Summary of Study ST003243

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 PR002013. The data can be accessed directly via it's Project DOI: 10.21228/M8DJ7S 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 IDST003243
Study TitleLipidomic analysis of serum from WT, liver-specific Gclc KO, liver-specific Nrf2 KO, and liver-specific Gclc-Nrf2 DKO mice.
Study SummaryThe 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 NameHarris
First NameIsaac
Address601 Elmwood Ave, Rochester, New York, 14642-0001, USA
Emailisaac_harris@urmc.rochester.edu
Phone8572348624
Submit Date2024-05-29
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2024-06-07
Release Version1
Isaac Harris Isaac Harris
https://dx.doi.org/10.21228/M8DJ7S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR002013
Project DOI:doi: 10.21228/M8DJ7S
Project Title:Glutathione synthesis in the mouse liver supports lipid abundance through NRF2 repression.
Project Summary:Cells rely on antioxidants to survive. The most abundant antioxidant is glutathione (GSH). The synthesis of GSH is non-redundantly controlled by the glutamate-cysteine ligase catalytic subunit (GCLC). GSH imbalance is implicated in many diseases, but the requirement for GSH in adult tissues is unclear. To interrogate this, we have developed a series of in vivo models to induce Gclc deletion in adult animals. We find that GSH is essential to lipid abundance in vivo. GSH levels are highest in liver tissue, which is also a hub for lipid production. While the loss of GSH does not cause liver failure, it decreases lipogenic enzyme expression, circulating triglyceride levels, and fat stores. Mechanistically, we find that GSH promotes lipid abundance by repressing NRF2, a transcription factor induced by oxidative stress. These studies identify GSH as a fulcrum in the liver's balance of redox buffering and triglyceride production.
Institute:University of Rochester Medical Center
Last Name:Harris
First Name:Isaac
Address:601 Elmwood Ave, Rochester, New York, 14642-0001, USA
Email:isaac_harris@urmc.rochester.edu
Phone:8572348624

Subject:

Subject ID:SU003362
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Male and female
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id Sample source Genotype
SA35336024-08-18-2022-POS-Isaac-S01Serum Liver-specific Gclc KO
SA35336127-08-18-2022-POS-Isaac-S04Serum Liver-specific Gclc KO
SA35336225-08-18-2022-POS-Isaac-S02Serum Liver-specific Gclc KO
SA35336326-08-18-2022-POS-Isaac-S03Serum Liver-specific Gclc KO
SA35336432-08-18-2022-POS-Isaac-S09Serum Liver-specific Gclc-Nrf2 DKO
SA35336531-08-18-2022-POS-Isaac-S08Serum Liver-specific Gclc-Nrf2 DKO
SA35336633-08-18-2022-POS-Isaac-S10Serum Liver-specific Gclc-Nrf2 DKO
SA35336730-08-18-2022-POS-Isaac-S07Serum Liver-specific Gclc-Nrf2 DKO
SA35336828-08-18-2022-POS-Isaac-S05Serum Liver-specific Gclc-Nrf2 DKO
SA35336929-08-18-2022-POS-Isaac-S06Serum Liver-specific Gclc-Nrf2 DKO
SA35337041-08-18-2022-POS-Isaac-S17Serum Liver-specific Nrf2 KO
SA35337142-08-18-2022-POS-Isaac-S18Serum Liver-specific Nrf2 KO
SA35337240-08-18-2022-POS-Isaac-S16Serum Liver-specific Nrf2 KO
SA35337339-08-18-2022-POS-Isaac-S15Serum Liver-specific Nrf2 KO
SA35337438-08-18-2022-POS-Isaac-S14Serum WT
SA35337536-08-18-2022-POS-Isaac-S12Serum WT
SA35337635-08-18-2022-POS-Isaac-S11Serum WT
SA35337737-08-18-2022-POS-Isaac-S13Serum WT
Showing results 1 to 18 of 18

Collection:

Collection ID:CO003355
Collection Summary:Mice were anesthetized with isoflurane, after which blood was collected via the retro-orbital venous sinus into BD microtainer tubes (BD #365967). Serum was isolated from the blood by centrifuging blood samples at 10000 xg for 5mins and stored at -80C.
Sample Type:Blood (serum)

Treatment:

Treatment ID:TR003371
Treatment Summary:WT (Gclc f/f), liver-specific Gclc KO (Gclc f/f), liver-specific Nrf2 KO (Nrf2 f/f) and liver-specific Gclc-Nrf2 DKO (Gclc f/f Nrf2 f/f) were induced by injecting mice via the tail vein with 2.5x1011 GC of AAV-TBG-Cre (Addgene, 107787-AAV8) in PBS. After 21 days, serum was isolated and stored at -80C.

Sample Preparation:

Sampleprep ID:SP003369
Sampleprep Summary:For serum for WT, liver-specific Gclc KO, liver-specific Nrf2 KO, and liver-specific Gclc-Nrf2 DKO, 20 µL of mouse serum was combined with 180 µL of chloroform:methanol extraction solvent (v:v=1:2) containing internal standards at the final concentrations: 5nM D7-Sphinganine (Avanti Polar Lipids Inc., Cat# 860658), 12.5nM D3-Deoxysphinganine (Avanti Polar Lipids Inc., Cat# 860474), and SPLASH LIPIDOMIX (1:1000, Avanti Polar Lipids Inc., Cat# 330707). After sonicating (1400 rpm, 20°C, 5 min), the extracts were cleared by centrifugation (17,000g, 20°C, 10 min), and the lipids in the supernatant were analyzed by LC-MS.

Combined analysis:

Analysis ID AN005313
Analysis type MS
Chromatography type Reversed phase
Chromatography system Thermo Vanquish
Column Perkin Elmer Brownlee SPP C18 (75 x 2.1mm, 2.7um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive HF hybrid Orbitrap
Ion Mode POSITIVE
Units Normalized to the median value of total lipid signals

Chromatography:

Chromatography ID:CH004017
Instrument Name:Thermo Vanquish
Column Name:Perkin Elmer Brownlee SPP C18 (75 x 2.1mm, 2.7um)
Column Temperature:50
Flow Gradient:0–2 min 35% B, 2–8 min from 35 to 80% B, 8–22 min from 80 to 99% B, 22–36 min 99% B, and 36.1–40 min from 99 to 35% B
Flow Rate:0.400 mL/min
Solvent A:100% water; 0.1% formic acid; 10mM ammonium acetate
Solvent B:50% acetonitrile/50% isopropanol; 0.1% formic acid; 10mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS005043
Analysis ID:AN005313
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:MS2 scan conditions were applied in positive mode, the scan range was from m/z 120–1000, resolution was 120,000 for MS, and 30,000 for DDMS2 (top 10), and the AGC target was 3E6 for full MS and 1E5 for DDMS2, allowing ions to accumulate for up to 200 ms for MS and 50 ms for MS/MS. For MS/MS, the following settings are used: isolation window width 1.2 m/z, stepped NCE at 10, 15, and 25 a.u., minimum AGC 5E2, and dynamic exclusion of previously sampled peaks for 8 s. Quality control (QC) samples were included to check the technical variability and were prepared by mixing an equal volume of lipid extract from each tissue or serum sample. QC samples were included in the analysis sequence every ten samples and monitored for changes in peak area, width, and retention time to determine the performance of the LC-MS/MS analysis. QC samples were subsequently used to align the analytical batches. The lipid peaks were identified, aligned, and exported using MS-DIAL. The data were further normalized to the median value of total lipid signals. Only lipids fully identified by MS2 spectra were included in the analysis.
Ion Mode:POSITIVE
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