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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
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.
| Study ID | ST002891 |
| Study Title | Glutamine metabolism improves left ventricular function but not macrophage-mediated inflammation following myocardial infarction |
| Study Type | untargeted metabolomics analysis |
| 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 |
| SIMONA.CODREANU@VANDERBILT.EDU | |
| Phone | 6158758422 |
| Submit Date | 2023-09-29 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-02 |
| Release Version | 1 |
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: | Mammals |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Gln Treatment |
|---|---|---|
| SA315459 | D1_1 | Day 1 |
| SA315460 | D1_6 | Day 1 |
| SA315461 | D1_5 | Day 1 |
| SA315462 | D1_2 | Day 1 |
| SA315463 | D1_3 | Day 1 |
| SA315464 | D1_4 | Day 1 |
| SA315465 | D3_6 | Day 3 |
| SA315466 | D3_5 | Day 3 |
| SA315467 | D3_3 | Day 3 |
| SA315468 | D3_4 | Day 3 |
| SA315469 | D3_1 | Day 3 |
| SA315470 | D3_2 | Day 3 |
| SA315471 | D7_5 | Day 7 |
| SA315472 | D7_4 | Day 7 |
| SA315473 | D7_2 | Day 7 |
| SA315474 | D7_1 | Day 7 |
| SA315475 | D7_3 | Day 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 |
