Summary of Study ST000822

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 PR000585. The data can be accessed directly via it's Project DOI: 10.21228/M8239Z This work is supported by NIH grant, U2C- DK119886.

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Study IDST000822
Study TitleMetabolomic profiling of IDH1 mutation (part II)
Study TypeMS analysis
Study SummaryWe are interested in using both spontaneous and stimulated Raman microscopy to visualize these metabolomic changes as spectral alterations. We have two isogneic cell lines of normal human astrocytes differing only by a point mutation in the IDH-1 gene. We will work with the metabolomics core to elucidate the changes in central metabolism and lipid synthesys in an effort to determine the precise biochemical alterations underlying observed spectral differences. We wil then use a selective inhibitor of the IDH1 R132H to demonstrate to attempt to return TCA metabolome and lipidome to WT phenotype. Lastly, we will use a cell-permeablized variant of 2HG (2R-octyl-alpha-hydroxyglutarate) to recaptitulate the R132H mutant phenotype in wild-type cells, providing strong evidence that 2HG accumulation uderlies the metabolomic (and thus, spectral) changes observed.
Institute
University of Michigan
DepartmentBiomedical Research Core Facilities
LaboratoryMetabolomics core
Last NameKachman
First NameMaureen
Address6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Emailmkachman@med.umich.edu
Phone(734) 232-8175
Submit Date2017-07-14
Num Groups4
Total Subjects9
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2018-02-07
Release Version1
Maureen Kachman Maureen Kachman
https://dx.doi.org/10.21228/M8239Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000585
Project DOI:doi: 10.21228/M8239Z
Project Title:Stimulated Raman Immunohistochemistry
Project Type:MS analysis
Project Summary:Profiling of WT, IDH1 R132H, WT + 2HG, Mut + AG!5198
Institute:University of Michigan
Department:Neurosurgery
Laboratory:Orringer Lab
Last Name:Orringer
First Name:Daniel
Address:Ann Arbor, MI
Email:dorringe@umich.edu
Phone:734-647-9222

Subject:

Subject ID:SU000848
Subject Type:HUMAN
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Mammals

Factors:

Subject type: HUMAN; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Cell Line Treatment
SA045836S00028830IDH1 R132H NORMAL CULTURE MEDIA
SA045837S00028828IDH1 R132H NORMAL CULTURE MEDIA
SA045838S00028829IDH1 R132H NORMAL CULTURE MEDIA
SA045839S00028833IDH1 WILD TYPE CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF
SA045840S00028832IDH1 WILD TYPE CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF
SA045841S00028831IDH1 WILD TYPE CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF
SA045842S00028826IDH1 WILD TYPE NORMAL CULTURE MEDIA
SA045843S00028827IDH1 WILD TYPE NORMAL CULTURE MEDIA
SA045844S00028825IDH1 WILD TYPE NORMAL CULTURE MEDIA
Showing results 1 to 9 of 9

Collection:

Collection ID:CO000842
Collection Summary:-
Sample Type:Astrocyte cells

Treatment:

Treatment ID:TR000862
Treatment Summary:-

Sample Preparation:

Sampleprep ID:SP000855
Sampleprep Summary:-
Sampleprep Protocol Filename:A037-2HG_cells_updated-20160721.pdf

Combined analysis:

Analysis ID AN001305
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent
Column Waters Acquity HSS T3 (50 x 2.1mm,1.8um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6490A QQQ
Ion Mode NEGATIVE
Units uM/ng protein

Chromatography:

Chromatography ID:CH000914
Instrument Name:Agilent
Column Name:Waters Acquity HSS T3 (50 x 2.1mm,1.8um)
Solvent A:100% water; 2 mM ammonium formate
Solvent B:100% acetonitrile; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS001198
Analysis ID:AN001305
Instrument Name:Agilent 6490A QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
Analysis Protocol File:A037-2HG_cells_updated-20160721.pdf
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