). We observed that livers of Aldh2-/-Sptbn1+/- mice had substantially higher levels of all investigated phospholipids, including Ć¢ĀĀ„ 2-fold increase in 26% of phosphatidylethanolamine (PE) lipid types and Ć¢ĀĀ„ 2-fold increase in 32% of phosphatidylserine (PS) lipid types, compared to livers of WT mice. Similarly, increased abundances of TGs and diacylglycerides (DGs) lipid types were also observed in the livers of Aldh2-/-Sptbn1+/- mice. These results demonstrated that Aldehydes altered lipid metabolism which may be implicated in the progression of liver MetS, MASLD/MASH in Aldh2-/-Sptbn1+/- mice.
Impact of high-fat diet enriched in cis or trans fatty acids and myriocin on hepatic lipidome in Ldlr-/- mice
STUDY_SUMMARY
We analyzed LPC, LPE, PC, PE, DG, TG, and CE in the liver of Ldlr-/- mice fed 1) Cis HFD, 2) Cis HFD + Myriocin, 3) Trans HFD, or 4) Trans HFD + Myriocin. We aimed to determine how cis and trans monounsaturated fatty alter the broader hepatic lipidome while pharmacologically inhibiting the initial rate-limiting enzyme of sphingolipid biosynthesis, serine palmitoyltransferase (SPT), via myriocin.
Deletion of Kcnj16 altered transcriptomic and metabolomic profiles of Dahl salt-sensitive rats
STUDY_TYPE
Untargeted metabolomis
STUDY_SUMMARY
The inwardly rectifying K+ Channel Kir5.1 (Kcnj16) is essential in renal salt handling and blood pressure control. However, the underlying mechanisms are not fully understood. Here, we integrated transcriptomics and metabolomics to comprehensively profile the changes in genes and metabolites in the Dahl salt-sensitive (SS) rat lacking Kcnj16 to identify potential mechanisms. Consistent with the phenotype of knock-out (KO) rats, the transcriptomic profile predicted reduced blood pressure, kidney damage, and increased ion transport. Canonical pathway analysis suggested activation of metabolic-related while suppression of immune responses-related pathways in KO rats. Untargeted metabolomic analysis revealed different metabolic profiles between WT and KO rats. Integration of transcriptomic and metabolomic profiles suggested altered tricarboxylic acid (TCA) cycle, amino acid, and reactive oxygen species (ROS) metabolism that are related to SS hypertension. In conclusion, besides increased ion transport, our data suggest suppressed immune responses-related and altered metabolic-related pathways of SS rats lacking Kir5.1.
Impact of high-fat diet enriched in cis or trans fatty acids and myriocin on newly synthesized sphingolipids from D2O in circulation in Ldlr-/- mice
STUDY_SUMMARY
We analyzed newly synthesized DHCer, Cer, and SM in plasma of Ldlr-/- mice fed 1) Cis HFD, 2) Cis HFD + Myriocin, 3) Trans HFD, or 4) Trans HFD + Myriocin. We aimed to determine how dietary cis and trans monounsaturated fatty acids impact hepatic secretion of newly synthesized sphingolipids into circulation while pharmacologically inhibiting the initial rate-limiting enzyme of sphingolipid biosynthesis, serine palmitoyltransferase (SPT), via myriocin.
Sphingolipid biosynthetic flux in Huh7 SPTLC3 KO cells
STUDY_SUMMARY
We analyzed 13C labeling on sphingolipids in Huh7 control and SPTLC3 KO cells treated with BSA-palmitate and the stable isotope tracers [13C]serine and [13C]glycine to measure intracellular sphingolipid biosynthetic flux.
Impact of high-fat diet enriched in cis or trans fatty acids and myriocin on the plasma lipoprotein profile of sphingolipids in Ldlr-/- mice
STUDY_SUMMARY
We analyzed sphingolipids in plasma lipoprotein fractions of Ldlr-/- mice fed 1) Cis HFD, 2) Cis HFD + Myriocin, 3) Trans HFD, or 4) Trans HFD + Myriocin. We aimed to determine how dietary cis and trans monounsaturated fatty acids impact sphingolipid enrichment in circulating lipoproteins while pharmacologically inhibiting the initial rate-limiting enzyme of sphingolipid biosynthesis, serine palmitoyltransferase (SPT), via myriocin.
Impact of high-fat diet enriched in cis or trans fatty acids and myriocin on plasma lipidome in Ldlr-/- mice
STUDY_SUMMARY
We analyzed LPC, LPE, PC, PE, DG, TG, and CE in the plasma of Ldlr-/- mice fed 1) Cis HFD, 2) Cis HFD + Myriocin, 3) Trans HFD, or 4) Trans HFD + Myriocin. We aimed to determine how dietary cis and trans monounsaturated fatty acids impact the broader plasma lipidome while pharmacologically inhibiting the initial rate-limiting enzyme of sphingolipid biosynthesis, serine palmitoyltransferase (SPT), via myriocin.
Annotation of Metabolites in Stable Isotope Tracing Untargeted Metabolomics via Khipu-web
STUDY_SUMMARY
Stable isotope tracing is a crucial technique for understanding the metabolic wiring of biological systems, determining meta-bolic flux through pathways of interest, and detecting novel metabolites and pathways. Despite the potential insights provid-ed by this technique, its application remains limited to a small number of targeted molecules and pathways. Because previ-ous software tools usually require chemical formulas to find relevant features, and the data are highly complex, especially in untargeted metabolomics and when the reactions and metabolites downstream the labeled substrates are poorly characterized. We report here Khipu version 2 and its new user-friendly web application. New functions are added to enhance analyzing stable isotope tracing data including metrics that evaluate peak enrichment in labeled samples, scoring methods to facilitate robust detection of intensity patterns and integrated natural abundance correction. We demonstrate that this approach can be applied to untargeted metabolomics to systematically extract isotope-labeled compounds and annotate the unidentified me-tabolites.
RMC-113 (dual-lipid kinase inhibitor, PIKfyve, PIP4K2C) alters the phosphoinositide regioisomer signature by advanced lipidomics analysis
STUDY_TYPE
lipid stduy of phosphoinostide lipids
STUDY_SUMMARY
To determine whether RMC-113 treatment impacts phosphoinositide abundance, lipid extracts derived from uninfected and SARS-CoV-2-infected A549-ACE2 cells were subject to lipidomic analysis. Employing Phosphoinositide Regioisomer Measurement by Chiral column chromatography and Mass Spectrometry (PRMC-MS), we comprehensively profiled all eight PI classes and their acyl chain variants (defined by the carbon number and the saturation level). Seven of the eight phosphoinositide classes (except for PI(3,4)P2) were detected in all tested conditions. Upon SARS-CoV-2 infection, the abundance of multiple PI classes was increased relative to uninfected samples, albeit with some variability across independent experiment. This increase was most pronounced with the abundant acyl chain (38:4), yet a similar trend was observed with other acyl chains. Notably, RMC-113 treatment in infected cells caused a 1.5-2-fold increase in the abundance of PI3P and PI5Pāthe substrates of PIKfyve and PIP4K2C, respectively6, 36ārelative to DMSO controls. No concomitant reduction in the levels of the respective phosphorylated products, PI(3,5)P2 and PI(4,5)P2, was detected in RMC-113- vs. DMSO-treated infected cells, likely due to intact activity of enzymes not targeted by RMC-113, such as PIP4K (1A/1B/1C). Nevertheless, the product-to-substrate ratios of PIKfyve and PIP4K2C were reduced in both uninfected and infected cells upon RMC-113 treatment relative to DMSO
INSTITUTE
Stanford University
DEPARTMENT
medicine
LABORATORY
Einav lab
LAST_NAME
Mishra
FIRST_NAME
Manjari
ADDRESS
300 pasteur drive, stanford
EMAIL
manjari1@stanford.edu
PHONE
6503849709
NUM_GROUPS
4 (uninfected and SARS-CoV-2 infected samples and RMC-113-treated and untreated samples)
Disruption of Glucose Homeostasis by Bacterial Infection Orchestrates Host Innate Immunity Through NAD+/NADH Balance
STUDY_SUMMARY
Metabolic reprogramming is crucial for activating innate immunity in macrophages, and the accumulation of immunometabolites is thought essential for effective defense against infection. The NAD+/NADH redox couple serves as a critical node that integrates metabolic pathways and signaling events, but how this metabolite couple engages macrophage activation remains unclear. Here, we showed that the NAD+/NADH ratio serves as a molecular signal that regulates proinflammatory responses and type I interferon (IFN) responses divergently. Salmonella Typhimurium infection led to a decreased NAD+/NADH ratio by inducing the accumulation of NADH. Further investigation showed that an increased NAD+/NADH ratio correlates to attenuated proinflammatory responses and enhanced type I IFN responses. Conversely, a decreased NAD+/NADH ratio is linked to intensified proinflammatory responses and restrained type I IFN responses. These results showed the NAD+/NADH ratio is an essential cell-intrinsic factor that orchestrates innate immunity, which enhanced our understanding of how metabolites fine-tune innate immunity.
In-depth profiling of biosignatures for Type 2 diabetes mellitus cohort utilizing an integrated targeted LC-MS platform
STUDY_TYPE
Observational study
STUDY_SUMMARY
A cohort of 200 healthy individuals and 100 newly diagnosed Type 2 diabetes mellitus patients from the northern region of China were conducted to profiling the metabolic abnormalities in the condition of diabetes.Then, the data by methods M1-M6 was merged and uploaded as a whole file. The overall differential analysis results were summarized, indicating an obvious metabolic disturbance in amino acid, fatty acids, lysophosphatidyl choline (LysoPC), and triacylglycerol (TAG) under T2DM.
INSTITUTE
The First Affiliated Hospital of Dalian Medical University
LABORATORY
Laboratory of Integrative Medicine
LAST_NAME
Ma
FIRST_NAME
Shurong
ADDRESS
No.222 Zhongshan Road, Xigang District, Dalian City, Liaoning Province
Impact of high-fat diet enriched in cis or trans fatty acids and myriocin on plasma sphingolipids in Ldlr-/- mice
STUDY_SUMMARY
We analyzed sphingolipids in the plasma of Ldlr-/- mice fed 1) Cis HFD, 2) Cis HFD + Myriocin, 3) Trans HFD, or 4) Trans HFD + Myriocin. We aimed to determine how dietary cis and trans monounsaturated fatty acids impact circulating sphingolipids while pharmacologically inhibiting the initial rate-limiting enzyme of sphingolipid biosynthesis, serine palmitoyltransferase (SPT), via myriocin.
Impact of high-fat diet enriched in cis or trans fatty acids and myriocin on hepatic sphingolipids in Ldlr-/- mice
STUDY_SUMMARY
We analyzed sphingolipids in the liver of Ldlr-/- mice fed 1) Cis HFD, 2) Cis HFD + Myriocin, 3) Trans HFD, or 4) Trans HFD + Myriocin. We aimed to determine how dietary cis and trans monounsaturated fatty acids impact hepatic sphingolipids while pharmacologically inhibiting the initial rate-limiting enzyme of sphingolipid biosynthesis, serine palmitoyltransferase (SPT), via myriocin.
Fatty acid oxygenated metabolites analysis of colon in Pla2g10 knockout mice.
STUDY_SUMMARY
We analyzed fatty acid oxygenated metabolites in the colons of WT and Pla2g10 knockout mice that were fed either low-fat diet (LFD) or high-fat diet (HFD).
Short chain fatty acids analysis of Feces and Plasma in Pla2g10 knockout mice.
STUDY_SUMMARY
We analyzed short-chain fatty acids, including acetic acid, propionic acid, and butyric acid, in the feces and plasma of WT and Pla2g10 knockout mice that were fed either a low-fat diet (LFD), high-fat diet (HFD), lard diet, or omega-3 diet.
Glutamine Tracing in Mice injected with HKJS001 and Malonic Acid
STUDY_SUMMARY
Mice were injected by IP with 13C515N2-glutamine solution. Each mice was injected with 15uL of 50mg/mL glutamine solution per gram of mouse. Mice were injected with DMSO as a control, HKJS001 compound, 1M malonic acid, or a combination of HKJS001 and Malonic Acid. The mice were sacrificed and the liver was harvested. Polar metabolites extraction performed on the liver tissue.
INSTITUTE
UMass Chan Medical School
LAST_NAME
UMass Chan
FIRST_NAME
Spinelli Lab
ADDRESS
55 Lake Avenue North, Worcester, Massachusetts, 01605, USA
Specific activation of the integrated stress response (ISR) uncovers regulation of lipid droplet biogenesis
STUDY_TYPE
Biology
STUDY_SUMMARY
U2OS cells were treated with Dimerizer-PERK for 0h,1h,2h,4h,8h and 24h. Lipidomics analysis using LC-MS was performed on these samples to understand the regulation of cellular lipidome upon ISR activation
Chemical Biology Meets Metabolomics: The Response of Barley Seedlings to 3,5-Dichloroanthranilic Acid, a Resistance Inducer
STUDY_TYPE
Plant metabolomics
STUDY_SUMMARY
Advances in combinatorial synthesis and high-throughput screening methods have led to renewed interest in synthetic plant immune activators as well as priming agents. 3,5-Dichloroanthranilic acid (3,5-DCAA) is a derivative of anthranilic acid that have shown potency in activating defence mechanisms in Arabidopsis and barley plants. Chemical biology which is the interface of chemistry and biology can make use of metabolomics approaches and tools to better understand molecular mechanisms operating in complex biological systems. Aim: Here we report on the untargeted metabolomics profiling of barley seedlings treated with 3,5-DCAA to gain deeper insights into the mechanism of action of this resistance inducer. Methodology: Hydro-methanolic extracts from different time periods (12, 24 and 36 h post-treatment) were analysed on ultra-high performance liquid chromatography hyphenated with a high-resolution mass spectrometer. Both unsupervised and supervised chemometric methods were used to reveal hidden patterns and highlight metabolite variables associated with the treatment. Results: Based on the metabolites identified, both the phenylpropanoid and octadecanoid pathways appear to be main route activated by 3,5-DCAA. Different classes of responsive metabolites were annotated with favonoids, more especially flavones, the most dominant. Given the limited understanding of this inducer, this study offers a metabolomics analysis of the response triggered by its foliar application in barley. This additional insight could help make informed decision for the development of more effective strategies for crop protection and improvement, ultimately contributing to agricultural sustainability and resilience.
A Study on the Mechanism of Perinatal BPS Exposure Promoting Obesity Based on Metabolomics and Microbiomics
STUDY_SUMMARY
Due to increasingly stringent regulations on bisphenol A (BPA) usage, its structurally similar counterpart, BPS, has become widely employed as the primary substitute in various industries such as food packaging. However, recent years have unveiled potential risks of obesity promotion and insulin resistance associated with BPS, particularly during early life stages, although its precise impact remains inadequately understood. Addressing these concerns, this study established a mouse model to investigate the effects of maternal BPS exposure during pregnancy and lactation, combined with offspring consumption of a high-fat diet. The research examined physiological indicators related to obesity and insulin resistance in offspring, evaluated pathological changes in vital organs including the heart, liver, pancreas, white adipose tissue (WAT), and brown adipose tissue (BAT), conducted metabolomics perturbation analysis across multiple organs, and performed microbiome analysis based on fecal samples. Finally, correlations between phenotype, metabolome, and microbiome were explored to unravel intergenerational effects and mechanisms under BPS exposure, aiming to identify potential biomarkers of its effects.
INSTITUTE
College of Marine Food and Biological Engineering, Jimei University
The double-edged role of FASII regulator FabT in Streptococcus pyogenes infection - Lipidomics
STUDY_SUMMARY
A FabT mutation impacts the membrane fatty acid (FA) composition. Lipids are composed of FA. The effects of the FabT mutation on membrane FA composition should also alter phospholipid metabolism. WT and mFabT strains were grown as 200 ml cultures in THY or THY-Tween until OD600 = 0.4 - 0.5. Lipid extractions and identifications were performed as described 1-4. Lipid separation was realized by normal phase HPLC (U3000 ThermoFisher Scientific) using a Inertsil Si 5Ī¼m column (150 x 2.1 mm I.D.) from GL Sciences Inc (Tokyo, Japan). Lipids were quantified using a Corona-CAD Ultra and identified by mass-spectrometry negative ionization and MS2/MS3 fragmentations (LTQ-Orbitrap Velos Pro). The adducts observed for molecular ions were: CH3COO- for MGDG, CH3COO- and H- for DGDG, H- for PG and CL. All raw data from MS/MS fragmentations can now be accessed in MZML open source format. This analysis was repeated on 3 independent experiments. 1-Kenanian, G. et al. Permissive Fatty Acid Incorporation Promotes Staphylococcal Adaptation to FASII Antibiotics in Host Environments. Cell Rep 29, 3974-3982 e3974, doi:10.1016/j.celrep.2019.11.071 (2019). 2-Abreu, S., Solgadi, A. & Chaminade, P. Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques. J Chromatogr A 1514, 54-71, doi:10.1016/j.chroma.2017.07.063 (2017). 3-Bligh, E. G. & Dyer, W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37, 911-917, doi:10.1139/o59-099 (1959). 4-Thedieck, K. et al. The MprF protein is required for lysinylation of phospholipids in listerial membranes and confers resistance to cationic antimicrobial peptides (CAMPs) on Listeria monocytogenes. Mol Microbiol 62, 1325-1339, doi:10.1111/j.1365-2958.2006.05452.x (2006).
The double-edged role of FASII regulator FabT in Streptococcus pyogenes infection - Metabolomics
STUDY_SUMMARY
In Streptococcus pyogenes, the type II fatty acid (FA) synthesis pathway FASII is feedback-controlled by the FabT repressor. A FabT mutation leads to bacterial growth defect in the presence of eukaryotic cells and their supernatant. We investigated a possible metabolic basis for the mFabT growth defect, which could reflect an incapacity to use cell-secreted products for growth. We used a metabolomics approach to assess metabolites that are differentially consumed by mFabT compared to the WT strain, cultivated in the presence of non-infected endometrial cells supernatants (HEC-1-A cells). HEC-1-A supernatants were inoculated or not with WT or mFabT strains during 8 or 16 h. The metabolite composition of these supernatants was analyzed by Proteigene (https://proteigene.com) using MxPĀ® Quant 500 kit (Biocrates) by two analytical methods, LC-MS/MS for small molecules and FIA-MS/MS for lipids. This analysis was repeated on 3 independent series of supernatants. These analyses have a defined detection threshold (LOD) for each family of metabolite. You can found in attachment the raw folders and sample informations.
143B cells that were cloned to have the SDHB(succinate dehydrogenase complex iron sulfur subunit B) KO and expression of the RquA enzyme were used to check for mitochondrial function of DHODH(dihydroorotate dehydrogenase). Cells were in culture for 4 days and then extracted for polar metabolites. Cells were treated with DMSO and brequinar. WT and RquA 143B cells were also used for comparison.
INSTITUTE
UMass Chan Medical School
LAST_NAME
UMass Chan
FIRST_NAME
Spinelli Lab
ADDRESS
55 Lake Avenue North, Worcester, Massachusetts, 01605, USA
Specific activation of the integrated stress response uncovers regulation of central carbon metabolism and lipid droplet biogenesis
STUDY_SUMMARY
The integrated stress response (ISR) enables cells to cope with a variety of insults, but its specific contribution to downstream cellular outputs remains unclear. Using a synthetic tool, we selectively activate the ISR without co-activation of parallel pathways and define the resulting cellular state with multi-omics profiling. We identify time- and dose-dependent gene expression modules, with ATF4 driving only a small but sensitive subgroup that includes amino acid metabolic enzymes. This ATF4 response affects cellular bioenergetics, rerouting carbon utilization towards amino acid production and away from the tricarboxylic acid cycle and fatty acid synthesis. We also find an ATF4-independent reorganization of the lipidome that promotes DGAT-dependent triglyceride synthesis and accumulation of lipid droplets. While DGAT1 is the main driver of lipid droplet biogenesis, DGAT2 plays an essential role in buffering stress and maintaining cell survival. Together, we demonstrate the sufficiency of the ISR in promoting a previously unappreciated metabolic state.
Deficiency in glutathione peroxidase 4 (GPX4) results in abnormal lens development and newborn cataract
STUDY_TYPE
lipidomics
STUDY_SUMMARY
This study aims to elucidate the role of GPX4 in lens plasma membrane stability during lens development using in vitro, ex vivo, and in vivo systems. We use lipidomics to analyze the profile of phospholipids and oxidized phospholipids in wild-type and GPX4 KO mice lenses at E18.5. E18.5 lenses were randomly pooled into 4 groups (10mg weight wet/sample) and were subjected to lipid extraction and mass spectrometry analysis. Following data collection, the analysis was conducted using LipidView and the LIPID MAPSĀ® Structure Database (LMSD) software. A notable decrease in the levels of numerous phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidic acids (PA), and phosphatidylglycerol (PG) species was observed in Gpx4 KO lenses compared to WT. For example, PE 34:1 exhibited a significant decrease (p=0.0479) in Gpx4 KO lenses compared to WT, while PE 36:1 and PE 33:2 displayed a trend of lower levels in Gpx4 KO lenses relative to WT, albeit not statistically significant. Conversely, elevated levels of oxidized phospholipids, particularly oxidized PE, were detected in Gpx4 KO lenses compared to WT. Notably, significant increases were observed in phospholipid oxidation products, including PE(16:0/18:2(O))Na (p=0.0259) and PE(16:0/18:2(OH)(OOH)) (p=0.0401), with a trending increase in PE(16:0/12:1(COOH)(9OH)Na (p=0.0635) in Gpx4 KO lenses relative to WT. These findings strongly indicate that GPX4 deficiency induces lipid peroxidation, consequently altering the composition of plasma membrane lipids.
Impact of giredestrant on the lipid profile of MCF-7 breast cancer cells
STUDY_SUMMARY
Based on some recent data from a giredestrant resistance/sensitivity screen, we have an evolving hypothesis that exposure to giredestrant might be altering levels of polyunsaturated fatty acids, and perhaps otherwise altering lipid homeostasis in cells. The sensitivity to ferroptosis results from peroxidation of PUFAs-PLs. We thus investigated whether giredestrant altered PL homeostasis with a time course analysis of MCF-7 cell lipid profile. MCF-7 cells were treated with 0.6 nM of giredestrant and DMSO respectively on Day 0 and collected on Day 2, Day 7 and Day 14 for lipid profile analysis. Giredestrant appeared to provoke, in MCF-7 cells, a broad remodelling of lipids towards PUFA-containing species, specifically after longer duration (7 and 14 day) treatment. We observed that giredestrant increases PUFA-containing phospholipids at the expense of MUFA-phospholipids.
Lipidomics Analysis of ER+ Breast Cancer Cells Treated with Giredestrant and Palbociclib
STUDY_SUMMARY
We observed in a previous experiment that giredestrant treatment had a profound impact on the lipid profile of MCF-7 cells, in particular through elevating PUFAs at the apparent expense of MUFAs. We hypothesize that this shift in PUFA/MUFA ratio underlies giredestrant-induced sensitivity to GPX4 inhibition. Emerging data from our lab and other suggests that palbociclib, a key combination partner for giredestrant in the clinic, also sensitizes to GPX4 inhibition, and that the combination of giredestrant and palbo may synergize to creating an even higher sensitivity to GPX4i. Here, we aim to directly compare the effects on the lipidome of giredestrant and palbociclib, in MCF-7 cells (the discovery cell line where we already have some lipidomics data), and also T-47D cells, in which giredestrant, palbociclib and their combination drive a very profound sensitization to GPX4i, that exceeds what was observed in MCF-7 cells. The cells were treated with DMSO, 1 nM giredestrant, 200 nM palbociclib, or 1nM giredestrant plus 200nM palbociclib in quadruplicates, and collected on Day 7 for lipid analysis. Despite their differences under basal conditions, drug treatments altered the lipid profiles of MCF7 and T47D cells in a similar manner, with 200nM palbociclib at the 7 day time point having a less pronounced effect than giredestrant, and with the greatest lipid changes occurring with the combination of palbociclib plus giredestrant. PUFA-ePLs were likewise elevated by giredestrant and/or palbociclib in both cell lines, with the combined action of both drugs generally driving greatest increases in individual PUFA-linked ePL species. Palbociclib combination with giredestrant enhance the accumulation of PUFAs-phospholipids compared to the single treatments.
LampreyDB: a spatial metabolomics dataset for lampreys
STUDY_SUMMARY
Lampreys are blood-sucking vampires in marine environments. However, the lamprey-specific metabolomics database is still missing. As such, we have established LampreyDB (https://www.lampreydb.com), a tissue-wide spatial lamprey metabolomics database that contains all the identified and annotated metabolites from our experiment. LampreyDB allows users to explore lamprey-specific metabolites with text-based searches, i.e., chemical formula, m/z value, or a list of MS/MS fragments.
The āmetabolic fingerprintā of cytotoxic gold drugs in cancer cells disclosed by NMR
STUDY_SUMMARY
NMR metabolomics is a powerful tool for characterizing changes in cancer cell metabolism induced by drug treatment. Here, this approach is used to elucidate the effects of five cytotoxic gold compounds in A2780 human ovarian cancer cells. Namely, two close analogues of auranofin (AF), AFCl and AFI, a gold(I) triphenylphosphine complex, (Ph3P)AuI, and two representative gold(III) compounds, AuL12 and Aubipyc, were investigated. Interestingly, the three gold(I) compounds were found to induce similar and pronounced metabolic changes in the lysates and growth media, whereas the two gold(III) compounds induced only minor changes. The results are analysed and discussed in the context of the existing knowledge on gold-based anticancer drugs, their modes of action and their effects on cellular metabolism. To this end, we included in our dataset four additional gold-based drugs, already individually investigated in previous studies. Statistical tools are used to highlight similarities and differences between the various compounds. Attempts are made to establish well-defined structure-function relationships within the set of tested gold compounds
Relative concentrations of acylcarnitines in BT-474 cells treated with FASN inhibitors TVB-2640 and TVB-3166
STUDY_TYPE
Intracellular metabolomics, medium metabolomics
STUDY_SUMMARY
Relative concentrations of acylcarnitines in cell extracts of BT-474 cells treated with TVB-2640 and TVB-3166 for 24 h assessed via triple-quadrupole precursor ion profiling of a fragment with m/z 85.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Analysis of lipid composition of TLCD1 KO cultured cells
STUDY_SUMMARY
Untargeted lipidomics experiments were conducted to analyze the differences in lipid composition between TLCD1 KO and isogenic control HeLa and U2OS cell lines. See a reference publication (Sheokand et al) for details.
Analysis of lipid composition of the mitochondria isolated from TLCD1 KO cell models
STUDY_SUMMARY
Untargeted lipidomics analysis was performed on whole cells and isolated mitochondria from control and TLCD1 knockout (KO) cell lines, both of which feature genetically tagged mitochondria.
INSTITUTE
University of Cambridge
LAST_NAME
Petkevicius
FIRST_NAME
Kasparas
ADDRESS
The Keith Peters Building, Cambridge, Cambridgeshire, CB2 0XY, United Kingdom
Analysis of lipid composition of control, YPR114w and YJR116w yeast mutants grown under exponential phase
STUDY_SUMMARY
Untargeted lipidomics experiments were conducted to analyze the differences in lipid composition between control, YPR114w and YJR116w yeast (Saccharomyces cerevisiae) mutants . See a reference publication (Sheokand et al) for details.
INSTITUTE
University of Cambridge
LAST_NAME
Petkevicius
FIRST_NAME
Kasparas
ADDRESS
The Keith Peters Building
EMAIL
kp416@mrc-mbu.cam.ac.uk
PHONE
07500233355
PUBLICATIONS
TRAMāLAG1āCLN8 family proteins are acyltransferases regulating phospholipid composition
BPA regulates the abundance of endogenous PPARa ligands and fatty acid oxidation
STUDY_SUMMARY
Given the regulation of PPARa(Peroxisome proliferator-activated receptor alpha)-dependent gene expression in the absence of ACLY, we postulated that endogenous PPARa ligand availability might be altered. Prior work has shown that phosphatidylcholines (PCs) can serve as PPARa ligands, and specifically, PC 16:0/18:1 has been established as an endogenous ligand. PC 16:0/18:1 abundance has also been found to be regulated in a circadian manner dependent on the fatty acid synthesis pathway. Since PCs as a class are suppressed in the liver ACLY KO mice, we asked if PC 16:0/18:1 is specifically reduced, finding that it its abundance is lower in the ACLY KO.
INSTITUTE
Salk Institute for Biological Studies
LAST_NAME
Kuna
FIRST_NAME
Ramya
ADDRESS
10010 N Torrey Pines Rd, La Jolla, California, 92037, USA
Impact of ACLY and ACSS2 on the development of MASLD on longterm Western diet
STUDY_SUMMARY
To further assess the impact of ACLY and ACSS2 on the development of MASLD on Western diet, we also carried out longer term experiments, in which mice remained on diet for 22 weeks. Prior work has shown that phosphatidylcholines (PCs) can serve as PPARa ligands, and specifically, PC 16:0/18:1 has been established as an endogenous ligand. PC 16:0/18:1 abundance has also been found to be regulated in a circadian manner dependent on the fatty acid synthesis pathway. Since PCs as a class are suppressed in the liver ACLY KO mice, we asked if PC 16:0/18:1 is specifically reduced, finding that it its abundance is lower in the ACLY and ACLY/ACSS2 KOs.
INSTITUTE
Salk Institute for Biological Studies
LAST_NAME
Kuna
FIRST_NAME
Ramya
ADDRESS
10010 N Torrey Pines Rd, La Jolla, California, 92037, USA
Impact of ACLY and ACSS2 on the development of MASLD on longterm Western diet,
STUDY_SUMMARY
To further assess the impact of ACLY and ACSS2 on the development of MASLD on Western diet, we also carried out longer term experiments, in which mice remained on diet for 22 weeks. Exacerbated steatosis was apparent in the liver ACLY KO and DKO mice, both histologically and by TAG quantification. Further lipid analysis by LC-MS revealed elevated DAGs and cholesterol esters (CEs) in ACLY-deficient and DKO livers, along with reduced abundance of phosphatidylcholines (PCs). Together the data indicate that ACLY deficiency results in perturbed lipid metabolism in the liver, with accumulation of TAGs, DAGs, and CEs.
INSTITUTE
Salk Institute for Biological Studies
LAST_NAME
KUNA
FIRST_NAME
RAMYA
ADDRESS
10010 N Torrey Pines Rd, La Jolla, California, 92037, USA
Analysis of lipid composition of CLN8 KO cultured cells
STUDY_SUMMARY
Untargeted lipidomics experiments were conducted to analyze the differences in lipid composition between CLN8 KO and isogenic control HeLa and U2OS cell lines. See a reference publication (Sheokand et al) for details.
INSTITUTE
University of Cambridge
LAST_NAME
Petkevicius
FIRST_NAME
Kasparas
ADDRESS
The Keith Peters Building
EMAIL
kp416@mrc-mbu.cam.ac.uk
PHONE
07500233355
PUBLICATIONS
TRAMāLAG1āCLN8 family proteins are acyltransferases regulating phospholipid composition
Targeted free fatty acids metabolomics studies on serum, cecal content, cecum tissue, chow diet, and T.mu cells.
STUDY_SUMMARY
Serum, cecal content, cecum tissue samples of murine, chow diet, and T.mu cells were collected to perform the targeted free fatty acids metabolome analysis. The aim of this study was to verify that whether T.mu could release free PUFA (especially ARA) to the intestinal tract of its host by comparing the PUFA concentration in the freshly isolated T.mu cells, chow diet, cecal content and serum of the host (Mus musculus).Based on the result of this targeted free fatty acids metabolomics studies, we found that T.mu could release ARA to the intestinal tract of its host and increase the concentration of ARA in its host's intestinal tract.
Intracellular and medium metabolomics of BT-474 cells treated with Fasnall that was manufactured by Enamine
STUDY_SUMMARY
BT-474 human breast cancer cells were treated with various concentrations of Fasnall for 24 h. The drug was manufactured by Enamine (1-benzyl-N-{5,6-dimethylthieno[2,3-d]pyrimidin-4-yl}pyrrolidin-3-amine, EN300-14327544).
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Metabolomics analysis of breast cancer cell lines treated with dimethylmalonate (DMM), GSK2194069, and their combination.
STUDY_SUMMARY
Seven breast cancer cell lines (MDA-MB-468, SUM159PT, MCF7, HCC1806, MDA-MB-231, HS578T, CAL120) were treated with 5 mM DMM, 1 uM GSK2194069, and their combination, for 24 h. Intracellular samples were collected for the metabolomics analysis.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Plasma concentrations of Fasnall in mice after a bolus of 10 mg/kg administered intraperitoneally.
STUDY_TYPE
Plasma metabolomics
STUDY_SUMMARY
A single dose of 10 mg/kg (drug/body weight) Fasnall was injected intraperitoneally (~50 ul of solution in 1:1 DMSO:PBS), and plasma samples were collected. Blood was collected through a cardiac puncture. EDTA was used as an anticoagulant.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Metabolomics analysis of zebrafish embryos treated with rotenone, Fasnall, TVB-2640, and GSK2194069
STUDY_SUMMARY
To compare the metabolic effects and toxicity of Fasnall with rotenone in vivo, zebrafish embryos 48 h post-fertilization were exposed to a drug-containing medium for 6 h. Rotenone at 25 nM is lethal to fish embryos, while 5 nM concentration leads to a ~15-fold lactate accumulation. Similarly, Fasnall treatment increases lactate content, although the magnitude of the effect is significantly lower. Unlike 5 nM rotenone, Fasnall treatment does not cause visible phenotypic changes in the yolk. The zebrafish model suggests that Fasnall acts as a Complex I inhibitor in vivo.
INSTITUTE
Wistar Institute
DEPARTMENT
Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
White adipose tissue remodeling in Little Brown Myotis (Myotis lucifugus) with white-nose syndrome
STUDY_SUMMARY
White-nose syndrome (WNS) is a fungal wildlife disease of bats that has caused precipitous declines in certain Nearctic bat species. A key driver of mortality is premature exhaustion of fat reserves, primarily white adipose tissue (WAT), that bats rely on to meet their metabolic needs during winter. However, the pathophysiological and metabolic effects of WNS have remained ill-defined. To elucidate metabolic mechanisms associated with WNS mortality, we infected a WNS susceptible species, the Little Brown Myotis (Myotis lucifugus), with Pseudogymnoascus destructans (Pd) and collected WAT biopsies for histology and targeted lipidomics. These results were compared to the WNS-resistant Big Brown Bat (Eptesicus fuscus). A similar distribution in broad lipid class was observed in both species, with total WAT primarily consisting of triacylglycerides. Baseline differences in WAT chemical composition between species showed that higher glycerophospholipids (GPs) levels in E. fuscus were dominated by unsaturated or monounsaturated moieties and n-6 (18:2, 20:2, 20:3, 20:4) fatty acids. Conversely, higher GP levels in M. lucifugus WAT were primarily compounds containing n-3 (20:5 and 22:5) fatty acids. Following Pd-infection, we found that perturbation to WAT reserves occurs in M. lucifugus, but not in the resistant E. fuscus. A total of 66 GPs (primarily glycerophosphocholines and glycerophosphoethanolamines) were higher in Pd-infected M. lucifugus, indicating perturbation to the WAT structural component. In addition to changes in lipid chemistry, smaller adipocyte sizes and increased extracellular matrix deposition was observed in Pd-infected M. lucifugus. This is the first study to describe WAT GP composition of bats with different susceptibilities to WNS and highlights that recovery from WNS may require repair from adipose remodeling in addition to replenishing depot fat during spring emergence.
INSTITUTE
Georgetown University
LAST_NAME
Pannkuk
FIRST_NAME
Evan
ADDRESS
3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Unbiased genetic screening and metabolomics identifies glial adenosine metabolism as a therapeutic target in Parkinsonās disease
STUDY_SUMMARY
Parkinsonās disease (PD) is the second most common neurodegenerative disorder and lacks disease-modifying therapies. We developed a Drosophila model for identifying novel glial- based therapeutic targets for PD. Human Ī±-synuclein is expressed in neurons and individual genes are independently knocked down in glia. We performed a large forward genetic screen, knocking down the entire Drosophila kinome in glia in Ī±-synuclein expressing flies. Among the top hits were five genes (Ak1, Ak6, Adk1, Adk2, and awd) involved in adenosine metabolism. Knockdown of each gene improved locomotor dysfunction, rescued neurodegeneration, and increased brain adenosine levels. We determined that the mechanism of neuroprotection involves adenosine itself, as opposed to a downstream metabolite. We dove deeper into the mechanism for one gene, Ak1, finding rescue of dopaminergic neuron loss, Ī±-synuclein aggregation, and bioenergetic dysfunction after glial Ak1 knockdown. We performed metabolomics in Drosophila and in human PD patients, allowing us to comprehensively characterize changes in purine metabolism and identify potential biomarkers of dysfunctional adenosine metabolism in people. These experiments support glial adenosine as a novel therapeutic target in PD.
Non-targeted metabolomic analysis of Enterobacter hormaechei_B17 (Eh_B17),Enterobacter sichuanensis_B36(Es_B36), and Enterobacter hormaechei_B56 (Eh_B56)
STUDY_SUMMARY
All samples were acquired by the LC-MS system according to the machine instructions. Firstly, all chromatographic separations were performed using an UPLC system (SCIEX). An XBridge BEH C18 column (3.5um,2.1mm Ć 100mm, Waters) was used for the reversed-phase separation. The column oven was maintained at 50Ā°C. The flow rate was 0.3 mL/min and the mobile phase consisted of solvent A (water containing 0.1% formic acid) and solvent B (acetonitrile containing 0.1% formic acid). The gradient conditions were set as follows: 0-0.5 min, 5% phase B; 0.5-5 min, 5% to 80% phase B; 5-7 min, 80%-100% phase B; 7-8 min,100% phase B; 8-8.1 min, 100%-5% phase B; 8.1-10 min, 5% phase B. The injection volume for each sample was 10 Ī¼L.A high-resolution tandem mass spectrometer TripleTOF5600 (SCIEX) was used to detect metabolites eluted from the column. The Q-TOF was operated in both positive ion and negative ion modes. For positive ion mode, the capillary voltages were set to 5 kV. For negative ion mode, the capillary voltages were set to -4.5 kV. Mass spectrometry data were acquired in IDA mode. The TOF mass range was 50 to 1200 Da. The top 8 precursors were fragmented for MS/MS detection. In addition, a quality control sample (pool of all samples) was acquired after every 10 samples to evaluate the stability of the LC-MS throughout the acquisition.
INSTITUTE
Institute of Zoology, Chinese Academy of Sciences
LAST_NAME
Zou
FIRST_NAME
Zhen
ADDRESS
1 Beichen West Road, Chaoyang District, Beijing, Beijing, 100101, China
Metabolomic analysis of plasma samples from mice fed low-folate or folate-replete diets
STUDY_SUMMARY
Metabolomic analysis was performed on samples from two cohorts of mice fed diets with different levels of folate (Vitamin B9). Mice were divided into four distinct groups, delineated by sex and diet. Blood was collected via cheek bleed and processed at weekly time points up to 6 weeks. Overall, there were 7 male and 7 female mice, with 3-4 mice in each diet cohort. Plasma was stored in the -80 until analysis. At the end of the experiment, the samples were thawed and pooled based on the group (male/female, low/replete folate diet) and week.
GNB1 and SCARB2 genes were knocked down in human subcutaneous adipocytes using RNA interference. Both proteomic and lipidomic analyses were performed using mass spectrometry. The proteomic results revealed significant changes in the expression levels of key proteins involved in lipid metabolism and adipogenesis. The lipidomic analysis identified significant changes in lipid species, including phosphatidylcholines, ceramides, and cholesterol esters. This integrated analysis provides insights into the molecular pathways and lipid metabolism regulated by GNB1 and SCARB2 in adipocyte function.
INSTITUTE
Hamamatsu University School of Medicine
LAST_NAME
Kitamoto
FIRST_NAME
Takuya
ADDRESS
1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Japan
Integration of Metabolomics and Transcriptomics Reveals Metabolic Characteristics and Potential Biomarker Involved in Radiation-Induced Liver Injury in a Rat Model.
STUDY_SUMMARY
Radiation-induced liver damage (RILD) is a disease characterized by a series of physiological and pathological changes in liver tissue following exposure to a certain dose of radiation, and it is also a common complication of liver cancer and abdominal tumor radiotherapy. To date, the pathogenesis of RILD remains unclear, and effective diagnostic and therapeutic approaches are lacking. Based on this, the present study established an animal model of radiation-induced liver disease (RILD) using whole-liver irradiated rats. Metabolomics and transcriptomics were integrated to analyze liver tissue samples collected 7 days post-irradiation. The involved metabolic disorders primarily include ammonia metabolism, amino acid metabolism, glutathione metabolism and lipid metabolism. Moreover, a panel of potential plasma metabolic markers was identified through correlation analysis between liver tissue and plasma metabolic characteristics. Subsequently, the levels of radiation injury within 7 days post irradiation were assessed. This study provides experimental evidence for the identification of early diagnostic markers for whole-liver irradiation and RILD, as well as for exploring the molecular and pathophysiological mechanisms of RILD.
INSTITUTE
Soochow University
LAST_NAME
Wang
FIRST_NAME
Chang
ADDRESS
Suzhou, No. 199, Renai Road, Suzhou Industrial Park
Gut Microbiota Regulates Stress Responsivity via the Circadian System - Mice Amygdala
STUDY_SUMMARY
Stress and circadian systems are interconnected through the hypothalamic-pituitary-adrenal (HPA) axis to maintain appropriate responses to external stimuli. Yet, the mechanisms of how such signals are orchestrated remains unknown. We now uncover the gut microbiota as a regulator of HPA-axis rhythmic function. Additionally, microbiota depletion markedly disturbs the brain transcriptome and metabolome in key stress response pathways in the hippocampus and amygdala across the day. This is coupled with a dysregulation of the master circadian pacemaker in the brain that resulted in perturbed glucocorticoid rhythmicity. The resulting hyper-activation of the HPA-axis at the sleep/wake transition drives time-of-day-specific impairments of the stress response and altered stress-sensitive behaviors. Finally, using microbiota transplantation we confirm that diurnal oscillations of gut microbes underlie altered glucocorticoid secretion and identify L. reuteri as a candidate strain for such effects. Together, our data offers compelling evidence that the microbiota regulates stress responsiveness in a circadian manner and is necessary to respond adaptively to psychological stressors throughout the day.
INSTITUTE
University College Cork
DEPARTMENT
Anatomy & Neuroscience
LABORATORY
Microbiome Gut Brain Axis
LAST_NAME
John
FIRST_NAME
Cryan
ADDRESS
4th Floor, Block E, Food Science Building, University College Cork, College Road, Cork, Ireland.
Gut Microbiota Regulates Stress Responsivity via the Circadian System - Mice Hippocampus
STUDY_SUMMARY
Stress and circadian systems are interconnected through the hypothalamic-pituitary-adrenal (HPA) axis to maintain appropriate responses to external stimuli. Yet, the mechanisms of how such signals are orchestrated remains unknown. We now uncover the gut microbiota as a regulator of HPA-axis rhythmic function. Additionally, microbiota depletion markedly disturbs the brain transcriptome and metabolome in key stress response pathways in the hippocampus and amygdala across the day. This is coupled with a dysregulation of the master circadian pacemaker in the brain that resulted in perturbed glucocorticoid rhythmicity. The resulting hyper-activation of the HPA-axis at the sleep/wake transition drives time-of-day-specific impairments of the stress response and altered stress-sensitive behaviors. Finally, using microbiota transplantation we confirm that diurnal oscillations of gut microbes underlie altered glucocorticoid secretion and identify L. reuteri as a candidate strain for such effects. Together, our data offers compelling evidence that the microbiota regulates stress responsiveness in a circadian manner and is necessary to respond adaptively to psychological stressors throughout the day.
INSTITUTE
University College Cork
DEPARTMENT
Anatomy & Neuroscience
LABORATORY
Microbiome Gut Brain Axis
LAST_NAME
John
FIRST_NAME
Cryan
ADDRESS
4th Floor, Block E, Food Science Building, University College Cork, College Road, Cork, Ireland.
An optimized plasmalogen modulating dietary supplement provides greater protection in a male than female mouse model of dilated cardiomyopathy
STUDY_SUMMARY
We previously reported that plasmalogens, a class of phospholipids, were decreased in a setting of dilated cardiomyopathy (DCM). Plasmalogen levels can be modulated via a dietary supplement called alkylglycerols (AG) which has demonstrated benefits in some disease settings. However, its therapeutic potential in DCM remained unknown. To determine whether an optimized AG supplement could restore plasmalogen levels and attenuate cardiac dysfunction/pathology, we placed a cardiac-specific transgenic DCM mouse model of both sexes on chow +/- 1.5% AG supplementation at ~10 weeks of age for 16 weeks. Cardiac function was assessed by echocardiography, tissues were collected for histological and molecular analyses including lipidomics and proteomics via liquid chromatography-mass spectrometry. AG supplementation increased total plasmalogens in DCM hearts and attenuated lung congestion of both sexes, but only prevented cardiac dysfunction in males. This was associated with attenuated cardiac and renal enlargement, a more favorable pro-cardiac gene expression profile, and a trend for lower cardiac fibrosis. By lipidomics, specific d18:1 ceramide species associated with cardiac pathology were lower in the DCM hearts from mice on the AG diet, and tetra-linoleoyl cardiolipin, a lipid crucial for mitochondria function was restored with AG supplementation. Proteomic analysis of hearts from male DCM mice receiving AG supplementation revealed enrichment in mitochondrial protein network, as well as upregulation of extracellular matrix binding proteins including agrin, a protein associated with cardiac regeneration. In summary, AG supplementation restored plasmalogens in DCM hearts, but showed greater therapeutic potential in males than females.
INSTITUTE
Baker Heart and Diabetes Institute
DEPARTMENT
Discovery and Preclinical Science
LABORATORY
Cardiac Hypertrophy
LAST_NAME
Belkin
FIRST_NAME
Teleah
ADDRESS
75 Commercial Road, Melbourne, Victoria, 3004, Australia
1H NMR metabolomics applied to assess the direct and transgenerational effects of simvastatin on the metabolism of the amphipod Gammarus locusta
STUDY_TYPE
1H NMR metabolomics to study the metabolic effects of direct exposure to simvastatin and transgenerational effects on the polar metabolome of amphipod Gammarus locusta
STUDY_SUMMARY
In this study, a comprehensive untargeted 1H NMR metabolomics strategy was applied to measure the metabolic impact of direct and transgenerational exposure (F0 and F3 generations, respectively) of amphipods G. locusta to simvastatin (SIM), a pharmaceutical widely prescribed for the treatment of hypercholesterolemia. To assess the direct and transgenerational effects of exposure to SIM on the metabolism of G. locusta, the following conditions were studied: i) direct exposure of parental generation (F0) of males and females, GM/GF-F0 (Exp); and ii) transgenerational effects, considering the first unexposed generation (F3) of males and females, to assess transgenerational effects, GM/GF-F3 (Tr). The obtained data added important knowledge, unveiling individual metabolic effects of direct exposure to simvastatin and its transgenerational effects, potentially contributing towards improving hazard and risk assessment of biologically active compounds.
INSTITUTE
University of Aveiro
DEPARTMENT
CICECO ā Aveiro Institute of Materials, Department of Chemistry
LABORATORY
Metabolomics Group
LAST_NAME
Rodrigues
FIRST_NAME
JoĆ£o A.
ADDRESS
University of Aveiro, Campus UniversitƔrio de Santiago, 3810-193 Aveiro, Portugal
Impact of Cattle Feeding Systems on Beef and Human Postprandial Metabolomics ā A Randomized Clinical Trial
STUDY_SUMMARY
Cattle feeding systems may have health implications for consumers of beef products. Organic grass-fed (GRA) and conventional (CON) cattle feeding systems may result in beef products with differing metabolite profiles, and therefore could impact the postprandial metabolomic response of consumers. This study aims to measure whole beef metabolomics and postprandial metabolomic response of consumers between GRA and CON beef to elucidate potential health implications. This study followed a double-blind, crossover design with healthy male and female subjects (n=10). Serum samples were taken at fasting (0) and postprandially for four hours after consumption of a steak from each condition. Untargeted metabolomic analysis of whole beef and human serum samples utilized LC/MS. Multivariate and pathway enrichment analysis in MetaboAnalyst was used to investigate metabolite and biochemical pathways that distinguished CON and GRA. Cattle feeding systems impacted both postprandial and whole beef steak metabolomic profiles. Metabolites that contributed to this variation included carnitine species (Proionylcarnitine), fatty acids, amino acids (L-Valine) and Calamendiol. These metabolites have been associated with oxidative stress, inflammation, and cardiovascular health. Functional pathway enrichment analysis revealed numerous amino acid degradation pathways, especially branched chain amino acids, and fatty acid degradation that changed throughout the postprandial time course. These findings suggest that CON and GRA cattle feeding systems differentially impact whole beef metabolomics, as well as consumer postprandial metabolic responses and the associated health implications.
Intracellular metabolome and Intracellular metabolic pathway prediction
STUDY_SUMMARY
The study used HepG2 cells as a model system to assess the individual impacts of endotoxin (LPS) and fatty acids on the intracellular global metabolome. This study elucidates the distinct effects of these two parameters on the overall metabolic balance of hepatocytes.
Gas chromatography - mass spectrometry (GC-MS) of liver hepatic extracts from adult male C57BL/6NCrl mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin
STUDY_SUMMARY
Epidemiological evidence suggests an association between dioxin and dioxin-like compound (DLC) exposure and human liver disease. The prototypical DLC, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), has been shown to induce the progression of reversible hepatic steatosis to steatohepatitis with periportal fibrosis and biliary hyperplasia in C57BL/6NCrl mice. Although the effects of TCDD toxicity are mediated by aryl hydrocarbon receptor (AHR) activation, the underlying mechanisms of TCDD-induced hepatotoxicity are unresolved. In the present study, male C57BL/6NCrl mice were gavaged every 4 days for 28 days with 0.03 - 30 Āµg/kg TCDD and evaluated for liver histopathology and gene expression as well as complementary 1-dimensional (1-D) 1H NMR urinary metabolic profiling. Urinary trimethylamine (TMA), trimethylamine N-oxide (TMAO), and 1-methylnicotinamide (1MN) levels were altered by TCDD at doses ā¤ 3 Āµg/kg; other urinary metabolites, like glycolate, urocanate, and 3-hydroxyisovalerate, were only altered at doses that induced moderate to severe steatohepatitis. Bulk liver RNA-seq data suggested altered urinary metabolites correlated with hepatic differential gene expression corresponding to specific metabolic pathways. In addition to evaluating whether altered urinary metabolites were liver-dependent, published single-nuclear RNA-seq (snRNA-seq), AHR ChIP-seq, and AHR knockout gene expression datasets provided further support of hepatic cell-type and AHR-regulated dependency, respectively. Overall, TCDD-induced liver effects were preceded by and occurred with changes in urinary metabolite levels due to AHR-mediated changes in hepatic gene expression.
INSTITUTE
Michigan State University
DEPARTMENT
Biochemistry and Molecular Biology
LABORATORY
Dr. Tim Zacharewski's
LAST_NAME
Sink
FIRST_NAME
Warren
ADDRESS
603 Wilson Rd Rm 212, East Lansing, MI 48823
EMAIL
sinkwarr@msu.edu
PHONE
6162953496
NUM_GROUPS
5
TOTAL_SUBJECTS
25
NUM_MALES
25
STUDY_COMMENTS
Adult male C57BL/6NCrl mice (PND 30) were gavaged every 4 days for 28 days with 1, 3, 10, or 30 Āµg/kg TCDD or sesame oil vehicle. Mice were euthanized and their liver tissue collected at the conclusion of the TCDD exposure (PND 58).
We assessed technical replicates of NIST Standard Reference Material (SRM 1950) human plasma specimens on different Biocrates MxP Quant 500 kit well-plates. In total, 9 different technical replicates on 9 different Biocrates MxP Quant 500 kit well-plates were measured. The resulting data is used as a showcase to demonstrate the utility of our MetaboSERV platform for selecting, exchanging, and visualizing metabolomics data with controlled data access.
INSTITUTE
Hannover Medical School
DEPARTMENT
Peter L. Reichertz Institute for Medical Informatics
Fatty acid analysis of differently sized MC38 tumors harvested 3 weeks post implantation from control-diet- or high-fat-diet-fed C57BL/6 mice revealed an increase in tumoral oleic acid in high-fat-diet-fed mice.
STUDY_SUMMARY
MC38 tumors were harvested 3 weeks post implantation from control-diet- and high-fat-diet-fed mice. At this point, tumors from high-fat-diet-fed mice were larger than that of control-diet-fed mice. These tumors were subject to fatty acid analysis via GC-FID. The resulting data revealed an increase in oleic acid in tumors from high-fat-diet-fed mice. Oleic acid was the only fatty acid to exhibit this trend.
Fatty acid analysis of similarly sized MC38 tumors from control-diet- or high-fat-diet-fed C57BL/6 mice revealed an increase in tumoral oleic acid in high-fat-diet-fed mice.
STUDY_SUMMARY
Similarly sized MC38 tumors from control-diet- and high-fat-diet-fed mice were subject to fatty acid analysis via GC-FID. These data revealed an increase in oleic acid in tumors from high-fat-diet-fed mice. Oleic acid was the only fatty acid to exhibit this trend.
Fatty acid analysis of human colorectal adenocarcinoma tumors of normal weight (BMI<25) and obese (BMI>30) patients revealed an increase in oleic acid in obese patients.
STUDY_SUMMARY
Frozen human colorectal adenocarcinoma tumors samples were acquired and analyzed for fatty acid composition via GC-FID. These data revealed an increase in several fatty acids, such as palmitic acid, stearic acid, oleic acid, etc., in obese patients. However, oleic acid was the most abundant fatty acid in tumors of obese patients and also the fatty acid that exhibited the most significant increase in obese patients compared to normal-weight patients.
Therapeutic potential of galactooligosachaccharides and Lactobacillus reuteri supplementation on dextran sulfate sodium-induced gut inflammation
STUDY_SUMMARY
Dextran sulfate sodium (DSS)-induced inflammatory bowel disease characterized by intestinal inflammation, barrier dysfunction, and dysbiosis, with limited treatment options available. This study systematically investigates the therapeutic potential of a synbiotic composed of galactooligosaccharides (GOS) and Limosilactobacillus reuteri in a murine model of colitis, revealing that GOS and L. reuteri synergistically protect against intestinal inflammation and barrier dysfunction by promoting the synthesis of pentadecanoic acid, an odd-chain fatty acid, from Bacteroides acidifaciens (B. acidifaciens). Intestinal segments and feces from DSS-treated mice were collected for cytokines detection and metabolomics. Notably, the synbiotic, B. acidifaciens, and pentadecanoic acid are each capable of suppressing intestinal inflammation and enhancing tight junction by inhibiting NF-ĪŗB activation. Furthermore, similar reduction in B. acidifaciens and pentadecanoic acid levels are also observed in the feces from both human ulcerative colitis (UC) patients and lipopolysaccharide-induced intestinal inflammation in pigs. Our findings elucidate the protective mechanism of the synbiotic and highlight its therapeutic potential, along with B. acidifaciens and pentadecanoic acid, for intestinal inflammatory disorders.
Tissue niche influences immune and metabolic profiles to Staphylococcus aureus biofilm infection
STUDY_SUMMARY
Infection is a devastating post-surgical complication, often requiring additional procedures and prolonged antibiotic therapy. This is especially relevant for craniotomy and prosthetic joint infections (PJI), both of which are characterized by biofilm formation on the bone or implant surface, respectively, with S. aureus representing a primary cause. The local tissue microenvironment likely has profound effects on immune attributes that can influence treatment efficacy, which becomes critical to consider when developing therapeutics for biofilm infections. However, the extent to which distinct tissue niches influence immune function during biofilm development remains relatively unknown. To address this, we compare the metabolomic, transcriptomic, and functional attributes of leukocytes in mouse models of S. aureus craniotomy and PJI complemented with patient samples from both infection modalities, which reveals profound tissue niche-dependent differences in nucleic acid, amino acid, and lipid metabolism with links to immune modulation. These signatures are both spatially and temporally distinct, differing not only between infection sites but evolving over time within a single model. Collectively, this demonstrates that biofilms elicit unique immune and metabolic responses that are heavily influenced by the local tissue microenvironment, which will likely have important implications when designing therapeutic approaches targeting these infections.
INSTITUTE
UNMC
DEPARTMENT
Department of pathology, microbiology and immunology
Tissue niche influences immune and metabolic profiles to Staphylococcus aureus biofilm infection (Extracellular data)
STUDY_SUMMARY
Infection is a devastating post-surgical complication, often requiring additional procedures and prolonged antibiotic therapy. This is especially relevant for craniotomy and prosthetic joint infections (PJI), both of which are characterized by biofilm formation on the bone or implant surface, respectively, with S. aureus representing a primary cause. The local tissue microenvironment likely has profound effects on immune attributes that can influence treatment efficacy, which becomes critical to consider when developing therapeutics for biofilm infections. However, the extent to which distinct tissue niches influence immune function during biofilm development remains relatively unknown. To address this, we compare the metabolomic, transcriptomic, and functional attributes of leukocytes in mouse models of S. aureus craniotomy and PJI complemented with patient samples from both infection modalities, which reveals profound tissue niche-dependent differences in nucleic acid, amino acid, and lipid metabolism with links to immune modulation. These signatures are both spatially and temporally distinct, differing not only between infection sites but evolving over time within a single model. Collectively, this demonstrates that biofilms elicit unique immune and metabolic responses that are heavily influenced by the local tissue microenvironment, which will likely have important implications when designing therapeutic approaches targeting these infections. This submission contains the extracellular metabolomic data for the larger project.
INSTITUTE
UNMC
DEPARTMENT
Department of pathology, microbiology and immunology
Metabolism of TNBC cell line MDA-MB-453 is altered by cytokines, TDO2 inhibition, or suspension growth conditions - cellular steady state metabolome and in vitro tracing with 13C11 tryptophan
STUDY_SUMMARY
TNBC cell line MDA-MB-453 was cultured in 13C11 tryptophan and experimental treatments were as follows: a) cytokine treatment (TNFĪ±+IL1Ī²); b) treated with TDO2/IDO1 dual inhibitor AT-0174 for 24 or 48 hours with an without labeled tryptophan; c) cultured in 13C11 tryptophan and suspension condition.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Tryptophan metabolite profiling of cell supernatants (i.e. cell culture media) from TNBC cell lines with either overexpression or stable knockdown of TDO2.
STUDY_SUMMARY
Indole-focused metabolomics analysis of cell supernatants (i.e. cell culture media) from TNBC cell lines with overexpression or knockdown of TDO2. BT549 cells were genetically manipulated with stable knockdown of TDO2 (shTDO2). TNBC cell line SUM159PT was generated with stable TDO2 overexpression (TDO2 OE) and treated with TDO2/IDO1 dual inhibitor AT-0174.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Tryptophan metabolite profiling of cell supernatants (culture media) from TNBC cell line BT549 grown in suspension with TDO inhibition.
STUDY_SUMMARY
Indole-focused metabolomics analysis of cell supernatants (i.e. cell culture media) from TNBC cell lines with pharmacological inhibition of TDO2 by AT-0174 or 680c91.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Tryptophan metabolite profiling of cell supernatants (i.e. cell culture media) from TNBC cell line BT549 grown in suspension with TDO2 knockdown and IDO1 compensation.
STUDY_SUMMARY
Conditioned media harvested from TNBC cell line BT549 with stable TDO2 knockdown (shTDO2) but harbor IDO1 compensation. The cells were treated with TDO2/IDO1 dual inhibitor AT-0174, IDO1 inhibitor epacadostat, or TDO2 inhibitor 680C91 for 48 hours.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Tryptophan pathway profiling of conditioned media from TNBC cell line MDA-MB-231 +/- inhibitors of the TDO/IDO pathway.
STUDY_SUMMARY
Conditioned media harvested from TNBC cell line MDA-MB-231 with and without treatment with TDO2/IDO1 dual inhibitor AT-0174 and IDO1 inhibitor epacadostat.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Metabolomic profiling of the secretome in normal primary human pulmonary artery smooth muscle cells
STUDY_SUMMARY
Growth medium of human pulmonary artery smooth muscle cells (PASMCs) was conditioned for 24 hours and analyzed by LC-MS, termed conditioned medium (CM). Comparing to fresh complete vascular smooth muscle cell growth medium (GM), LC-MS profiling screened 138 metabolites and identified that three branched chain alpha-ketoacids, namely KIC, KIV, and KMV are the most abundant metabolites in CM of human PASMCs.
INSTITUTE
Peking University
DEPARTMENT
Department of Toxicology
LAST_NAME
Xiao
FIRST_NAME
Wusheng
ADDRESS
38 Xueyuan Rd, Haidian District, Beijing, Beijing, 100191, China
Metabolic analysis of ADSL deficiency patients' EBV-LCLs.
STUDY_SUMMARY
Adenylosuccinate lyase (ADSL) deficiency (ADSLd) is a rare autosomal recessive defect of purine metabolism associated with a wide range of clinical manifestations. Despite ADSL implication in purine synthesis, no additional molecular alterations have been identified as a cause of ADSLd besides the accumulation of toxic substrates. Here we uncover a novel association between ADSLd and mitochondrial dysfunction, which is characterized by an increase in fragmentation and reduction in respiration and ATP production. The extent of this mitochondrial dysfunction is directly proportional to the pathological manifestations of ADSLd, which are predominantly observed in tissues that rely heavily on mitochondria. Our analysis also unveils a striking defect in mitochondrial dynamics and transport, which are associated with the suppression of ERK2 and AKT function. Remarkably, the mitochondrial phenotype can be rescued in part by overexpression of a constitutive form of ERK2 or through the administration of purine intermediates. This scenario provides an alternative explanation of ADSLd onset, reorienting research towards developing innovative therapeutic strategies based on the restoration of mitochondrial metabolism.
A UHPLC-MS/MS Method for Profiling of Urinary Mercapturic Acids using Positive Ion Mode
STUDY_SUMMARY
We report the first application of a UHPLC-MS/MS method using positive ion mode detection for the unbiased characterization of mercapturic acids. The proposed method utilizes a neutral loss monitoring paradigm to monitor for two diagnostic fragmentation pathways for this class of compound. Using a cohort of 20 nonsmokers and 20 smokers, we detected 180 putative mercapturic acid signatures that exhibited a high degree of reproducibility from the complex urine metabolome background. Following a combination of multivariate and univariate statistics, we found 33 putative mercapturic acids associated with smoking status.
TREM2 expression level is critical for microglial state, metabolic capacity and efficacy of TREM2 agonism
STUDY_SUMMARY
Triggering receptor expressed on myeloid cells 2 (TREM2) is a central regulator of microglial activity and sequence variants are major risk factors for late onset Alzheimerās disease (LOAD). To better understand the molecular and functional changes associated with TREM2 signalling, we generated a TREM2 reporter mouse model and observed a gradual upregulation of reporter expression with increasing plaque proximity. Isolated microglia were sorted based on reporter expression and their transcriptomic profiles acquired in both wildtype and APP transgenic animals, allowing us to disentangle TREM2 versus pathology-specific effects. Bulk RNA sequencing highlighted TREM2 level-dependent changes in major immunometabolic pathways, with enrichment of genes in oxidative phosphorylation and cholesterol metabolism in microglia with increased TREM2 expression. To confirm these findings, we next analysed uptake of fluorodeoxyglucose (FDG) and examined metabolomic and lipidomic profiles. Again, independent of AĪ² pathology, TREM2 expression correlated with uptake of FDG as well as increased cellular redox, energetics, and cholesterol homeostasis. Finally, we performed chronic treatment with a brain penetrant TREM2 agonist and identified a window of TREM2 expression where microglia are most responsive. Thus, our data provide novel insights into TREM2-mediated regulation of microglial metabolic function and informs current efforts to bring TREM2 agonists into clinical application.
INSTITUTE
Denali Therapeutics
LAST_NAME
Suh
FIRST_NAME
Jung
ADDRESS
161 Oyster Point Blvd, South San Francisco, California, 94080, USA
Compound lipids comprise a diverse group of metabolites present in living systems, and metabolic- and environmentally-driven structural distinctions across this family is increasingly linked to biological function. However, methods for deconvoluting these often isobaric lipid species are lacking or require specialized instrumentation. Notably, acyl-chain diversity within cells may be influenced by nutritional states, metabolic dysregulation, or genetic alterations. Therefore, a reliable, validated method of quantifying structurally similar even-, odd-, and branched-chain acyl groups within intact compound lipids will be invaluable for gaining molecular insights into their biological functions. Here we demonstrate the chromatographic resolution of isobaric lipids containing distinct combinations of straight-chain and branched-chain acyl groups via ultra-high-pressure liquid chromatography (UHPLC)-mass spectrometry (MS) using a C30 liquid chromatography column. Using metabolically-engineered adipocytes lacking branched-keto acid dehydrogenase A (Bckdha), we validate this approach through a combination of fatty acid supplementation and metabolic tracing using monomethyl branched-chain fatty acids and valine. We observe resolution of numerous isobaric triacylglycerols and other compound lipids, demonstrating the resolving utility of this method. This approach strengthens the ability to quantify and characterize the inherent diversity of acyl chains across the lipidome.
Methionine-SAM metabolism-dependent ubiquinone synthesis is crucial for ROS accumulation in ferroptosis induction
STUDY_SUMMARY
Ferroptosis is a cell death modality in which iron-dependent lipid peroxides accumulate on cell membranes. Cysteine, a limiting substrate for the glutathione system that neutralizes lipid peroxidation and prevents ferroptosis, can be converted by cystine reduction or synthesized from methionine. However, accumulating evidence shows methionine-based cysteine synthesis fails to effectively rescue intracellular cysteine levels upon cystine deprivation and is unable to inhibit ferroptosis. Here, we report that methionine-based cysteine synthesis is tissue-specific. Unexpectedly, we find that rather than inhibiting ferroptosis, methionine in fact plays an essential role during cystine deprivation-induced ferroptosis. Methionine-derived S-adenosylmethionine (SAM) contributes to methylation-dependent ubiquinone synthesis, which leads to lipid peroxides accumulation and subsequent ferroptosis. Moreover, SAM supplementation synergizes with imidazole ketone erastin in a tumor growth suppression mouse model. Inhibiting the enzyme that converts methionine to SAM protects heart tissue from doxorubicin-induced and ferroptosis-driven cardiomyopathy. This study broadens our understanding about the intersection of amino acid metabolism and ferroptosis regulation, providing insight into the underlying mechanisms and suggesting the methionine-SAM axis is a promising therapeutic strategy to treat ferroptosis-related diseases. To test, we firstly performed a metabolomics analysis of HT1080 cells that were cultured in medium lacking cystine or lacking both cystine and methionine. The metabolomics results containing full profile of the metabolites was subjected to cluster assay, and the triplicates of each group were automatically clustered together, indicating better homogeneity of the data in each condition. Then, we filtrated the metabolomics data with the threshold fold change (FC) ā„2 and Pā¤ 0.05.Only the RP Pos method results are used/reported in this study. The results showed that the cellular cysteine and GSH levels were expectedly depressed upon cystine limitation; meanwhile, once methionine was restricted, cellular SAM level was significantly reduced. This highlights the decisive regulatory role of methionine on the metabolite levels of SAM.
INSTITUTE
Northeast Normal University
LAST_NAME
xia
FIRST_NAME
chaoyi
ADDRESS
5268 Renmin Street, changchun, ji lin, 130024, China
Investigation of polyamine biosynthesis and metabolism in gut microbiome by stale isotope resolved metabolomics
STUDY_SUMMARY
Polyamines are important gut microbial metabolites known to affect host organs, yet the mechanisms behind their microbial production remain poorly understood. In this study, we used a stable isotope-resolved metabolomic (SIRM) approach to track polyamine biosynthesis in the fecal microbiome. Viable microbial cells were extracted from fresh human and mouse feces and incubated anaerobically with 13C-labeled inulin (tracer). Liquid chromatography-high resolution mass spectrometry analysis revealed distinct 13C enrichment profiles for spermidine (SPD) and putrescine (PUT), with the arginine-agmatine-SPD pathway predominating over the well acknowledged spermidine synthase pathway (PUT aminopropylation) for SPD biosynthesis. Furthermore, significant species differences were observed in the 13C enrichments of polyamines and related metabolites between the human and mouse microbiome. Further investigations using single-strain SIRM analyses (Bacteroides) identified the key microbial genes and gut microbes responsible for the polyamine biosynthesis. Taken together, this study expands our understanding of polyamine biosynthesis in the gut microbiome and will facilitate the development of precision therapies to target polyamine-associated diseases.
Comparison of serum and interstitial fluid from patients with breast cancer-related lymphedema and healthy control subjects with NMR-based metabolomics
STUDY_SUMMARY
Diagnosing and treating diseases such as breast cancer-related lymphedema (BCRL) is challenging due to a limited understanding of the underlying mechanisms. Despite recent advancements, BCRL significantly impacts patients' quality of life, as current treatments only manage symptoms. Leveraging modern high-throughput omics technologies, particularly metabolomics, holds potential to address these challenges. Metabolomics offers insights into dynamic changes influenced by internal and external factors, aiding in understanding the tissue physiology and detecting pathological conditions. The investigation of metabolomic biomarkers holds promise for early lymphedema diagnosis and personalized treatment. The deposited dataset represent high-resolution nuclear magnetic resonance (NMR) data for patients' blood serum and interstitial fluid, obtained after breast cancer treatment and with diagnosed BCRL, as well as control samples. Simple statistical analysis yielded increased concentrations of pyruvate, citrate, 2-ketoisovalerate, ketoleucine, 3-methyl-2-oxovalerate, tryptophan, and ascorbate in serum samples from patients with lymphedema. This dataset can aid in identifying early-stage lymphedema biomarkers and deepen insights into lymphatic system function thus leading to the development of effective diagnostic and therapeutic tools.
A NRF2/Ī²3-adrenoreceptor axis drives a sustained antioxidant and metabolic rewiring through the pentose-phosphate pathway to alleviate cardiac stress
STUDY_SUMMARY
WT TAC vs TG TAC vs WT baseline vs TG baseline Background: Cardiac Ī²3-adrenergic receptors (Ī²3AR) are upregulated in diseased hearts and mediate antithetic effects to those of Ī²1AR and Ī²2AR. Ī²3AR agonists were recently shown to protect from myocardial remodeling in preclinical studies and to improve systolic function in patients with severe heart failure. The underlying mechanisms, however, remain elusive. Methods: To dissect functional, transcriptional and metabolic effects, hearts and isolated ventricular myocytes from mice harboring a moderate, cardiac-specific expression of a human ADRB3 transgene (Ī²3AR-Tg) and subjected to transverse aortic constriction (TAC) were assessed using echocardiography, RNAseq, PET scan, metabolomics, seahorse and metabolic flux analysis. Subsequently, signaling and metabolic pathways were investigated further in vivo in Ī²3AR-Tg and in vitro in neonatal rat ventricular myocytes adenovirally infected to express Ī²3AR and subjected to neurohormonal stress. These results were completed with an analysis of single nucleus RNAseq data from human cardiac myocytes from heart failure patients. Results: Compared with WT littermate, Ī²3AR-Tg mice were protected from hypertrophy after transaortic constriction (TAC), while systolic function was preserved. Ī²3AR-expressing hearts displayed enhanced myocardial glucose uptake under stress in absence of increased lactate levels. Instead, metabolomic and metabolic flux analyses in stressed hearts revealed an increase in intermediates of the Pentose-Phosphate Pathway (PPP) in Ī²3AR-Tg, an alternative route of glucose utilization, paralleled with increased transcript levels of NADPH-producing and rate-limiting enzymes of the PPP, without fueling the hexosamine metabolism. The ensuing increased content of NADPH and of reduced glutathione decreased myocyte oxidant stress, while downstream oxidative metabolism assessed by oxygen consumption was preserved with higher glucose oxidation in Ī²3ARTg post-TAC compared to WT, together with increased mitochondrial biogenesis. Unbiased transcriptomics and pathway analysis identified NRF2 (NFE2L2) as upstream transcription factor which was functionally verified in Ī²3AR- expressing cardiac myocytes where its translocation and nuclear activity was dependent on Ī²3AR activation of nitric-oxide synthase (NOS) NO production. Conclusion: Moderate expression of cardiac Ī²3AR, at levels observed in human cardiac myocardium, exerts antioxidant effects through activation of the PPP and NRF2 pathway, thereby preserving myocardial oxidative metabolism, function and integrity under pathophysiological stress.
Associations Between Maternal Microbiome, Metabolome and Incidence of Low Birth Weight in Guatemalan Participants from the Women First Trial
STUDY_SUMMARY
Low birth weight (babies born at less than 2,500 grams) affects approximately 15 to 20 percent of global births annually and is associated with poor child development. The goal of this study was to examine relationships between maternal microbial taxa, fecal metabolites, and maternal anthropometry on incidence of LBW in resource-limited settings. This was a secondary analysis of the Women First trial conducted in a semi-rural region of Guatemala. Maternal weight was measured at 12 and 34 weeks (wk) of gestation. Infant anthropometry measures were collected within 48 h of delivery. Maternal fecal samples at 12 and 34 wk were used for microbiome (16S rRNA gene amplicon sequencing) and metabolomics analysis (34 wk only). Linear mixed models using the MaAslin2 package were utilized to assess changes in microbiome associated with LBW. Predictive models using gradient boosted machines (XGBoost) were developed using the H2o.ai engine. No differences in Ī²-diversity were observed at either time point between mothers with LBW infants relative to normal weight (NW) infants. Simpson diversity at 12 and 34 wk was lower in mothers with LBW infants. Notable differences in genus-level abundance between LBW and NW mothers (p< 0.05) were observed at 12 weeks with increasing abundances of Barnesiella, Faecalibacterium, Sutterella, and Bacterioides. At 34 weeks, there were lower abundances of Magasphaera, Phascolarctobacterium, and Turicibacter and higher abundances of Bacteriodes, and Fusobacterium in mothers with LBW infants. Fecal metabolites related to bile acids, tryptophan metabolism and fatty acid related metabolites changed in mothers with LBW infants. Classification models to predict LBW based on maternal anthropometry and predicted microbial functions showed moderate performance. Collectively, the findings indicate that less beneficial gut microbes and circulating metabolites of the mother is associated with low birth weight infants compared to normal weight. Future research should target functional and predictive roles of the maternal gut microbiome in infant birth outcomes including birthweight.
3-Hydroxybutyrate Suppresses Colon Cancer Growth through Metabolic Reprogramming and Monocarboxylate Transporter-Mediated Lactate Accumulation
STUDY_SUMMARY
Cancer cachexia (CAC) remains a challenging complication in colon cancer, often leading to poor clinical outcomes. This study investigates the therapeutic potential of 3-hydroxybutyrate (3-HB) in CAC by exploring its effects on tumor growth and cellular metabolism using a colon CAC mouse model and CT26 colon cancer cells. Through NMR-based metabolomics and molecular biology approaches, we reveal that 3-HB slows tumor growth in CAC mice, potentially by increasing lactate accumulation within tumor tissues, modulating key metabolic pathways. Additionally, 3-HB induced oxidative stress and apoptosis in CT26 cells, characterized by elevated reactive oxygen species (ROS) and caspase-3 activation. Mechanistically, 3-HB appears to compete with lactate for monocarboxylate transporters (MCTs), leading to intracellular lactate accumulation, cellular acidification, and tumor suppression. These findings suggest that 3-HB holds promise as a therapeutic agent for colon cancer cachexia, offering new insights into metabolic targeting strategies for cancer treatment.
3-Hydroxybutyrate Suppresses Colon Cancer Growth through Metabolic Reprogramming and Monocarboxylate Transporter-Mediated Lactate Accumulation
STUDY_SUMMARY
Cancer cachexia (CAC) remains a challenging complication in colon cancer, often leading to poor clinical outcomes. This study investigates the therapeutic potential of 3-hydroxybutyrate (3-HB) in CAC by exploring its effects on tumor growth and cellular metabolism using a colon CAC mouse model and CT26 colon cancer cells. Through NMR-based metabolomics and molecular biology approaches, we reveal that 3-HB slows tumor growth in CAC mice, potentially by increasing lactate accumulation within tumor tissues, modulating key metabolic pathways. Additionally, 3-HB induced oxidative stress and apoptosis in CT26 cells, characterized by elevated reactive oxygen species (ROS) and caspase-3 activation. Mechanistically, 3-HB appears to compete with lactate for monocarboxylate transporters (MCTs), leading to intracellular lactate accumulation, cellular acidification, and tumor suppression. These findings suggest that 3-HB holds promise as a therapeutic agent for colon cancer cachexia, offering new insights into metabolic targeting strategies for cancer treatment.
Lipidomic analysis of Malassezia globosa at different growth stages and the dynamics of uptake and secreted lipids with growth media
STUDY_SUMMARY
Malassezia is one of the most abundant genera found on human skin; specifically, M. globosa is one of the yeast species dominant in this organ as it has been associated with several skin diseases. Malassezia cannot synthesize fatty acids. In response, the yeast cell intakes external fatty acids from the host or the growth media for survival. Several studies have focused on investigating the identity of lipids and enzymes in M. globosa to understand its lipid metabolism and the biology of the yeast cell-host interaction. In this work, we performed a supernatant lipidomic analysis on the mDixon media and the supernatant and on the M. globosa at early and late stationary phase (72h and 90h, respectively) to determine the lipid dynamics (lipids consumed vs. lipids secreted) between the growth media and the two stages of growth. We were able to identify 87 lipids within 17 classes of lipids; during the analysis, the increment of several lipids increased throughout time concerning the growth media, suggesting a secretion pattern from the cell to the media; some lipids found in this group were conjugated Sterols (ST) such as Glycochenodeoxycholic acid (GCDCA), Glycerophospholipids (GP), specifically phosphocholine's (PCs), Cardiolipins (CL), in particular those with chains of (47 to 54 carbons) and Sphingolipids (SP) such as Cer-PI which might have some role in pathogenicity. Likewise, the increment of some lipids decreased, but some only reduced at the late stationary phase (90h) only when the nutrients available was minimal. Finally, we observed a third pattern in which the amount of some lipids decreased throughout time (starting in the early stationary phase and finishing in the late stationary phase), hinting at a distinctive consumption pattern. The principal lipids consumed wereSterols (ST) bile acids, cholic acid, and its derivates, some phosphocholines (PCs), Fatty acyls (FA), and cardiolipins (CL). The consumption of these lipids was associated with different metabolic roles of the lipids in the cell as it lacks production of these lipids in M. globosa.
Exogenous L-serine supply protects against retinopathy of prematurity in a murine model
STUDY_SUMMARY
Pathological retinal angiogenesis with irregular and fragile vessels (also termed as neovascularization, a response to hypoxia and dysmetabolism) is a leading cause of vision loss in all age groups driven in part by unmet metabolic demand from retinal neurons. Sustaining neural retinal metabolism with an adequate nutrient supply may prevent vision-threatening neovascularization. Low circulating serine levels are associated with neovascularization in macular telangiectasia and altered serine/glycine metabolism is suggested in retinopathy of prematurity. Here we assessed the role of serine metabolism in suppressing hypoxia-driven retinal neovascularization in mice. Serine or vehicle control was supplemented systemically via intraperitoneal injection. Systemic serine supplementation decreased retinal neovascularization. Metabolomic analysis showed serine treatment increased metabolites involved in fatty acid oxidation and phospholipid synthesis.
Highly reliable LC-MS lipidomics database for efficient human plasma profiling based on NIST SRM 1950
STUDY_SUMMARY
Liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS)-based methods have become the gold standard methodology for the comprehensive profiling of the human plasma lipidome. However, both the complexity of lipid chemistry and LC-HRMS-associated data pose challenges to the characterization of this biological matrix. In accordance with the current consensus of quality requirements for LC-HRMS lipidomics data, we aimed to characterize the NISTĀ® Standard Reference Material for Human Plasma (SRM 1950) using an LC-ESI(+/ā)-MS method compatible with high-throughput lipidome profiling. We generated a highly curated lipid database with increased coverage, quality, and consistency, including additional quality assurance procedures involving adduct formation, within-method m/z evaluation, retention behavior of species within lipid chain isomers, and expert-driven resolution of isomeric and isobaric interferences. As a proof-of-concept, we showed the utility of our in-house LC-MS lipidomic database āconsisting of 592 lipid entriesā for the fast, comprehensive, and reliable lipidomic profiling of the human plasma from healthy human volunteers. We are confident that the implementation of this robust resource and methodology will have a significant impact by reducing data redundancy and the current delays and bottlenecks in untargeted plasma lipidomic studies.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
Chemistry and Biochemistry
LABORATORY
CEMBIO
LAST_NAME
MartĆnez
FIRST_NAME
Sara
ADDRESS
UrbanizaciĆ³n MonteprĆncipe, 28660, Boadilla del Monte, Madrid, Spain
Untargeted Metabolomics of 3xTg-AD Neurotoxic Astrocytes
STUDY_SUMMARY
Alzheimer's disease (AD) is the most common form of dementia, affecting approximately 47M people worldwide. Histological features and genetic risk factors, among other evidence, supported the amyloid hypothesis of the disease. This neuronocentric paradigm is currently undergoing a shift, considering evidence of the role of other cell types, such as microglia and astrocytes, in disease progression. Previously, we described a particular astrocyte subtype obtained from the 3xTg-AD murine model that displays neurotoxic properties in vitro. We continue here our exploratory analysis through the lens of metabolomics to identify potentially altered metabolites and biological pathways. Cell extracts from neurotoxic and control astrocytes were compared using HRMS-based metabolomics. Around 12% of metabolic features demonstrated significant differences between neurotoxic and control astrocytes, including alterations in the key metabolite glutamate. Consistent with our previous transcriptomic study, the present results illustrate many homeostatic and regulatory functions of metabolites, suggesting that neurotoxic 3xTg-AD astrocytes exhibit alterations in the Krebs cycle as well as the prostaglandin pathway. This is the first metabolomic study performed in 3xTg-AD neurotoxic astrocytes. These results provide insight into metabolic alterations potentially associated with neurotoxicity and pathology progression in the 3xTg-AD mouse model and strengthen the therapeutic potential of astrocytes in AD.
INSTITUTE
Instituto de Investigaciones BiolĆĀ³gicas Clemente Estable (IIBCE)
LAST_NAME
Carvalho
FIRST_NAME
Diego
ADDRESS
Isidoro de MarĆĀa 1614, Montevideo, Montevideo, 11800, Uruguay
Macrophages are activated towards phagocytic Lymphoma Cell Clearance by Pentose Phosphate Pathway Inhibition
STUDY_SUMMARY
Macrophages in the B-cell lymphoma microenvironment represent a functional node in progression and therapeutic response. We assessed metabolic regulation of macrophages in the context of therapeutic antibody-mediated phagocytosis. Pentose phosphate pathway (PPP) inhibition induced increased phagocytic lymphoma cell clearance by macrophages in vitro, in primary human Chronic lymphocytic leukemia (CLL) patient co-cultures, and in mouse models. Addition of the PPP inhibitor S3 to antibody therapy achieved significantly prolonged overall survival in an aggressive B-cell lymphoma mouse model. PPP inhibition induced metabolic activation and pro-inflammatory polarization of macrophages while it decreased macrophagesĀ“ support for survival of lymphoma cells empowering anti-lymphoma function. As mechanism of macrophage repolarization, the link between PPP and immune regulation was identified. PPP inhibition causes decreased glycogen level and subsequent modulation of the immune modulatory UDPG-Stat1-Irg1-Itaconate axis. Thus, we hypothesize the PPP as key regulator and targetable modulator of macrophage activity in lymphoma to improve efficacy of immunotherapies and prolong survival.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
QuĆmica y BioquĆmica
LAST_NAME
VillaseƱor
FIRST_NAME
Alma
ADDRESS
UrbanizaciĆ³n MonteprĆncipe, n/s, Boadilla del Monte, Madrid, 28003, Spain
Identify key lipid species in CAF-PDAC crosstalk under hypoxia
STUDY_SUMMARY
This study ran lipidomic profiling to identify the lipids enriched in CAF-conditioned media and depleted in PDAC cell culture under hypoxia at 72 hours.
Effect of insluin on metabolism of ex vivo ischemia heart
STUDY_SUMMARY
Ex vivo hearts were pre-perfused with or without insulin, followed by ischemia induced by the cessation of perfusion. After 25 minutes of ischemia, hearts underwent 30 minutes of reperfusion. Right atrial tissue was then collected for IC-MS analysis. Three groups were included: a control group without ischemia, a group with 25 minutes of ischemia followed by reperfusion, and a group pre-perfused with insulin for 30 minutes prior to ischemia and reperfusion.
INSTITUTE
Mayo Clinic
LAST_NAME
Qiu
FIRST_NAME
Huiliang
ADDRESS
5951 E Mayo Blvd, RE 1-408, Zhu Lab, Phoenix, AZ 85054
Identification of Plasma Metabolomic Biomarkers of Juvenile Idiopathic Arthritis
STUDY_TYPE
Clinical
STUDY_SUMMARY
This study utilizes plasma metabolomic profiling to identify biomarkers associated with juvenile idiopathic arthritis (JIA) by analyzing samples from treatment-naĆÆve JIA patients and non-JIA controls. Significant metabolic alterations were detected, with sphingosine metabolites and fatty acid ethanolamides showing notable increases in JIA patients, while specific compounds such as sarcosine were decreased. The research highlights 11 highly discriminatory metabolites, including sphinganine-1-phosphate, demonstrating potential for improved JIA diagnosis and treatment through targeted metabolic profiling.
INSTITUTE
University of Kansas
DEPARTMENT
Center for Computational Biology
LABORATORY
Funk
LAST_NAME
Kumar
FIRST_NAME
Amar
ADDRESS
Multidisciplinary Research Bldg. 2030 Becker Drive Lawrence, KS 66047
EMAIL
amarkumar@ku.edu
PHONE
18723016225
NUM_GROUPS
2
TOTAL_SUBJECTS
210
NUM_MALES
82
NUM_FEMALES
128
STUDY_COMMENTS
Although the raw dataset initially included 210 subjects, only 207 were included in the final analysis due to consent-related exclusions. These three subjects were removed to ensure compliance with ethical standards.
Metabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii
STUDY_TYPE
Biomedical research
STUDY_SUMMARY
This untargeted metabolomics study investigated the synergistic antibacterial activity of polymyxin B and Leu10-teixobactin, a depsipeptide inhibitor of cell wall biosynthesis. Checkerboard microdilution assays revealed a significant synergy against polymyxin-susceptible and -resistant A. baumannii, excluding lipopolysaccharide-deficient variants. Time-kill assays confirmed bactericidal synergy, reducing bacterial burden by approximately 4-6-log10CFU/mL. The combination (2xMIC polymyxin B and 0.5xMIC Leu10-teixobactin) prevented bacterial regrowth after 24 h, indicating sustained efficacy against the emergence of resistant mutants. The analysis of A. baumannii ATCCTM 19606 metabolome demonstrated that the polymyxin BāLeu10-teixobactin combination produced more pronounced perturbation compared to the individual antibiotics across all time points (1, 3 and 6 h). Pathway analysis revealed that lipid metabolism, cell envelope biogenesis, and cellular respiration were predominantly impacted by the combination, and to a lesser extent by polymyxin B monotherapy. Leu10-teixobactin treatment alone had only a minor impact on the metabolome, primarily at the 6 h time point. Peptidoglycan assays confirmed the combinationās concerted deleterious effects on bacterial cell envelope integrity. Electron microscopy further substantiated these findings, revealing pronounced cell envelope damage, membrane blebbing, and vacuole formation. These findings highlight the potential of the polymyxin BāLeu10-teixobactin combination as an effective treatment in preventing resistance in A. baumannii.
INSTITUTE
Monash University
DEPARTMENT
Pharmacology
LABORATORY
Velkov
LAST_NAME
HUSSEIN
FIRST_NAME
MAYTHAM
ADDRESS
9 Ancora Imparo Way, Building 13E, Monash University
EMAIL
maytham.hussein.old@monash.edu
PHONE
+61406574736
PUBLICATIONS
Metabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii
Untargeted mass spectrometry analysis of neutral lipids in MC38 tumors and visceral adipose tissues of control-diet and high fat diet-fed C57BL/6 mice
STUDY_SUMMARY
Both tumors and the visceral adipose tissues were collected from Control Diet (CD) and High Fat Diet (HFD)-fed C57BL/6 (B6) mice. Mice were fed a HFD for 2 months. For tumor studies, MC38 tumors were subcutaneously injected after 2 months of feeding mice the HFD. Tumors were allowed to grow for 3 weeks, at which point, tumors from CD and HFD-fed mice were harvested and snap frozen. Visceral adipose tissues were also collected from these mice and snap frozen. Subsequently, samples were sent to the UC Davis Metabolomics Core for analysis of neutral lipids. Some triglycerides, including TG(54:3), were found to be significantly more abundant in the adipose tissue and tumors of HFD-fed mice compared to CD-fed mice.
Metabolic and Proteomic Divergence is Present in Spleens and Livers from Berkeley Sickle Cell Anemia and beta-thalassemia mice
STUDY_SUMMARY
Steady state metabolomics of liver tissue from the Berkeley sickle cell disease (Berk-SS), heterozygous B1/B2 globin gene deletion (HbbTh3/+) a known -Thalassemia model, and wildtype (WT, C57/Bl6) murine models.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Metabolic and Proteomic Divergence is Present in Spleens from Berkeley Sickle Cell Anemia and beta-thalassemia mice
STUDY_SUMMARY
Steady state metabolomics of spleen tissue from the Berkeley sickle cell disease (Berk-SS), heterozygous B1/B2 globin gene deletion (HbbTh3/+) a known -Thalassemia model, and wildtype (WT, C57/Bl6) murine models.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Combining antibiotics alters the longitudinal maturation of gut microbiota and its short chain fatty acid metabolites in extremely and very preterm infants
STUDY_SUMMARY
Antibiotics are routinely prescribed to extremely and very premature infants as a pre-emptive and prophylactic treatment to reduce the risk of acute neonatal illness (i.e. necrotizing enterocolitis, NEC) associated with morbidity. To investigate the effects of antibiotic types, combinations, and duration on the preterm gut microbiome and metabolome, we analyzed the microbiome compositions of 123 stool samples collected at 3 timepoints (postnatal day 1, 28 and 56) from extremely- and very-low-birthweight infants treated with 14 different antibiotics spanning across 5 classes. Targeted metabolomics were performed on 47 samples available, allowing us to quantify 649 metabolites including amino acids, bile acids, fatty acids, and lipids. As a result, we found that antibiotics exerted the most profound disruptive impact on the gut microbiota, while antibiotics and breastfeeding highly influence the gut metabolome. Short chain fatty acids were reduced in both antibiotic-treated and NEC group. Finally, we revealed that cephalosporins negatively impact conjugated bile acids due to a positive correlation with bile salt hydrolase-producing Staphylococcus.
Lipidomics profiling of livers from Alb-Cre +/- SART-/- (KO) versus age matched wild type WT mouse littermates to characterize progression to hepatic steatosis and hepatocellular carcinoma
STUDY_SUMMARY
Mice bearing hepatocyte- specific deletion of the gene SART1 (Alb-Cre+/- SART-/-) develop hepatic steatosis at age 6 months and spontaneous liver tumors at age 18 months. To determine the changes in lipid composition of the SART1-deficient livers compared to age-matched wild- type livers, Liver lipidomics profiling of Male animals at different age groups were performed: Age 6 months: KO versus WT livers, Age 12 months: KO versus WT livers, Age 18 months: KO liver tumors versus WT livers. Flash-frozen livers from male Alb-Cre +/- SART1 +/+ (control) and Alb-Cre +/- SART-/- (KO) were subjected to lipidomics profiling.
Measurement of itaconic acid in liver of male mice treated with TCDD
STUDY_TYPE
chromatograms
STUDY_SUMMARY
In this study, we tested the hypothesis that the dose-dependent disruption of propionyl-CoA metabolism produces toxic intermediates that contribute to TCDD hepatotoxicity and progression of steatosis to steatohepatitis with fibrosis. Our results suggest TCDD dose-dependently reduced cobalamin (Cbl aka vitamin B12) levels compromising methylmalonyl-CoA mutase (MUT) activity and limiting the metabolism of propionyl-CoA to succinyl-CoA using the canonical Cbl-dependent carboxylation pathway. More recently, lower Cbl levels have been linked to itaconate, a cis-aconitate metabolite produced in large quantities by activated macrophages. In the current study, targeted metabolomics analysis of hepatic extracts detected a dose-dependent increase in itaconic acid. Itaconate can be activated to itaconyl-CoA which can then interact with the 5-deoxyadenosyl moiety of AdoCbl to form an uncharacterized adduct that disrupts auxiliary repair protein interactions, inactivates AdoCbl, and reduces Cbl levels that inhibit MUT activity.
Microbiome and metabolome association network analysis identifies Clostridium_sensu_stricto_1 and Paraprevotella as putative keystone genera in the gut of common marmosets
STUDY_SUMMARY
The common marmoset (Callithrix jacchus), a nonhuman primate species, is a model organism that has garnered interest in recent years for its potential translational value in a variety of research settings including the field of microbiomics. While the composition of the marmosetās gut microbiome has been described in captivity, little is known about how gut microbiota interact with each other over time and how they relate to metabolite productions. To help answer this, we characterized interactions in the gut microbiome of the common marmoset by calculating the Spearman correlation coefficient between 16S rDNA-derived relative genera abundance data and targeted metabolomics data collected longitudinally from 10 marmosets (6 males and 4 females). Association network graphs were used to visualize significant correlations and identify genera and metabolites that exhibit a high degree of associations, marking them as more influential within the microbiome. Clostridium_sensu_stricto_1, among the highest-degree genera for bacterial and metabololomic associations, also had a high relative betweenness centrality and negatively associated with high-degree Paraprevotella, indicating that it potentially plays a gatekeeping role within the bacteria-bacteria interaction and communication network. Corresponding metabolites with more numerous bacterial associations, including bile acids and taurine, are known regulators of bacterial growth that provide a potential mechanism through which Clostridium_sensu_stricto_1 and others exert their influence. To further characterize microbiome interactions, we performed hierarchical clustering on significant within-dataset associations and developed a new āKeystone Candidate Scoreā metric that identified Clostridium_sensu_stricto_1 and Paraprevotella as the most influential bacteria (so-called candidate keystone genera) in the marmoset gut microbiome.
Micropeptide hSPAR, a glutamine regulator, suppresses tumor growth via TRIM21-P27KIP1-mTOR pathway - human MDA-MB-231 breast cancer cell
STUDY_SUMMARY
hSPAR, a microprotein, is capable of specifically inhibiting the mTOR signaling activity and cell proliferation in MDA-MB-231 cells. Glutamine, an essential amino acid, plays a crucial role in regulating the mTOR signal. Our data indicate that in MDA-MB-231 cells, overexpressing hSPAR inhibits glutamine uptake, consequently suppressing the activation of the mTOR signaling.
INSTITUTE
University of Science and Technology of China
LAST_NAME
Wang
FIRST_NAME
Wei
ADDRESS
Division of Life Sciences and Medicine, 443 Huangshan Road, Hefei city, Anhui Province
Micropeptide hSPAR, a glutamine regulator, suppresses tumor growth via TRIM21-P27KIP1-mTOR pathway - HEK293T human embryonic kidney cells
STUDY_SUMMARY
The microprotein hSPAR can specifically regulate the mTOR signaling activity and cell proliferation in breast cancer cells. However, such regulatory effects of hSPAR are not applicable in HEK293T cells. Metabolic data of amino acids indicate that overexpressing hSPAR does not affect the content of amino acids in HEK293T cells, including glutamine. The metabolic variations in amino acids triggered by hSPAR in different cell types may be the underlying molecular mechanism for the distinct regulatory effects of hSPAR.
INSTITUTE
University Of Science And Technology Of China
LAST_NAME
Wang
FIRST_NAME
Wei
ADDRESS
443 Huangshan Road, Hefei city, Anhui Province, 230022, China
Nutrient intervention protects against retinal dysfunction caused by disrupted peroxisomal fatty acid oxidation
STUDY_SUMMARY
Dyslipidemia is a significant contributor to many eye diseases, but the lipid processing pathways are not fully understood. Peroxisomes oxidize very long-chain fatty acids and generate docosahexaenoic acid. Mutations in peroxisomal genes can result in severe neural retinal dysfunction. However, therapeutic approaches for peroxisomal diseases remain scarce. In this study, we found impaired retinal function and altered retinal metabolism in mice lacking the first enzyme of peroxisomal fatty acid oxidation (ACOX1). Supplementation of the depleted metabolite pyruvate or the depleted omega-3 fatty acid docosahexaenoic acid mitigated the progression of retinal dysfunction in Acox1 KO mice. Nutrient intervention may therefore offer a promising therapeutic approach for peroxisomal diseases.
Arsenic-induced enhancement of diazotrophic recruitment and nitrogen fixation in Pteris vittata rhizosphere
STUDY_SUMMARY
Heavy metal contamination poses an escalating global challenge to soil ecosystems. Hyperaccumulators play a crucial role in environmental remediation and resource recovery. The enrichment of diazotrophs and resulting nitrogen accumulation promoted hyperaccumulator growth and facilitated phytoremediation. Nonetheless, the regulatory mechanism of hyperaccumulator biological nitrogen fixation has remained elusive. Here, we report the mechanism by which arsenic regulates biological nitrogen fixation in the arsenic-hyperaccumulator Pteris vittata. Field investigations and greenhouse experiments, based on multi-omics approaches, reveal that elevated arsenic stress induces an enrichment of key diazotrophs, enhances plant nitrogen acquisition, and thus improves plant growth. Metabolomic analysis and microfluidic experiments further demonstrate that the upregulation of specific root metabolites plays a crucial role in recruiting key diazotrophic bacteria. These findings highlight the pivotal role of nitrogen-acquisition mechanisms in the arsenic hyperaccumulation of Pteris vittata, and provide valuable insights into the plant stress resistance.
INSTITUTE
Zhejiang University
DEPARTMENT
College of Environmental and Resource Sciences
LABORATORY
Institute of Soil and Water Resources and Environmental Science
LAST_NAME
Lin
FIRST_NAME
Jiahui
ADDRESS
866 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China
Probiotics and their metabolite spermidine enhance IFN-Ī³+CD4+ T cell immunity to inhibit hepatitis B virus
STUDY_SUMMARY
The therapeutic potential of commensal microbes and their metabolites is promising in the functional cure of chronic hepatitis B virus (HBV) infection, which is defined as HBsAg loss. Here, using both specific-pathogen-free and germ-free mice, we report that probiotics significantly promote the decline of HBsAg and inhibit HBV replication by enhancing intestinal homeostasis and provoking intrahepatic IFN-Ī³+CD4+ T cell immune response. Depletion of CD4+ T cells or blockage of IFN-Ī³ abolishes probiotics-mediated HBV inhibition. Specifically, probiotics-derived spermidine accumulates in gut and transports to liver, where it exhibits a similar anti-HBV effect. Mechanistically, spermidine enhances IFN-Ī³+CD4+ T cell immunity by autophagy. Strikingly, administration of probiotics in HBV patients reveals a preliminary trend to accelerate the decline of serum HBsAg. In conclusion, probiotics and theirs derived spermidine promote HBV clearance via autophagy-enhanced IFN-Ī³+CD4+ T cell immunity, highlighting the therapeutic potential of probiotics and spermidine for the functional cure of HBV patients.
Metabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3_3 genes enhancing soybean resistance to cyst nematodes
STUDY_SUMMARY
The Gretchen Hagen 3 genes maintain endogenous hormone homeostasis by conjugating excess hormones with amino acids. Herein, we identified the members of the GH3 family in soybeans and analyzed their phylogeny, gene duplication, structure, domains, conserved motifs, cis-elements in promoter regions for stress responses, and functional characteristics. We found that GH3 genes are induced by pathogens in Group-II. Furthermore, 8 out of 16 Group-II genes responded to cyst nematode infection. Through functional analysis of eight GmGH3 genes via overexpression, they play a negative role in soybean resistance to cyst nematode. In addition, our metabolomic analysis showed that overexpression of Glyma.02G125600, Glyma.17G165500, and Glyma.13G284600 affected the content of salicylic acid and jasmonic acid. Our findings clarify the functional features of GH3 genes and reveal their involvement in plant hormone signaling pathways. This provides valuable insights into the complex molecular mechanisms underlying the interaction between soybeans and cyst nematode.
Metabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3_2 genes enhancing soybean resistance to cyst nematodes
STUDY_SUMMARY
The Gretchen Hagen 3 genes maintain endogenous hormone homeostasis by conjugating excess hormones with amino acids. Herein, we identified the members of the GH3 family in soybeans and analyzed their phylogeny, gene duplication, structure, domains, conserved motifs, cis-elements in promoter regions for stress responses, and functional characteristics. We found that GH3 genes are induced by pathogens in Group-II. Furthermore, 8 out of 16 Group-II genes responded to cyst nematode infection. Through functional analysis of eight GmGH3 genes via overexpression, they play a negative role in soybean resistance to cyst nematode. In addition, our metabolomic analysis showed that overexpression of Glyma.02G125600, Glyma.17G165500, and Glyma.13G284600 affected the content of salicylic acid and jasmonic acid. Our findings clarify the functional features of GH3 genes and reveal their involvement in plant hormone signaling pathways. This provides valuable insights into the complex molecular mechanisms underlying the interaction between soybeans and cyst nematode.
Metabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3_1 genes enhancing soybean resistance to cyst nematodes
STUDY_SUMMARY
The Gretchen Hagen 3 genes maintain endogenous hormone homeostasis by conjugating excess hormones with amino acids. Herein, we identified the members of the GH3 family in soybeans and analyzed their phylogeny, gene duplication, structure, domains, conserved motifs, cis-elements in promoter regions for stress responses, and functional characteristics. We found that GH3 genes are induced by pathogens in Group-II. Furthermore, 8 out of 16 Group-II genes responded to cyst nematode infection. Through functional analysis of eight GmGH3 genes via overexpression, they play a negative role in soybean resistance to cyst nematode. In addition, our metabolomic analysis showed that overexpression of Glyma.02G125600, Glyma.17G165500, and Glyma.13G284600 affected the content of salicylic acid and jasmonic acid. Our findings clarify the functional features of GH3 genes and reveal their involvement in plant hormone signaling pathways. This provides valuable insights into the complex molecular mechanisms underlying the interaction between soybeans and cyst nematode.
Metabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3_61 genes enhancing soybean resistance to cyst nematodes
STUDY_SUMMARY
The Gretchen Hagen 3 genes maintain endogenous hormone homeostasis by conjugating excess hormones with amino acids. Herein, we identified the members of the GH3 family in soybeans and analyzed their phylogeny, gene duplication, structure, domains, conserved motifs, cis-elements in promoter regions for stress responses, and functional characteristics. We found that GH3 genes are induced by pathogens in Group-II. Furthermore, 8 out of 16 Group-II genes responded to cyst nematode infection. Through functional analysis of eight GmGH3 genes via overexpression, they play a negative role in soybean resistance to cyst nematode. In addition, our metabolomic analysis showed that overexpression of Glyma.02G125600, Glyma.17G165500, and Glyma.13G284600 affected the content of salicylic acid and jasmonic acid. Our findings clarify the functional features of GH3 genes and reveal their involvement in plant hormone signaling pathways. This provides valuable insights into the complex molecular mechanisms underlying the interaction between soybeans and cyst nematode.
Metabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3_62 genes enhancing soybean resistance to cyst nematodes
STUDY_SUMMARY
The Gretchen Hagen 3 genes maintain endogenous hormone homeostasis by conjugating excess hormones with amino acids. Herein, we identified the members of the GH3 family in soybeans and analyzed their phylogeny, gene duplication, structure, domains, conserved motifs, cis-elements in promoter regions for stress responses, and functional characteristics. We found that GH3 genes are induced by pathogens in Group-II. Furthermore, 8 out of 16 Group-II genes responded to cyst nematode infection. Through functional analysis of eight GmGH3 genes via overexpression, they play a negative role in soybean resistance to cyst nematode. In addition, our metabolomic analysis showed that overexpression of Glyma.02G125600, Glyma.17G165500, and Glyma.13G284600 affected the content of salicylic acid and jasmonic acid. Our findings clarify the functional features of GH3 genes and reveal their involvement in plant hormone signaling pathways. This provides valuable insights into the complex molecular mechanisms underlying the interaction between soybeans and cyst nematode.
Dysregulated Follicular Fluid Metabolism in Women with Unexplained Infertility
STUDY_TYPE
HILIC and RPLC based Untargeted LC-MS/MS
STUDY_SUMMARY
Research Question: Are there specific metabolomic alterations in the Follicular fluid (FF) of women with unexplained infertility (UI)? Study Design: This case-control study included 20 women undergoing in-vitro fertilization (IVF), comparing 10 women diagnosed with UI to 10 control women whose male partners had abnormal semen parameters. FF samples were collected during oocyte retrieval and analysed using hydrophilic interaction liquid chromatography and reversed-phase liquid chromatography, coupled with tandem mass spectrometry (MS/MS) on a Q-TOF mass spectrometer. Metabolites were identified using XCMS Online and MetaboAnalyst, followed by pathway enrichment analysis via the KEGG database. Statistical analyses including OPLS-DA and ROC analysis assessed their diagnostic potential. Metabolite levels were correlated with clinical parameters, including embryo development rates, oocyte maturation, and IVF outcomes. Results: In women with UI, 12 metabolites, including Diacylglycerols, Phosphatidic acids, Vitamin D3 glucosiduronate, 1Ī±-hydroxy-2Ī²-(5-hydroxypentoxy) vitamin D3, Asparginyl-Asparagine, Lithocholic acid, Leu-Pro-Ala-Ser-Phe, Triacylglycerols, Phosphatidyl choline, Phosphatidylethanolamine and Lactosyl ceramide were significantly decreased, while Ile-Lys-Val-Val was significantly increased compared to controls. These metabolites were linked to glycerophospholipid metabolism, glycerolipid metabolism, and steroid synthesis pathways. PLS-DA, OPLS-DA, and ROC analysis indicated high diagnostic performance, with AUC values exceeding 0.8. Additionally, Vitamin D3 glucosiduronate levels negatively correlated with embryo development rates, while Asparginyl-Asparagine levels demonstrated a positive correlation with the MII oocyte rate. Conclusion: This study constitutes the first comprehensive characterization of metabolic dysregulation in the FF of women with UI, offering novel insights into the underlying mechanisms contributing to this condition and advocates for routine assessment of vitamin D3 levels in serum/FF of these women.
INSTITUTE
ICMR - National Institute for Research in Reproductive and Child Health
DEPARTMENT
Gamete Immunobiology
LAST_NAME
Panchal
FIRST_NAME
Durva
ADDRESS
ICMR - NIRRCH, J.M.Street, Parel, Mumbai, India
EMAIL
durva_gib@nirrh.res.in
PHONE
9619565514
NUM_GROUPS
2
TOTAL_SUBJECTS
20
NUM_FEMALES
20 (10 per group)
STUDY_COMMENTS
This case-control study included 20 women undergoing in-vitro fertilization (IVF), comprising 10 women diagnosed with unexplained infertility (UI) and 10 control women whose male partners exhibited abnormal semen parameters. The aim was to investigate metabolic differences in the follicular fluid of these groups, contributing to a deeper understanding of factors influencing reproductive outcomes.
Improved Soil Health and Pasture Phytochemical Richness Underlies Improved Beef Nutrient Density in Southern US Grass-Finished Beef Systems
STUDY_SUMMARY
With growing concerns about beef production's impact on health and environment, there is increasing interest in practices that may improve the soil-plant-animal-human health continuum. We compared three Southern US grass-fed beef systems to grain-fed control, examining effects on soil health in terms of physicochemical properties whereas feed and beef samples were tested for metabolomic profile using orbitrap mass spectrometer. Pasturelands displayed healthier soils compared to paired feedcrop lands, showing 1.4-fold more organic matter and 1.7 to 3.0-fold higher potassium, phosphorus, calcium, and zinc levels (all, p<0.05). Of 784 compounds analyzed, 165 differed between grass-fed and grain-fed beef (all, p<0.05). Grass-fed beef had 3.1-fold higher phytochemical antioxidants, tied to 118.2-fold higher precursor levels in pasture samples compared to total mixed rations (p<0.05). This resulted in reduced oxidative stress in grass-fed animals, with 0.35- and 0.37-fold lower levels of homocysteine and 4-hydroxynonenal glutathione, respectively (both, p<0.05). Findings highlight potential linked benefits across the soil-plant-animal-human nutrition continuum in beef production.
INSTITUTE
Utah State University
DEPARTMENT
Department of Nutrition, Dietetics and Food Sciences
Analysis of the fate of docosahexaenoic acid in HCT116 colorectal cancer cells cultured at pH 7.4 or 6.5.
STUDY_SUMMARY
Cancer cells in acidic tumor regions exhibit a shift from glucose metabolism to the preferential uptake and use of fatty accids. This study aimed to determine the fate of docosahexaenoic acid (DHA) in colon HCT116 cancer cells maintained at pH 7.4 or pH 6.5. The incorporation of triglycerides and/or phospholipids was analysed together with associated changes in the proportion of saturated and monounsaturated fatty acids. We found that DHA heightened the elevated levels of triacylglycerol (TAG) induced by acidic conditions. However, while the abundance of the most prominent Mono-unsaturated Fatty Acid (MUFA), i.e., C18:1 oleate, was directly proportional to the TAG increase in acid-exposed cancer cells, DHA exposure did not further increase the oleate contribution in TAG. The increase in total TAG levels in acidic cancer cells mostly resulted from the incorporation of C22:6 DHA. A more detailed analysis of the composition of TAG revealed that DHA emerges as the predominant constituent of TAG, with 45% of them incorporating at least one DHA, while the TAG content in C18:1 oleate decreased from 46% to 22% in acid-exposed cancer cells. Importantly, while acidic pH and DHA exposure independently increased the amounts of TAG to a similar extent, the PUFA/MUFA ratio in the neutral lipid (NL) fraction was increased by 5-fold in the DHA condition. The PUFA/MUFA ratio eventually peaked in the NL but also phospholipid (PL) fractions of acid-exposed cancer cells when subjected to DHA treatment. Altogether these data indicate that the preferred accumulation of Poly-unsaturated Fatty Acid (PUFA) in acidic cancer cells sensitizes them to lipid peroxidation and associated ferroptosis. In a parallel set of experiments, we showed that this is further supported by the PUFA-induced downregulation of SCD1, an enzyme supporting MUFA synthesis.
Metabolomics of Aldh7a1-/- mice treated with lysine alpha-oxidase from Trichoderma viride
STUDY_SUMMARY
Metabolite analysis of brain, liver, and plasma isolated from Aldh7a1-/- (n=3/each) fed diets consisting of 0.9% w/w lysine and 18 ppm pyridoxine administered with PBS or 150 U/kg lysine alpha-oxidase (LOX) from Trichoderma viride. Tissues were cryo-homogenized using a liquid nitrogen cooled mortar and pestle into a fine powder and approximately 25-50 mg of each tissue was transferred to a pre-weighed homogenization tube to which ~20 2.3 mm zirconia beads and 1 mL of 80% ice cold methanol were added. Tissues were homogenized using a bead mill and cleared by centrifugation at 4C, where each of the 3-4 cycles consisted of a 60 second homogenization at 6 m/s followed by a 30 second pause. A volume equivalent to 25 mg of tissue extract was transferred to a new tube, and 250 ul of 80% methanol containing 1 ul of stable isotope-labeled internal amino acid standard mix was added. Metabolites were concentrated using a SpeedVac until dry. For plasma metabolite analysis, 10 Āµl of plasma was added to 250 Āµl of 80% methanol containing 1 Āµl of stable isotope-labeled internal amino acid standard mix, vortexed at 4C for 10 minutes, and centrifuged. 0.9 ml of supernatant was transferred to a new tube and concentrated using a SpeedVac until dry. Metabolites were reconstituted into 25-50 Āµl of water, vortexed, centrifuged, and transferred to vials for analysis by LCMS. LCMS was performed using a ZIC-pHILIC LC column coupled to a Vanquish LC and a flow gradient consisting of 10 mM ammonium carbonate in water and pure acetonitrile. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k. Data files are labeled with brain, liver, and plasma followed by a number. The number represents a mouse. Samples 1-3 are from PBS-treated mice and samples 4-6 are from LOX-treated mice.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Metabolomics analysis of kidney, brain, liver, and plasma from Aldh7a1-/- and Aldh7a1+/+ mice.
STUDY_SUMMARY
Metabolite analysis of kidney, brain, liver, and plasma isolated from Aldh7a1+/+ and Aldh7a1-/- (n=3/each) fed diets consisting of 0.9% w/w lysine and 18 ppm pyridoxine. Tissues were cryo-homogenized using a liquid nitrogen cooled mortar and pestle into a fine powder and approximately 25-50 mg of each tissue was transferred to a pre-weighed homogenization tube to which ~20 2.3 mm zirconia beads and 1 mL of 80% ice cold methanol were added. Tissues were homogenized using a bead mill and cleared by centrifugation at 4C, where each of the 3-4 cycles consisted of a 60 second homogenization at 6 m/s followed by a 30 second pause. A volume equivalent to 25 mg of tissue extract was transferred to a new tube, and 250 ul of 80% methanol containing 1 ul of stable isotope-labeled internal amino acid standard mix was added. Metabolites were concentrated using a SpeedVac until dry. For plasma metabolite analysis, 10 Āµl of plasma was added to 250 Āµl of 80% methanol containing 1 Āµl of stable isotope-labeled internal amino acid standard mix, vortexed at 4C for 10 minutes, and centrifuged. 0.9 ml of supernatant was transferred to a new tube and concentrated using a SpeedVac until dry. Metabolites were reconstituted into 25-50 Āµl of water, vortexed, centrifuged, and transferred to vials for analysis by LCMS. LCMS was performed using a ZIC-pHILIC LC column coupled to a Vanquish LC and a flow gradient consisting of 10 mM ammonium carbonate in water and pure acetonitrile. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k. Data files are labeled with brain, liver, kidney, and plasma followed by a number. The number represents a mouse. Samples 1-3 are from Aldh7a1-/- mice and samples 4-6 are from Aldh7a1+/+ mice.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Stable-isotope lysine tracing in primary mouse astrocytes and HEK293 cells.
STUDY_SUMMARY
Stable-isotope tracing using unlabeled, 13C6-, alpha-15N-, or epsilon-15N- labeled L-lysine in primary mouse astrocytes and HEK293 cells. Primary mouse astrocytes were isolated from wild-type C57BL/6J mice or Aldh7a1+/- (Het) or Aldh7a1-/- (Hom) mice and cultured for 24-hours in DMEM containing one of the above tracers and 10% dialyzed FBS. Parental, sgTomato, or sgALDH7A1 HEK293 cells were cultured for 24-hours in DMEM containing one of the above tracers and 10% dialyzed FBS. Metabolites were extracted and analyzed by LCMS.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Metabolomics analysis of N-methyl-arginine-treated HEK293 control (sgTomato) and ALDH7A1-deficient (sgALDH7A1) cells.
STUDY_SUMMARY
Metabolite analysis of HEK293 cells treated with N-methyl-arginine (NMA), an inhibitor of lysine/arginine transport. Cells were treated for 20-24 hours with varying concentration of NMA, briefly washed with ice cold 0.9% saline, and quenched with 1 mL of ice cold 80% methanol containing 1 ul of stable-isotope-labeled internal amino acid standard mix. Metabolites were concentrated using a SpeedVac until dry and analyzed by LCMS. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Investigation of age-dependent changes in the gut of honeybee workers using targeted metabolomics for SCFA determination
STUDY_SUMMARY
To determine changes in the metabolic activity of the gut microbiota, we measured the concentrations of seven short-chain fatty acids (SCFA) and lactate. All SCFA showed low concentrations in newly emerged workers. We found that acetate, propionate and butyrate are the main SCFA in the honeybee gut and peak in 5-day old and in-hive workers. Lactate showed an inverse trend with highest concentrations in 1-day old workers and a significant decrease on all subsequent days (Kruskal Wallis test, p <0.05).
Investigation of age-dependent changes in the gut of honeybee workers using targeted metabolomics for amino acid and biogenic amine determination
STUDY_SUMMARY
In the honeybee guts we measured the concentrations of 20 amino acids and 6 biogenic amines. Tryptophan and its downstream metabolite kynurenine followed the same trends across the worker development, with a significant decrease in their concentrations from day 3 onwards. Serotonin, on the other hand, was significantly increased from day 1. Tyramine and octopamine showed significant increases in concentrations (octopamine for all age groups apart from Day 1; tyramine for all age groups), while their precursor tyrosine showed significantly decreased concentrations for all age groups except experienced foragers.
Investigation of age-dependent changes in the brain of honeybee workers using targeted metabolomics for amino acid and biogenic amine determination
STUDY_SUMMARY
In the honeybee guts we measured the concentrations of 21 amino acids and 6 biogenic amines. Using a k-means clustering, the metabolites were grouped by their trends into six clusters. Tyramine and tyrosine are involved in the octopamine and dopamine biosynthesis. We found that all four metabolites were part of the same cluster, with their concentrations being significantly decreased in comparison to newly emerged workers from day 1 onwards for the entire duration of the study.
Metabolomics analysis of conditioned media from N-methyl-arginine-treated HEK293 control (sgTomato) and ALDH7A1-deficient (sgALDH7A1) cells.
STUDY_SUMMARY
Metabolite analysis of HEK293 cells treated with N-methyl-arginine (NMA), an inhibitor of lysine/arginine transport. Conditioned media was collected from cells that were treated for 20-24 hours with varying concentration of NMA, centrifuged at 21,000xg to remove cell debris, and 5 ĆĀµL was added to 250 ĆĀµL of ice cold 80% methanol containing 1 ĆĀµL of stable-isotope-labeled internal amino acid standard mix. Metabolites were concentrated using a SpeedVac until dry and analyzed by LCMS. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Metabolomics analysis of kidney, brain, liver, and plasma from Aldh7a1-/- mice administered PBS or N-methyl-arginine (80 mg/kg) via a single intraperitoneal injection.
STUDY_SUMMARY
Metabolite analysis of kidney, brain, liver, and plasma isolated from Aldh7a1-/- mice fed diets consisting of 0.9% w/w lysine and 18 ppm pyridoxine. Tissues were cryo-homogenized using a liquid nitrogen cooled mortar and pestle into a fine powder and approximately 25-50 mg of each tissue was transferred to a pre-weighed homogenization tube to which ~20 2.3 mm zirconia beads and 1 mL of 80% ice cold methanol were added. Tissues were homogenized using a bead mill and cleared by centrifugation at 4C, where each of the 3-4 cycles consisted of a 60 second homogenization at 6 m/s followed by a 30 second pause. A volume equivalent to 25 mg of tissue extract was transferred to a new tube, and 250 ul of 80% methanol containing 1 ul of stable isotope-labeled internal amino acid standard mix was added. Metabolites were concentrated using a SpeedVac until dry. For plasma metabolite analysis, 10 Āµl of plasma was added to 250 Āµl of 80% methanol containing 1 Āµl of stable isotope-labeled internal amino acid standard mix, vortexed at 4C for 10 minutes, and centrifuged. 0.9 ml of supernatant was transferred to a new tube and concentrated using a SpeedVac until dry. Metabolites were reconstituted into 25-50 Āµl of water, vortexed, centrifuged, and transferred to vials for analysis by LCMS. LCMS was performed using a ZIC-pHILIC LC column coupled to a Vanquish LC and a flow gradient consisting of 10 mM ammonium carbonate in water and pure acetonitrile. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Competitive inhibition of N-methyl-arginine uptake in HEK293 cells treated with cationic and neutral amino acids.
STUDY_SUMMARY
N-methyl-arginine uptake analysis was performed by supplementing cells with 0.5 mM of N-methyl-arginine and a vehicle control (PBS) or 5 mM of one of the following amino acids: L-lysine, D-lysine, L-ornithine, L-arginine, L-leucine, or L-alanine. In a subset of experiments, the cells were pretreated with 0.2 mM of N-ethylmaleimide (NEM) for 15-minutes to alkylate SLC7 transporters. After a 1-hour treatment with N-methyl-arginine, cells were quickly washed with ice cold saline following by quenching and metabolite extraction using 1 mL of 80% ice cold methanol containing 1 ul of stable isotope-labeled internal amino acids standards. After vortexing at 4C for 10 minutes and centrifuging, 0.9 ml of supernatant was transferred to a new tube and concentrated using a SpeedVac until dry. Metabolites were reconstituted into 25-50 Āµl of water, vortexed, centrifuged, and transferred to vials for analysis by LCMS. LCMS was performed using a ZIC-pHILIC LC column coupled to a Vanquish LC and a flow gradient consisting of 10 mM ammonium carbonate in water and pure acetonitrile. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Metabolomics and N-methyl-arginine uptake in control (siNTC) and cationic amino acid transporter knockdown HEK293 cells.
STUDY_SUMMARY
Metabolite analysis of HEK293 cells transfected with a non-targeting siRNA (siNTC) or a siRNA targeting either SLC7A1 (CAT1), SLC7A2 (CAT2), or SLC7A6 (y+LAT2). Analysis was performed four days after transfection, and knockdown was confirmed using qPCR for each experiment. Each independent experiment was performed on an independent batch of cells transfected with siRNAs. For metabolomics, cells were plated at day 2, and the media was changed to DMEM containing 10% dialyzed FBS at day 3. For NMA uptake assays, cells were plated at day 2 and NMA uptake was assessed by supplementing cells with PBS or PBS containing 0.5 mM NMA for 1 hour at day 4. All samples were processed by extracting metabolites using 1 mL of ice cold 80% methanol containing internal, isotope-labeled amino acid standards. Metabolites were concentrated using a SpeedVac until dry. For plasma metabolite analysis, 10 Āµl of plasma was added to 250 Āµl of 80% methanol containing 1 Āµl of stable isotope-labeled internal amino acid standard mix, vortexed at 4C for 10 minutes, and centrifuged. 0.9 ml of supernatant was transferred to a new tube and concentrated using a SpeedVac until dry. Metabolites were reconstituted into 25-50 Āµl of water, vortexed, centrifuged, and transferred to vials for analysis by LCMS. LCMS was performed using a ZIC-pHILIC LC column coupled to a Vanquish LC and a flow gradient consisting of 10 mM ammonium carbonate in water and pure acetonitrile. The LC was coupled to an Exploris 240 mass spectrometer operated in a polarity switching data-dependent Top 5 mode. Full MS scan parameters for both positive and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were collected at a resolution of 30k.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry & Molecular Biology
LABORATORY
Parker laboratory
LAST_NAME
Parker
FIRST_NAME
Seth
ADDRESS
950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Poplar leaf bud resin metabolomics: Seasonal profiling of leaf bud chemistry in Populus trichocarpa provides insight into resin biosynthesis
STUDY_SUMMARY
Trees in the genus Populus synthesize sticky and fragrant resins to protect dormant leaf buds during winter. These resins contain diverse phenolic metabolites, in particular hydroxycinnamate esters and methylated flavonoids. P. trichocarpa leaf bud resin is characterized by methylated dihydrochalcone aglycones. To determine how the resin profile is influenced by seasonal changes, P. trichocarpa lateral leaf bud extracts and secreted surface resin were collected monthly over a one-year cycle. THE Chemical changes monitored using nontargeted metabolomics by UPLC-HRMS. The results indicate that the dihydrochalcone content changes over the seasons and that biosynthesis occurs concomitant with bud development in the summer months. Non-targeted metabolomics data confirmed a pattern of dramatic changes in the summer, and further suggested additional periods of substantive biochemical change in the resin. While overall patterns of surface-extracted resin matched that of whole bud extracts, some of the dynamics were shifted in the surface resin samples. This study provides the basis for the use of dihydrochalcones and other identified resin components as metabolic markers for more detailed investigations of resin biosynthesis, secretion and movement to the bud surface.
Multiple, redundant carboxylic acid transporters support mitochondrial metabolism in Plasmodium falciparum
STUDY_SUMMARY
The mitochondrion of the deadliest human malaria parasite, Plasmodium falciparum, is an essential source of cellular acetyl-CoA during the asexual blood-stage of the parasite life cycle. Indeed, blocking mitochondrial acetyl-CoA synthesis leads to a hypoacetylated proteome and parasite death. We previously determined that mitochondrial acetyl-CoA is primarily synthesized from glucose-derived pyruvate by ĆĀ±-ketoacid dehydrogenases. Here, we asked if inhibiting the import of glycolytic pyruvate across the mitochondrial inner membrane would affect acetyl-CoA production and, thus, could be a potential target for novel antimalarial drug development. We selected the two predicted mitochondrial pyruvate carrier proteins (MPC1 and MPC2) for genetic knockout and isotopic metabolite tracing via HPLC-MS metabolomic analysis. Surprisingly, we observed that asexual blood-stage parasites could survive the loss of either or both proteins with only minor asexual blood-stage growth defects, despite a substantial reduction in the amount of glucose-derived isotopic labelling into acetyl-CoA. Furthermore, genetic deletion of two additional mitochondrial carboxylic acid transporters Ć¢ĀĀ DTC (di/tricarboxylic acid carrier) and YHM2 (a putative citrate/ĆĀ±-ketoglutarate carrier protein) Ć¢ĀĀ only mildly affected asexual blood-stage replication, even in the context of MPC-deficiency. Although we observed no added impact on the incorporation of glucose carbon into acetyl-CoA in these quadruple knockout mutants, we noted a large decrease in glutamine-derived label in tricarboxylic acid cycle metabolites, suggesting that DTC and YHM2 both import glutamine derivatives into the mitochondrion. Altogether, our results expose redundant routes used to fuel the mitochondria of blood-stage malaria parasites with imported carbon from two major carbon sources Ć¢ĀĀ glucose and glutamine.
Metabolic changes in imbibed seeds of malting barley produced under drought or elevated temperature
STUDY_TYPE
Barley seed phenotyping and GC-MS based metabolomic analysis
STUDY_SUMMARY
Plants of a āHana-typeā landrace (B1) were taller, flowered earlier and produced heavier, larger and more vigorous seeds that resisted ageing longer compared to a semi-dwarf breeding line (B2). Drought significantly reduced seed yield in both genotypes, and elevated temperature reduced seed size. Genotype B2 showed partial thermodormancy that was alleviated by drought and elevated temperature, in line with lower abundance of the TF ABI5, a key regulator of seed dormancy and vigour. Metabolite profiling revealed clear differences between the embryos of B1 and B2. Drought, but not elevated temperature, affected the metabolism of amino acids, organic acids, osmolytes and nitrogen assimilation, in the seeds of both genotypes.
Metabolomics of Papanicolaou Tests for the Discovery of Ovarian Cancer Biomarkers
STUDY_TYPE
Metabolomics Studies on Papanicolaou Tests
STUDY_SUMMARY
Ovarian cancer (OC) remains one of the most lethal cancers among female populations due to the vast majority of cases going undiagnosed until later stages (stage III, IV). Early detection and treatment of this malignancy provides the best prognosis, but the lack of accurate and sensitive screening tools combined with the presence of ambiguous symptoms hinders these diagnoses. In contrast, screening for cervical cancer via Papanicolaou (Pap) tests is a widespread practice with use spanning the past several decades, reducing this cancerĆ¢ĀĀs morbidity and mortality rates. Two types of Pap smear tests exist: a conventional method where cells from the ectocervix are stained on a glass slide and a liquid-based cytology method. Interestingly, previous studies show evidence of OC cells in Pap tests, suggesting that lipids shed from ovarian tumors may end up in the cervix. The goal of this study is to evaluate the practicality of using liquid-based Pap tests as biospecimens for OC screening-related metabolic studies. Cell pellets, extracted from residual Pap test fluid (RPF), from liquid-based Pap tests from 29 healthy women were analyzed via ultra-high performance liquid chromatography mass spectrometry (UHPLC-MS). This approach facilitated the detection and annotation of 453 unique lipids across 20 lipid subclasses, including ceramides, triacylglycerols, and hexosylceramides. These results demonstrated the feasibility of using an MS-based approach to analyze residual Pap test samples for the discovery of OC-related lipid biomarkers with the goal of detecting OC at early stages of the disease.
Metaboloomics analysis of the antimalarial compound WEHI-1888504 (aka compound 59) in Plasmodium falciparum (3D7) infected red blood cells
STUDY_SUMMARY
Emerging resistance to frontline antimalarials is resulting in them becoming increasingly ineffective, highlighting the need for new antimalarials. To discover new antimalarials a screen of the Janssen Jumpstarter library against the asexual stage parasite uncovered an N-acetamide indole hit class. The structure-activity relationship of this chemotype was defined by generating optimized frontrunner analogs including WJM664 with potent asexual stage activity and high metabolic stability. To determine the mechanism of action, resistance was selected and whole genome sequencing revealed mutations in PfATP4. PfATP4 was validated as the target of the N-acetamide indole class showing cross-resistance to PfATP4 drug-resistant strains, and a metabolomic signature consistent with the PfATP4 inhibitor KAE609. The indole acetamide class exhibited a fast-to-moderate rate of kill and a vacuolized asexual blood stage phenotype, phenocopying known PfATP4 inhibitors. Moreover, N-acetamide indole analogs increased cytosolic Na+ demonstrating on-target inhibition of PfATP4. N-Acetamide indole derivatives inhibited male and female gamete development and blocked transmission from infected blood to the mosquito. WJM664 exhibited low oral efficacy in an asexual stage P. berghei mouse model which may be attributed to the low potency against PbATP4, and the low aqueous solubility and Caco-2 permeability. Further optimization of these attributes is required for the N-acetamide indole class to be considered for development in a curative or transmission blocking combination therapy.
INSTITUTE
Monash University
LAST_NAME
Giannangelo
FIRST_NAME
Carlo
ADDRESS
381 Royal Parade, Parkville, Victoria, 3052, Australia
Unraveling cysteine deficiency-associated rapid weight loss
STUDY_SUMMARY
Forty percent of the US population and 1 in 6 individuals worldwide are obese, with the incidence surging globally. Various dietary interventions, including carbohydrate, fat and more recently amino acid restriction, have been explored to combat this epidemic. We investigated the impact of removing individual amino acids on the weight profiles of mice. Here, we show that conditional cysteine restriction resulted in the most dramatic weight loss when compared to essential amino acid restriction, amounting to 30% within one week, which was readily reversed. We found that cysteine deficiency activated the integrated stress response and oxidative stress response, which amplify each other, leading to the induction of GDF15 and FGF21, which partly explained the phenotype. Surprisingly, we observed lower tissue coenzyme A (CoA), which has been considered to be extremely stable10, resulting in reduced mitochondrial functionality and metabolic rewiring. This results in energetically inefficient anaerobic glycolysis and defective TCA cycle, with sustained urinary excretion of pyruvate, orotate, citrate, Ī±-ketoglutarate, nitrogen rich compounds, and amino acids In summary, our investigation reveals that cysteine restriction, by depleting GSH and CoA, exerts a maximal impact on weight loss, metabolism, and stress signaling compared to other amino acid restrictions. These findings suggest novel strategies for addressing a range of metabolic diseases and the growing obesity crisis.
Enoyl-acyl carrier protein reductase (ZmENR1) expression level is critical for de novo fatty acid biosynthesis and anther cuticle and pollen exine formation in maize.
STUDY_SUMMARY
Lipid metabolism is very important for plant male reproduction. Many lipid metabolism genes, male sterility (GMS) genes play a role in the endoplasmic reticulum of anther tapetum, but little is known about GMS genes involved in the de novo synthesis of fatty acids in anther tapetum. Using gene mapping, we cloned a new GMS gene, enoyl-acyl carrier protein (ACP) reductase (ZmENR1). enr1 (mutation of enoyl-ACP reductase gene) has early tapetum degradation, anther epidermis and pollen exine defects. In order to further study the function of ZmENR1, we compared the changes of lipid content in anthers of wild-type (WT) and enr1 mutants at maturity. The expression of ZmENR1 significantly affected the contents of cutin, wax and total fatty acids (TFA) in anthers. In addition, enr1 mutant reduced the expression of cutin/wax and sporopollen related genes to inhibit the formation of anther epidermis and pollen exine. Therefore, our researches provide a new insight into the synthesis of fatty acids mediated by ZmENR1, and provide an insight into the molecular mechanism of ZmENR1 regulating maize male sterility.
INSTITUTE
University of Science and Technology Beijing
LAST_NAME
Zhang
FIRST_NAME
Shaowei
ADDRESS
30 XUEYUAN ROAD, HAIDIAN DISTRICT, BEIJING 100083, CHINA
Differentially regulated metabolites in intestinal epithelial cells from Hmga1flox/flox and Hmga1ā³IEC mice
STUDY_SUMMARY
Dysregulations of cell metabolism, such as elevated aerobic glycolysis and increased fatty acid metabolism, play a critical role in the tumorigenesis of colorectal cancer (CRC). To determine whether HMGA1 promotes CRC through regulating cell metabolism, we performed an untargeted metabolomics using intestinal epithelial cells (IECs) from Hmga1flox/flox and Hmga1ā³IEC mice. In comparison with the Hmga1flox/flox mice, the Hmga1ā³IEC mice displayed 187 up-regulated metabolites and 120 down-regulated metabolites in IECs. Enrichment analysis of the metabolites showed that these differential metabolites mainly enriched in 20 pathways. Fatty acid synthesis was the most significantly deregulated signal pathway in HMGA1 deficient IECs. We separated the altered metabolites into up-regulated and down-regulated groups, and showed the top five classes of metabolites in up-regulated and down-regulated groups in heatmaps. The results showed that fatty acyl metabolites were mainly down-regulated in HMGA1-deficient IECs.
INSTITUTE
Henan University
LAST_NAME
Xu
FIRST_NAME
Zhi-Xiang
ADDRESS
Jinming Road, Kaifeng, Henan province, 475000, China
Differentially regulated metabolites in colorectal cancer from Rosa26Hmga1/+ and Hmga1IEC-OE/+mice
STUDY_SUMMARY
Dysregulations of cell metabolism, such as elevated aerobic glycolysis and increased fatty acid metabolism, play a critical role in the tumorigenesis of colorectal cancer (CRC). To determine whether HMGA1 promotes CRC through regulating cell metabolism, we performed an untargeted metabolomics using intestinal epithelial cells (IECs) from Hmga1flox/flox and Hmga1ā³IEC mice firstly. The results showed that fatty acyl metabolites were mainly down-regulated in HMGA1-deficient IECs. Further, untargeted metabolomics analysis on tumors derived from the Hmga1IEC-OE/+ mice also showed that fatty acyl metabolites were mainly up-regulated by HMGA1 overexpression. FA is a major component of structurally complex lipids and is one of the most basic categories of bio-lipids. We analyzed all lipids-related metabolites and found the free fatty acids were significantly up-regulated in tumors derived from the Hmga1IEC-OE/+ mice. Thus, untargeted metabolomics measurement of IECs (CRCs) from both Hmga1 conditional knockout (Hmga1ā³IEC) and knock-in (Hmga1IEC-OE/+) mice demonstrates a positive correlation between expression of HMGA1 and activation of fatty acid synthesis.
INSTITUTE
Henan University
LAST_NAME
Xu
FIRST_NAME
Zhi-Xiang
ADDRESS
Jinming Road, Kaifeng, Henan province, 475000, China
Hepatocytes transform fructose into polar metabolites that are selectively utilized by cancer cells
STUDY_SUMMARY
The goal of this project was to determine if hepatocytes can metabolically transform fructose into polar metabolites that CaSki cancer cells can uptake and metabolize. The CaSki cells do not metabolize fructose directly to much extent, but the liver robustly metabolizes fructose. Therefore, we cultured primary hepatocytes from C57BL/6 mice in [U-13C] fructose as the only sugar source for 24 hours to form hepatocyte-conditioned media. We then transferred this media to CaSki cells for 24 hours to form CaSki-conditioned media. LC-MS profiling of polar 13C-labeled metabolites of hepatocyte-conditioned media and CaSki-conditioned media revealed that 13C-labeled glucose and 13C-labeled aspartate are released from hepatocytes and are up taken by CaSki cells.
Hepatocytes transform fructose into lipids that can metabolized by cancer cells
STUDY_SUMMARY
Fructose is known to be metabolized by the liver to produce lipids that can circulate in the systemic circulation. Metabolism of fructose by CaSki cancer cells, on the other hand, is minimal. Therefore, we wished to understand if fructose metabolism in hepatocytes resulted in the release of lipids that CaSki cells could then uptake and metabolize to support their growth. Therefore, we formed fresh media which contained fructose as the only sugar source in the media. We cultured primary hepatocytes from C57BL/6 mice in this fresh media to form hepatocyte-conditioned media. We then transferred this hepatocyte-conditioned media to CaSki cells to form CaSki-conditioned media. LC-MS analysis on reverse-phase chromatography of fresh media, hepatocyte-conditioned media, and CaSki-conditioned media revealed that several lysophosphatidylcholine species are released from hepatocytes and up taken by CaSki cells.
Dietary sugar supplementation in mice enhances the availability of many circulating lipids including lysophosphatidylcholines
STUDY_SUMMARY
Sugar consumption is known to enhance de novo lipogenesis, but it is still not comprehensively known which lipid species are altered secondary to sugar consumption. To understand the effects of sugar consumption on circulating lipids, we supplemented C57BL/6 mice on a modified chow diet that contained no free sugars with either normal water, 10% glucose water, 10% fructose water, or 20% HFCS water. A comparative analysis of the serum between the four groups revealed 128 lipids that had statistically significant changes among all four conditions. We observed that LPCs were a particularly altered lipid class among the lipids we detected. Notably, 16:1 and 18:1 LPCs were increased severalfold in either the 10% glucose, 10% fructose, or 20% HFCS conditions relative to the normal water condition.
Lipidomic Profiling of a Preclinical Model of Streptozotocin induced Diabetic Cardiomyopathy
STUDY_SUMMARY
Lipidomic profiling was conducted on plasma from ~22 week old, male FVB/NJ mice that were subjected to five consecutive daily intraperitoneal (i.p.) injections of citrate buffer vehicle (non-diabetic) or STZ (55 mg/kg in 0.02 mol/L citrate buffer, pH 4.5) to induce diabetes at 6 weeks of age. Mice were monitored for 16 weeks of untreated diabetes.
Comparison of the capillary and venous blood lipidomes- validation of the Tasso SST capillary blood collection device for circulating lipid biomarker analysis
STUDY_SUMMARY
Venipuncture of the upper extremities continues to be the standard method of whole blood collection used for lipidomics, but self-administered Tasso+TM capillary sampling is now an accepted clinical method. Wide adoption of the capillary blood test method for lipid biomarker measurements would require distinguishing between the universal global lipid levels variations in venous versus capillary blood. As lipids are biomarkers for a number of disorders, and because they may provide valuable information on nutritional and toxicological status, we view this study as an example of broader biomarker assay consistency. Because exchange of blood components including lipids occurs in capillaries, the capillary and venous blood lipidomes might be different. Here we compared the lipidomes of Tasso+ drawn capillary blood plasma to venous blood plasma by high-resolution mass spectrometry based lipidomics, in 10 human subjects. While we found substantial inter individual variability when comparing lipid profiles, comprehensive statistical analysis with cross validation and multiple testing adjustments show that the lipid composition of paired Tasso+ and venous blood samples do not significantly differ (adjusted p-value > 0.05). A linear regression model with Spearman correlation analysis also showed a significant-to-perfect level (r = 0.95-0.99) of concordance between the matched samples. With the exception of monoacylglycerols (MG) and cardiolipins (CL), every class of lipid biomarkers also demonstrated substantially high degree of correlation (0.9-0.99) between paired venous and capillary blood. These findings demonstrate that capillary and venous blood plasma lipidomes are similar; indicating that self-administered capillary sampling could be used interchangeably with standardized venipuncture in clinical research settings.
INSTITUTE
University of Arkansas for Medical Sciences
DEPARTMENT
Arkansas Children Nutrition Center, University of Arkansas for Medical Sciences
Multi āomics indicate depth-discrete partitioning of nitrogen metabolism in a toxic Planktothrix rubescens bloom in the winter water column
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Limnological sampling efforts have traditionally focused on the summer water column of larger lakes. For example, hundreds of researchers visit prominent systems like the Laurentian Great Lakes to investigate summer cyanobacterial blooms. Concurrently, the recent advancement of meta-omics techniques has led to a surge in molecular studies within these systems. However, these approaches have created several knowledge gaps: smaller, regional lakes remain largely unstudied, the winter ecology of lakes is poorly understood, and meta-omics techniques often fail to make meaningful scientific contributions when used in isolation. To address these gaps, we conducted a combined metagenomic and metabolomic field study focused on a local community concern: the winter pink discoloration of Meadās Quarry (Knoxville, TN). This case study was designed and executed by seven graduate students at the University of Tennessee-Knoxville enrolled in the course MICR 669: Advanced Techniques in Field Microbiology (Spring 2023). Utilizing a $3,000 budget, the team carried out in-depth meta-omics research. Our metagenomic analyses and phylogenetic confirmations identified a bloom of the pink cyanobacterium Planktothrix rubescens as the primary cause. Physiochemical measurements revealed that the bloom was concentrated in the light-limited (<1 Ī¼mol photons mā»Ā² sā»Ā¹) metalimnion of the cold (<14Ā°C) water column. Additionally, microcystin-LR levels exceeded the U.S. EPA limit for recreational exposure (8 Ī¼g Lā»Ā¹). Untargeted metabolomics indicated that P. rubescens communities were differentially partitioning nitrogen metabolism strategies by depth (0.5 m vs. 2.0 m). This study highlights the previously underappreciated roles of arginine biosynthesis and pyrimidine metabolism in the ecological success of P. rubescens. We propose a novel ecological hypothesis regarding the proliferation of this harmful cyanobacterial bloom former in the light-limited metalimnion. Overall, our findings suggest that meaningful meta-omics research can be conducted locally with limited funding as part of a course-based curriculum involving at least seven graduate students. Furthermore, we demonstrate that the synergistic use of multiple meta-omics techniques is feasible on a small scale while yielding significant scientific impact.
NRF2 supports non-small cell lung cancer growth independently of CBP/p300-enhanced glutathione synthesis
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
Nuclear factor erythroid 2-related factor 2 (NRF2) is a stress responsive transcription factor that is mutationally activated in a subset (~25%) of clinically-aggressive non-small cell lung cancers (NSCLC). Mechanistic insight into drivers of the NRF2 dependency remains poorly understood. Here, we defined a novel NRF2 target gene set linked to NRF2-dependency in cancer cell lines, and observed that a significant portion of these genes is devoid of promoter-proximal NRF2. Using integrated genomic analyses, we characterized extensive NRF2-dependent enhancer RNA (eRNA) synthesis and NRF2-mediated H3K27ac deposition at proximal and distal enhancer regions regulating these genes. While CBP/p300 is a well-validated direct interaction partner of NRF2 with prominent functions at enhancers, we report that this interaction is not required for NRF2-dependent NSCLC cell growth, indicating that NRF2 can sustain sufficient transcriptional activity in the absence of CBP/p300 coactivation. Broad metabolic profiling established a primary role for CBP/p300 in NRF2-dependent accumulation of glutathione and glutathione-related metabolites. While redox homeostasis via enhanced glutathione production is commonly associated with the normal physiological role of NRF2, collectively our results suggest that NRF2-dependent cancer cell growth does not require this enhanced glutathione production.
Metabolomic profiling of normal and fibrotic mouse lungs
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
C57BL/6 wild-type (8-10-week old) mice were i.t. injected with saline or bleomycin. 1 to 4 weeks after injection, bronchoalveolar lavage fluid (BALF) was collected with a saline solution. The BALFs samples were processed for untargeted metabolomics.
Glucagon is a major controller of metabolism and malignancy in pancreatic neuroendocrine tumour (pNET) cells
STUDY_SUMMARY
Cancer metabolism research has studied the relationship between cellular bioenergetics, biosynthesis, and tumour progression. However, the impact of systemic metabolism and diet on tumour evolution is less understood. This study delves into the role of glucagon in the regulation of cancer cell features and the metabolic remodelling in the presence and absence of glucose, in pancreatic neuroendocrine tumours (pNETs). To fulfil the experiments pNET cell lines (BON-1 and QGP-1) and non-malignant pancreatic Ī±-TC1 cell line were used as models. Results showed that pNET cells responded differently to glucose deprivation than Ī±-TC1 cells. Specifically, pNET cells upregulated GCGR in the absence of glucose, while Ī±-TC1 cells did so in high-glucose conditions. Glucagon activated the MAPK/ERK pathway, especially in pNET cells without glucose and Ī±-TC1 cells with high glucose conditions. Glucagon enhanced metabolic viability, proliferation, and migration in pNET cells under glucose-deprived conditions and hyperglucagonaemia. Meanwhile, in Ī±-TC1 cell line, glucagon modulated these features under high-glucose conditions and physiological glucagon levels. Metabolic analysis using NMR revealed differences in amino acid levels and metabolic processes based on glucose availability between cell lines. Furthermore, gene expression analysis showed that glucose conditions largely influenced gene expression patterns in both pNETs and Ī±-TC1. Interestingly, QGP-1 and Ī±-TC1 produced glucose in no-glucose conditions, and glucagon upregulated glucose production in Ī±-TC1. Hence gluconeogenesis may be beneficial for some pNET subsets, and it may open new perspectives to novel metabolism-based strategies to clinically manage pNETs. Furthermore, the production of glucose by Ī±-cells was not described so far and is a step forward in endocrinology and systemic metabolism. Lastly, an association between GCGR expression and malignancy was observed, despite the decreased expression in tumours comparing with the normal tissue. A negative correlation between GCGR and GLP-1R expression was observed, highlighting the putative biological role of glucagon in pNETs malignancy.
INSTITUTE
ITQB NOVA
LAST_NAME
GonƧalves
FIRST_NAME
LuĆs
ADDRESS
Avenida Republica, Oeiras, Not USCanada, 2780-157 Oeiras, Portugal
Glutaminase 1 (GLS1)-based therapy targets glutamine reliance in glioblastoma (GBM)- cell line assays supernants
STUDY_SUMMARY
Glioblastomas (GBMs) are the most lethal central nervous system (CNS) neoplasms, accounting for this the inexistence of specific therapies. Glutamine plays a key role in CNS physiology, and metabolic adjustments have been reported in GBM, which are considered glutamine-addicted. Therefore, our goal was to affect GBM cells by decreasing the systemic bioavailability of glutamine with a glutaminase (GLS1)-based therapy. In order to understand how Glutaminase treatment affects the metabolism of GBM cell lines (U251 and U-87MG), we exposed these cell lines to GLS1 for 24 h and the conditioned media (supernatants) and cell extracts (aqueous and organic phase) were further analyzed by 1H-NMR. The in vitro results showed that the GBM cell lines were indeed glutamine-dependent, triggering a metabolic remodeling upon the presence/absence of glutamine. GLS1 treatment increased cell death, decreased cell proliferation, and induced U-87MG invasion, due to glutamine scarcity. This intervention triggered a metabolic shift, evident in altered gene expression and metabolite profiles in cell extracts and conditioned media. Results from the in vivo orthotopic xenograft mouse model of GBM showed an increase in overall survival (OS) with the GLS1 treatment, along with lower cachexia. Moreover, metabolomicsā multivariate analysis of serum collected during tumor induction and the therapeutic course showed statistically different metabolic profiles of GLS1-treated mice midst treatment, which could help monitor the therapy response.
INSTITUTE
ITQB NOVA
LAST_NAME
GonƧalves
FIRST_NAME
LuĆs
ADDRESS
Avenida Republica, Oeiras, Not USCanada, 2780-157 Oeiras, Portugal
To examine whether a molecule associated with metabolic liver zonation has an effect on systemic metabolism, we first administer adenoviruses expressing Rspo3 or LacZ (control) into the murine liver. Then, by performing metabolome analyses, we compare the results of Rspo3-mice with those of LacZ-mice.
Comparison of Machine Learning Models for Metabolomic-Based Clinical Prediction of Preterm Birth
STUDY_SUMMARY
Machine learning (ML), with advancements in algorithms and computations, is seeing an increased presence in life science research. This study investigated several ML models' efficacy in predicting preterm birth using untargeted metabolomics from serum collected during the third trimester of gestation. Samples from 48 preterm and 102 term delivery mothers (1:2 ratio) from the All Our Families Cohort (Calgary, AB) were examined. Selected ML applications were used to examine the small-scale clinical dataset for both model performance and metabolite interpretation. Model performance was evaluated based on confusion matrices, receiver operating characteristic curves, and feature importance rankings. Conventional linear models, like Partial Least Squares Discriminant Analysis (PLS-DA) and linear logistic regression, showed moderate predictive potential with AUC-ROC around 0.60. Non-linear models, including Extreme Gradient Boosting (XGBoost) and artificial neural networks, had marginally improved predictive accuracy and strength. Resampling by bootstrapping was also examined. Among all MLs, bootstrap resampling enhanced XGBoost's performance the most, improving AUC-ROC (0.85, 95% CI:0.574-0.995, p<0.001) for the best fitted model. Feature importance analysis by Shapley Additive Explanations analysis consistently identified acylcarnitines and amino acid derivatives as significant metabolites. Findings underscored the complexity of modeling preterm birth prediction, suggesting a trial-and-error approach for model selection.
Mitochondria complex III is essential for IL-10 secretion in macrophages independent of respiration
STUDY_SUMMARY
Mitochondrial electron transport chain (ETC) function is linked to macrophage function, however, mechanisms underlying mitochondrial ETC control of macrophage immune responses are not fully understood. We used genetic tools to examine the necessity of mitochondrial electron transport chain (ETC)-dependent respiration and the production of reactive oxygen species (mtROS) in macrophage immune responses. Here we report that mitochondrial ETC complex III (CIII)-deficient mouse macrophages, which have impaired macrophage respiration and mtROS production, exhibit increased susceptibility to influenza A virus and LPS-induced endotoxic shock. Mitochondrial CIII-deficient bone marrow-derived macrophages (BMDMs) produced IL-10 but exhibit dampened release following TLR3 or TLR4 stimulation in vitro. Surprisingly, restoring mitochondrial respiration without generating mtROS in mitochondrial CIII-deficient macrophages with Ciona intestinalis alternative oxidase (AOX) is not sufficient to reverse the increased vulnerability to LPS-induced endotoxic shock or rescue IL-10 release in vitro. However, activation of PKA, a mtROS-responsive pathway, was sufficient to rescue BMDM IL-10 release following lipopolysaccharide (LPS) stimulation. Additionally, mitochondrial CIII impairment in macrophages does not affect canonical responses to interleukin-4 (IL-4) stimulation. Thus, our results highlight the necessity of mitochondrial ROS but not respiration in the release of IL-10.
INSTITUTE
Northwestern University, Feinberg School of Medicine
Metabolomics of Drosophila melanogaster under manganese (Mn) exposure (head)
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
Parkinsonās disease (PD) is the most common motor disorder, for which environmental factors such as manganese (Mn) exposure may play a major etiologic role. In this study, we aim to identify metabolic pathways related to Mn-induced parkinsonism and discover therapeutic targets for manganism and potentially for PD. We developed an in vivo model of acute Mn toxicity in male Drosophila melanogaster (0, 1, 10, and 30 mM of Mn2+ for 10 days) to recapitulate PD hallmarks, spanning behavior (climbing, locomotor assay), histology (H&E staining of neurons, MitoTracker assay), and mitochondrial function (Seahorse assay). High-coverage metabolomics of flies were conducted at an earlier point of dosage (Day 5), to identify early biochemical modulators before motor symptoms surfaced. Fly heads and bodies were extracted and analyzed using liquid chromatographyāquadrupole-orbitrap mass spectrometry, following informatics approaches for compound identification and pathway analysis. The uploaded is the untargeted metabolomics component which contains raw data, alignment tables, and associated method details.
INSTITUTE
Columbia University
DEPARTMENT
Department of Environmental Health Sciences
LABORATORY
The Miller Lab
LAST_NAME
Lai
FIRST_NAME
Yunjia
ADDRESS
630 West 168th Street, P&S 16-421, New York, NY 10032
EMAIL
yunjia.lai@outlook.com
PHONE
9194805489
NUM_GROUPS
3
TOTAL_SUBJECTS
150; five samples per group; 10 subjects per sample
NUM_MALES
150; five samples per group; 10 subjects per sample
Metabolomics of Drosophila melanogaster under manganese (Mn) exposure (body)
STUDY_TYPE
untargeted metabolomics
STUDY_SUMMARY
Parkinsonās disease (PD) is the most common motor disorder, for which environmental factors such as manganese (Mn) exposure may play a major etiologic role. In this study, we aim to identify metabolic pathways related to Mn-induced parkinsonism and discover therapeutic targets for manganism and potentially for PD. We developed an in vivo model of acute Mn toxicity in male Drosophila melanogaster (0, 1, 10, and 30 mM of Mn2+ for 10 days) to recapitulate PD hallmarks, spanning behavior (climbing, locomotor assay), histology (H&E staining of neurons, MitoTracker assay), and mitochondrial function (Seahorse assay). High-coverage metabolomics of flies were conducted at an earlier point of dosage (Day 5), to identify early biochemical modulators before motor symptoms surfaced. Fly heads and bodies were extracted and analyzed using liquid chromatographyāquadrupole-orbitrap mass spectrometry, following informatics approaches for compound identification and pathway analysis. The uploaded is the untargeted metabolomics component which contains raw data, alignment tables, and associated method details.
INSTITUTE
Columbia University
DEPARTMENT
Department of Environmental Health Sciences
LABORATORY
The Miller Lab
LAST_NAME
Lai
FIRST_NAME
Yunjia
ADDRESS
630 West 168th Street, P&S 16-421, New York, NY 10032
EMAIL
yunjia.lai@outlook.com
PHONE
9194805489
NUM_GROUPS
3
TOTAL_SUBJECTS
150; five samples per group; 10 subjects per sample
NUM_MALES
150; five samples per group; 10 subjects per sample
Trifluoroacetate reduces plasma lipid levels and the development of atherosclerosis in mice
STUDY_SUMMARY
Trifluoroacetate (TFA) has been assumed to be an innocuous counterion (to cationic amino acid side chains) present in countless synthetic bioactive peptides and a few FDA-approved therapeutics. We show here that TFA is in fact bioactive and causes dramatic biological effects in multiple strains of mice and cultured human and rat liver cells. In high-fat diet (HFD)-fed low-density lipoprotein receptor-null (LDLr-/-) mice, TFA reduces the levels of plasma cholesterol, triglycerides, and the development of atherosclerotic lesions following either oral or intraperitoneal administration. These physiological effects were observed with TFA alone, or with TFA present as a counterion of a variety of short, unrelated synthetic peptide sequences. Mechanistic investigations including RNA-seq, confocal microscopy, western blotting, metabolomics, proteomics, pharmacokinetics, and biochemical assays indicated that TFA induces peroxisome proliferation by activating peroxisome proliferator-activated receptor (PPAR)-alpha. We confirmed that TFA also caused peroxisome proliferation and downstream phenotypic effects in cultured human and rat liver cells, wild-type C57/Bl mice, and apolipoprotein E-null (apoE-/-) mice, leading to anti-atherosclerotic effects in the latter strain. Given that TFA is a counterion in many peptides employed in early research and development settings, these findings raise the possibility that TFA may be confounding or contributing to phenotypic changes observed in many studies involving peptides. Although our studies suggest that TFA or its analogues might have therapeutic applications, it should be noted that TFA is also a persistent environmental contaminant that is found at high levels in humans relative to other polyfluoroalkyl substances (PFAS), and is a major metabolite following treatment of patients with common inhaled anesthetics, suggesting that the biological effects reported here could have other implications for human health.
INSTITUTE
Scripps Research
LAST_NAME
Tang
FIRST_NAME
Wei
ADDRESS
10550 North Torrey Pines Road, La Jolla, California, 92037, USA
An Organism-Level Quantitative Flux Model of Energy Metabolism in Mice
STUDY_SUMMARY
Mammalian tissues feed on nutrients in the blood circulation. At the organism-level, mammalian energy metabolism comprises of oxidation, storage, interconverting, and releasing of circulating nutrients. Though much is known about the individual processes and nutrients, a holistic and quantitative model describing these processes for all major circulating nutrients is lacking. Here, by integrating isotope tracer infusion, mass spectrometry, and isotope gas analyzer measurement, we developed a framework to systematically quantify fluxes through these metabolic processes for 10 major circulating energy nutrients in mice, resulting in an organism-level quantitative flux model of energy metabolism. This model revealed in wildtype C57BL/6J mice that circulating nutrients have more dominant metabolic cycling fluxes than their oxidation fluxes, with distinct partition between cycling and oxidation flux for individual circulating nutrients. Applications of this framework in obese mouse models showed on a per animal basis extensive elevation of metabolic cycling fluxes in ob/ob mice, but not in diet-induced obese C57BL/6J mice. Thus, our framework describes quantitatively the functioning of energy metabolism at the organism-level, valuable for revealing new features of energy metabolism in physiological and disease conditions.
Development of Food Material Source Technology for Future Alternative Meats (Including Cultured Meat)
STUDY_SUMMARY
The growing demand for sustainable food sources has accelerated the development of cultured meat as an alternative to traditional meat products. This study aims to predict the safety and nutritional equivalence of cultured meat compared to conventional meat using a comprehensive metabolomics approach. In this study, we conducted a comparative metabolomic analysis of conventional chicken meat, muscle satellite cells, and differentiated cells. Our findings reveal that while the overall metabolic profiles of cultured and original meats are comparable, significant differences are observed in specific metabolites. Notably, metabolites associated with nutrient metabolism and synthesis display substantial variations among the samples. These differences suggest that the nutritional content of cultured meat may differ from that of traditional meat, potentially affecting its dietary value. Despite these differences in metabolic profiles, our analysis indicates that there is no significant impact on the safety of cultured meat itself. The safety of cultured meat remains within acceptable limits, and no adverse health risks were identified in the context of this study. However, the observed variations in nutrient-related metabolites highlight the need for further investigation to fully understand their implications. Our study contributes to the ongoing evaluation of cultured meat as a viable and safe alternative in the pursuit of sustainable food sources.
Rapid modulation of gut microbiota composition by hypothalamic circuits in mice
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Whether the hypothalamus is able to influence gut microbiota composition remains enigmatic. Here, we present a proof-of-concept study designed to unravel this challenging question. To this aim, we centrally administered leptin or ghrelin to male mice. Subsequently, we conducted microbiota composition analysis throughout the gut using 16S rRNA gene sequencing. Our results showed that these brain interventions significantly changed the gut microbiota in an anatomical and short-term (two to four hours) fashion. Metabolomics analysis was performed on the intestinal content of the duodenum 4h after leptin injection, revealing changed metabolite levels.
Comparison of metabolic responses in murine bone marrow derived macrophages upon LPS treatment versus following the engulfment of live or dead E coli.
STUDY_SUMMARY
Global metabolomics and 13C-tracing analysis of murine bone marrow derived macrophages exposed to live or heat-killed uniformly 13C-labelled E. coli in a trans-well system for 6h or 18h. Global metabolomics was performed in parallel on murine BMDMs treated with 500 ng/ml LPS for 18h.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Investigation of global metabolites and E coli-derived metabolites following engulfment of dead bacteria by wild type or RagAGTP-expressing bone marrow derived macrophages.
STUDY_SUMMARY
Global metabolomics and 13C-tracing analysis of wild type or RagAGTP-expressing bone marrow derived macrophages exposed to heat-killed uniformly 13C-labelled E. coli in a trans-well system for 6h or 18h.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Global metabolomics and tracing of E coli-derived metabolites following engulfment of heat-killed bacteria by wild type or RagAGTP-deficient bone marrow derived macrophages.
STUDY_SUMMARY
Global metabolomics and 13C-tracing analysis of wild type or RagA-deficient bone marrow derived macrophages exposed to heat-killed uniformly 13C-labelled E. coli in a trans-well system for 6h or 18h.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Global metabolomics and tracing of E coli-derived metabolites following engulfment of live or heat-killed bacteria by bone marrow derived macrophages expressing constitutively active mutants of Kras or Hras
STUDY_SUMMARY
We sought to identify possible mechanisms governing the recycling of microbially derived nutrients produced by the degradation of killed E coli in macrophages. Recent studies have demonstrated that extracellular protein utilization supports the cellular metabolism of cancer cells expressing activated Ras pathways. Here we tested the involvement of Kras and Hras pathway in metabolic recycling of microbial nutrients upon phagocytosis by macrophages using global and 13C-tracing metabolomics in bone marrow derived macrophages exposed to 13C-labeled E coli.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Global metabolomics and tracing of 13C-labeled metabolites of bone marrow derived macrophages following engulfment of dead 13C-labeled Citrobacter rodentium, Listeria innocua, Staphilococcus aureus, or Pseudomonas aeruginosa.
STUDY_SUMMARY
Global metabolomics and 13C-tracing analysis of bone marrow derived macrophages exposed to heat-killed uniformly 13C-labelled Citrobacter rodentium, Listeria innocua, Staphilococcus aureus or Pseudomonas aeruginosa for 4h.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
13C-tracing metabolomics of peritoneal macrophages 1hour and 2hours after engulfment of 13C-labeled heat-killed E coli
STUDY_SUMMARY
13C-tracing analysis at 1h and 2h of peritoneal macrophages isolated from C57BL6/N mice injected with uniformly 13C-labelled and DAPI labeled heat-killed E. coli.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
13C-tracing metabolomics of peritoneal macrophages 1hour after engulfment of 13C-labeled heat-killed E coli vs untreated peritoneal macrophages
STUDY_SUMMARY
13C-tracing analysis of peritoneal resident macrophages isolated from C57BL6/N mice injected or not with uniformly 13C-labelled heat-killed E. coli, 1 hours post-infection.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Candida auris planktonic and dispersed cells Chromatographic analysis using GC-MS
STUDY_SUMMARY
The emerging Candida auris (C. auris) has caused several outbreaks globally. While several studies explored the resistant biofilm formed by C. auris, little is known regarding the cells dispersed following biofilm formation. Herein, I investigated and characterized the cells dispersed from C. auris biofilms. Cells dispersed from biofilm developed in 96 well plate were isolated and counted. GC-MS analysis indicated that dispersed cells exhibit altered metabolic profile that enhance cells survivability under stress and nutrient limit condition. The presented study is the first to explore C. auris dispersed cells and indicated that they are not able to revert to the planktonic mode once released from the biofilm.
SLC7A5 is required for cancer cell growth in arginine-limited conditions
STUDY_TYPE
LCMS quantitative analysis
STUDY_SUMMARY
Polar metabolite abundance studies from A375m human melanoma cells containing native levels of SLC7A5 (WT), CRISPR-Cas9 induced knockout of SLC7A5 (KO) or lentiviral induced overexpression of SLC7A5 (OE). Studies used RPMI media with no arginine. In place of arginine was 110uM citrulline.
Kupffer cells control neonatal hepatic glucose metabolism via Igf1 signaling - lipidomics analysis of postnatal day 0 murine livers after macrophages depletion using Spi1 conditional KO
STUDY_TYPE
lipidomics analysis of P0 livers from mice
STUDY_SUMMARY
During perinatal development, liver metabolism is tightly regulated to ensure energy supply for the newborn. Before birth, glycogen is stored in hepatocytes and later metabolized to glucose, meeting the energy demands of the neonate. Shortly after birth, lipogenesis begins, driven by the transcriptional activation of enzymes involved in fatty acid oxidation. These processes are thought to be largely regulated by systemic insulin and glucagon levels. However, the role of liver-derived local factors in neonatal hepatocyte metabolism remains unexplored. Kupffer cells (KCs), the liverĆ¢ĀĀs resident macrophages, colonize the fetal liver early in embryogenesis and support liver metabolism in adulthood. Yet, whether KCs influence neonatal hepatocyte metabolism is unknown. Here, using conditional knockout mouse models targeting macrophages, we demonstrate that yolk sac-derived KCs play a critical role in hepatocyte glycogen storage and function by regulating the TCA cycle - a role that monocyte-derived KC-like cells cannot substitute. in order to check the lipid levels after birth, Newborn pups were collected and the different lipid species were assessd by lipidomics analysis using mass spectrometer.
INSTITUTE
University of Bonn
DEPARTMENT
Developmental Biology of the Immune System, The Life & Medical Sciences Institute (LIMES)
Kupffer cells control neonatal hepatic glucose metabolism via Igf1 signaling - C13 glucose traced metabolites analysis of postnatal day 0 livers after macrophages depletion
STUDY_TYPE
metabolites analysis of P0 livers from mice
STUDY_SUMMARY
During perinatal development, liver metabolism is tightly regulated to ensure energy supply for the newborn. Before birth, glycogen is stored in hepatocytes and later metabolized to glucose, meeting the energy demands of the neonate. Shortly after birth, lipogenesis begins, driven by the transcriptional activation of enzymes involved in fatty acid oxidation. These processes are thought to be largely regulated by systemic insulin and glucagon levels. However, the role of liver-derived local factors in neonatal hepatocyte metabolism remains unexplored. Kupffer cells (KCs), the liverĆ¢ĀĀs resident macrophages, colonize the fetal liver early in embryogenesis and support liver metabolism in adulthood. Yet, whether KCs influence neonatal hepatocyte metabolism is unknown. Here, using conditional knockout mouse models targeting macrophages, we demonstrate that yolk sac-derived KCs play a critical role in hepatocyte glycogen storage and function by regulating the TCA cycle - a role that monocyte-derived KC-like cells cannot substitute. n this experiment, C13-labeled glucose is administered to track glucose metabolism. The labeled glucose is metabolized, and its incorporation into downstream metabolites is analyzed using techniques such as mass spectrometry . This approach enables the identification and quantification of metabolic pathways involving glucose, such as glycolysis, gluconeogenesis, and the TCA cycle, providing insights into glucose utilization and related metabolic processes.
INSTITUTE
University of Bonn
DEPARTMENT
Developmental Biology of the Immune System, The Life & Medical Sciences Institute (LIMES)
Non-Targeted Analysis of the Chemical Exposome During Pregnancy Using High-Resolution Mass Spectrometry
STUDY_TYPE
Research
STUDY_SUMMARY
The widespread use of human-made chemicals in daily life leads to chronic exposure to complex mixtures of potentially harmful substances. To investigate chemical exposures among pregnant women in New York City, we applied a non-targeted analysis (NTA) workflow to 83 paired prenatal placenta and serum samples (including 30 preterm birth pairs) collected through the New York University Childrenās Health and Environment Study. Our objectives were to: i) characterize chemical exposures in this population, ii) compare chemical profiles between placenta and serum samples as well as between preterm and term births, and iii) examine potential associations between exogenous chemicals and endogenous metabolites.
INSTITUTE
NYU Grossman School of Medicine
DEPARTMENT
Division of Environmental Pediatrics, Department of Pediatrics
LAST_NAME
Xiaowen
FIRST_NAME
Ji
ADDRESS
NYU Medical Science Building, 6th Floor, 540 1st Ave., New York, NY 10016
A multi-omic census reveals obesity-associated microRNA miR-let-7 as novel instigator of adipose mitochondrial dysfunction and of intergenerational metabolic decline.
STUDY_SUMMARY
We here describe that obesity and weight loss in male mice causes reversible abnormalities in glucose and lipid metabolism, serum metabolomes, and white adipose tissue alongside reversible reductions in activity of genes controlling mitochondrial energy dissipation. When mating obese male mice with lean females, we observed concordant reductions in mitochondrial gene expression and translation in offspring that resemble those observed in the paternal (F0) generation. When mapping miRNA responses across somatic and gametic cell types and generations, we found that obesity and weight loss reversible affected miRNA abundance, and observed miR-let7 isoforms induced in obese adipose tissues of F0 and offspring (F1) generations, as well as in sperm of obese F0 mice. Mechanistically, when overexpressing miR-let-7 in adipocytes, we found it to silence DICER1, a cellular rheostat required for adipose tissue adaptation in obesity as evidenced by functional deficiency in mitochondrial functioon following DICER1 loss. Delivery of miR-let-7 into oocytes elicited glucose intolerance and impairments in adipose mitochondrial gene expression in mice sired from miRNA-injected embryos, phenocopying aspects of paternal obesity. When performing single-cell RNA-Seq of embryos, we found that miR-let7 impaired mitochondrial gene expression, suggesting altered energy metabolism following sperm-mediated changes in zygotic miRNAs. When studying miRNA alterations in human semen, we lifestyle-induced weight loss to downregulate MIR-LET-7 in human subjects, suggesting similar roles for human MIR-LET-7 in gametic epigenomes and embryogenesis.
INSTITUTE
University of Southern Denmark
LAST_NAME
Kornfeld
FIRST_NAME
Jan-Wilhelm
ADDRESS
University of Southern Denmark (SDU), Campusvej 55, 5230 Odense M, Denmark
NRF2 supports non-small cell lung cancer growth independently of CBP/p300-enhanced glutathione synthesis: Global metabolomics analysis on A549 cells at different NRF2 status (Part 1 of 3)
STUDY_TYPE
Untargeted Metabolomics
STUDY_SUMMARY
This study aims at discovering metabolic changes in A549 cancer cells in the presence and absence of NRF2, and a mutant NRF2 genotypes. Metabolites were extracted from cell pellets by using an LLE method with Methanol/Chloroform/water. The aqueous layer was analyzed by HILIC-HRMS. An in-house RT library was used to identify metabolites. Statistical analyses was performed to identify statistically significant changes in the metabolism. 35 metabolites presented differential abundance between NRF2 knockdown and wildtype conditions. In particular, GSH and several glutamate dipeptides were significantly depleted upon NRF2 knockdown, in line with the prevailing role of NRF2 in controlling GSH biosynthesis. Disruption of additional metabolites involved in the PPP (sedoheptulose-7-phosphate, S7P), nucleotide (CMP, IMP) and amino acid (kynurenine, homocitrulline) metabolism were also observed upon NRF2 knockdown.
NRF2 supports non-small cell lung cancer growth independently of CBP/p300-enhanced glutathione synthesis: Absolute Quantification of NADP+ and NADPH in A549 cells (Part 2 of 3)
STUDY_SUMMARY
This study aims at directly quantifying NADPH, NADP+, and ratios of NADPH/NADP+ and its association with NRF2 to reveal NRF2 function in redox homeostasis. Briefly, cells were extracted by using acidified ACN/MeOH/water mixture with 15N-labeled ADP followed by adjusting pH to 8. Standard curve was established to quantify NADPH and NADP+ by using LC-HRMS. The result shows that when NRF2 is absent, both NADPH and NADP+ levels decrease.
NRF2 supports non-small cell lung cancer growth independently of CBP/p300-enhanced glutathione synthesis: Using 1,2-13C-glucose to measure PPP Flux in A549 cells (Part 3 of 3)
STUDY_SUMMARY
The study aims at validating the hypothesis that NRF2 mediates the redox homeostasis through regulating oxidative pentose phosphate pathway. The study uses 1,2-13C-glucose to label A549 cells with or without NRF2 and the oxidative PPP can be measured by looking at ratios of [M+1] and [M+1]+[M+2] lactates. Briefly, metabolites were extracted from cells and media with cold ACN/MeOH/water mixture and analyzed by LC-HRMS to determine glucose uptake rate and lactate isotopologues. The result shows that PPP flux is reduced in the absence of NRF2.
Individual glycemic responses to carbohydrates vary and reflect underlying metabolic physiology (Lipidomics)
STUDY_SUMMARY
We measured PPGRs using continuous glucose monitoring (CGM) in 55 well-phenotyped participants challenged with seven different carbohydrates administered in replicate under standardized conditions. Plasma sample were collected at baseline visit for metabolomics. The ClinicalTrials.gov registration identifier is NCT03919877.
INSTITUTE
Stanford University
LAST_NAME
Michael
FIRST_NAME
Snyder
ADDRESS
300 Pasteur Drive, M-344A Stanford, California 94305
1H-NMR-based Metabolomic Analysis of Hypersalinity-Induced Oviparity in Brine Shrimp
STUDY_SUMMARY
This study investigated the mechanisms by which high salinity conditions stimulate adult Artemia females to produce diapaused cysts. We used a 1H-NMR-based metabolomic approach to elucidate the metabolic regulation between ovoviviparity and oviparity in Artemia exposed to different salinities. At a salinity of 80 parts per thousand (ppt), 100% of females produced diapaused cysts, compared to 20% at 50 ppt. Metabolic profiling revealed significant alterations in a range of metabolites, including 5,6-dihydrouracil, betaine, and malate, in females undergoing oviparity at 80 ppt compared to ovoviviparity at 30 ppt. Multivariate statistical analyses indicated clear separation between the two reproductive strategies. The up-regulated metabolites in oviparity were involved in significant metabolic pathways, such as Ī²-alanine metabolism and the citrate cycle, highlighting substantial metabolic differences between the two reproductive strategies. These identified metabolic pathways might play crucial roles in the maternal response to high salinity, facilitating embryo protection and enhancing the survival and reproductive success of brine shrimp. These findings provide a basis for further research into the molecular mechanisms underlying Artemia adaptation to high salinity environments.
INSTITUTE
National Kaohsiung University of Science and Technology
DEPARTMENT
Department and Graduate Institute of Aquaculture
LABORATORY
Lab. of Live Feeds Aquaculture
LAST_NAME
Chiu
FIRST_NAME
Kuohsun
ADDRESS
142 Hai-tuang Rd., Kaohsiung, Not USCanada, 811, Taiwan
NMR metabolomics confirms PRPS homooligomerization in PRPS2/PRPSAP1/PRPSAP2 triple knockout mouse embryonic fibroblasts decreases flux through the nucletide biosynthesis pathways
STUDY_SUMMARY
Phosphoribosyl pyrophosphate synthetase (PRPS) is a highly conserved enzyme that conducts the chokepoint reaction of nucleotide biosynthesis by converting ribose-5-phosphate (R-5-P) to phosphoribosyl pyrophosphate (PRPP). Due to the presence of multiple isoforms and related PRPS-associated proteins in opisthokont species, the precise nature of the PRPS enzyme in cells and tissues remains uncertain. Using proteomics approaches and biochemistry in mammalian cells, we demonstrate that these individual components assemble together to form a heterogeneous megadalton complex comprising of PRPS1 (P1), PRPS2 (P2), and two PRPS associated proteins ā PRPSAP1 (AP1) and PRPSAP2 (AP2). Employing CRISPR-Cas9 knockout (KO) strategy in NIH3T3 cells (mouse embryonic fibroblasts) to generate all viable genetic knockout combinations, we observed that PRPS1-only expressing cells (P2/AP1/AP2 triple KO) has severe cellular proliferation defects as compared to other knockout lines. To pinpoint the metabolic basis for decreased proliferative capacity of PRPS1-only expressing cells (P2/AP1/AP2 KO), we used NMR based 13C isotopic tracing to compare the metabolic state of wildtype and P2/AP1/AP2 KO NIH3T3 cells. The results demonstrate that P2/AP1/AP2 KO cells exhibit reduced pentose phosphate pathway flux and nucleotide production, alongside alterations in glycolysis, the creatine phosphate pathway, and choline metabolism. These changes reflect decreased PRPP synthesis rates, likely due to suboptimal PRPS1 homotypic configurations. Collectively, this study revealed that PRPS1 as a standalone enzyme in cells is insufficient to sustain the metabolic flux necessary to keep up with the demands of proliferation.
Intracellular and supernatant metabolomes of ferroptotic Pfa1 cells using HPLC-MS/MS
STUDY_SUMMARY
In order to investigate metabolites both secreted from and enriched in cells upon ferroptotic cell death, we compared metabolomes from supernatants and lysates of ferroptotic to nonferroptotic Pfa1 cells after 24 h(cells are alive), 30 h (ferroptosis is present) as well as supernatants after 72 h (no cells left). In Pfa1 cells, ferroptosis is induced upon treatment with tamoxifen which induces knock-out of GPX4, causing massive ferroptosis. Ferroptosis, in turn, was rescued using lipid ros scavenger FER-1 (Ferrostatin1).
Individual glycemic responses to carbohydrates vary and reflect underlying metabolic physiology (metabolomics)
STUDY_SUMMARY
We measured PPGRs using continuous glucose monitoring (CGM) in 55 well-phenotyped participants challenged with seven different carbohydrates administered in replicate under standardized conditions. Plasma sample were collected at baseline visit for metabolomics. The ClinicalTrials.gov registration identifier is NCT03919877.
INSTITUTE
Stanford University
LAST_NAME
Michael
FIRST_NAME
Snyder
ADDRESS
300 Pasteur Drive, M-344A Stanford, California 94305
Multilevel Plasticity and Altered Glycosylation Drive Aggressiveness in Hypoxic and Glucose-Deprived Bladder Cancer Cells
STUDY_TYPE
MS quantitative analysis
STUDY_SUMMARY
Bladder tumours with aggressive characteristics often present microenvironmental niches marked by low oxygen levels (hypoxia) and limited glucose supply due to inadequate vascularization. The molecular mechanisms facilitating cellular adaptation to these stimuli remain largely elusive. Employing a multi-omics approach, we discovered that hypoxic and glucose-deprived cancer cells enter a quiescent state supported by mitophagy, fatty acid Ī²-oxidation, and amino acid catabolism, concurrently enhancing their invasive capabilities. Reoxygenation and glucose restoration efficiently reversed cell quiescence without affecting cellular viability, highlighting significant molecular plasticity in adapting to microenvironmental challenges. Furthermore, cancer cells exhibited substantial perturbation of protein O-glycosylation, leading to simplified glycophenotypes with shorter glycosidic chains. Exploiting glycoengineered cell models, we established that immature glycosylation contributes to reduced cell proliferation and increased invasion. Our findings collectively indicate that hypoxia and glucose deprivation trigger cancer aggressiveness, reflecting an adaptive escape mechanism underpinned by altered metabolism and protein glycosylation, providing grounds for clinical intervention.
INSTITUTE
Portuguese Oncology Institute of Porto (IPO-Porto)
The Spatial Transcriptional Activity of Hepatic TCF7L2 Regulates Zonated Metabolic Pathways that Contribute to Liver Fibrosis
STUDY_SUMMARY
Background and Aims: The molecular mechanisms regulating the zonal distribution of metabolism in liver are incompletely understood. Here we used single nuclei genomics techniques to examine the spatial transcriptional function of the transcription factor 7-like 2 (TCF7L2) in rodent liver. We also determined the consequences of TCF7L2 transcriptional inactivation on the metabolic architecture of the liver, and on the function of key zonated metabolic pathways that influence the development of fibrotic liver diseases. Methods: Multimodal 10X single nuclei RNA- and ATAC-Seq were used to dissect the cell-type specific expression and DNA binding activity of TCF7L2 and transcriptional co-regulators across the liver lobule. The transcriptional activity of TCF7L2 was targeted by excising exon 11 of Tcf7l2, which encodes part of the DNA binding domain (DBD). The consequences of TCF7L2 inactivation on zonation was investigated with a focus on the disruption to zonated metabolic pathways, including those linked to ammonia detoxification and glutamine metabolism, cholesterol homeostasis and bile acid synthesis. The susceptibility of Hep-TCF7L2ĆĀDBD mice to hepatic fibrosis under dietary stress was investigated using the Gubra Amylin Nash (GAN) and choline-deficient amino acid-defined high fat (CDAHFD) diets. Results: Tcf7l2 mRNA expression was ubiquitous across the liver lobule, but accessibility of the consensus TCF/LEF DNA binding motif was restricted to pericentral (PC) hepatocytes in zone 3. PC hepatocyte-specific gene expression was lost in Hep-TCF7L2ĆĀDBD mice, which we link to alterations in the transcriptional activity of zonal repressors Tbx3 and Tcf7l1. The absence of a classic zone 3 gene expression program in TCF7L2 mutant mice led to hepatic cholesterol accumulation, impaired bile acid synthesis, and significant disruptions to glutamine and glutamate homeostasis. In human liver biopsy tissue, TCF7L2 expression declined as the severity of fibrosis progressed, and Hep-TCF7L2ĆĀDBD mice developed more severe hepatic fibrosis when fed the CDAHFD. Conclusion: TCF7L2 is key transcriptional regulator of the PC hepatocyte gene expression program in zone 3, and regulates multiple zonated metabolic pathways that may contribute to the development of fibrotic liver diseases.
Hexosamine Biosynthesis Disruption Impairs GPI Production and Arrests Plasmodium falciparum Growth at Schizont Stages
STUDY_SUMMARY
UDP-N-acetylglucosamine (UDP-GlcNAc) is a crucial sugar nucleotide for glycan synthesis in eukaryotes. In Plasmodium falciparum, UDP-GlcNAc is synthesized via the hexosamine biosynthetic pathway (HBP) and is essential for glycosylphosphatidylinositol (GPI) anchor production, the most prominent form of protein glycosylation in this parasite. In this study, we explore a conditional knockout of glucosamine-6-phosphate N-acetyltransferase (PfGNA1), a key HBP enzyme. PfGNA1 depletion led to significant disruptions in HBP metabolites, impairing GPI biosynthesis and causing mislocalization of the merozoite surface protein 1 (MSP1), the most abundant GPI-anchored protein in the parasite. As a result, parasites were arrested at the schizont stage, exhibiting severe segmentation defects and an incomplete rupture of the parasitophorous vacuole membrane (PVM), preventing egress from host red blood cells. Our findings demonstrate the critical role of HBP and GPI biosynthesis in P. falciparum asexual development and underscore the potential of targeting these pathways as a therapeutic strategy against malaria.
Metabolite analysis for WT and BnaMYB52 mutants by LC-MS/MS
STUDY_SUMMARY
BnaA09.MYB52 directly targets the BnaBAN promoters and promotes BnaBAN expression in Brassica napus. BAN, encoding anthocyanidin reductase that converts anthocyanidins to 2,3- cis-flavan-3-ols compounds (proanthocyanidins starter units), is involved in the flavonoid biosynthesis pathway. Thus, Metabolite analysis was conducted to detect the content of flavonoid in WT (Wild-type), OE (BnaA09.MYB52 overexpression lines in the genetic background Westar) and mutants (four homologous genes of BnaMYB52 knocked out) plants. About 0.1 g mature seeds were collected from WT, OE and mutant plants. Metabolites analysis demonstrated that BnaMYB52 positively regulated the content of several metabolites (such as L-phenylalanine, p-coumaric acid, grosvenorine and astragalin) in flavonoid pathway.
INSTITUTE
Huazhong Agricultural University
LAST_NAME
Jiang
FIRST_NAME
Ye
ADDRESS
Shi Zishan Street 1th, Wuhan, Hubei, 430070, China
Skin-specific double knockout (DKO) of acetyl-CoA synthesizing enzymes, ACLY and ACSS2, induces skin barrier dysfunction and systemic fat loss in mice that can be partially rescued by olive oil supplementation. In this study, lipidomics was performed for epidermis derived from DKO versus wild-type mouse skin with or without olive oil supplementation to examine the changes in epidermal lipid profile in response to limited acetyl-CoA availability and to dietary lipid intervention.
Targeting the c-MYC/ELOVL6 Pathway Alters Cell Membrane Mechanics and Enhances Chemotherapeutic Efficacy in Pancreatic Cancer
STUDY_SUMMARY
Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease, marked by a survival rate of only 12%. Consequently, the exploration of novel therapeutic strategies becomes a critical clinical imperative. Among the genetic alterations contributing to PDAC, c-MYC (an oncogenic transcription factor) overexpression arises due to upstream mutations, amplifications, and copy number alterations. c-MYC serves as a key regulator in the tumor's metabolic reprogramming, playing a pivotal role in proliferation, migration, and metastasis. This study delves into the investigation of the role of the elongase ELOVL6 in c-MYC-induced cell transformation and its potential as a therapeutic target in PDAC. Here, we demonstrate that c-MYC regulates lipid elongation to promote cell transformation, offering a new avenue for therapeutic intervention. Initially, we show the direct regulation of ELOVLs expression by c-MYC in various PDAC mouse models and cell lines, elucidating its upregulation during transformation and tumor progression. Genetic or chemical inhibition of ELOVL6 results in decreased proliferation and migration, accompanied by alterations in fatty acid elongation. These changes in fatty acid composition led to modifications in membrane rigidity, permeability, and thickness, which collectively affect micropinocytosis and macropinocytosis. Importantly, we observe an increase in Abraxane uptake and a synergistic effect when combined with ELOVL6 interference in vitro. In vivo validation demonstrates that ELOVL6 inhibition significantly reduces tumor growth and enhances the response to Abraxane, thereby increasing overall survival. Altogether, these results position ELOVL6 as a promising therapeutic target in the treatment of PDAC.
INSTITUTE
Universidad Francisco de Vitoria - Hospital 12 de Octubre
Targeted mass spec-based metabolomic and clinical analyte analyses of liver and plasma samples from rats with and without hepatic knockdown of ChREBP expression.
STUDY_SUMMARY
The transcription factor Carbohydrate Response Element-Binding Protein (ChREBP) activates genes of glucose, fructose and lipid metabolism in response to carbohydrate feeding. Integrated transcriptomic and metabolomic analyses in rats with GalNac-siRNA-mediated suppression of ChREBP expression (GalNac-siChREBP treatment) in liver revealed novel ChREBP functions relative to rats treated with a GalNac vector expressing a non-targeting siRNA (GalNac-siCtrl treatment). GalNac-siChREBP treatment reduced expression of genes involved in coenzyme A (CoA) biosynthesis, and lowered CoA and short chain acyl CoA levels. Despite suppression of pyruvate kinase, pyruvate levels were maintained, possibly via increased expression of pyruvate and amino acid transporters. In addition, expression of multiple anaplerotic enzymes was decreased by GalNac-siChREBP treatment, affecting TCA cycle intermediates. Finally, GalNAc-siChREBP treatment suppressed late steps in purine and NAD synthesis, with increases in precursors and lowering of end products in both pathways. In sum, our studies have revealed functions of ChREBP beyond its canonical roles in carbohydrate and lipid metabolism to include regulation of substrate transport, mitochondrial function, and cellular energy balance.
Analysis of the role of PNPLA6 in the mouse retina
STUDY_SUMMARY
We generated PNPLA6 flox/flox CreER(+) mice, knocked out PNPLA6 by treatment with tamoxifen and isolated their retinal pigment epithelium. Lipid analysis of the retinal pigment epithelial cells was subsequently performed.
Untargeted metabolomics of Human embryonic kidney cells (HEK293) infected by Human adenovirus serotype 5 during early and late infection.
STUDY_SUMMARY
Human adenoviruses overtake the DNA replication machinery of the infected host, rewiring mitotic events. Infection leads to the elevations in glucose and glutamine consumption rates, while also increasing lactate production rate. Fibroblast lineages are normally quiescent cells that display a repertoire of responses to certain agonists. Data on the shifts in fibroblast metabolism in response to human adenoviral infection are lacking. Specifically, knowledge pertaining to metabolic shifts aside from those involved in glycolytic metabolism after human adenoviral infection remains sparse. We used an untargeted metabolomic approach to better understand the dynamic metabolic changes to fibroblasts in response to 3 viral dosages, across 4 time points post-infection. Cells were infected for the following periods: 6, 12, 24, and 36 hours. Noninfected cells cultured for 24 hours were used as a negative control. Cells were subjected to the following viral dosages: 0.5, 1.0, and 2.0 multiplicity of infection (MOI) and 6 replicates were used for each experimental condition. Cellular pellets were collected at each time point following the removal of media. Samples were sent to the UC Davis Metabolomics Core for analysis of primary carbons using GC-TOF-MS. Profound shifts were seen in cysteine, purine, and unsaturated fatty acid metabolites. This analysis provides a global perspective and highlights previously underappreciated aspects of how human adenovirus alters host metabolism.
Investigation of the effects of 3-hydroxybutyrate on myoblast proliferation and differentiation using NMR-based metabolomics
STUDY_SUMMARY
This study aims to further elucidate the molecular mechanisms by which 3-HB promotes the proliferation and differentiation of C2C12 myoblasts. By performing NMR-based metabolomic analysis on myoblasts, we explored the dual roles of 3-HB functioning as a metabolic substrate and a signaling molecule in myoblasts, and uncovered the underlying molecular mechanisms. Our results may significantly contribute to the development of 3-HB as a potential therapeutic agent against skeletal muscle atrophy.
Biguanides antithetically regulate tumor properties by the dose-dependent mitochondrial reprogramming-driven c-Src pathway
STUDY_SUMMARY
The biguanide metformin attenuates mitochondrial oxidation and has been proposed as an anti-cancer therapy. However, recent clinical studies suggested increased proliferation and fatty acid beta-oxidation (FAO) in a subgroup of breast cancer (BC) patients after metformin therapy. Considering that FAO can activate Src kinase in aggressive triple-negative BC (TNBC), we hypothesized that a low-dose biguanides-driven AMPK-ACC-FAO signaling may activate the Src pathway in TNBC. The low bioavailability of metformin in TNBC xenografts mimicked metformin's in vitro low-dose effect. Pharmacological or genetic inhibition of FAO significantly enhanced the anti-tumor properties of biguanides. Lower doses of biguanides induced and higher doses suppressed Src signaling. Dasatinib and metformin synergistically inhibited TNBC patient-derived xenograft growth but not in high-fat diet-fed mice. This combination also suppressed TNBC metastatic progression. A combination of biguanides with Src inhibitors provides synergy to target metastatic TNBC suffering with limited treatment options.
Multiplatform Metabolomic Profiling of the Unilateral Ureteral Obstruction Murine Model of CKD
STUDY_SUMMARY
In chronic kidney disease (CKD) research, animal models provide invaluable insights into the diseaseās etiopathogenesis and progression, particularly through the evaluation of renal tissue. The unilateral ureteral obstruction (UUO) rodent model stands out for its widespread use in CKD studies, due to its advantages to generate renal fibrosis and accelerated mimicry of obstructive nephropathy in humans. Despite its extensive use, the molecular underpinnings driving kidney disease progression remain incompletely understood. Given the crucial interplay between metabolism and fibrosis in CKD, a thorough examination of the UUO renal tissue through metabolomics is required. Untargeted multiplatform analysis enables a comprehensive measurement of the sample metabolic profile, ensuring a maximum coverage of metabolite diversity to yield extensive insights into the metabolism of this renal injury model. Therefore, in this study, murine kidney tissue from the UUO model underwent analysis using three separation techniquesāliquid chromatography (LC), gas chromatography (GC), and capillary electrophoresis (CE)ācoupled with mass spectrometry (MS). The findings reveal metabolic changes associated with tubulointerstitial fibrosis, impacting essential pathways such as the TCA cycle, urea cycle, polyamine metabolism, amino acids, one-carbon metabolism, purine catabolism, and NAD+ synthesis, among others. Furthermore, fibrosis significantly influences the renal tissue's lipidomic profile, characterized by a general decrease in most lipid classes and an increase in glycerophospholipids with ether substituents, hexosylceramides, and cholesterol esters compared to the control. These results underscore the relevance of the untargeted multiplatform approach to obtain a comprehensive overview of the alterations within the renal metabolic map, paving the way for further exploration of the molecular mechanisms underlying CKD.
Marine community metabolomes in the eastern tropical North Pacific Oxygen Deficient Zone
STUDY_SUMMARY
Oxygen deficient zones (ODZs) are subsurface marine systems that harbor distinct microbial communities, including populations of the picocyanobacteria Prochlorococcus that can form a secondary chlorophyll maxima (SCM), and low-oxygen tolerant strains of the globally abundant heterotroph Pelagibacter (SAR11). Yet, the small labile molecules (metabolites) responsible for maintaining these ODZ communities are unknown. Here, we compared the metabolome of an ODZ to that of an oxygenated site by quantifying 87 metabolites across depth profiles in the eastern tropical North Pacific ODZ and the oxygenated waters of the North Pacific Gyre. We further use transcriptomes to identify taxa involved in production and subsequent transformation of glycine betaine (GBT), a metabolite we suggest is involved in microbial interdependencies in this community, and elsewhere in the ocean.
Rapid and Reagent-free Analysis of Dried Blood Spot by Paper Spray Mass Spectrometry Reveals Sex: Implications in Forensic Investigations.
STUDY_SUMMARY
Identifying sex from an unknown dried blood spot (DBS), especially when the corpse remains undiscovered, often provides valuable evidence in forensic casework. While DNA-based sex determination is a reliable method in forensic settings, it requires ex-pensive reagents and is time-consuming. To develop a rapid reagent-free blood test for sex, we employed paper spray ionization mass spectrometry (PSI-MS) to capture sex-discriminatory lipid profiles from 200 DBS samples comprising 100 males and 100 females. We conducted a supervised machine learning (ML) analysis on all detected lipid signals to hunt biomarkers of sex within the dataset. This analysis unveiled significant differences in specific sphingomyelin and phospholipid species levels between male and female DBS samples. Using the parsimonious set of 60 lipid signals, we constructed a classifier that achieved 100% overall accuracy in predicting sex from DBS on paper. Additionally, we assessed three-day-old air-exposed DBS on glass and granite surfaces, simulating the typical blood samples available for forensic investigations. Consequently, we achieved ~92% overall sex prediction accuracy from the holdout test dataset of DBS on glass and granite surfaces. As a highly sensitive detection tool, PSI-MS combined with ML has the potential to revolutionize forensic methods by rapidly analyzing blood molecules encoding personal information.
INSTITUTE
Indian Institute of Science Education and Research Tirupati
DEPARTMENT
Chemistry
LABORATORY
SBlab
LAST_NAME
Mondal
FIRST_NAME
Supratim
ADDRESS
Venkatagiri Road, Yerpedu Mandal, Tirupati, Andhra Pradesh - 517619, India
Lactate-coated polyurea-siRNA dendriplex: a gene therapy-directed and metabolism-based strategy to impair glioblastoma (GBM)
STUDY_SUMMARY
Glioblastoma (GBM) is a highly lethal disease with limited treatment options due to its infiltrative nature and the lack of efficient therapy able to cross the protective blood-brain barrier (BBB). GBMs are metabolically characterized by increased glycolysis and glutamine dependence. This study explores a novel metabolism-based therapeutic approach using a polyurea generation 4 dendrimer (PUREG4) surface functionalized with lactate (LA) (PUREG4-LA12), to take advantage of glucose-dependent monocarboxylate transporters (MCTs) overexpression, loaded with selenium-chrysin (SeChry) and temozolomide (TMZ) or complexed with anti-glutaminase (GLS1) siRNAs to abrogate glutamine dependence. The anti-GLS1 siRNA dendriplex with PUREG4-LA12 (PUREG4-LA12-anti-GLS1-siRNA) knocked down GLS1 in the GBM cell lines. In two in vitro BBB models, these dendriplexes successfully crossed the BBB, decreased GLS1 expression and altered the exometabolome of GBM cell lines, concomitantly with autophagy activation. Our findings highlight the potential of targeting glucose and glutamine pathways in GBM using dendrimer-based nanocarriers, overcoming the BBB and disrupting key metabolic processes in GBM cells.
INSTITUTE
ITQB NOVA
LAST_NAME
GonĆĀ§alves
FIRST_NAME
LuĆĀs
ADDRESS
Avenida Republica, Oeiras, 2780-157 Oeiras, Portugal
Functional analysis of retinal pigment epithelial cells with PNPLA6 knockdown
STUDY_SUMMARY
PNPLA6 was knocked down using siRNA on ARPE-19, a representative cultured cell line of human retinal pigment epithelial cells. Lipid analysis of the constituent phospholipids was performed on these cells, comparing them to a group treated with scramble siRNA. We also overexpressed PNPLA6 in human retinal pigment epithelial cells ARPE-19 and performed the same lipid analysis.
The native small molecule interaction landscape of transcription factors and essential enzymes in E. coli
STUDY_TYPE
untargeted metabolite-protein interactions
STUDY_SUMMARY
Knowledge of proteināmetabolite interactions can enhance mechanistic understanding and chemical probing of biochemical processes, but the discovery of endogenous ligands remains challenging. Here, we combined rapid affinity-purification with precision mass spectrometry and highresolution molecular docking to precisely map the physical associations of 296 chemically diverse small molecule metabolite ligands with 69 distinct essential enzymes and 45 transcription factors in the Gram-negative bacterium Escherichia coli. We then conducted systematic metabolic pathway integration, pan-microbial evolutionary projections, and independent in-depth biophysical characterization experiments to define the functional significance of novel ligand interfaces. This effort revealed principles governing functional crosstalk on a network-level, divergent patterns of binding pocket conservation, and scaffolds for designing selective chemical probes. This structurally resolved ligand interactome mapping pipeline can be scaled to illuminate the native small molecule networks of complete cells and potentially entire microbial communities.
Kingdom-specific lipid unsaturation calibrates sequence evolution in membrane arm subunits of eukaryotic respiratory complexes
STUDY_SUMMARY
Sequence evolution of protein complexes (PCs) is constrained by protein-protein interactions (PPIs). PPI-interfaces are predominantly conserved and hotspots for disease-related mutations. How do lipid-protein interactions (LPIs) constrain sequence evolution of membrane-PCs? We explore Respiratory Complexes (RCs) as a case study as these allow to compare sequence evolution in subunits exposed to both lipids in inner-mitochondrial membrane (IMM) and lipid-free aqueous matrix. We find that lipid-exposed surfaces of the IMM-subunits but not of the matrix subunits are populated with non-PPI disease-causing mutations signifying LPIs in stabilizing RCs. Further, IMM-subunits including their exposed surfaces show high intra-kingdom sequence conservation but remarkably diverge beyond. Molecular Dynamics simulation suggests contrasting LPIs of structurally superimposable but sequence-wise diverged IMM-exposed helices of Complex I (CI) subunit Ndufa1 from human and Arabidopsis depending on kingdom-specific unsaturation of cardiolipin fatty acyl chains. in cellulo assays consolidate inter-kingdom incompatibility of Ndufa1-helices due to the lipid-exposed amino acids. Plant-specific unsaturated fatty acids in human cells also trigger CI-instability. Taken together, we posit that altered LPIs calibrate sequence evolution at the IMM-arms of eukaryotic RCs.
INSTITUTE
CSIR-Centre for Cellular & Molecular Biology
LAST_NAME
Raychaudhuri
FIRST_NAME
Swasti
ADDRESS
Habsiguda, Uppal Road, Hyderabad, 500007, Telangana, India
Lipidomics facilitates the discovery of diagnostic biomarkers in patients with chronic total occlusion during the perioperative period
STUDY_SUMMARY
Chronic total occlusion (CTO) is a subtype of cardiovascular disease associated with high mortality and an increased risk of ventricular arrhythmia. This study aimed to investigate lipidomic changes in CTO patients undergoing percutaneous coronary intervention (PCI) using a tandem-lipidomic strategy. We first applied a global lipidomic approach to identify the serum lipidomes of CTO-PCI patients during the perioperative period, successfully separating and identifying over 1,500 lipids. Based on these results, a Multiple Reaction Monitoring (MRM) quantification method was developed and employed for targeted lipidomic analysis. Using a high-throughput MRM tandem liquid chromatography-mass spectrometry approach, 613 lipids were successfully quantified in CTO-PCI patients and control donors. PA 18:2/11:0 emerged as a potential biomarker for distinguishing CTO patients from those suspected of having the condition. Notably, patients with different prognostic outcomes exhibited significantly distinct serum lipidomes in both pre- and post-CTO-PCI samples. This finding suggests that lipidomic data hold significant potential not only for monitoring postoperative prognosis but also for predicting surgical outcomes
Metabolic and Morphometric Analysis of Allometric and Total Liver Growth in Post-Hatch Chickens
STUDY_SUMMARY
This study was designed to characterize the liver and plasma metabolome of Ross 708 broiler chickens. Liver and plasma samples were obtained at necropsy on days 4,6,8,10,12,14,16,18 and 20 post-hatch and frozen in liquid nitrogen. Samples were submitted to the University of Carlifornia Davis campus for metabolomic analysis. Analysis revealed three distinct metabolic periods, A (days 4,6,8), B (days 10,12,14) and C (days 16,18,20). Period A corresponded to metabolism driven by yolk nutrients. Period B was a transition period during which yolk resources were being exhausted, and nutrients from feed were metabolized. Period C corresponded to the metabolism of a mature liver.
Tumour interstitial fluid-enriched phosphoethanolamine suppresses T cell function.
STUDY_SUMMARY
Nutrient stress represents a significant barrier for antitumor immunity, and tumor interstitial fluid (TIF) often contains metabolites that hinder immune function. However, it is difficult to isolate the effects of tumor nutrient stress from other suppressive factors. Thus, we employed a chemically-defined cell culture medium based on the metabolomic profile of TIF: Tumor Interstitial Fluid Medium (TIFM). Culture of CD8+ T cells in TIFM limited cell expansion and impaired CD8+ T cell effector functions upon restimulation, suggesting tumor nutrient stress alone is sufficient to drive T cell dysfunction. We identified phosphoethanolamine (pEtn), a phospholipid intermediate, as a driver of T cell dysfunction. pEtn dampened T Cell Receptor (TCR) signaling by depleting T cells of diacylglycerol required for TCR signal transduction. Reduction of pEtn accumulation in tumors improved intratumoral T cell function and tumor control, suggesting pEtn accumulation plays a dominant role in TME immunosuppression.
Conditioned media from SaGA-derived leader and follower subpopulations (Khatib 2023, Konen 2017) were collected and analyzed using mass spectrometry-based proteomics to assess differences in cell secretion profiles. Label-free quantification was employed to compare spectra against the 2020 human UniProtKB/Swiss-Prot database (20,379 target sequences). This analysis identified 1,174 proteins. Our findings revealed that the follower subpopulation produces and secretes TGFB1 and TGFB-related factors. Notably, the leader subpopulation secretes FN1 and ALCAM, which are downstream effectors of TGFB-mediated signaling.
Advanced Lipidomics Using UHPLC-ESI-QTOF-MS/MS Reveals Novel Lipids in Hibernating Syrian Hamsters
STUDY_SUMMARY
Mammalian hibernation offers a unique model for exploring neuroprotective mechanisms relevant to neurodegenerative diseases. In this study, we employed untargeted lipidomics with iterative tandem mass spectrometry (MS/MS) to profile the brain lipidome of Syrian hamsters across different hibernation stages: late torpor, arousal, and euthermia (control). Previously, a lipid species identified as methyl-PA(16:0/0:0) showed a significant increase during torpor, but its precise structure was unresolved due to technological constraints. Leveraging iterative MS/MS and advanced lipid annotation tools (LipidAnnotator and MS-DIAL), we accurately annotated 377 lipid species, including the re-identification of methyl-PA(16:0/0:0) as methylated lysophosphatidic acid (PMeOH 16:0/0:0). This reannotation led to the discovery of two additional lipids during torpor: PMeOH 18:0/0:0 and PMeOH 18:1/0:0. Verification involved manual inspection of MS/MS spectra and Kendrick Mass Defect plots. The lipid alterations observed during torpor suggest biochemical adaptations to maintain membrane fluidity and protect against oxidative stress under hypothermic conditions. Elevated levels of PMeOH lipids and their lyso-forms may play roles in cell survival signalling. Additionally, a decrease in phosphatidic acid species and an increase in diacylglycerol species imply a metabolic shift favouring diacylglycerol production, potentially activating protein kinase C signalling pathways. The increased levels of monogalactosyl diglyceride lipids during torpor suggest a role in neuroprotection by enhancing oligodendrocyte function and myelination. Our comprehensive lipidomic profiling provides detailed insights into lipid dynamics associated with hibernation and underscores the potential of advanced MS/MS methodologies in lipidomics for developing therapeutic strategies against neurodegenerative diseases.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
Centro de MetabolĆ³mica y BioanĆ”lisis (CEMBIO)
LAST_NAME
GonzƔlez
FIRST_NAME
Carolina
ADDRESS
km 0, Universidad CEU-San Pablo UrbanizaciĆ³n MonteprĆncipe. M-501
Identification of plasma metabolites responding to oxycodone exposure in rats
STUDY_SUMMARY
Metabolomics studies of prescription opioid medications including oxycodone can provide insights into biochemical mechanisms of the addiction cycle and prognosis prediction. Although oxycodone has an elevated abuse liability profile compared to other opioids, many human and rodent metabolomics studies have not been specifically focused on oxycodone. In this study, we investigated metabolomics changes associated with oxycodone exposure using plasma samples from 16 rats at pre-exposure and intoxication time points. Among the 798 metabolites with low levels of missingness, 364 showed significant changes be-tween pre-exposure and intoxication (FDR < 0.01) including succinate, oleamide, and sarcosine. We identified four pathways including tryptophan metabolism that were nominally enriched among the metabolites that change with oxycodone exposure (p < 0.05). Furthermore, we identified sev-eral metabolites that showed nominal correlations with the Addiction Index (composite of ox-ycodone behaviors), 17 at pre-exposure and 8 at intoxication.
Metabolomic and lipoproteomic differences and similarities among different types of pneumonia
STUDY_TYPE
NMR-based metabolomics
STUDY_SUMMARY
An 1H NMR analysis of the plasma of patients with non-COVID-19 interstitial (24 individuals) or lobar (21 individuals) pneumonia, all matched by age, gender and disease severity has been performed to compare the metabolomic and lipoproteomics profiles of different types of pneumonia. The analysis is based on 24 metabolites and 114 lipoprotein parameters. Several common traits are observed between the two groups, with the most dysregulated molecules in the lobar group (same trends of changes but higher fold changes); some peculiar features characteristic of each group could be also identified.
Polar Metabolite Profiles Distinguish Between Early and Severe Sub-maintenance Nutritional States of Wild Bighorn Sheep
STUDY_SUMMARY
Understanding the metabolic adaptations of wild bighorn sheep (Ovis c. canadensis) to nutritional stress is crucial for their conservation. This study employed 1H nuclear magnetic resonance (NMR) metabolomics to investigate the biochemical responses of these animals to varying sub-maintenance nutritional states. Serum samples from 388 wild bighorn sheep collected between 2014 and 2017 during December through March across Wyoming and Montana were analyzed. Multivariate statistics and machine learning analyses were employed to identify characteristic metabolic patterns and metabolic interactions between early and severe sub-maintenance nutritional states. Significant differences were observed in the levels of 15 of the 49 quantified metabolites, including formate, thymine, glucose, choline, and others, pointing to disruptions in one-carbon, amino acid, and central carbon metabolic pathways. These metabolites may serve as indicators of critical physiological processes such as nutritional intake, immune function, energy metabolism, and protein catabolism, essential for understanding how wild bighorn sheep adapt to nutritional stress. The study has generated valuable insights into molecular networks underlying the metabolic resilience of wild bighorn sheep, highlighting the potential for using specific biochemical markers to evaluate nutritional and energetic states in free-ranging ungulates. These insights may help wildlife man-agers and ecologists compare populations across different times in seasonal cycles, providing information to assess the adequacy of seasonal ranges and support conservation efforts. This research strengthens our understanding of metabolic adaptations to envi-ronmental stressors in wild ruminants, offering a foundation for improving management practices to maintain healthy bighorn sheep populations.
Ketogenic diet suppresses colorectal cancer through the gut microbiome long chain fatty acid stearate - untargeted LCMS data from SPF experiment
STUDY_SUMMARY
Mouse fecal pellets from an AOM/DSS CRC mouse experiment performed with specific pathogen-free mice were analyzed by a LC-MS approach to characterize the fecal metabolome of mice from different treatment groups. Metabolites were extracted from mouse fecal samples as described in the method file. Resulting samples were used for a LC-MS untargeted metabolomics screen using a Vanquish UHPLC (ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer (ThermoFisher Scientific).
Ketogenic diet suppresses colorectal cancer through the gut microbiome long chain fatty acid stearate - untargeted LCMS data from CMT experiment
STUDY_SUMMARY
Germ-free mice received cecal content from mice having received a ketogenic diet (KC) or from mice having received a standard diet (SC) and were subjected to AOM/DSS treatment as described in Tsenkova et al. (2025). Mouse fecal pellets were analyzed by a LC-MS approach to characterize the fecal metabolome of mice from different treatment groups. Metabolites were extracted from mouse fecal samples as described in the method file. Resulting samples were used for LC-MS untargeted metabolomics screen using a Vanquish UHPLC (ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer (ThermoFisher Scientific).
INSTITUTE
University of Luxembourg
LAST_NAME
Letellier
FIRST_NAME
Elisabeth
ADDRESS
6, avenue du Swing, Belval, Esch, 4367, Luxembourg
Ketogenic diet suppresses colorectal cancer through the gut microbiome long chain fatty acid stearate - untargeted LCMS data from human fecal samples
STUDY_SUMMARY
Stool samples from healthy human donors were analyzed by a LC-MS. Metabolites were extracted from human stool samples as described in the method file. Resulting samples were used for LC-MS untargeted metabolomics screen using a Vanquish UHPLC (ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer (ThermoFisher Scientific).
INSTITUTE
University of Luxembourg
LAST_NAME
Letellier
FIRST_NAME
Elisabeth
ADDRESS
6, avenue du Swing, Belval, Esch, 4367, Luxembourg
Ketogenic diet suppresses colorectal cancer through the gut microbiome long chain fatty acid stearate - untargeted LCMS data from mouse plasma samples
STUDY_SUMMARY
Mouse plasma samples from the SD/KD-fed SPF mice were analyzed by a LC-MS. Metabolites were extracted from human plasma samples as described in the method file. Resulting samples were used for LC-MS untargeted metabolomics screen using a Vanquish UHPLC (ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer (ThermoFisher Scientific).
INSTITUTE
University of Luxembourg
LAST_NAME
Letellier
FIRST_NAME
Elisabeth
ADDRESS
6, avenue du Swing, Belval, Esch, 4367, Luxembourg
Investigation of C. elegans adah-1 gene for potential adenosine deaminase activity via quantification of predicted substrate and product in control animals and animals with knockdown in expression of adah-1
STUDY_SUMMARY
We have previously demonstrated that the C. elegans pnp-1 gene encodes a purine nucleoside phosphorylase that catalyzes the conversion of inosine to hypoxanthine. We hypothesize that the C. elegans adah-1 (ADA homolog) gene encodes the adenosine deaminase that acts upstream of PNP-1 to produce inosine from adenosine. To investigate the hypothesis that ADAH-1 functions as a canonical adenosine deaminase, we used RNAi to knockdown expression of ADAH-1, and used LC-MS to compare relative levels of adenosine, inosine, and hypoxanthine in the knockdown animals and an RNAi control (L4440). As a control we also confirmed the activity of PNP-1, by comparing levels of adenosine, inosine, and hypoxanthine in control (N2) and the loss of function mutant pnp-1(jy90). As expected we found that levels of hypoxanthine were reduced and levels of inosine were increased in pnp-1 mutants relative to control. In contrast, RNAi against adah-1 led to significantly increased levels of adenosine, and decreased levels of inosine and the downstream metabolite hypoxanthine. We conclude that ADAH-1 is a bona fide adenosine deaminase.
Characterization of the transpulmonary metabolome at the intersection of pulmonary vascular disease and exercise
STUDY_SUMMARY
Pathologic implications of dysregulated pulmonary vascular metabolism to pulmonary arterial hypertension (PAH) are increasingly recognized, but their clinical applications have been limited. We hypothesized that metabolite quantification across the pulmonary vascular bed in connective tissue disease (CTD)-associated PAH would identify transpulmonary gradients of pathobiologically relevant metabolites, in an exercise stage-specific manner. 63 CTD patients with established or suspected PAH underwent exercise right heart catheterization. Using mass spectrometry-based metabolomics, metabolites were quantified in plasma samples simultaneously collected from the pulmonary and radial arteries at baseline and during resistance-free wheeling, peak exercise, and recovery. We identified uptake and excretion of metabolites across the pulmonary vascular bed, unique and distinct from single vascular site analysis. We demonstrated the physiological relevance of metabolites previously shown to promote disease in animal models and end-stage human lung tissues, including acylcarnitines, glycolytic intermediates, and tryptophan catabolites. Notably, pulmonary vascular metabolite handling was exercise stage-specific. Transpulmonary metabolite gradients correlated with hemodynamic endpoints largely during free-wheeling. Glycolytic intermediates demonstrated physiologic significance at peak exercise, including net uptake of lactate in those with more advanced disease. Contribution of pulmonary vascular metabolism to CTD-PAH pathogenesis and therapeutic candidacy of metabolism modulation must be considered in the context of physiologic stress.
INSTITUTE
University of Colorado Anschutz Medical Campus
LAST_NAME
Haines
FIRST_NAME
Julie
ADDRESS
12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Metabolomics studies on mouse cecum samples on a Western diet
STUDY_SUMMARY
Targeted metabolomics was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS).to measure polar metabolites in both positive and negative ionization mode on cecummice tissue acquired after a 30 week dietary intervention.
Impact of High Fat diet-induced metabolic dysfunction-associated steatotic liver disease (MASLD) on Heart, Kidney and Skeletal Muscle Metabolomes in Wild-type Mice
STUDY_TYPE
1H NMR metabolomics to study the effects high-fat diet-induced MASLD on extrahepatic tissues metabolomes, namely the heart, kidney and skeletal muscle, in C57BL6J wild-type mice.
STUDY_SUMMARY
Excessive caloric intake is a primary driver of metabolic dysfunction-associated steatotic liver disease (MASLD), and this has been recapitulated in mice fed a high-fat diet. In 2023, the global prevalence of MASLD was estimated at 30%, with high incidences affecting wealthy urbanised countries. This implication of hypercaloric diets can also perturb metabolism and function of extrahepatic tissues such as heart, kidney and skeletal muscle. These effects that can take place in extrahepatic tissues are still poorly understood in terms of metabolic alterations and physiology, and represent an important point of improvement in the knowledge gap that connects early stage MASLD with other obesity related comorbidities, such as type 2 diabetes, insulin resistance, cardiovascular and renal complications, and overall, with the so known metabolic syndrome. In this study, we aimed to evaluate the potential of using metabolomics to unravel the effects and interactions taking place in a diet-induced MASLD model related to the development of the disorder. Black-6 mice were subjected to either a control diet or a high-fat diet for 18 weeks, from which at the end their heart, kidney and skeletal muscle metabolites were extracted. The metabolites, divided into aqueous and lipophilic fractions, were acquired by 1H-NMR, and then processed using a untargeted Metabolomics and Lipidomics analysis approach, to identify key changes occurring between control and high-fat diet in these models. These results added important information to better understand the link between early onset MASLD and the Metabolic Syndrome and its comorbidities, though several metabolic changes in the extrahepatic tissues, namely in ectopic fat deposition and alterations to Randle cycle and gut microbiota activity.
INSTITUTE
Center for Innovative Biomedicine and Biotechnology (CIBB UC)
DEPARTMENT
Institute of Interdisciplinary Research
LABORATORY
Metabolic Modelling and Systems Biology
LAST_NAME
Silva
FIRST_NAME
JoĆ£o
ADDRESS
Rua Larga - Faculdade de Medicina, 1Āŗandar - POLO I Universidade de Coimbra
EMAIL
jgsilva@cnc.uc.pt
PHONE
(+351) 239 820 190
NUM_GROUPS
2
TOTAL_SUBJECTS
23
NUM_MALES
23
STUDY_COMMENTS
Full NMR sample preparation and analysis procedures are available in the accompanying document entitled 1. MASLD Extrahepatic Metabolomics experimental procedure. The normalized data that was used in uni- and multivariate analysis is available in the accompanying files: 4. MASLD Extrahepatic Metabolomics results data.txt The raw fid as well as 1r file can be found in 5. MASLD Extrahepatic Metabolomics 1H NMR Raw Data.zip
Kupffer cells control neonatal hepatic glucose metabolism via Igf1 signaling - lipidomics analysis of postnatal day 0 murine livers after macrophages depletion using Csf1r conditional KO
STUDY_TYPE
lipidomics analysis of P0 livers from mice after macrophages depletion by Csf1r conditional KO
STUDY_SUMMARY
During perinatal development, liver metabolism is tightly regulated to ensure energy supply for the newborn. Before birth, glycogen is stored in hepatocytes and later metabolized to glucose, meeting the energy demands of the neonate. Shortly after birth, lipogenesis begins, driven by the transcriptional activation of enzymes involved in fatty acid oxidation. These processes are thought to be largely regulated by systemic insulin and glucagon levels. However, the role of liver-derived local factors in neonatal hepatocyte metabolism remains unexplored. Kupffer cells (KCs), the liverĆ¢ĀĀs resident macrophages, colonize the fetal liver early in embryogenesis and support liver metabolism in adulthood. Yet, whether KCs influence neonatal hepatocyte metabolism is unknown. Here, using conditional knockout mouse models targeting macrophages (Csf1r-flox Tnfrsf11a-Cre), we demonstrate that yolk sac-derived KCs play a critical role in hepatocyte glycogen storage and function by regulating the TCA cycle - a role that monocyte-derived KC-like cells cannot substitute. in order to check the lipid levels after birth, Newborn pups were collected and the different lipid species were assessd by lipidomics analysis using mass spectrometer.
INSTITUTE
University of Bonn
DEPARTMENT
Developmental Biology of the Immune System, The Life & Medical Sciences Institute (LIMES)
NAD Depletion in Skeletal Muscle does not Compromise Muscle Function or Accelerate Aging
STUDY_SUMMARY
NAD is a ubiquitous electron carrier essential for energy metabolism and the posttranslational modification of numerous regulatory proteins. Perturbation of NAD metabolism is considered detrimental to health, with NAD depletion commonly thought to promote aging. However, the extent to which cellular NAD concentration can be decreased without deleterious repercussions is unclear. We generated a mouse model where nicotinamide phosphoribosyltransferase (NAMPT)-mediated NAD+ biosynthesis is disrupted in adult skeletal muscle. The resulting 85% decrease in muscle NAD+ abundance was associated with preserved tissue integrity and functionality, as demonstrated by its unchanged morphology, contractility, and exercise tolerance. This lack of defects was corroborated by intact mitochondrial respiratory capacity and unaffected muscle transcriptomic and proteomic profiles. Furthermore, lifelong NAD depletion did not accelerate muscle aging or impair whole-body metabolism. Collectively, these findings indicate that NAD depletion does not contribute to agerelated declines in skeletal muscle function.
INSTITUTE
University of Copenhagen
LAST_NAME
Treebak
FIRST_NAME
Jonas Thue
ADDRESS
Blegdamsvej 3B, MƦrsk TĆ„rnet, 7. sal 2200 KĆøbenhavn N.
Hyperglycemia leads to BMSC(bone marrow mesenchymal stromal cells) impaired osteogenesis, enhanced adipogenesis, and altered metabolism
STUDY_TYPE
metabolomics
STUDY_SUMMARY
Diabetes, a major risk factor for osteoporosis, negatively impacts bone health, but the mechanisms underlying the effects of hyperglycemia on bone marrow mesenchymal stromal cells (BMSC) are not fully understood. This study investigates how high glucose conditions influence BMSC differentiation, proliferation, viability, and metabolism. Our results demonstrate that high glucose inhibits osteogenesis, as evidenced by significantly reduced alkaline phosphatase activity, impaired calcium deposition, and downregulation of key osteogenic genes (RUNX2, Osteocalcin, Osteopontin). Additionally, high glucose conditions promote adipogenesis, characterized by increased lipid accumulation, larger fat droplets, and upregulation of adipogenic genes (PPARĪ³2, CEBPĪ±, Leptin, AdipoQ), suggesting a shift towards fat cell differentiation. Furthermore, BMSCs cultured in high glucose show decreased proliferation, elevated DNA damage, increased oxidative stress, enhanced apoptosis and senescence, particularly in later passages, highlighting the negative impact of hyperglycemia on BMSC viability. Metabolomic profiling of osteogenic and adipogenic differentiation in normal and high glucose conditions revealed key metabolic shifts, with Nicotinamide Adenine Dinucleotide (NAD+) and L-glutamate/Ī±-ketoglutarate (Ī±-KG) identified as critical metabolites driving these processes. Supplementation with NAD+ and Ī±-KG in high glucose conditions significantly enhanced ALP activity and upregulated osteogenic markers, suggesting that these metabolites play a vital role in restoring osteogenic potential under hyperglycemic conditions. Overall, our findings suggest that high glucose promotes adipogenesis at the expense of osteogenesis, exacerbates cellular damage, and accelerates aging in BMSC. The identification of NAD+ and Ī±-KG as key regulators in this process provides new insights into the metabolic mechanisms behind impaired bone health in diabetes and highlights potential therapeutic avenues to counteract these detrimental effects. Further research into the role of NAD+ and Ī±-KG in BMSC differentiation under high glucose may offer novel strategies for managing diabetes-related bone diseases.
INSTITUTE
University of Adelaide
DEPARTMENT
Health and Medical Sciences
LABORATORY
MSC Lab
LAST_NAME
Shirazi
FIRST_NAME
Suzanna
ADDRESS
Adelaide University, North Terrace, Adelaide, SA, 5000, Australia
Reprogramming the metabolome of Centella asiatica (L.) Urban callus: Profiling of newly synthesized cryptic anthocyanins triggered by LED light exposure
STUDY_TYPE
Plant metabolomics
STUDY_SUMMARY
In addition to the pentacyclic triterpenoid centelloids, Centella asiatica also synthesizes hydroxycinnamic acid conjugates as well as flavonoids. The latter is the major class of secondary plant metabolites and comprises of various subclasses, including anthocyanidins. Anthocyanins are rarely reported in extracts from C. asiatica and differ structurally due to a flavylium (2-phenylchromenylium) ion that carries a positive charge at the oxygen atom of the C-ring of the basic flavonoid structure. Callus of C. asiatica was initiated and propagated on synthetic media and subjected to different light regimes. White callus resulted from white fluorescent illumination, while purple callus developed in response to white light emitting diode (LED) illumination. In order to profile the metabolites responsible for the intense purple colouration, methanolic extracts were prepared from the two cell lines. Total phenolic, flavonoid and anthocyanin content were determined and indicated (i) very low levels of flavonoids and anthocyanins in white callus and (ii) that anthocyanins dominate the flavonoid content of the purple callus. Extracts were subjected to untargeted ultra high-performance liquid chromatography coupled to high-definition mass spectrometry to profile newly synthesised anthocyanins. Metabolite annotation was based on accurate mass determination and characteristic fragmentation patterns. We report on the reprogramming of the metabolome of white C. asiatica callus due to LED illumination and profile the cryptic anthocyanins as well as putative flavonoid and caffeoylquinic acid co-pigments in purple callus. This study will provide contribute to the knowledge this understudied class of metabolites and will bring light into the role of LED in the activation their production in vitro.
Phytochemical analysis of Quercus ilex nuts revealed high variability, nutritional and nutraceutical value.
STUDY_TYPE
Untargeted metabolomics
STUDY_SUMMARY
In recent years, new feeding systems based on non-conventional crops have been gaining in popularity. Holm oak is a non-domesticated forest species and very dominant in the Mediterranean Basin. Its fruit, the acorn, is under study due to its high nutritional and nutraceutical value. However, the Quercus ilex species presents a great natural variability, producing fruits with very different qualities. Therefore, in this work, 14 individuals distributed in different locations have been selected to characterise their metabolomic profile and to identify bioactive compounds of interest in different industries and compounds that have been previously associated with specific flavours. The acorns have not been previously classified as there are no specialised tasters. The results obtained may be of great value in laying the foundations for the nutritional potential of acorns and may be complemented by other chemical analyses and future studies looking for bittersweet acorns.
INSTITUTE
University of Cordoba
DEPARTMENT
Department of Biochemistry and Molecular Biology
LABORATORY
AGR-164
LAST_NAME
Tienda
FIRST_NAME
Marta
ADDRESS
Campus de Rabanales, Edificio C6 Severo Ochoa, Ctra. Madrid, Km 396. 14071 CĆ³rdoba, Spain
Integrating Metagenomics and Metabolomics to Study the Gut Microbiome and Host Relationships in Sports Across Different Energy Systems
STUDY_SUMMARY
This study explored the role of the gut microbiome in modulating host metabolism among Colombian athletes by comparing elite weightlifters (n = 16) and cyclists (n = 13) through integrative omics analysis. Fecal and plasma samples collected one month before an international event underwent metagenomic, metabolomic, and lipidomic profiling. Metagenomic analysis via bioBakery tools revealed significant microbial pathways, including L-arginine biosynthesis III and fatty acid biosynthesis initiation. Key metabolic pathways, such as phenylalanine, tyrosine, and tryptophan biosynthesis; arginine biosynthesis; and folate biosynthesis, were enriched in both athlete groups. Plasma metabolomics and lipidomics revealed distinct metabolic profiles and a separation between athlete types through multivariate models, with lipid-related pathways such as lipid droplet formation and glycolipid synthesis driving the differences. Notably, elevated carnitine, amino acid, and glycerolipid levels in weightlifters suggest energy system-specific metabolic adaptations. These findings underscore the complex relationship between the gut microbiota composition and metabolic responses tailored to athletic demands, laying the groundwork for personalized strategies to optimize performance. This research highlights the potential for targeted modulation of the gut microbiota as a basis for tailored interventions to support specific energy demands in athletic disciplines.
INSTITUTE
Universidad del Rosario
LAST_NAME
Aya
FIRST_NAME
Viviana
ADDRESS
calle 13 a sur # 7a 28, Bogota, Bogota, 110411, Colombia
(p)ppGpp and DksA play crucial role in reducing the efficacy of -lactam antibiotics by modulating bacterial membrane permeability
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
The key signaling molecules in the bacterial stress sensing pathway, the alarmone (p)ppGpp and transcription factor DksA, help in survival during nutritional deprivation and exposure to xenobiotics by modulating cellular metabolic pathways. In Vibrio cholerae, (p)ppGpp metabolism is solely linked with the functions of three proteins: RelA, SpoT, and RelV. At threshold or elevated concentrations of (p)ppGpp, the level of cellular metabolites and proteins in the presence and absence of DksA in V. cholerae and other bacteria has not yet been comprehensively studied. We engineered the genome of V. cholerae to develop DksA null mutants in the presence and absence of (p)ppGpp biosynthetic enzymes. We observed a higher sensitivity of the (p)ppGpp0ĪdksA V. cholerae mutant to different -lactam antibiotics compared to the wild-type (WT) strain. Our whole-cell metabolomic and proteome analysis revealed that the cell membrane and peptidoglycan biosynthesis pathways are significantly altered in the (p)ppGpp0, ĪdksA, and (p)ppGpp0ĪdksA V. cholerae strains. Further, the mutant strains displayed enhanced inner and outer membrane permeability in comparison to the WT strains. These results directly correlate with the tolerance and survival of V. cholerae to -lactam antibiotics. These findings may help in the development of adjuvants for -lactam antibiotics by inhibiting the functions of stringent response modulators.
INSTITUTE
Translational Health Science And Technology Institute (THSTI)
Lipidomics analysis of mouse PDAC cell lines treated with alpha-eleostearic acid
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Mouse pancreatic cancer epithelial cells (mPDAC1) were cultured in RPMI, TIFM (tumor interstitial fluid medium), or TIFM + arginine and treated with 50 uM alpha-Eleostearic acid (a-ESA) or vehicle for 24 hours and analyzed for lipidomic changes. Cells that were arginine-deprived (TIFM) were found to exhibit a greater increase in alpha-eleostearic acid incorporation, particularly in triglycerides and phosphatidylethanolamines. Arginine deprived cells also showed a strong decrease in the abundance of monounsaturated fatty acids in triglycerides and phosaphtidylethanolamines, whereas these changes were not observed in cells cultured in RPMI or TIFM + arginine.
Lipidomics analysis of mouse PDAC cell lines treated with tung oil
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Mouse pancreatic cancer epithelial cells (mPDAC1) cultured in RPMI, TIFM (tumor interstitial fluid medium), or TIFM + arginine were treated with 200 ug/mL tung oil-bsa conjugate or fatty acid free BSA as a control for 24 hours and analyzed for lipidomic changes. We found that arginine depleted cells (TIFM) exhibited greater incorporation of 18:3 fatty acids (the dominant fatty acid in tung oil) than cells cultured in RPMI or TIFM + arginine.
Lipidomics analysis of mice treated with tung or safflower oil by oral gavage.
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
Lyz2(-/-)-Cre;Arg1(fl/fl) or Lyz2(+/+)-Cre;Arg1(fl/fl) mice were injected with orthotopic pancreatic cancer cell tumors and mice were gavaged with tung oil (PUFA high) or safflower oil (control) for 4 weeks. After treatment, tumors were dissected and sorted for pancreatic cancer cells and lipidomics was performed to determine how the lipidome of tumors changed. Lyz2(-/-)-Cre;Arg1(fl/fl) tumors are arginine deprived, while Lyz2(+/+)-Cre;Arg1(fl/fl) tumors have plasma levels of arginine, which allowed us to assay how arginine deprivation affects PUFA treatment in vivo. We found that arginine low tumors were more strongly affected by tung oil administration than tumors with plasma levels of arginine. This was reflected in the cellular peroxidation index of triglycerides in this dataset, which was calculated from this dataset.
Lipidomics analysis of mouse pancreatic cancer cells cultured RPMI, TIFM, or TIFM + arginine under lipid deprivation
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
KrasG12D mutant mouse pancreatic ductal adenocarcinoma cell lines (mPDAC1) were cultured in lipid replete or lipid deplete medium and analyzed for lipidomic changes. Cell lines were cultured in RPMI-1640, TIFM (https://elifesciences.org/articles/81289), or TIFM + arginine (1.149 mM). To do this, cells in each of the three mediums were deprived of lipids for 24 hours and their lipidomes were analyzed for changes. We find that mPDAC1 cells cultured in arginine-deprived conditions (TIFM) were unable to maintain their pools of monounsaturated and saturated fatty acids, whereas cells cultured in RPMI or in TIFM + arginine were able to maintain monounsaturated and saturated fatty acid pools upon lipid deprivation.
INSTITUTE
University of Chicago
DEPARTMENT
Ben May Department for Cancer Research
LABORATORY
Muir Lab
LAST_NAME
Jonker
FIRST_NAME
Patrick
ADDRESS
929 E 57th St. Chicago IL, 60637
EMAIL
pbjonker@uchicago.edu
PHONE
6162884547
NUM_GROUPS
10
STUDY_COMMENTS
Lipidomics analysis performed at Van Andel Institute (Grand Rapids, MI). Thanks to Dr. Ryan Sheldon and the Mass Spec Core for running and processing these samples.
Metabolomics of plasma and tumor interstitial fluid (TIF) of tumors from Lyz2-Cre(+/+);Arg1(fl/fl) mice.
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Mice or the genotype Lyz2-Cre(-/-);Arg1(fl/fl) or Lyz2-Cre(+/+);Arg1(fl/fl) were implanted with PDAC tumors and allowed to grow for 4 weeks. After 3 weeks of growth, mice were placed on 8% deuterated water for fatty acid synthesis analysis. To confirm that Lyz2-Cre(+/+);Arg1(fl/fl) had plasma concentrations of arginine, we performed metabolomics analysis on the plasma and tumor interstitial fluid of tumors from each mouse genotype. We found that arginine concentrations rose to plasma levels in Lyz2-Cre(+/+);Arg1(fl/fl) mice. This confirmed that this mouse model allows us to assay arginine deprivation in vivo.
RNA modification profiles in archaeal and non-archaeal species
STUDY_SUMMARY
Total RNA from five archaea, E. coli, S. cerevisiae and universal human reference RNA were digested to nucleosides and subjected to UHPLC-QqQ analysis. Each species was searched against a panel of 76 nucleoside standards.
Identification and quantification of a m7G modification in T. kodakarensis ribosomal RNA
STUDY_SUMMARY
To investigate the activity of the protein encoded by TK0008, we conducted a comprehensive nucleoside analysis comparing the total RNA from the TS559 wild-type strain with that of the TK0008-deletion strain.
INSTITUTE
New England Biolabs
LAST_NAME
Tsai
FIRST_NAME
Yueh-Lin
ADDRESS
44 Dunham Ridge, Beverly, Massachusetts, 01915, USA
Honey bee egg composition changes seasonally and after acute maternal virus infection.
STUDY_TYPE
Metabolomics and lipidomics
STUDY_SUMMARY
Honey bee (Apis mellifera) colonies depend on the reproductive output of their queens, which in turn is contingent on the care provided by worker bees. In this study, we collected eggs from honey bee queens in natural field conditions and found that egg collection date strongly influenced egg composition, likely reflecting seasonal variations in pollen resources. These findings highlight that while viral infections can induce transgenerational effects on egg proteomes under short-term experimental conditions, such effects are less apparent in natural settings and can be overshadowed by seasonal and other ecological factors.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry and Molecular Biology, Life Sciences Institute
Virus infection of honey bee queens alters lipid profiles and indirectly suppresses a retinue pheromone component via reducing ovary mass
STUDY_SUMMARY
In this study, we investigated the effects of viral infection on honey bee queens by conducting controlled laboratory infections and performing lipidomic profiling. Our analysis revealed that virus infections specifically reduce the queen retinue pheromone (QRP) component methyl oleate, a key signal in colony communication. Additionally, lipidomics data suggest that viral infection leads to a decrease in triacylglycerol abundances, potentially impacting queen energy metabolism and pheromone production.
INSTITUTE
University of British Columbia
DEPARTMENT
Biochemistry and Molecular Biology, Life Sciences Institute
Proteomics and metabolomics analysis of Mongolian acupuncture therapy for type 2 diabetes mellitus with atherosclerosis
STUDY_SUMMARY
The results of serum biochemical and H&E staining analysis suggest that acupuncture therapy can improve cellular inflammatory factors, kidney function, serum lipid function and upper carotid artery tissue lesions in rat of T2DM with atherosclerosis, which is an effective treatment method. In addition, we applied proteomics and metabolomics to comprehensively demonstrate that Mongolian acupuncture therapy in T2DM with atherosclerosis mainly through regulation of fatty acid metabolism.
Altered Omega-6/Omega-3 PUFA Ratios and Phospholipid Profiles in CFTR-Mutant PANC-1 Cells Reveal Novel Links Between CFTR Function and Lipid Metabolism
STUDY_SUMMARY
CFTR mutations have been implicated in pancreatitis and pancreatic cancer. CFTR is known to affect lipid metabolism and immune regulation. In this study, we employed untargeted lipidomics with iterative tandem mass spectrometry (MS/MS) to characterize the differences in PANC-1 cells with different CFTR states (overexpression, empty vector, and mutations). Our study found that three PANC-1 cells carrying CFTR mutations (p.L88X, p.E681V, p.C1355Y) showed a significantly higher ratio of FA 20:4 (omega-6 PUFA) and FA 22:6 (omega-3 PUFA) compared to the CFTR wild-type (CFTR OE) PANC-1 cells. Among the arachidonoyl species, the compounds PC16:0_20:4 and PC18:0_20:4 showed higher normalized intensities in CFTR-mutated PANC-1 cells compared to CFTR OE. The compound PS18:0_20:4 exhibited a difference only between CFTR p.L88X PANC-1 cells and CFTR OE. The compounds PI16:0_20:4 and PI18:0_20:4 showed differences comparing CFTR p.L88X and p.C1355Y cells with CFTR OE. In the docosahexaenoyl species, the compounds PC18:0_22:6 and PI18:0_22:6 displayed higher normalized intensities in CFTR-mutated PANC-1 cells compared to CFTR OE. The compound PC16:0_22:6 exhibited differences comparing CFTR p.L88X and p.C1355Y cells with CFTR OE.
The Chromosome-Scale Assembly and Multi-Omics Analysis Reveal Adaptive Evolution and Nitrogen Utilization Mechanisms in Edible Grass (Rumex patientia L.ĆĀ Rumex tianschanicus A. LOS)
STUDY_SUMMARY
Edible grass (Rumex patientia L.ĆĀ Rumex tianschanicus A. LOS), a perennial herbaceous plant from the Polygonaceae family, boasts a high protein content and rapid growth rate, making it a promising solution to feed shortages as a forage protein source. In this study, we utilized the PacBio sequencing platform and integrated methods including Hi-C to achieve a chromosomal-scale assembly of the R. patientia genome. The assembled genome spans 2.19 Gb with an N50 of 18.84 Mb, and 93.61% (2.05 Gb) of the assembly has been allocated to 30 pseudochromosomes. Comparative genomic analysis has revealed significant expansion of gene families involved in nitrogen metabolism and D-glutamine and D-glutamate metabolism pathways, which are responsible for the plant's strong nitrogen utilization capabilities and high protein content. Additionally, expansions in gene families associated with the Wnt signaling pathway, ubiquitin-mediated proteolysis, Toll and Imd signaling pathways, TGF-ĆĀ² signaling pathway, protein processing in the endoplasmic reticulum, photosynthesis-antenna proteins, circadian rhythm, and cell cycle pathways are closely related to the rapid growth and development of R. patientia. We have also identified the rhizosphere microbiome of R. patientia and, by integrating metabolomic data from root tissues and soil, found that during rapid growth phases, the plant secretes various apigenin-like compounds into the soil, enhancing the symbiotic nitrogen-fixing capabilities and potentially providing nitrogen sources to the leaves through symbiotic nitrogen fixation. Our research provides crucial insights into the genetic basis of R. patientia 's utility as a forage protein source.
INSTITUTE
Hunan Agricultural University
LAST_NAME
li
FIRST_NAME
zhu
ADDRESS
1 Nongda Road, Changsha City, Hunan Province, Changsha, Hunan, 410128, China
PILRA regulates microglial neuroinflammation and lipid metabolism as a candidate therapeutic target for Alzheimerās disease
STUDY_TYPE
In vitro study
STUDY_SUMMARY
The Alzheimerās disease (AD) human genetic landscape identified microglia as a key disease-modifying brain cell type. A common loss-of-function coding variant in paired immunoglobulin-like type 2 receptor alpha (PILRA) associated with reduced AD risk, was found enriched in a cohort of healthy centenarians and rescued APOE4 AD risk, however the mechanisms underlying protection are undefined. Here we identify biological functions of PILRA, an immunoreceptor tyrosine-based inhibitory motif (ITIM)-containing receptor, in human iPSC-derived microglia (iMG) and chimeric AD mice. CRISPR-mediated PILRA knockout (KO) in iMG rescued ApoE4-induced immunometabolic deficits. Moreover, loss of PILRA confers a signature of metabolic resilience in microglia with increased mitochondrial capacity in tandem with elevated antioxidants, reduced ROS and toxic lipid species. Additionally, PILRA KO iMG exhibit improved lysosomal degradation, enhanced migration, and attenuated cytokine responses. We show PPAR and STAT1/3 act as master regulators that mediate downstream signaling and regulate PILRA-dependent microglial functions. Importantly, AD mice transplanted with human PILRA KO microglia showed reduced amyloid pathology and rescued levels of synaptic markers. Finally, we identify a high-affinity human PILRA-specific antagonist antibody that phenocopies PILRA KO. Together, these findings suggest a therapeutic approach to modulate microglial immunometabolism by inhibiting PILRA, thus identifying a pharmacologically tractable target for AD.
INSTITUTE
Denali Therapeutics
LAST_NAME
Suh
FIRST_NAME
Jung
ADDRESS
161 Oyster Point Blvd, South San Francisco, CA 94080
Impact of Trans-Fatty acids on renal health in mice with partial TgfĪ²3 deletion: a Metabolomic and Lipidomic analysis
STUDY_TYPE
Multiplatform non-targeted metabolomics
STUDY_SUMMARY
Trans-fatty acids, prevalent in Western diets, contribute to the onset and progression of renal damage. This study investigates the effects of trans-fat consumption on kidney health using a comprehensive approach encompassing metabolomics, lipidomics, molecular, and cellular analyses. We analyzed both wild-type mice and mice with partial TgfĪ²3 deletion, a model for pre-existing renal damage. In control animals, the trans-fat diet induced significant dysregulation in renal lipid metabolism. Omic analyses revealed a reduction in triglycerides and an increase in plasmalogens, which are associated with cellular protection and signaling. Furthermore, upregulation of TCA cycle metabolites, malic and fumaric acids, indicated mitochondrial dysfunction. In mice with partial TgfĪ²3 deletion, trans-fat consumption exacerbated renal fibrosis, and further depleted triglycerides and plasmalogens. Despite similar lipid profiles in both groups, TgfĪ²3-deleted mice exhibited larger lipid droplets and worsened renal damage, including a notable reduction in taurine levels. This study underscores the value of detailed metabolic profiling to uncover mechanisms driving renal dysfunction linked to harmful dietary habits and genetic mutations.
INSTITUTE
Universidad Rey Juan Carlos
DEPARTMENT
Basic Sciences of Health
LABORATORY
Lipobeta
LAST_NAME
Lanzon
FIRST_NAME
Borja
ADDRESS
Avenida de Atenas S/N
EMAIL
borja.lanzon@urjc.es
PHONE
663692554
NUM_GROUPS
4
TOTAL_SUBJECTS
28
NUM_MALES
28
STUDY_COMMENTS
Study is composed of 4 groups of mice: wild-type on control diet (WT-CD), wild-type on trans diet (WT-HFD), heterozygous TGFBeta 3 on control diet (HZ-CD) and heterozygous TGFBeta 3 on trans diet (HZ-HFD)
Cardiac metabolome signatures of hepato-cardiac FGF21 signaling in pressure overload-induced cardiac hypertrophy
STUDY_SUMMARY
This study investigates the early (3 days) mid- (2 weeks) and late (8 weeks) cardiac metabolome changes driven by FGF21 signaling in the context of pressure overload-induced cardiac hypertrophy. Targeted NMR-based metabolomic profiling was performed on cardiac tissue from control mice at 3 days, 2 weeks, and 8 weeks following transverse aortic constriction (TAC), as well as from hepatocyte-specific (HEP-FGF21) and cardiomyocyte specific (CM-FGF21) Fgf21 knockout mice at the 8-week post-TAC timepoint. Sham-operated wild type mice and mice expressing Cre in hepatocytes or cardiomyocytes served as controls for each timepoint. The early and mid-timepoints were selected to capture transient and potentially triggering metabolic adaptations that precede overt cardiac remodeling, while the late timepoint allowed assessment of established metabolic reprogramming in advanced hypertrophy. Inclusion of the knockout models at 8 weeks enabled dissection of the respective contributions of endocrine (hepatocyte-derived) and autocrine (cardiomyocyte-derived) FGF21 signaling to the cardiac metabolome phenotype. The analysis revealed time-dependent alterations in the metabolic profile, with lower glucose and glucose derived metabolites in early timepoints and accumulation of Branched-Chain Amino Acids when pathological hypertrophy has fully developed (8 weeks). Ablation of Fgf21 in either the hepatocytes or cardiomyocytes mitigated maladaptive metabolic remodeling at 8 weeks, highlighting distinct and cooperative roles of hepatic and cardiac FGF21 in the progression of hypertrophy.
INSTITUTE
University of Cincinnati College of Medicine
LAST_NAME
Siokatas
FIRST_NAME
Georgios
ADDRESS
231 ALBERT SABIN WAY, University of Cincinnati, Cincinnati, OH, 45267-2827
Screen potential marker proteins influencing spermatogenesis in the testes of Shandong Black Cattle bulls via multi-omics integrated analysis.
STUDY_SUMMARY
This study was designed to identify candidate marker proteins that influence the growth and development of Shandong Black Cattle bull testes through a multi-omics joint analysis, thereby providing a certain theoretical basis for testis growth and development as well as bull selection. Eight 12-month-old Shandong Black Cattle bulls were selected, and testes tissues were collected. The testes were categorized into two groups based on their morphological characteristics: Group 1 (weight > 120 g) and Group 2 (weight < 120 g). Group 2 was employed as the control group to construct a protein and metabolite library for joint analysis to screen candidate marker proteins that affect testis growth and development. The results revealed that 1553 differential proteins (DEPs) were differentially expressed between the large and small testes of Black Bleykett bulls, with 1219 being upregulated and 334 being downregulated. The KEGG enrichment results manifested that the upregulated DEPs were primarily involved in the cell cycle (CDK1, CCNB, MCM4), DNA replication (MCM3, MCM4), etc. The downregulated DEPs were mainly associated with metabolic pathways (ACSM1, IMPDH1), etc. The GO enrichment results disclosed that the DEPs were significantly enriched in the categories of cytoskeleton movement. The weighted gene co-expression analysis suggested that the testis weight was significantly correlated with MCM, STRADA, and SEC31B. After integrating the DEPs, a PPI analysis was performed, and 10 key regulatory proteins were identified, including MCM3, MCM4, CDK1, and CDK2. Metabolomics demonstrated that 59 upregulated metabolites were enriched in the glucose metabolism pathway (uridine diphosphate glucose), and 14 downregulated metabolites were significantly enriched in metabolic pathways (hypoxanthine).
Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures embryonic radial glial cell integrity - part 1
STUDY_SUMMARY
Since fragmentation of the Golgi apparatus is associated with defects in phosphoinositide (PIP) metabolisms, we measured PI in E11.5-E13.5 cortices by using liquid chromatography-mass spectrometry (LC-MS). In Mboat7 KO mice, PI with arachidonic acid (38:4 PI), which is the major species in Mboat7 heterozygous mice, significantly decreased, and instead, PI with monounsaturated fatty acid (34:1, 34:2, 36:1, 36:2 PI) and PI with docosapentaenoic acid (DPA) or docosahexaenoic acid (DHA) (38:6, 40:5, 40:6, 40:7 PI) increased. On the other hand, the composition of molecular species of phosphatidylserine (PS), phosphatidylcholine (PC), and phosphatidylethanolamine (PE) showed little or no change in the cortices of E11.5-E13.5 Mboat7 KO mice compared with Mboat7 heterozygous mice.
Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures embryonic radial glial cell integrity - part 2
STUDY_SUMMARY
Since fragmentation of the Golgi apparatus is associated with defects in phosphoinositide (PIP) metabolisms, we measured PIPs in E11.5-E13.5 cortices by supercritical fluid chromatography-mass spectrometry (SFC-MS). In Mboat7 KO mice, PI4P with arachidonic acid (38:4 PI4P), which is the major species in Mboat7 heterozygous mice, significantly decreased, and instead, PI4P with monounsaturated fatty acid (34:1, 34:2, 36:1, 36:2 PI) and PI4P with docosapentaenoic acid (DPA) or docosahexaenoic acid (DHA) (38:6, 40:5, 40:6, 40:7 PI4P) increased. PI(4,5)P2 also showed changes in the molecular species similar to those observed in PI4P. Interestingly, the total amount of PI(4,5)P2 started to decrease at E12.5 in Mboat7 KO mice, whereas the total amount of PI4P was unchanged. This is because the decrease of arachidonic acid-containing PI(4,5)P2 species was not fully compensated by the increase of other PI(4,5)P2 species. To rule out the possibility that changes in systemic lipid metabolism in global Mboat7 KO mice affect radial glial cells in a non-cell-autonomous manner, we generated telencephalon-specific Mboat7 KO mice (hereafter, Mboat7-Emx1-KO) by crossing Mboat7fl/fl mice with Emx1-Cre transgenic mice, in which Cre expression starts at E10.5 in the dorsal telencephalon. The number of cleaved caspase-3-positive cells increased in E13.5 and E14.5 cortices of Mboat7-Emx1-KO mice but not as much as in Mboat7 KO mice. Moreover, whereas p-H3-positive RGCs were located at the ventricular wall at E12.5, some p-H3-positive RGCs were located apart from the ventricular zone from E13.5 cortices of Mboat7-Emx1-KO mice (Fig 8C and D). Taken together, these results indicate that telencephalon-specific LPLAT11 deficiency caused the apical detachment and apoptosis of RGCs. In Mboat7-Emx1-KO mice, the change in the fatty acyl composition of PI and PIPs was delayed compared to Mboat7 KO mice, probably because Emx1-Cre drives the expression of Cre recombinase only from E10.5. Therefore, we generated neural-specific Mboat7 KO mice (hereafter, Mboat7-Sox1-KO) by crossing Mboat7fl/fl mice with Sox1-Cre transgenic mice, which drives the expression of Cre recombinase before E8.5 in the neuroepithelium. Detachment of RGCs from the ventricular wall was not observed in the E12.5 cortices of Mboat7-Sox1-KO. On the other hand, E13.5 cortices of Mboat7-Sox1-KO showed detachment from the ventricular wall and apoptosis to the same extent as those of Mboat7 KO mice. Also, Mboat7-Sox1-KO showed rounding of the Golgi apparatus at E12.5 when the detachment of RGCs from the ventricular wall was not observed, which is consistent with the phenotype observed in E12.0 cortices of Mboat7 KO mice. Moreover, E-cadherin overexpression failed to rescue the detachment of RGCs from the apical ventricular wall. In the Mboat7-Sox1-KO cortices, a significant change in the fatty acyl composition of PI and PIPs was observed. The total amount of PI(4,5)P2 started to decrease at E12.5 in the Mboat7-Sox1-KO cortices as well as in the Mboat7 KO cortices. Taken together, these results indicate that Mboat7-Sox1-KO mice mimicked almost all the phenotypes observed in the brain of Mboat7 KO mice.
Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures embryonic radial glial cell integrity - part 3
STUDY_SUMMARY
To further examine whether PI(4,5)P2 is needed for the proper morphology of the Golgi apparatus in RGCs, we treated E12.5 hemispheres with UNC3230, a PIPKIĪ³ inhibitor. UNC3230 significantly reduced the total amount of PI(4,5)P2, primarily affecting PI(4,5)P2 species containing arachidonic acid and its analogous fatty acid, Mead acid (20:3), while exerting minimal effect on other PI(4,5)P2 molecular species. UNC3230 treatment caused rounding of the Golgi apparatus, as observed in Mboat7 KO mice. We then administered UNC3230 into the lateral ventricle at E12.5 and immunostained E13.5 cortices with antibodies against PI(4,5)P2, GM130, E-cadherin, Sox2, and p-H3. UNC3230 administration caused a decreased level of PI(4,5)P2, a rounding of the Golgi apparatus, and reduced apical expression of E-cadherin in the ventricular zone, whereas the detachment of RGCs from the ventricular wall was not apparent. These results suggest that reduced PI(4,5)P2 levels cause rounding of the Golgi apparatus and decreased E-cadherin expression at the apical surface. We also treated E12.5 Mboat7 heterozygous hemispheres with Sevenin-1, an LPLAT11 inhibitor. Sevenin-1 treatment caused rounding of the Golgi apparatus in the Mboat7 heterozygous hemispheres, whereas it had no effect on the morphology of the Golgi apparatus in the Mboat7 KO hemispheres, highlighting the importance of LPLAT11 activity. SFC-MS analysis confirmed that Sevenin-1 decreased the amount of PI with arachidonic acid and the total amount of PI(4,5)P2.
Comprehensive Lipidomic Analysis Identifies Critical Lipid and Metabolic Pathway Shifts in Alport Syndrome
STUDY_SUMMARY
Alport syndrome (AS) is a hereditary kidney disease caused by COL4A3-5 gene mutations, leading to glomerular basement membrane abnormalities. While the genetic and structural aspects of AS are well established, the mechanisms linking collagen IV defects to podocyte injury remain incompletely understood. Emerging evidence suggests that lipotoxicity and lipid dysregulation may play a pivotal role in mediating podocyte damage in AS, akin to its established role in diabetic kidney disease (DKD). We sought to identify plasma and urine lipid alterations in autosomal dominant and X-linked AS compared with DKD and healthy controls. Using liquid chromatography coupled to mass spectrometry (LC-MS), we annotated 580 and 203 lipid species in plasma and urine, respectively. Volcano plot and ROC analyses (AUC ā„ 0.80) identified key lipids, including urinary HexCer 18:0(3O)/24:0(2OH) and CAR 12:0. These analyses highlighted the most relevant lipotoxic pathways, which may warrant deeper investigation for drug development in AS. Compared to controls, AS exhibited unbalanced sphingolipid catabolism, ceramide overload, and impaired fatty acid Ī²-oxidation, alongside phospholipid and cholesterol imbalances suggestive of compromised ABCA1-mediated lipid efflux and mitochondrial dysfunction. Comparisons with DKD indicated a shared lipotoxic environment with ceramide elevation and disrupted fatty acid metabolism. However, disease-specific adaptations emerged, with severe ABCA1 dysfunction and marked phospholipid/cholesterol derangements in DKD, whereas AS showed pronounced sphingomyelin depletion. These findings demonstrate that AS involves distinct lipidomic disruptions and underscore shared lipotoxic mechanisms. This improved understanding of disease-specific lipid imbalances provides new potential therapeutic targets to mitigate podocyte injury and slow progression of AS.
INSTITUTE
Universidad CEU San Pablo
DEPARTMENT
Centro de MetabolĆ³mica y BioanĆ”lisis (CEMBIO)
LAST_NAME
GonzƔlez
FIRST_NAME
Carolina
ADDRESS
km 0, Universidad CEU-San Pablo UrbanizaciĆ³n MonteprĆncipe. M-501
Untargeted lipidomics of gemcitabine-resistant PDAC cells
STUDY_SUMMARY
In this work we developed gemcitabine (GEMC) resistant (GEMR) PDAC cell lines from Panc1 and MiaPaCa2 cells. This dataset contains results and raw data for untargeted lipidomics of Panc1 cells, including control (CON), and GEMR cells alongside internal quality controls (iQC) and blank samples for background subtraction (as described under sample prep). Untargeted lipidomics enabled the quantification of more than 400 distinct molecular lipid species within Panc1 & MiaPaCa2 control (CON) & GEMR cells from 12 different classes of lipid, with 77 molecular species being differentially abundant between Panc1 control and GEMR cells following unpaired t test. Data were analysed using MS-DIAL 4.9.
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010 Australia
Untargeted lipidomics of gemcitabine-resistant cells
STUDY_SUMMARY
In this work we developed gemcitabine (GEMC) resistant (GEMR) PDAC cell lines from Panc1 and MiaPaCa2 cells. This dataset contains results and raw data for untargeted lipidomics of Panc1 cells, including control (CON), and GEMR cells alongside internal quality controls (iQC) and blank samples for background subtraction (as described under sample prep). Untargeted lipidomics enabled the quantification of more than 400 distinct molecular lipid species within Panc1 & MiaPaCa2 control (CON) & GEMR cells from 12 different classes of lipid, with 79 molecular species being differentially abundant between MiaPaCa2 control and GEMR cells following unpaired t test. Data were analysed using MS-DIAL 4.9.
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW, 2010, Australia
Analysis of metabolite levels in the circulation of pancreatic tumor bearing mice with or without depletion of myeloid cells
STUDY_SUMMARY
In this study, we assessed the impact of myeloid cells on the levels of circulating metabolites in the plasma of mice bearing tumors. To do so, murine pancreatic ductal adenocarcinoma (PDAC) cells were implanted into C57Bl6J CD11b-DTR host animals. Myeloid cells were then ablated with 25 ng/kg diptheria toxin (DT). Serum from DT treated mice was collected for analysis. Additionally, serum was collected from PDAC bearing mice that were vehicle treated. This data allows for identification of how myeloid cells in tumor bearing mice contribute to levels of circulating metabolites.
Untargeted lipidomics of combination gemcitabine/paclitaxel attenuated (CombAT) PDAC cells
STUDY_SUMMARY
In this work produced combination gemcitabine/paclitaxel attenuated (CombAT) Panc1 and MiaPaCa3 cells, and used untargeted lipidomics to profiled changes in their lipidome. This dataset includes CombAT cells and control (CON) cells, alongside internal quality controls (iQC) and blank samples for background subtraction (as described under sample prep). These data revealed extensive increases across 166 lipid species in Panc1 CombAT cell lines compared with CON, including increases in many triacylglycerol (TG) species and total TG. Increases were also seen in several TG species in CombAT MiaPaCa2 cells, with 30 lipid species increased in total when compared with CON. Data were analysed using MS-DIAL v4.9.
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, Australia
Fatty acids profiling of gemcitabine based chemotherapy resistant PDAC cells
STUDY_SUMMARY
In this work produced combination gemcitabine/paclitaxel attenuated (CombAT) Panc1 and MiaPaCa2 cells. We profiled AMP-derivatised fatty acids from both cell lines and compared them to control (CON) cells. Also included are a reference standard produced from AMP-derivatised fatty acid methyl esters (FAMEs), internal quality controls (iQC) made from pooled samples, and a blank sample for background subtraction. This study found increased 16:1n-7 and decreased 16:1n-10 in Panc1 CombAT cells, with no change in MiaPaCa2 CombAT cells. Also apparent were changes in polyunsaturated fatty acids in Panc1 CombAT cells consistent with a decrease in fatty acid desaturase 2 (FADS2) activity. Data were analysed using Agilent Profinder 10.0 with fatty acids identified using a Personal Compound Databases and Libraries (PCDL) developed from the FAMEs reference standard.
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, Australia
Stable-label palmitate tracing of gemcitabine-based chemotherapy in PDAC cells
STUDY_SUMMARY
In this work we developed both gemcitabine resistant (GEMR) Panc1 and MiaPaCa2 cell lines and combination gemcitabine/paclitaxel attenuated (CombAT) Panc1 and MiaPaCa2 cells. We found significant upregulation of de novo lipogenesis in both cell lines following GEMR or CombAT. In this study, we traced uniformly 13C labelled palmitate into all cell lines and compared it with controls (CON) to derived relative activity of stearoyl-CoA desaturase 1 (SCD1) and fatty acid desaturase 2 (FADS2). We found upregulation of SCD1 in Panc1 GEMR through increased ratio 16:1n-7/16:0, with no change in SCD1 activity with CombAT in either cell line. FADS2 activity was substantially suppressed in Panc1 CombAT cells through decreased 16:1n-10/16:0. This study contains the following groups: source - cell lines Panc1 or MiaPaCa2, blank or fatty acid methyl ester reference (FAMEs) group - control (CON), gemcitabine resistant (GEMR), or combination gemcitabine/paclitaxel attenuated (CombAT). matched_CON - GEMR or CombAT each had their own matched control cells plated at the same time label - cells cultured in either unlabelled (UL) or labelled (13C) palmitate (12.5 uM) conjugated to bovine serum albumin (2% w/v).
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, Australia
Effect of SCD1 or FADS2 knockdown on fatty acid profile of PDAC cells.
STUDY_SUMMARY
In this work we sought to characterise the impact of knockdown of stearoyl-CoA desaturase 1 (SCD1) and fatty acid desaturase 2 (FADS2) on Panc1 and MiaPaca2 PDAC cells by small interfering RNA (siRN). Panc1 cells exhibit significant Ī6 desaturase activity on saturated fatty acids producing n-10 monounsaturated fatty acids, whereas MiaPaCa2 exhibit little to no FADS2 activity. Knockdown of SCD1 (siSCD) in MiaPaCa2 cells produces a significant decrease in 16:1n-7 when compared with an siRNA negative control (siNEG). In Panc1 cells, siSCD results in a large increase in FADS2-derived 16:1n-10, whereas siFADS2 results in a large increase in SCD1-derived 16:1n-7.
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, Australia
Profiling of fatty acid double bond positing ing gemcitabine-resistant pancreatic ductal adenocarcinoma cells
STUDY_SUMMARY
In this work we developed gemcitabine (GEMC) resistant (GEMR) PDAC cell lines from Panc1 and MiaPaCa2 cells. This dataset contains ozone-induced fatty acid discovery (OzFAD) data, which shows differences in carbon-carbon double bond position between Panc1 and MiaPaCa2 cells. Panc1 cells contain significant amounts of 16:1n-10, which is an unusual isomeric fatty acid produced by Ī6 desaturation of palmitic acid by fatty acid desaturase 2 (FADS2). Panc1 GEMR cells show increased stearoyl-CoA desaturase 1 (SCD1)-derived 16:1n-7 alongside decreased 16:1n-10.
INSTITUTE
Victor Chang Cardiac Research Institute
LAST_NAME
Hancock
FIRST_NAME
Sarah
ADDRESS
Level 7 Lowy Packer Building, 405 Liverpool Street, Darlinghurst, NSW 2010, Australia
The Chromosome-Scale Assembly and Multi-Omics Analysis Reveal Adaptive Evolution and Nitrogen Utilization Mechanisms in Edible Grass (Rumex patientia L.Ć Rumex tianschanicus A. LOS)
STUDY_SUMMARY
Edible grass (Rumex patientia L.Ć Rumex tianschanicus A. LOS), a perennial herbaceous plant from the Polygonaceae family, boasts a high protein content and rapid growth rate, making it a promising solution to feed shortages as a forage protein source. In this study, we utilized the PacBio sequencing platform and integrated methods including Hi-C to achieve a chromosomal-scale assembly of the R. patientia genome. The assembled genome spans 2.19 Gb with an N50 of 18.84 Mb, and 93.61% (2.05 Gb) of the assembly has been allocated to 30 pseudochromosomes. Comparative genomic analysis has revealed significant expansion of gene families involved in nitrogen metabolism and D-glutamine and D-glutamate metabolism pathways, which are responsible for the plant's strong nitrogen utilization capabilities and high protein content. Additionally, expansions in gene families associated with the Wnt signaling pathway, ubiquitin-mediated proteolysis, Toll and Imd signaling pathways, TGF-Ī² signaling pathway, protein processing in the endoplasmic reticulum, photosynthesis-antenna proteins, circadian rhythm, and cell cycle pathways are closely related to the rapid growth and development of R. patientia. We have also identified the rhizosphere microbiome of R. patientia and, by integrating metabolomic data from root tissues and soil, found that during rapid growth phases, the plant secretes various apigenin-like compounds into the soil, enhancing the symbiotic nitrogen-fixing capabilities and potentially providing nitrogen sources to the leaves through symbiotic nitrogen fixation. Our research provides crucial insights into the genetic basis of R. patientia 's utility as a forage protein source.
Characterization of braināderived extracellular vesicle lipids in Alzheimer's disease
STUDY_SUMMARY
Lipid dyshomeostasis is associated with the most common form of dementia, Alzheimer's disease (AD). Substantial progress has been made in identifying positron emission tomography and cerebrospinal fluid biomarkers for AD, but they have limited use as frontāline diagnostic tools. Extracellular vesicles (EVs) are released by all cells and contain a subset of their parental cell composition, including lipids. EVs are released from the brain into the periphery, providing a potential source of tissue and disease specific lipid biomarkers. However, the EV lipidome of the central nervous system is currently unknown and the potential of braināderived EVs (BDEVs) to inform on lipid dyshomeostasis in AD remains unclear. The aim of this study was to reveal the lipid composition of BDEVs in human frontal cortex, and to determine whether BDEVs have an altered lipid profile in AD. Using semiāquantitative mass spectrometry, we describe the BDEV lipidome, covering four lipid categories, 17 lipid classes and 692 lipid molecules. BDEVs were enriched in glycerophosphoserine (PS) lipids, a characteristic of small EVs. Here we further report that BDEVs are enriched in etherācontaining PS lipids, a finding that further establishes ether lipids as a feature of EVs. BDEVs in the AD frontal cortex offered improved detection of dysregulated lipids in AD over global lipid profiling of this brain region. AD BDEVs had significantly altered glycerophospholipid and sphingolipid levels, specifically increased plasmalogen glycerophosphoethanolamine and decreased polyunsaturated fatty acyl containing lipids, and altered amideālinked acyl chain content in sphingomyelin and ceramide lipids relative to CTL. The most prominent alteration was a twoāfold decrease in lipid species containing antiāinflammatory/proāresolving docosahexaenoic acid. The inādepth lipidome analysis provided in this study highlights the advantage of EVs over more complex tissues for improved detection of dysregulated lipids that may serve as potential biomarkers in the periphery.
INSTITUTE
The University of Melbourne
DEPARTMENT
The Florey Institute of Neuroscience and Mental Health
Untargeted metabolomics using LC-MS/MS was used to identify and classify differentially abundant metabolites in inv(16) AML model mice (n=3) compared to controls (n=3).
Investigation of Hepatic Lipid Alterations Following Micro- and Nanoplastic-Ingestion
STUDY_SUMMARY
To investigate the metabolic consequences of micro- and nanoplastic (MNP) ingestion, a comprehensive lipidomics analysis of liver tissue was performed. Mice were orally administered 50 nm (nanoplastic) or 500 nm (microplastic) polystyrene particles weekly for 12 weeks. Following the treatment period, liver samples were collected, and lipid profiles were analyzed using lipidomics techniques. The results were compared to a control group of mice that received phosphate-buffered saline (PBS) under identical conditions, allowing for a clear assessment of MNP-induced alterations in hepatic lipid metabolism Following MNP ingestion, lipid class analysis revealed alterations in abundance. While triacylglycerols (TAG) and odd-chain triacylglycerols (TAGodd) levels were reduced in MNP-exposed livers, phosphatidylethanolamine (PE), lysophosphatidylethanolamine (LPE), and sphingomyelin (SM) levels were increased.
INSTITUTE
University of Bonn
DEPARTMENT
Developmental Biology of the Immune System, The Life & Medical Sciences Institute (LIMES)
LABORATORY
Mass Lab
LAST_NAME
Makdissi
FIRST_NAME
Nikola
ADDRESS
Carl Troll straĆe 31, Bonn, nordRhein westfalen, 53115, Germany
Separation and Characterization of a Clinically-Derived Staphylococcus epidermidis Strain HE23: Revealing Its Antibiotic Resistome and Metabolic Potential
STUDY_SUMMARY
Neonatal incubators, while essential for infant care, are high-risk environments for nosocomial infections due to microbial colonization. Staphylococcus epidermidis, a biofilm-forming opportunistic pathogen, frequently contaminates medical devices, yet its metabolic adaptation mechanisms and virulence potential in incubator settings remain poorly understood. This study integrates genomics and metabolomics to comprehensively profile a multidrug-resistant S. epidermidis HE23 strain isolated from neonatal incubators, aiming to reveal its antibiotic resistance determinants and toxic metabolite production linked to clinical pathogenicity. A bacterial strain (HE23) was isolated from a neonatal incubator and identified as Staphylococcus epidermidis by 16S rRNA sequencing. Whole genome sequencing was performed using Illumina and Oxford Nanopore platforms, and a high-quality genome was obtained by hybrid assembly (Unicycler). And metabolomic analysis was performed by liquid mass spectrometry detection. Metabolomic profiling of Staphylococcus epidermidis HE23 revealed 47 differentially expressed metabolites (15 upregulated, 32 downregulated; VIP > 1.0, p < 0.05). Among upregulated metabolites, two clinically significant toxins were identified:Harmaline (m/z 446.2529 ), Cinobufagin (m/z 460.2686). Pathway enrichment highlighted glutathione metabolism (VIP = 2.1, p = 0.008) and branched-chain amino acid degradation (VIP = 1.8, p = 0.015), suggesting redox stress adaptation and nutrient scavenging strategies.
INSTITUTE
Anhui Medical University
DEPARTMENT
College of Life Sciences
LABORATORY
College of Life Sciences, Anhui Medical University
Human to mouse microbiota transfer model demonstrates disease-modifying effects of the short-chain fatty acid biotherapy modified microbiota
STUDY_TYPE
Biomedical research
STUDY_SUMMARY
We used a human-to-mouse fecal microbiota transfer model to investigate whether the functional changes in the human microbiota present after the Short-chain fatty acid biotherapy (SCFA-biotherapy) could modify gut microbiome and thus stool metabolome in pups. Oral delivery of a SCFA-yielding biotherapy in adults with T1D was followed by increased SCFAs, altered gut microbiota and immunoregulation, as well as delaying diabetes in preclinical models. Fecal microbiota transfer demonstrated that the microbiota of SCFA-responders modified gut bacteria metabolism in humanized gnotobiotic mice.
Ryanodine receptor 1 Ca2+ leak is a signal in skeletal muscle, but chronic leak can underlie pathology. Here we show that in healthy male mouse, limb-girdle muscle presents higher sympathetic input, elevated ryanodine receptor 1 basal phosphorylation, Ca2+ leak and mitochondrial Ca2+ content compared to distal leg muscles. These regional differences are consistent with heat generation in resting muscle to maintain core temperature. The dysferlin-null mouse develops severe pathology in the limb-girdle but not leg muscles. Absence of dysferlin disrupts dihydropyridine receptorsā inhibitory control over ryanodine receptor 1 leak, synergistically increasing leak through the already phosphorylated receptor of limb-girdle muscle, altering Ca2+ handling and distribution leading to reactive oxygen species production prior to disease onset. With age, oxidation of Ca2+ -handling proteins in dysferlin-null limb-girdle muscle exacerbated basal Ca2+ movements. Our results show that muscle-specific pathology in dysferlin-null mice is triggered by increased ryanodine receptor 1 Ca2+ leak.
INSTITUTE
University of Queensland
DEPARTMENT
School of Biomedical Sciences
LAST_NAME
Launikonis
FIRST_NAME
Bradley
ADDRESS
School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, 4072, Australia
Sex differences in lipid profiles of visceral adipose tissue with obesity and gonadectomy
STUDY_TYPE
Lipidomics
STUDY_SUMMARY
In obesity, adipose tissue (AT) expansion is accompanied by chronic inflammation. Altered lipid composition in the visceral or gonadal white AT (GWAT) directly drive AT macrophage (ATM) accumulation and activation, switching immune cells to a proinflammatory phenotype. Sex steroid hormones modulate sex differences in visceral vs. subcutaneous lipid accumulation that correlates with metabolic syndrome, especially in men and post-menopausal women who are more prone to truncal obesity. Prior studies demonstrated sex differences in lipid species in HFD-fed mice but there is a gap in understanding the role of sex hormones in these lipid species. We hypothesized that sex hormone alterations with gonadectomy (GX) would further impact lipid composition in the obese GWAT. We performed untargeted lipidomics on GWAT from castrated (CAS) male (M) mice and ovariectomized (OVX) female (F) mice. The control groups are named M SHAM HFD and F SHAM HFD, and the experimental groups are named M CAS HFD (or M GX HFD in text) and F OVX HFD (or F GX HFD in text). Untargeted lipidomics of obese GWAT identified sex differences in phospholipids, sphingolipids, sterols, fatty acyls, saccharo-lipids and prenol- lipids. Males had significantly higher content of key precursor fatty acids (palmitic, oleic, linoleic and arachidonic acid) when compared to female and GX mice. Bulk RNA sequencing of sorted GWAT ATMs highlighted sex and diet differences in PUFA metabolism genes and oxylipin genes. These findings of sexual dimorphism in stored lipid species emphasize sex-differences in GWAT lipid metabolism pathways that alter lipid composition driving inflammation responses and metabolic disease risk.
Pre-treatment untargeted cerebrospinal fluid metabolomic profiling in tuberculous meningitis reveals multiple pathways associated with mortality
STUDY_SUMMARY
Background Dysregulation of cerebrospinal fluid (CSF) tryptophan metabolism contributes to the high mortality of tuberculous meningitis (TBM). We aimed to identify novel metabolic pathways associated with TBM mortality through untargeted metabolome-wide analysis. Methods We measured 619 metabolites using untargeted liquid chromatography-mass spectrometry in pre-treatment CSF from adults with TBM from Indonesia (n=388; 34 HIV-positive) and Vietnam (n=679; 250 HIV-positive). Sixty-day mortality was modelled using Cox regression, adjusting for age and HIV-status. Metabolites were ranked in a screening subset (n=194, Indonesia), and validated in the same cohort (n=194) and externally (n=679, Vietnam). Secondary analysis included variable selection, clustering to classify associated metabolites into subgroups, comparison with non-infectious controls, and correlation with patient characteristics, CSF cytokines, CSF protein, and serum metabolite concentrations. Findings Sixty-day mortality was 21.6% and was associated with the concentration of ten CSF metabolites, including tryptophan. The strongest association was with 3-hydroxyoctanoate (FA 8:0;3OH), part of a cluster of hydroxylated fatty acids, further including hydroxy-isocaproate (FA 6:0;OH), hydroxyisobutyrate (FA 4:0;OH), and C4-OH-carnitine. These fatty acids correlated weakly with CSF TNF-Ī±, IL-6, leukocyte counts, bacterial load and CSF protein. Mediation analysis showed that the variation in fatty acids was linked directly to mortality rather than through disease severity. Conclusion We identified and validated nine new metabolites associated with TBM mortality, independent of HIV-status, disease severity, and tryptophan. These metabolites suggest that altered fatty acid beta-oxidation is linked to TBM associated mortality. Interventions targeting cerebral fatty acid metabolism may improve survival from TBM.
Immune checkpoint blockade (ICB) has transformed cancer treatment, but success rates remain limited. Recent research suggests dietary fiber enhances ICB response in mice and patients through microbiome-dependent mechanisms. Yet, the robustness of this effect across cancer types and dietary contexts remains unclear. Specifically, prior literature compared grain-based chow (high fiber) to low-fiber purified diet, but these diets differ also on other dimensions including phytochemicals. Here we investigated, in mice fed grain-based chow or purified diets with differing quantities of isolated fibers (cellulose and inulin), the gut microbiome, metabolite levels and ICB activity in multiple tumor models. Isolated fibers shifted multiple microbial taxa toward chow-like levels, although diet-type (chow vs. purified) had a stronger impact on gut-microbiome composition. Metabolomic profiles were relatively similar between mice fed high- and low-fiber purified diets, but differed massively between mice fed purified diets or chow, identifying the factor as diet type, independent of fiber. Tumor growth studies in multiple murine models revealed that fiber has a weaker impact on ICB (anti-PD-1) efficacy than previously reported. While diet impacted ICB activity in some models, the effect was not directionally consistent. In no model did we observe the pattern expected if fiber controlled ICB efficacy: strong efficacy in both chow and high-fiber purified diet but low efficacy in low-fiber purified diet. Thus, dietary fiber appears to have limited or inconsistent effect on ICB efficacy in mouse models, and other dietary factors that correlate with fiber intake may underlie the clinical correlations between fiber consumption and immunotherapy outcomes.
INSTITUTE
Princeton University
LAST_NAME
Roichman
FIRST_NAME
Asael
ADDRESS
Lewis-Sigler Institute for Integrative Genomics Princeton University Carl Icahn Laboratory, Washington Road Princeton, NJ 08544
Immune checkpoint blockade (ICB) has transformed cancer treatment, but success rates remain limited. Recent research suggests dietary fiber enhances ICB response in mice and patients through microbiome-dependent mechanisms. Yet, the robustness of this effect across cancer types and dietary contexts remains unclear. Specifically, prior literature compared grain-based chow (high fiber) to low-fiber purified diet, but these diets differ also on other dimensions including phytochemicals. Here we investigated, in mice fed grain-based chow or purified diets with differing quantities of isolated fibers (cellulose and inulin), the gut microbiome, metabolite levels and ICB activity in multiple tumor models. Isolated fibers shifted multiple microbial taxa toward chow-like levels, although diet-type (chow vs. purified) had a stronger impact on gut-microbiome composition. Metabolomic profiles were relatively similar between mice fed high- and low-fiber purified diets, but differed massively between mice fed purified diets or chow, identifying the factor as diet type, independent of fiber. Tumor growth studies in multiple murine models revealed that fiber has a weaker impact on ICB (anti-PD-1) efficacy than previously reported. While diet impacted ICB activity in some models, the effect was not directionally consistent. In no model did we observe the pattern expected if fiber controlled ICB efficacy: strong efficacy in both chow and high-fiber purified diet but low efficacy in low-fiber purified diet. Thus, dietary fiber appears to have limited or inconsistent effect on ICB efficacy in mouse models, and other dietary factors that correlate with fiber intake may underlie the clinical correlations between fiber consumption and immunotherapy outcomes.
INSTITUTE
Princeton University
LAST_NAME
Roichman
FIRST_NAME
Asael
ADDRESS
Lewis-Sigler Institute for Integrative Genomics Princeton University Carl Icahn Laboratory, Washington Road Princeton, NJ 08544
Associations between lipoprotein subclasses and cardiovascular health in ageing
STUDY_SUMMARY
Abnormal lipoprotein profiles have been frequently described in association with atherogenic cardiovascular diseases. The relevance of lipoproteins in human cardiovascular health with ageing, which commonly manifests as heart failure in older adults, is less often investigated. Capitalizing on 1H NMR spectroscopy which identifies lipoprotein subclasses with great detail, we explored in-depth associations between 112 lipoproteins and cardiac function of N=474 unique older adults, hypothesizing that specific lipoproteins may be associated with cardiac ageing.Decreases in intermediate density LDLs (1.034-1.037 g/mL) and increases in intermediate density VLDLs (0.961-0.995 g/mL) were observed among older adults with poorer cardiac function. There was heterogeneous distribution of low density (0.950-0.961 g/mL) VLDL fractions, which became more prominent in patients with poor cardiac function. Prominent apolipoprotein-A increases specific to those with poorer cardiac function were observed among those taking lipid lowering medications, localized to specific lower density HDL fractions (1.063-1.112 g/mL).
Genetic architecture for maize leaf and kernel metabolome in CUBIC population
STUDY_SUMMARY
Maize is one of the most important crops worldwide, and enhancements of yield and quality are primary breeding targets for its genetic improvement. Here, we explored the metabolome profiling for maize leaf and kernel from a synthetic population CUBIC (Complete-diallel plus Unbalanced Breeding-derived Inter-Cross) of 1404 maize progenies derived from 24 diverse founders. 67 metabolite from leaf and 68 from kernel confirmed by authentic standardswith gas chromatography-mass spectroscopy (GC-MS) for metabolic profiling were selected for further genetic exploration. Using single-locus and haplotype-based genome-wide association study (sGWAS and hGWAS, respectively), we investigated the genetic basis for primary metabolite accumulation in maize leaves and kernels at a genome-wide scale with 1373 mQTL identified. We identified three genes (ZmASN2, ZmMIPS1 and ZmMGL) to regulate kernel filling and a variety of metabolites, affecting the nutritional value of maize kernels and a novel gene, ZmAVT1A-1, which regulates amino acid levels and enhances maize NUE-related traits through the allocation of nitrogen from vegetative organs to kernels. Our study provided a valuable resource for understanding maize primary metabolism and offered novel insights into the metabolic organization at the genetic and molecular levels that can enable crop genetic improvement.
A lipidomic exploration of the effects of high-intensity interval exercise in healthy men after metformin intake
STUDY_SUMMARY
Changes in the plasma levels of individual lipids could play an important role in the response to exercise under metformin intake. This study explores the changes in the plasma lipidome of nine healthy individuals for 12 h after metformin intake followed by either high-intensity interval exercise (HIIE) or rest, followed by food intake in both cases. We observed several variations in the lipid profiles due to HIIE. Most affected lipid classes are fatty acids, acyl carnitines, phosphatidylcholines, sphingomyelins, and triglycerides, of which most were increased upon HIIE, but only some (mainly fatty acids and triglycerides) upon food intake.
Glucose-6-phosphate-dehydrogenase on old peroxisomes maintains self-renewal of epithelial stem cells after asymmetric cell division
STUDY_SUMMARY
Peroxisomes play a crucial role in cellular metabolism. Glucose-6-phosphate dehydrogenase (G6PD), the gatekeeper enzyme of the pentose phosphate pathway, is primarily localized in the cytosol. However, studies have reported its presence in peroxisomes as well. This project aims to determine the function of G6PD on the peroxisomal membrane. In this study, we overexpressed G6PD in either the cytosol or on the peroxisomal membrane of mammary epithelial stem-like cells (hMECs). By comparing their lipidomic profiles, we found that peroxisomal membrane-associated G6PD provides NADPH, which feeds into peroxisomal ether lipid synthesis.
Longitudinal multi-omics analysis of cord blood and childhood serum in Autism Spectrum Disorder (Negative mode)
STUDY_TYPE
LC MS/MS quantitative analysis
STUDY_SUMMARY
Untargeted Metabolomic analysis was performed on cord blood plasma from 22 children diagnosed with Autism before age 5, and 44 neurotypical controls from the Cork Baseline Birth Cohort.
Longitudinal multi-omics analysis of cord blood and childhood serum in Autism Spectrum Disorder (Positive mode)
STUDY_TYPE
LC MS/MS quantitative analysis
STUDY_SUMMARY
Untargeted Metabolomic analysis was performed on cord blood plasma from 22 children diagnosed with Autism before age 5, and 44 neurotypical controls from the Cork Baseline Birth Cohort.
Characterisation of the gut microbiome in hypertensive and type II diabetic populations in different regions of Xinjiang
STUDY_SUMMARY
In this study, the diversity of gut microbial profiles and their associations with dietary habits and metabolites in different ethnic groups and disease states were revealed through gut microbiome and metabolomic analyses of 192 healthy and diseased people (including hypertension, type II diabetes and their co-morbidities) in Xinjiang.It was found that the dominant genus in the Hui group was Faecalibacterium, while the Uyghur group was dominated by Prevotella, which is different from the previously reported distribution of enterotypes.The abundance of Prevotella was significantly higher in hypertensive patients and positively correlated with a high-salt diet, and the abundance of Bifidobacterium adolescentis was significantly higher in patients with type II diabetes.By integrative analysis of multi-omics data, we further found that changes in the proportion of specific microorganisms (e.g. Coriobacteriales_bacterium and Dorea_formicigenerans) in disease-comorbid states were strongly associated with significant differences in urinary metabolites (e.g. L-carnitine and hydroxycinnamic acid).In addition, metabolic pathway analyses revealed significant alterations in glycolysis/glycolysis, phenylalanine metabolism, and other pathways in the disease state.This study systematically reveals for the first time the gut microbiome and metabolome characteristics of healthy and diseased populations of different ethnic groups in Xinjiang region, which provides a new perspective for understanding the role of gut microbes in metabolic diseases and potential scientific basis for early diagnosis and personalised treatment of diseases. Future studies will further combine multi-omics technology and longitudinal design to comprehensively reveal the interactions among factors and specific mechanisms.
INSTITUTE
Xinjiang University
LABORATORY
Laboratory of Synthetic Biology
LAST_NAME
Yue
FIRST_NAME
Haitao
ADDRESS
Yushugou Street, Shuimogou District, Urumqi City, Xinjiang Province
Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle - metabolomics from CD36 x PdgfBcre vs WT mice on control or SAAR diet
STUDY_SUMMARY
Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle Ī²-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability. We performed metabolomics analysis on the extensor digitorum longus (EDL), soleus (Sol), and serum to investigate the systemic effects of genotype and diet on muscle and circulatory metabolites. After one week of dietary treatment, significant changes in a wide range of metabolites were not observed. Interestingly, only minimal differences were detected between the WT and CD36 Ć PDGFb KO models.
Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle - extensor digitorum longus (EDL) versus soleus (Sol) on Con and SAAR diet metabolomics
STUDY_SUMMARY
Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle Ī²-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability. Here we performed metabolomics using EDL and Sol to evaluate the systemic effects of the diet on muscle metabolites. One week of dietary treatment was not sufficient to significantly alter a wide range of metabolites. However, known metabolites downstream of the methionine cycle, such as taurine, were significantly affected. Additionally, ophthalmic acidāwhich has been reported to compensate for glutathione loss under protein restriction (MacArthur et al., 2022; https://doi.org/10.1016/j.celrep.2022.111187)āexhibited significant upregulation, highlighting the robustness of the data.
INSTITUTE
Princeton University
DEPARTMENT
Department of Chemistry
LABORATORY
Prof. Dr. Joshua Rabinowitz
LAST_NAME
Mann
FIRST_NAME
Charlotte Greta
ADDRESS
Joseph Stelzmann Strasse 26, 50931 Cologne, Germany
Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle - extensor digitorum longus (EDL) versus soleus (Sol) on Con and SAAR diet lipidomics
STUDY_SUMMARY
Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle Ī²-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability. Here we performed lipidomics using EDL and Sol to evaluate the systemic effects of the diet on muscle lipid composition. One week of dietary treatment was not sufficient to significantly alter a wide range of lipid species in the muscle, prompting us to investigate changes in functional utilization rather than lipid storage.
Background: Ovarian cancer (OC) ranks as the most lethal gynecological malignancy worldwide, with early diagnosis being crucial yet challenging. Current diagnostic methods like transvaginal ultrasound and blood biomarkers show limited sensitivity/specificity. This study aimed to identify and validate serum metabolic biomarkers for OC diagnosis using the largest cohort reported to date. Methods: We constructed a large-scale OC-associated cohort of 1,432 subjects, including 662 OC, 563 benign ovarian disease, and 207 healthy control subjects, across retrospective (n=1,073) and set-aside validation (n=359) cohorts. Serum metabolic fingerprints (SMFs) were recorded using nanoparticle-enhanced laser desorption/ionization mass spectrometry (NELDI-MS). A diagnostic panel was developed through machine learning of SMFs in the discovery cohort and validated in independent verification and set-aside validation cohorts. The identified metabolic biomarkers were further validated using liquid chromatography MS and their biological functions were assessed in OC cell lines. Findings: We identified a metabolic biomarker panel including glucose, histidine, pyrrole-2-carboxylic acid, and dihydrothymine. This panel achieved consistent areas under the curve (AUCs) of 0.87-0.89 for distinguishing between malignant and benign ovarian masses across all cohorts, and improved to AUCs of 0.95-0.99 when combined with risk of ovarian malignancy algorithm (ROMA). In vitro validation provided initial biological context for the metabolic alterations observed in our diagnostic panel. Interpretation: Our study established a reliable serum metabolic biomarker panel for OC diagnosis with potential clinical translations. The NELDI-MS based approach offers advantages of fast analytical speed (~30 seconds/sample) and low cost (~2-3 dollars/sample), making it suitable for large-scale clinical applications.
Irisin promotes bone tissue repair and intestinal protection in osteoporotic mice
STUDY_SUMMARY
In this study, we constructed a mouse model of osteoporosis by performing ovariectomies and administering irisin injections. The impact of ovariectomy was assessed based on changes in body and uterine weights. Bone microstructure analysis, HE staining, MASSON staining, and Goldner staining were employed to observe alterations in bone structure. Post-irisin injection, mice showed recovery in weight and restoration of bone structure, including the formation of new bone. The presence of bone-forming factors (ALP, BGP), bone resorption factors (CTX-1, TRACP-5b), and a bone protective protein (OPG) indicated that irisin could promote bone cell growth in bone tissue. Additionally, HE staining and immunohistochemical analysis on intestinal sections revealed that irisin restored the mucosal barrier of the intestinal wall and alleviated enlargement of intestinal epidermal cells. Analysis of IL-1Ī², IL-6, and TNF-Ī± levels demonstrated irisin's role in reducing inflammation. Furthermore, sequencing and analysis of intestinal flora 16S microbial sequencing and blood metabolome in mice revealed that irisin mitigated the dysbiosis caused by osteoporosis and promoted flora growth, while enrichment analysis showed that irisin induced changes in basal metabolic levels. This study highlights the therapeutic potential of irisin in treating osteoporosis and its beneficial effects on the intestinal barrier and flora, suggesting that irisin enhances intestinal protection to foster bone tissue repair.
INSTITUTE
Huizhou Central People's Hospital
DEPARTMENT
Department of Traumatology and Orthopaedic Surgery, Orthopaedic Institute
LAST_NAME
Zhang
FIRST_NAME
Zhiwen
ADDRESS
No. 41, Eling North Road, Huicheng District, Huizhou City, Guangdong Province
Irisin Attenuates Postmenopausal Osteoporosis by Upregulating Phosphatidylcholine Metabolism via the Wnt/Ī²-catenin Pathway
STUDY_SUMMARY
Irisin can ameliorate postmenopausal osteoporosis (PMOP) in ovariectomized (OVX) mice by promoting osteogenesis, inhibiting adipogenesis, and regulating the bone-fat balance. Similarly, irisin enhances the osteogenic differentiation of human bone marrow mesenchymal stem cells (BMSCs) in vitro while reducing their adipogenic differentiation. Metabolomic analysis of mouse plasma revealed that the therapeutic effects of irisin are associated with pathways such as glycerophospholipid metabolism, wherein the metabolite phosphatidylcholine (PC) plays a significant role. Irisin can upregulate the metabolism of PC in OVX mice, thereby improving OP. We further investigated the metabolic role of PC and its mechanisms. By administering PC to BMSCs, we observed its effects on BMSCs and the underlying mechanisms. It was found that PC activates the canonical Wnt/Ī²-catenin signaling pathway, promoting the osteogenic differentiation of BMSCs while inhibiting their adipogenic differentiation.
INSTITUTE
Huizhou Central People's Hospital
DEPARTMENT
Department of Traumatology and Orthopaedic Surgery, Orthopaedic Institute
LAST_NAME
Zhang
FIRST_NAME
Zhiwen
ADDRESS
No. 41, Eling North Road, Huicheng District, Huizhou City, Guangdong Province, Guangdong Province, Huizhou City, Guangdong Province, 516001, China
Lactate activates trained immunity by fueling the tricarboxylic acid cycle and regulating histone lactylation
STUDY_SUMMARY
Using 13C-based metabolic tracers, we investigate the role of physiologic carbon sources (PCSs) on glucose metabolism in trained immunity. Ī²-glucan treatment led to the rapid conversion of 13C-glucose into lactate and intracellular TCA cycle intermediates (e.g., citrate and fumarate) within the first 8 hours in monocytes. As expected, glucose and glutamine were consumed within 12-24 hrs during the trained immunity process, and PCS treatment mildly affected glucose and glutamine consumption in Ī²-glucan treated monocytes. A slower decrease of other carbon sources, including Ī²OHB, citrate, and pyruvate, was observed in the PCS-supplemented medium compared to the glucose-supplemented medium.
INSTITUTE
Wuhan University
LAST_NAME
Liu
FIRST_NAME
Shi
ADDRESS
Luojia Hill, Wuchang District, Wuhan, Hubei, 430072, China
metabolomics of wild-type and TauT-/- leukemia cells
STUDY_SUMMARY
C57BL/6J mice conditioned with irradiation were transplanted with wild type and TauT-/- leukemia stem cells. The mice were sacrificed on days 11-13 post-transplant. Spleens from leukemic mice were quickly dissected and dissociated in Hanksā balanced salt solution (Gibco) with 5% fetal bovine serum and 2mM EDTA at 4C. The leukemia cells were stained for Lin+ (CD3Īµ-, CD4-, CD8-, Gr1-, CD11b/Mac-1-, TER119-, CD45R/B220- and CD19-) markers and were magnetically depleted using LD columns (Milteny Biotec). Lin- leukemia stem cell fraction was washed with PBS containing 5mM glucose and centrifuged at 3000g for 1 min, snap frozen, and processed for untargeted metabolomics
We traced 13C taurine in K562 human leukemia cells. Briefly K562 cells were cultured in serum free RPMI-1640 supplemented with 200uM taurine (1,2 13C2, 98%) or no taurine for 24 h. Untargeted metabolomic analysis was carried out to determine direct incorporation of taurine into any cellular metabolite.
Taurochenodeoxycholic acid levels are associated with scar hypertrophy in the rabbit ear hypertrophic scar model.
STUDY_SUMMARY
New Zealand white rabbits were used to establish the rabbit ear hypertrophic scar model. On postoperative day 28, the rabbits were divided into scar-susceptible (group H) and scar-insusceptible (group L) groups according to macroscopic scar thickness. Using fecal microbiota transplantation (FMT), we exchanged the gut microbiota of group H and group L. Untargeted metabolomics was performed before and after FMT. We found that in the rabbit ear hypertrophic scar model, scar hypertrophy was more severe in group H and was associated with fecal microbiota and their metabolites. Gut microbiota components and key genera were transferred between group H and group L by FMT, accompanied by altered levels of key metabolites such as TCDCA (bile acid taurochenodeoxycholic acid). FMT relieved scar hypertrophy in group H and vice versa. TCDCA were decreased in group H rabbit and its relative abundance was successfully reversed by heterochronic FMT.
INSTITUTE
Shenzhen Center for Chronic Disease Control,
DEPARTMENT
Plastic surgery, Shenzhen Institute of Dermatology
LAST_NAME
Liang
FIRST_NAME
Zhengyun
ADDRESS
No. 2021, Buxin Road, Luohu District, Shenzhen 518020, China
Shift in the urinary metabolome associated with 2,3,7,8-tetrachlorodibenzo-p-dioxin activation of the hepatic aryl hydrocarbon receptor
STUDY_SUMMARY
Epidemiological evidence suggests an association between dioxin and dioxin-like compound (DLC) exposure and human liver disease. In rodents, the prototypical DLC, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), has been shown to induce the progression of reversible hepatic steatosis to steatohepatitis with periportal fibrosis and biliary hyperplasia. Although the effects of TCDD toxicity are mediated by aryl hydrocarbon receptor (AHR) activation, the underlying mechanisms of TCDD-induced liver toxicity are not well understood. In the present study, male C57BL/6NCrl mice were gavaged every 4 days for 28 days with 0.03 - 30 Āµg/kg TCDD or sesame oil vehicle and evaluated for liver histopathology and gene expression as well as complementary 1-dimensional proton magnetic resonance (1-D 1H NMR) urinary metabolic profiling. Urinary trimethylamine (TMA), trimethylamine N-oxide (TMAO), and 1-methylnicotinamide (1MN) levels were altered by TCDD at doses ā¤ 3 Āµg/kg; other urinary metabolites, like glycolate, urocanate, and 3-hydroxyisovalerate, were only altered at doses that induced moderate to severe steatohepatitis. Hepatic differential gene expression of rate-limiting enzymes of choline, gloxylate, and amino acid metabolism coincided with the altered urinary metabolites. Published single-nuclear RNA-seq (snRNA-seq), AHR ChIP-seq, and AHR knockout gene expression datasets provided further support for hepatic cell-type and AHR-regulated dependency for the affected metabolic pathways.
Hepatic Coenzyme Q (CoQ) Deficiency Drives Reverse Electron Transport and Disrupts Hepatic Metabolic Homeostasis in Obesity
STUDY_SUMMARY
Mitochondrial reactive oxygen species (mROS) play a crucial role in physiology. While excessive mROS production has been associated with several disease states, its precise sources, regulation, and mechanisms of generation in vivo remain unclear, limiting translational efforts. mROS production is not a single process; at least 11 distinct sites associated with the electron transport chain and matrix substrate oxidation can partially reduce oxygen to superoxide and hydrogen peroxide. These sites exhibit substrate specificity and differ in their capacities to generate mROS. Here, we show that in obesity, hepatic coenzyme Q (CoQ) synthesis in vivo is impaired, leading to decreased CoQ9 levels and an increased CoQHā/CoQ ratio. These alterations in CoQ metabolism drive excessive mROS production via reverse electron transport (RET) at site IQ of complex I. Using multiple complementary genetic and pharmacological in vivo models, we demonstrate that RET is critical for maintaining metabolic health. Furthermore, we show that in patients with steatosis, hepatic CoQ biosynthesis is also suppressed, and the CoQHā/CoQ ratio positively correlates with steatosis grade. Our data identify a highly selective mechanism of pathological mROS production in obesity, which can be targeted to preserve metabolic homeostasis.
Differential acquisition of extracellular lipid correlates with pancreatic cancer subtype and metastatic tropism
STUDY_TYPE
Metabolomics
STUDY_SUMMARY
Pancreatic ductal adenocarcinoma (PDAC) stands as an aggressive disease, ranking at the third leading cause of cancer-related death with a 5-year overall survival rate of less than 5%. Recent studies have identified two major subtypes of pancreatic cancer, basal and classical, which predict patient prognosis, with basal tumors corresponding to more aggressive disease. While transcriptional signatures define basal and classical tumors and cell lines, little is known regarding the metabolic vulnerabilities linked to the basal versus classical state. Using unbiased computational interrogation to uncover metabolic genes correlating with known markers of classical PDAC, we identified high expression of Proprotein Convertase Subtilisin/Kexin type-9 (PCSK9) ā a negative regulator of receptor mediated uptake of cholesterol-containing low-density lipoprotein (LDL) ā as correlating with several established markers of classical PDAC. In contrast, PCSK9 expression is suppressed in basal PDAC. Accordingly, basal PDAC cells uptake high levels of LDL, are sensitive to LDL depletion and show higher dependence on cholesterol uptake, relative to classical PDAC. As lipoproteins are produced in the liver, we hypothesized that basal PDAC cells might possess a greater propensity to seed and thrive within liver tissues. Conversely, basal cells demonstrated a robust ability to grow within the liver but faced challenges in invading the lungs whereas classical cells harbour the opposite phenotype. Moreover, we observed that patients with liver metastasis alone had weaker PCSK9 staining at the primary tumor site compared to those with lung metastasis only. This observation suggests a potential influence of cholesterol metabolism in the primary tumor on the tropism of metastatic sites. Modulating PCSK9 levels in pancreatic cancer cells with low expression of this protein resulted in reduced LDL uptake and a shift towards the cholesterol biosynthesis pathway. More importantly, overexpressing PCSK9 not only reduced the liver metastasis burden but also increased lung metastasis in liver-tropic cell lines. Lastly, we found that PCSK9 expression and protein abundance remained low in liver metastatic lesions and high in lung metastasis lesions. This observation suggests that cancer cells maintain distinct cholesterol metabolism profiles in metastatic lesions. Ongoing studies will interrogate the role of cholesterol uptake and utilization in liver metastasis , and uncover unique features and vulnerabilities of the most aggressive variant of PDAC.
INSTITUTE
University of California, San Francisco
DEPARTMENT
Anatomy
LABORATORY
Perera
LAST_NAME
Rademaker
FIRST_NAME
Gilles
ADDRESS
513 Parnassus avenue, San Francisco, CA, 94143, USA
Occurrence of white flesh color and refreshing flavor following phytoene synthase 2A gene variation in loquat fruit
STUDY_SUMMARY
The white fleshed loquat (Eriobotrya japonica) fruit are more preferentially chosen by consumers than the orange fleshed ones. However, the metabolic basis that the white-fleshed loquat are more favorable is not known. The project carried out widely targeted metabolomes on 18 representative cultivars, and identified 1420 metabolites with 223 differentially accumulated metabolites between the two groups. The metabolite comparison in this project demonstrated that low levels of bitter or astringent compounds, such as flavonoids, lignans and coumarins, phenolic acids, nucleotides, alkaloids, and terpenoids, may confer to refreshing flavor of the white fleshed loquats. Furthermore, 18 metabolic biomarkers were identified by machine learning to distinguish loquat flesh colors.
INSTITUTE
Fujian Academy of Agricultural Science
DEPARTMENT
Fruit Research Institute
LAST_NAME
Su
FIRST_NAME
Wenbing
ADDRESS
Pudang 104, Jin'an District, Fuzhou, Fujian, 350013, China
Metabolomic Screening of Biomaterials for Mesenchymal stem/stromal cells (MSC) Culture
STUDY_SUMMARY
Mesenchymal stem/stromal cells (MSCs) have transformative healthcare potential, but to achieve therapeutic cell numbers, MSCs must be culture expanded. On tissue culture poly(styrene), however, MSC expansion is constrained by replicative senescence, and cells experience progressive loss of therapeutic potency with continued expansion. The lack of large numbers of reliably potent therapeutic cells is considered one of the most critical roadblocks to the success of MSC clinical trials. However, screening of improved MSC culture conditions has been hampered by the fact that there are no characterization tools that might allow prediction of cell fitness early in culture. Metabolism is the most dynamic level of cellular response, and metabolomics may enable the identification of early signatures associated with optimal cell product quality. Therefore, it is hypothesized that early metabolomics data from MSCs cultured on biomaterials will allow prediction of cell fitness later in culture and thus promote development of materials to promote a desired cell phenotype during expansion. The objective of this application is to determine the relationship between biomaterial carrier properties, cell metabolism, and maintenance of MSC fitness during expansion. This objective will be approached through the following specific aims: 1) Evaluate the effects of altering ligand type and stiffness of the biomaterial substrate on metabolism and replicative senescence of human MSCs during expansion in serum, and 2) Evaluate the effects of the substrate heparin content and pattern of initial seeding on metabolism and replicative senescence of human MSCs during expansion in serum-free conditions. The proposed work is innovative because it uses early cellular metabolic responses to design material substrates that will promote cell expansion without senescence in a range of media compositions. Results from these studies are expected to have an important positive impact because they will lead to more efficacious expansion protocols for MSCs and thus produce more effective cell-based therapies for a wide variety of diseases. From this study, we found that various triglycerides were elevated in older/highly senescent MSC, with longer chain lengths and more unsaturations in the fatty acid chains specifically correlating to higher senescence. We also observed an increased abundance of trihexosylceramides in higher senescent MSC samples.
Lipidomic signatures of hepato-cardiac FGF21 signaling in pressure overload- induced cardiac hypertrophy
STUDY_SUMMARY
This study investigates the early (3 days) mid- (2 weeks) and late (8 weeks) cardiac lipidome changes driven by FGF21 signaling in the context of pressure overload-induced cardiac hypertrophy. Untargeted LC-MS/MS lipidomic profiling was performed on cardiac tissue from mice at 3 days, 2 weeks, and 8 weeks following transverse aortic constriction (TAC), as well as from hepatocyte-specific (HEP-FGF21) and cardiomyocyte-specific (CM-FGF21) Fgf21 knockout mice at the 8-week post-TAC timepoint. Sham-operated wild type mice and mice expressing Cre in hepatocytes or cardiomyocytes served as controls at each stage. The early timepoints were chosen to capture transient and potentially initiating cardiac lipidome adaptations prior to overt cardiac remodeling, while the late timepoint provided insight into established lipidome reprogramming associated with advanced hypertrophy. Inclusion of the knockout models at 8 weeks enabled the dissection of endocrine (hepatocyte-derived) versus autocrine (cardiomyocyte-derived) FGF21 contributions to cardiac lipid metabolism. The analysis revealed time-dependent shifts in the cardiac lipidome, with an early increase in triglyceride species observed at 3 days post-TAC, followed by progressive accumulation of phospholipidsāincluding phosphatidylcholines (PC), phosphatidylethanolamines (PE), and phosphatidylserines (PS)at 2 and 8 weeks. Ablation of Fgf21 in either hepatocytes or cardiomyocytes significantly attenuated this phospholipid accumulation at 8 weeks, suggesting that both hepatic and cardiac FGF21 contribute to maladaptive lipidome remodeling during hypertrophic progression.
INSTITUTE
University of Cincinnati College of Medicine
LAST_NAME
Siokatas
FIRST_NAME
Georgios
ADDRESS
231 ALBERT SABIN WAY, University of Cincinnati, Cincinnati, OH, 45267-2827
Exploring Metabolomic Profiles in Vitamin D Deficient and Obese Individuals
STUDY_TYPE
LC/MS/MS
STUDY_SUMMARY
Cardiovascular diseases (CVDs) continue to pose a significant global health challenge, with vitamin D (Vit D) deficiency and obesity emerging as prominent risk factors. Arterial stiffness is recognized as a pivotal predictor and an independent risk element for CVDs. This study explores the metabolomic profiling of blood samples obtained from individuals afflicted with early arterial stiffness along with Vit D deficiency and obesity, compared with a healthy control group with normal Vit D and non-obese individuals. The study comprised nine participants with Vit D deficiency (Vit D level of 20 ng/ml or lower), and obese participants (BMI ā„ 30), along with eleven control participants (with Vit D levels above 20 ng/ml and normal BMI). Eleven metabolites exhibited statistically significant differences in patients with Vit D deficiency and obesity. Of these, eight metabolites demonstrated increased levels (FC > 2, p < 0.05), including Nutriacholic acid, N-Acetylputrescine, Succinylacetone, Adenosine monophosphate, Elaidic acid, Niacinamide, DUMP, and 2-Pyrrolidinone, while three metabolites exhibited decreased expression: PC (18:1(9Z)/18:1(9Z)), Trimethylamine, and Imidazole. The enrichment analysis revealed that top metabolic pathways predicted to be altered in the context of dysfunctional enzymes, encompassing processes such as nicotinamide acid uptake, fatty-acyl-CoA synthase (n-C18:0CoA), and extracellular NADP nucleosidase, among others. The ROC analysis showed that the blood metabolite concentrations can reliably differentiate with acceptable accuracy (AUC >0.8, p<0.05) between individuals at risk of arterial stiffness with Vit D deficiency and obesity from healthy controls showing the highest discriminator effect for DUMP and Niacinamide with AUC > 0.9 and p<0.00005. A combination metabolites model calculated with the linear SVM model achieved the highest performance of AUC=0.996 (95% CI: 0.97-1) and a predictive accuracy of 94.5% with the combination of seven metabolites. Integration of metabolomics and previous transcriptomics data identifies among the Vit D deficient and obese group disturbances in metabolic and regulatory pathways potentially related to vascular health such as inositol phosphate metabolism. The average PWV value relative to the participant's age as previously reported was 19.4 Ā± 4.7 m/s in the group with both Vit D deficiency and obesity, compared to 14.7 Ā± 2.1 m/s in the healthy control group (p < 0.05), indicating increased arterial stiffness.
INSTITUTE
Sharjah Institute for Medical Research
DEPARTMENT
Research institute of medical and health science
LABORATORY
Biomarker Discovery Group
LAST_NAME
Facility
FIRST_NAME
Core
ADDRESS
M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah, UAE, Sharjah, 000, United Arab Emirates
Mammary gland metabolism and its relevance to the fetoplacental expression of cytokine signaling in Caveolin-1 null mice
STUDY_SUMMARY
Mice lacking Caveolin-1 (Cav1), a major protein of the lipid raft of plasma membrane, show dysregulated cellular proliferation of mammary gland and an abnormal fetoplacental communication during pregnancy. The aim of this study is to better understand the functional links of mammary gland metabolism with gene expression of the placenta and fetus. Untargeted metabolomics analysis was performed to examine changes in mammary gland metabolism due to the absence of Cav1. The metabolomics analysis detected a total of 141 metabolites. We identified 81 metabolites that showed differential rank order in the level of expression in the mammary glands of Cav1-null compared to control mice. Differential metabolomics analysis identified 11 metabolites that were significantly changed in the mammary gland due to the absence of Cav1. Integrative metabolomics and transcriptomics analyses were applied to untangle functional links of metabolic pathways of the mammary gland with the gene expression changes of the placenta and fetus. The findings of this study show that metabolism and gene expression of the mammary gland are significantly impacted due to the loss of Cav1. Genes associated with specific metabolic and signaling pathways show coordinated expression changed in the placenta, mammary gland and fetal brain in Cav1-null mice. The cytokine signaling pathway emerges as a key player of the molecular crosstalk among the mammary gland, placenta and fetal brain. By interrogating the single-nuclei gene expression data of placenta and fetal brain previously generated from Cav1-null mice, the study further reveals that these metabolic and signaling genes are differentially regulated in specific cell types of the placenta and fetal brain. The findings of this study expand our understanding about the role of mammary gland metabolism in the regulation of fetoplacental communication in mammalian pregnancy.
Spatial lipidomics with in situ segmentation for profiling the effect of the drug doxorubicin on hepatoma carcinoma cell spheroids
STUDY_TYPE
Mammal
STUDY_SUMMARY
This study developed a simple method to analyze lipid changes in 3D liver cancer cell spheroids after treatment with the drug doxorubicin (DOX). The changes of spatial lipid molecules in 3D Hep-G2 cell spheroids model before and after DOX treatment were analyzed by MALDI-MSI. Using high-resolution MALDI mass spectrometry imaging (MALDI-MSI) and tissue analysis, the researchers mapped spatial lipid distribution across different regions of the spheroids: the outer proliferative zone, intermediate quiescent zone, and inner necrotic core. They identified distinct lipid alterations in each region (71, 60, and 56 lipids, respectively), with glycerophospholipid metabolism being the most affected pathway. These changes, particularly in PA, PC, PI, and SM lipids, may inhibit necrotic core formation. This spatial lipidomics approach provides insights into the effects of DOX and could help optimize personalized cancer therapies using tumor models.
INSTITUTE
Shanxi Medical University
DEPARTMENT
School of Basic Medical Sciences, Academy of Medical Sciences, Research Institute of Circadian Rhythm and Disease
LABORATORY
Shanxi Provincial Key Laboratory of Nanoimaging and Drug-Loaded Preparations
LAST_NAME
Zhao
FIRST_NAME
Huifang
ADDRESS
56 Xinjian South Road, Yingze District, Taiyuan, Shanxi Province, China
Metabolomics study on Xenopus tropicalis tadpoles overexpressing human IMPDH2 pathogenic variant
STUDY_SUMMARY
Xenopus tropicalis embryos were injected with 1 ng of mRNA encoding either WT human IMPDH2 or a gain-of-function variant of human IMPDH2, S160del, at the two-cell stage and raised until stage 41. Whole tadpoles lysates were analyzed by LC-MS to assess the consequences of IMPDH2 dysregulation on purine nucleotide metabolism. We find that overexpression of WT human IMPDH2 does not significantly alter purine nucleotide metabolism of the developing tadpole, but S160del overexpression significantly increases the levels of guanine nucleotides and uric acid, a degredation product of purine nucleotides. These results suggest increased activity through the guanine nucleotide biosynthetic pathway caused by IMPDH2 hyperactivity.
Evaluation of FASP protocol for a mass spectrometry based multiomics analysis of urine sample
STUDY_SUMMARY
Integrative multi-omics analysis of biological specimens such as tissues and biofluids (e.g., plasma and urine) is a powerful approach for gaining comprehensive understanding of the complex biological systems as well as in the identification of disease biomarkers. The great majority of omics datasets collected thus far resulted from a single omic (e.g. proteomics, metabolomics, lipidomics and others) study, which represents a challenge for any subsequent multiomics integration analysis. Filter Aided Sample Preparation (FASP) is a rapid and well-established technique for facilitating bottom-up proteomics preparation. However, FASP has only been employed in proteomics analysis and its utility for simultaneously isolating other biomolecules remains largely unexplored. This study assesses the performance of FASP as a convenient protocol to isolate protein and metabolite fraction from the same urine sample. Here FASP based LC-MS/MS analysis resulted in the identification of 3163 non-redundant peptides, 957 of which were unique protein groups. In parallel, LC-Qtof-MS and GC-MS/MS analysis of metabolites fraction obtained from urine solvent based extraction, FASP filtrate, FASP residue (concentrated protein) detected 145 metabolites by LC-MS and 139 metabolites by GC-MS. Our study demonstrates that the outcomes from FASP filtrate are comparable to those from solid phase extraction or FASP residue, in terms of both qualitative and quantitative analysis. Arguing that the proposed multiomics- FASP protocol should be considered for a multiomics single-step sample preparation analysis.
Assessment of the effects of Faecal microbiota transplantation with anti-inflammatory diet (FMT-AID) on faecal metabolome in patients with ulcerative colitis.
STUDY_SUMMARY
The study aims to understand FMT-AID driven recuperation of faecal metabolome in patients with UC.
INSTITUTE
All India Institute of Medical Sciences
DEPARTMENT
Gastroenterology
LABORATORY
IBD Research Group
LAST_NAME
Bajaj
FIRST_NAME
Aditya
ADDRESS
AIIMS Campus, Ansari Nagar East, Delhi, Delhi, 110092, India
EMAIL
adityabajaj93@gmail.com
PHONE
+91-9718405090
NUM_GROUPS
3 (Non-IBD Controls, Patients with UC before FMT and Patients with UC after FMT)
Impact of Exogenous Arginine Addition on the Bacterial Metabolome
STUDY_SUMMARY
The interaction between bacterial metabolic state modulation and antibiotic susceptibility has gained increasing attention as a strategy to combat antibiotic resistance. This study investigated the mechanism by which L-arginine enhanced the bactericidal efficiency of antibiotics. The metabolites in the exogenous Arg group and the control group were analyzed using LC-MS. KEGG enrichment analysis identified three significantly altered pathways: phenylalanine metabolism, degradation of aromatic compounds, and the TCA cycle. The results confirmed that arginine can change multiple metabolic pathways of bacteria, providing theoretical support for the study of arginine to change the bactericidal efficiency of antibiotics.
Fatty-acid oxidation is a metabolic hallmark of human proliferative retinopathy
STUDY_TYPE
comparison of vitreous humor from subjects with proliferative diabetic retinopathy to epiretinal membrane (control)
STUDY_SUMMARY
Healthy blood vessels supply neurons to preserve metabolic function. In blinding proliferative retinopathies (PRs), pathological neovascular tufts often emerge in lieu of needed physiological revascularization. Here we show that metabolic shifts in the neovascular niche define angiogenic fate. Fatty acid oxidation (FAO) metabolites accumulated in human and murine retinopathy samples. Neovascular tufts with a distinct single-cell transcriptional signature highly expressed FAO enzymes. The deletion of Sirt3, an FAO regulator, shifted the neovascular niche metabolism from FAO to glycolysis and suppressed tuft formation. This metabolic transition increased Vegf expression in astrocytes and reprogrammed pathological neovessels to a physiological phenotype, hastening vascular regeneration of the ischemic retina and improving vision. Hence, strategies to change the metabolic environment of vessels could promote a regenerative phenotype in vascular diseases. In this experiment, we used nontargeted metabolomics to measure metabolite profiles of vitreous humor from subjects with proliferative diabetic retinopathy. Vitreous samples were collected adjacent to leaky pathological neovascular tufts that characterize PR. Amongst the top 25 most significantly dysregulated metabolites, we observed an accumulation of acylcarnitines in human neovascular PR samples compared to control subjects with epiretinal membranes. Fatty acylcarnitines are by-products of fatty-acid beta-oxidation (FAO), the process of oxidizing fatty acids in mitochondria to produce acetyl-CoA (Fig. 1b). 5 of the 6 most enriched metabolic pathways in human PR pertained to FA metabolism, including long and short-chain saturated FAO pathways.
Lipids are essential for tumours because of their structural, energetic, and signaling roles. While many cancer cells upregulate lipid synthesis, growing evidence suggests that tumours simultaneously intensify the uptake of circulating lipids carried by lipoproteins. Which mechanisms promote the uptake of extracellular lipids, and how this pool of lipids contributes to cancer progression, are poorly understood. Here, using functional genetic screens, we find that lipoprotein uptake confers resistance to lipid peroxidation and ferroptotic cell death. Lipoprotein supplementation robustly inhibits ferroptosis across numerous cancer types, an effect largely driven by lipoprotein delivery of Ī±-tocopherol, the most abundant form of vitamin E. Mechanistically, cancer cells take up lipoproteins through a pathway dependent on sulfated glycosaminoglycans (GAGs) linked to cell-surface proteoglycans. Tumour GAGs are a major determinant of the uptake of both low and high density lipoproteins. Impairment of glycosaminoglycan synthesis or acute degradation of surface GAGs decreases the uptake of lipoproteins, sensitizes cells to ferroptosis and reduces tumour growth in mice. We also find that human clear cell renal cell carcinomas, a distinctively lipid-rich tumour type, display elevated levels of the GAG chondroitin sulfate and lipoprotein-derived antioxidants compared to non-malignant human kidney. Altogether, our work identifies lipoprotein uptake as an essential anti-ferroptotic mechanism for cancer cells to overcome lipid oxidative stress in vivo, and reveals GAG biosynthesis as a critical mediator of this process.
A human milk oligosaccharide alters the microbiome, circulating hormones, and metabolites in a randomized controlled trial of older individuals
STUDY_SUMMARY
Aging-related immune dysfunction is linked to cancer, atherosclerosis, and neurodegenerative diseases. This 6-week randomized controlled trial evaluated whether 2ā²-fucosyllactose (2ā²-FL), a human breast milk oligosaccharide with established benefits in infants and animal models, could improve gut microbiota and immune function in 89 healthy older adults (mean age 67.3 years).. While the primary endpoint of cytokine response change wasn't met, 2ā²-FL supplementation increased gut Bifidobacterium levels and elevated serum insulin, HDL cholesterol, and FGF21 hormone. Bifidobacterium ārespondersā experienced additional metabolic and proteomic changes and also performed better on a cognitive test of visual memory. Nonresponders were more likely to lack Bifidobacterium in their gut microbiota at the start of the intervention. Multi-omics analysis indicated a systemic response to 2ā²-FL, which could be detected in blood and urine, showcasing the potential of this prebiotic to provide diverse benefits for healthy aging. Trial registered at ClinicalTrials.gov (NCT03690999). We found three metabolites that had statistically significant interaction effects in our LME modeling: octanoylcarnitine, glutamate and taurine (P = 0.0011, P = 0.042, P = 0.049, respectively, LME, Benjamini-Hochberg corrected, Table S5). Octanoylcarnitine is a medium-chain fatty acid covalently linked to carnitine which undergoes fatty acid oxidation in the mitochondria. Post hoc analysis revealed that octanoylcarnitine decreased significantly in the High Dose 2ā²-FL group during the intervention (P = 0.015, Wilcoxon rank-sum test, Figure 4A), but not in the Low Dose 2ā²-FL group or Placebo group (P = 0.27, P = 0.39, respectively, Wilcoxon rank-sum test). Glutamate and taurine significantly increased in abundance in the Placebo group during the intervention (P = 0.028 for glutamate, P = 0.003 for taurine, Wilcoxon rank-sum tests). Levels of these metabolites also increased in the Low Dose 2ā²-FL (P = 0.14 for glutamate, P = 0.10 for taurine, Wilcoxon rank-sum tests) and High Dose 2ā²-FL groups (P = 0.12 for glutamate, P = 0.13 for taurine, Wilcoxon rank-sum tests), albeit not in a statistically significant manner.
Exploration of AGMO-dependent changes in the lipid mediator profile of polarized BMDM cells (M0, M1 and M2 phenoptypes)
STUDY_SUMMARY
Murine BMDMs isolated from wildtype (WT) or Agmo knockout (KO) mice were polarized to the M0, M1 or M2 phenotypes, and then incubated in PBS + Ca2+ (1 mM) for 3 h. Lipid mediators were extracted and analyzed by UPLC-MS/MS.
Integrative spatially-resolved multi-omics analysis reveals metabolic evolution of the transition to invasive breast cancer
STUDY_SUMMARY
We used MSTI (Mass Spectrometry imaging and spatially-resolved Transcriptomics Integrator) framework identified the intricate interplay between sphingolipid metabolism and prosaposin (PSAP), highlighting the transition from ductal carcinoma in situ (DCIS) to invasive breast cancer (IBC) within the tumor microenvironment. To investigate the impact of PSAP and sphingolipid metabolism on DCIS evolution, we performed PSAP knockdown in 4T1 cells and conducted LC-MS analysis to compare the metabolic profiles of wild-type (WT) and PSAP-knockdown cells. LC-MS results revealed significant metabolic alterations: compared to 4T1-shCtrl cells, 4T1-shPSAP cells exhibited a marked increase in various ceramides, a significant upregulation of hexosylceramides (HexCer), and a notable decrease in other sphingolipids. These findings highlight the key role of PSAP in modulating sphingolipid metabolism during tumor progression.
INSTITUTE
Xiamen university
LAST_NAME
Shu-hai
FIRST_NAME
Lin
ADDRESS
No. 4221 Xiang'an South Road, Xiang'an District, Xiamen City, Fujian Province, Xiamen, Fujian, 361100, China
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC Tissues
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolG D256A mutation in NSCLC. Using in vivo (GEMMs), we characterized the metabolic profile of NSCLC harboring this mutation compared to controls. Here, we show that PolG mutation causes mitochondrial dysfunction, impacting serine/glycine metabolism as well as energy metabolism through glycolysis. This study was conducted in animals divided into two age groups. The "young" group consisted of animals in which NSCLC was initiated at 3 months of age, while the "old" group included animals whose tumors were initiated at 10 months of age. WT: Wild Type animals KP: NSCLC conditional animals PolG: Animals with PolG mutation PGKP: NSCLC conditional animals with PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC Em tumor derived cell lines (TDCLs)
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolGD256A mutation in NSCLC tumor derived cell lines (TDCLs). We characterized the metabolic profile of TDCLs harboring this mutation compared to controls. KP: NSCLC TDCLs generated from conditional animals PGKP: NSCLC TDCLs generated from conditional animals bearing PolG mutation. Here, we show that PolG mutation causes mitochondrial dysfunction, impacting serine/glycine metabolism as well as energy metabolism through glycolysis due intensive usage of glucose over time.
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC tumor derived lines (TDCLs)
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolGD256A mutation in NSCLC. Using in vitro models (TDCLs), we characterized we collected conditioned medium to mesure glucose and serine consumption. Here we found that mitochondria impairment cause more use of glucose and less use of serine from the culture medium. KP: NSCLC TDCLs generated from conditional animals PGKP: NSCLC TDCLs generated from conditional animals bearing PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC TDCLs
STUDY_SUMMARY
This study explores the impact of mtDNA mutation burden induced by the PolG š· 256 š“ D256A mutation in NSCLC. Using in vitro models (TDCLs) cultured in either standard RPMI medium or RPMI without serine and glycine, we characterized the [U-Ā¹Ā³C]D-glucose metabolism in NSCLC cells harboring this mutation compared to controls. Here we found that mitochondria impairment cause more use of glucose to synthesize serine. Due the lack of carbons from glucose to TCA (Tricarboxylic acid cycle) the PGKP cells have a energetic imbalance. KP: NSCLC TDCLs generated from conditional animals PGKP: NSCLC TDCLs generated from conditional animals bearing PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC conditioned medium
STUDY_SUMMARY
This study explores the impact of mtDNA mutation burden induced by the PolG š· 256 š“ D256A mutation in NSCLC. Using in vitro models (TDCLs) cultured in either standard RPMI medium or RPMI without serine and glycine, we characterized the [U-Ā¹Ā³C]D-glucose metabolism in NSCLC conditioned medium. Here we found that mitochondria impairment cause more use of glucose to synthesize serine. Due the lack of carbons from glucose to TCA (Tricarboxylic acid cycle) the PGKP cells have a energetic imbalance. KP: NSCLC TDCLs generated from conditional animals PGKP: NSCLC TDCLs generated from conditional animals bearing PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC Tissues_Muscle
STUDY_SUMMARY
Here we found that mitochondria impairment causes more use of glucose to synthesize serine in NSCLC tumors but not in other tissues (lungs, liver, plasma, and muscle. Due the lack of carbons from glucose to TCA (Tricarboxylic acid cycle) the PGKP cells have an energetic imbalance. WT: Wild Type animals KP: NSCLC conditional animals PolG: Animals with PolG mutation PGKP: NSCLC conditional animals with PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC Tissues_Lung
STUDY_TYPE
In vivo Isotope Tracing
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolGD256A mutation in NSCLC. Using in vivo, we characterized the metabolic profile of NSCLC harboring this mutation compared to controls after submitting the animals to 12 weeks of special diet and 2h30 of [U-13C]D-Glucose infusion. Here we found that mitochondria impairment causes more use of glucose to synthesize serine in NSCLC tumors but not in other tissues (lungs, liver, plasma, and muscle. Due the lack of carbons from glucose to TCA (Tricarboxylic acid cycle) the PGKP cells have an energetic imbalance. WT: Wild Type animals KP: NSCLC conditional animals PolG: Animals with PolG mutation PGKP: NSCLC conditional animals with PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC Tissues_Liver
STUDY_TYPE
In vivo Isotope Tracing
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolGD256A mutation in NSCLC. Using in vivo, we characterized the metabolic profile of NSCLC harboring this mutation compared to controls after submit the animals to 12 weeks of special diet and 2h30 of [U-13C]D-Glucose infusion. Here we found that mitochondria impairment causes more use of glucose to synthesize serine in NSCLC tumors but not in other tissues (lungs, liver, plasma, and muscle. Due the lack of carbons from glucose to TCA (Tricarboxylic acid cycle) the PGKP cells have an energetic imbalance. WT: Wild Type animals KP: NSCLC conditional animals PolG: Animals with PolG mutation PGKP: NSCLC conditional animals with PolG mutation
Respiration defects limit serine synthesis required for lung cancer growth and survival - Effect of Polg mutation in NSCLC Tissues_Plasma
STUDY_TYPE
In vivo Isotope Tracing
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolGD256A mutation in NSCLC. Using in vivo, we characterized the metabolic profile of NSCLC harboring this mutation compared to controls after submitting the animals to 12 weeks of special diet and 2h30 of [U-13C]D-Glucose infusion.
Respiration defects limit serine synthesis required for lung cancer growth and survival - NSCLC rewires glycolytic metabolism to synthesize more serine from glucose (Tumors)
STUDY_TYPE
In vivo Isotope Tracing
STUDY_SUMMARY
This study investigates the impact of mtDNA mutation burden induced by the PolGD256A mutation in Non-Small cell lung cancer (NSCLC). Using in vivo, we characterized the metabolic profile of NSCLC harboring this mutation compared to controls after submit the animals to 12 weeks of special diet and 2h30 of [U-13C]D-Glucose infusion. Here we found that mitochondria impairment causes more use of glucose to synthesize serine in NSCLC tumors but not in other tissues (lungs, liver, plasma, and muscle. Due the lack of carbons from glucose to TCA (Tricarboxylic acid cycle) the PGKP cells have an energetic imbalance. WT: Wild Type animals KP: NSCLC conditional animals PolG: Animals with PolG mutation PGKP: NSCLC conditional animals with PolG mutation
Glycosaminoglycan-mediated lipoprotein uptake protects cancer cells from ferroptosis
STUDY_SUMMARY
Lipids are essential components of cancer cells due to their structural and signaling roles and many cancers take up extracellular lipids to meet metabolic demands. How these lipids contribute to cancer growth and progression remains poorly understood. Using functional genetic screens, lipoprotein uptakeāthe primary mechanism for lipid transport in circulationāis identify as a key determinant of ferroptosis sensitivity in cancer. Lipoprotein supplementation robustly inhibits ferroptosis across numerous cancer types, an effect largely driven by lipoprotein delivery of Ī±-tocopherol. Mechanistically, cancer cells take up lipoproteins through a pathway dependent on sulfated glycosaminoglycans (GAGs) linked to cell-surface proteoglycans. Disrupting GAG biosynthesis or acutely degrading surface GAGs reduces lipoprotein uptake, sensitizes cancer cells to ferroptosis, and impairs tumor growth in mice. Notably, human clear cell renal cell carcinomas (ccRCC), a lipid-rich malignancy, exhibit elevated levels of chondroitin sulfate and increased lipoprotein-derived Ī±-tocopherol compared to normal kidney tissue. Altogether, this work establishes lipoprotein uptake as a critical anti-ferroptotic mechanism in cancer and implicates GAG biosynthesis as a therapeutic target.
INSTITUTE
University of Texas Southwestern Medical Center at Dallas