Summary of Study ST001952

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 PR001239. The data can be accessed directly via it's Project DOI: 10.21228/M8JT51 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001952
Study TitleGLS2KO vs WT mouse hepatocytes
Study TypeGenotype
Study SummaryTest 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.
Institute
Pfizer
Last NameClasquin
First NameMichelle
Address1 Portland St., Cambridge, MA 02139
Emailmichelle.clasquin@pfizer.com
Phone6174487289
Submit Date2021-10-22
Num Groups2
Total Subjects6
Num Males6
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailLC-MS
Release Date2021-11-08
Release Version1
Michelle Clasquin Michelle Clasquin
https://dx.doi.org/10.21228/M8JT51
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Subject:

Subject ID:SU002030
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
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