Summary of Study ST003623

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 PR002208. The data can be accessed directly via it's Project DOI: 10.21228/M86Z6P 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	ST003623
Study Title	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.
Institute	Genentech Inc.
Last Name	Wang
First Name	Mike
Address	1 DNA Way, South San Francisco, CA 94080, USA
Email	wang.mike@gene.com
Phone	6502457991
Submit Date	2024-12-06
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2025-01-02
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP003760
Sampleprep Summary:	Cells were trypsinized, washed 3x with ice cold PBS and flash frozen in liquid nitrogen for metabolic quenching. All metabolite extraction procedures were kept on ice. Briefly, 350μL of cold methanol containing in-house metabolomics Recovery IS (Stable Isotope Labeled Internal Standards) mixture and 200μL of cold chloroform were added to each sample. Samples were vortexed for 10 seconds, homogenized with beads for 2 minutes, and centrifuged at 4000 RPM for 5 minutes. Next, supernatants were transferred and 200μl cold water was added to perform liquid–liquid phase separation. Samples were mixed and centrifuged again. The top layer aliquots were transferred, dried and reconstituted in 100μl acetonitrile:water (8:2, v/v) containing the in-house metabolomics Global IS (Stable Isotope Labeled Internal Standards) mixture and submitted for metabolomics analysis.

Sampleprep ID:

SP003760

Sampleprep Summary:

Cells were trypsinized, washed 3x with ice cold PBS and flash frozen in liquid nitrogen for metabolic quenching. All metabolite extraction procedures were kept on ice. Briefly, 350μL of cold methanol containing in-house metabolomics Recovery IS (Stable Isotope Labeled Internal Standards) mixture and 200μL of cold chloroform were added to each sample. Samples were vortexed for 10 seconds, homogenized with beads for 2 minutes, and centrifuged at 4000 RPM for 5 minutes. Next, supernatants were transferred and 200μl cold water was added to perform liquid–liquid phase separation. Samples were mixed and centrifuged again. The top layer aliquots were transferred, dried and reconstituted in 100μl acetonitrile:water (8:2, v/v) containing the in-house metabolomics Global IS (Stable Isotope Labeled Internal Standards) mixture and submitted for metabolomics analysis.