{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000821","ANALYSIS_ID":"AN001304","VERSION":"1","CREATED_ON":"August 2, 2017, 9:48 am"},

"PROJECT":{"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"},

"STUDY":{"STUDY_TITLE":"Metabolomic profiling of IDH1 mutation","STUDY_TYPE":"MS analysis","STUDY_SUMMARY":"We 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","DEPARTMENT":"Biomedical Research Core Facilities","LABORATORY":"Metabolomics core","LAST_NAME":"Kachman","FIRST_NAME":"Maureen","ADDRESS":"6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714","EMAIL":"mkachman@med.umich.edu","PHONE":"(734) 232-8175","NUM_GROUPS":"2","TOTAL_SUBJECTS":"3","STUDY_COMMENTS":"The IDH-1 R132H mutation in low grade gliomas preceptitates oncogenesis through a neomorphic enzyme function: the reduction of alpha-ketoglutarate (aKG) to 2-hydroxygutarate (2HG). 2HG accumukates to extremely high (~100mM) concentrations in IDH1 mutant cells and has substantial, predictable metabolic effects, including inhibition of a-KG dependent dioxygenases and hypermethylation of histones and chromatin."},

"SUBJECT":{"SUBJECT_TYPE":"HUMAN","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"SU0016656",
"Sample ID":"S00026530",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016657",
"Sample ID":"S00026531",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016658",
"Sample ID":"S00026532",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016656",
"Sample ID":"S00026533",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016657",
"Sample ID":"S00026534",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016658",
"Sample ID":"S00026535",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016656",
"Sample ID":"S00026536",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016657",
"Sample ID":"S00026537",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016658",
"Sample ID":"S00026538",
"Factors":{"IDH1_MUTATION_STATUS":"WT"}
},
{
"Subject ID":"SU0016656",
"Sample ID":"S00026539",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016657",
"Sample ID":"S00026540",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016658",
"Sample ID":"S00026541",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016656",
"Sample ID":"S00026542",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016657",
"Sample ID":"S00026543",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016658",
"Sample ID":"S00026544",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016656",
"Sample ID":"S00026545",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016657",
"Sample ID":"S00026546",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
},
{
"Subject ID":"SU0016658",
"Sample ID":"S00026547",
"Factors":{"IDH1_MUTATION_STATUS":"R132H Mutant"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"-"},

"TREATMENT":{"TREATMENT_SUMMARY":"-"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"-","SAMPLEPREP_PROTOCOL_FILENAME":"A037-2HG_cells_updated-20160721.pdf"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent","COLUMN_NAME":"Waters Acquity HSS T3 (50 x 2.1mm, 1.8um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"A037-2HG cells updated-20160721.pdf"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 6490A QQQ","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"NEGATIVE"},

"MS_METABOLITE_DATA":{
"Units":"uM/ng protein",

"Data":[{"Metabolite":"(2R)-2-HYDROXYPENTANEDIOIC ACID","S00026530":"4.071306714","S00026531":"3.098099464","S00026532":"4.703858735","S00026533":"4.798879275","S00026534":"3.086003529","S00026535":"4.044164366","S00026536":"2.321686412","S00026537":"2.332264382","S00026538":"2.737507717","S00026539":"43.54915156","S00026540":"38.63892668","S00026541":"24.66584537","S00026542":"38.18522914","S00026543":"42.39913148","S00026544":"41.66765007","S00026545":"34.76693065","S00026546":"33.5109968","S00026547":"32.91367874"},{"Metabolite":"(2S)-2-HYDROXYPENTANEDIOIC ACID","S00026530":"7.295459461","S00026531":"5.89119898","S00026532":"5.925238318","S00026533":"6.187509603","S00026534":"6.209511497","S00026535":"5.454204664","S00026536":"5.148028685","S00026537":"5.1653177","S00026538":"5.473715581","S00026539":"6.299511709","S00026540":"4.89508229","S00026541":"3.822237079","S00026542":"4.885550348","S00026543":"6.014301899","S00026544":"4.834213193","S00026545":"4.660787068","S00026546":"4.835995756","S00026547":"4.299657892"}],

"Metabolites":[{"Metabolite":"(2R)-2-HYDROXYPENTANEDIOIC ACID","moverz_quant":"","ri":"","ri_type":"","pubchem_id":"439391","inchi_key":"","kegg_id":"","other_id":"","other_id_type":""},{"Metabolite":"(2S)-2-HYDROXYPENTANEDIOIC ACID","moverz_quant":"","ri":"","ri_type":"","pubchem_id":"439939","inchi_key":"","kegg_id":"","other_id":"","other_id_type":""}]
}

}