Summary of Study ST002891

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001804. The data can be accessed directly via it's Project DOI: 10.21228/M8HX5R This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002891
Study TitleGlutamine metabolism improves left ventricular function but not macrophage-mediated inflammation following myocardial infarction
Study Typeuntargeted metabolomics analysis
Study SummaryGlutamine 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
DepartmentChemistry
LaboratoryCenter for Innovative Technology
Last NameCodreanu
First NameSimona
Address1234 STEVENSON CENTER LANE, NASHVILLE, TN, 37235, USA
EmailSIMONA.CODREANU@VANDERBILT.EDU
Phone6158758422
Submit Date2023-09-29
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2024-04-02
Release Version1
Simona Codreanu Simona Codreanu
https://dx.doi.org/10.21228/M8HX5R
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR001804
Project DOI:doi: 10.21228/M8HX5R
Project Title:Glutamine metabolism improves left ventricular function but not macrophage-mediated inflammation following myocardial infarction
Project Type:Untargeted Metabolomics analysis
Project 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
Email:SIMONA.CODREANU@VANDERBILT.EDU
Phone:6158758422

Subject:

Subject ID:SU003004
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:15-20 week old
Gender:Male
Species Group:C57BL/6J

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Gln Treatment
SA315459D1_1Day 1
SA315460D1_6Day 1
SA315461D1_5Day 1
SA315462D1_2Day 1
SA315463D1_3Day 1
SA315464D1_4Day 1
SA315465D3_6Day 3
SA315466D3_5Day 3
SA315467D3_3Day 3
SA315468D3_4Day 3
SA315469D3_1Day 3
SA315470D3_2Day 3
SA315471D7_5Day 7
SA315472D7_4Day 7
SA315473D7_2Day 7
SA315474D7_1Day 7
SA315475D7_3Day 7
Showing results 1 to 17 of 17

Collection:

Collection ID:CO002997
Collection Summary:MI was induced in adult (15-20 week old) male C57BL/6J mice as previously described.2,4,6 Mice were anesthetized (2% isoflurane) and laid supine on a heated stage (37°C), intubated and connected to a mouse ventilator (Harvard Apparatus; 300µL stroke volume, 300 breaths/min). The chest was incised to expose the ribs, and the heart exposed between the 3rd and 4th ribs. The pericardial layer was gently removed, and the left coronary artery was ligated ~1mm below the left atrium using 8-0 suture. Ischemia was confirmed by blanching of the LV. For glutamine administration, mice were injected intraperitoneally with glutamine (0.75-1g/kg) or saline vehicle beginning at either day 1 post-MI (for day 3 time-point) or beginning at day 3 (for day 7 time-point). A separate cohort of mice was injected 2 hr after MI and studied 24 hr later (day 1). To inhibit glutamine metabolism, mice were administered BPTES (12.5mg/kg; 10% in DMSO/corn oil), a glutaminase-1 inhibitor, beginning at either day 1 or day 3.
Sample Type:Macrophages

Treatment:

Treatment ID:TR003013
Treatment Summary:For glutamine administration, mice were injected intraperitoneally with glutamine (0.75-1g/kg) or saline vehicle beginning at either day 1 post-MI (for day 3 time-point) or beginning at day 3 (for day 7 time-point). A separate cohort of mice was injected 2 hr after MI and studied 24 hr later (day 1). To inhibit glutamine metabolism, mice were administered BPTES (12.5mg/kg; 10% in DMSO/corn oil), a glutaminase-1 inhibitor, beginning at either day 1 or day 3.

Sample Preparation:

Sampleprep ID:SP003010
Sampleprep Summary:Frozen samples were stored at -80°C until analyzed by LC-MS-based metabolomics in the Vanderbilt Center for Innovative Technology (CIT). Isotopically labeled phenylalanine-D8 and biotin-D2 were added to 200 μL of culture supernatant per sample, and protein was precipitated by addition of 800 μL of ice-cold methanol followed by overnight incubation at −80°C. Precipitated proteins were pelleted by centrifugation (15,000 rpm, 15 min), and extracted metabolites were dried down in vacuo and stored at −80°C. Individual samples were reconstituted in 100 μL of reconstitution buffer (acetonitrile/water, 90:10, vol/vol) containing tryptophan-D3 and inosine-4N15. Equal volumes of individual samples were pooled to create a quality control (QC) pooled sample used for column conditioning, retention time alignment, to assess instrument reproducibility, and for batch acceptance.
Processing Storage Conditions:-80℃
Extract Storage:-80℃

Combined analysis:

Analysis ID AN004750
Analysis type MS
Chromatography type HILIC
Chromatography system Vanquish UHPLC
Column ACQUITY UPLC BEH Amide HILIC
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive HF hybrid Orbitrap
Ion Mode NEGATIVE
Units time_m/z

Chromatography:

Chromatography ID:CH003582
Chromatography Summary:Metabolite extracts were separated on ACQUITY UPLC BEH Amide HILIC 1.7 μm, 2.1 × 100 mm column (Waters Corporation, Milford, MA) held at 30°C. Liquid chromatography was performed at a 200 μL min using solvent A (5 mM Ammonium formate in 90% water, 10% acetonitrile, and 0.1% formic acid) and solvent B (5 mM Ammonium formate in 90% acetonitrile, 10% water, and 0.1% formic acid) with a gradient length of 30 min.
Instrument Name:Vanquish UHPLC
Column Name:ACQUITY UPLC BEH Amide HILIC
Column Temperature:30
Flow Gradient:Linear gradient of 30 min
Flow Rate:0.20mL/min
Solvent A:90% water/10% acetonitrile; 5mM ammonium formate; 0.1% formic acid
Solvent B:10% water/90% acetonitrile; 5mM ammonium formate; 0.1% formic acid
Chromatography Type:HILIC

MS:

MS ID:MS004496
Analysis ID:AN004750
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:The acquired raw data were imported, processed, normalized and reviewed using Progenesis QI v.3.0 (Non-linear Dynamics, Newcastle, UK). All MS and MS/MS sample runs were aligned against a QC pool reference run, and unique ions (retention time and m/z pairs) were deadducted and deisotoped to generate unique "features" (retention time and m/z pairs). Data were normalized to all features using Progenesis QI. Experimental data annotations were assigned based on consistent retention time and MS2 fragmentation pattern matches with reference standards.
Ion Mode:NEGATIVE
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