#METABOLOMICS WORKBENCH TEXT OUTPUT STUDY_ID:	ANALYSIS_ID:	
VERSION             	1
CREATED_ON          	2017-07-14
#PROJECT
PR:PROJECT_TITLE    	Stimulated Raman Immunohistochemistry
PR:PROJECT_TYPE     	MS analysis
PR:PROJECT_SUMMARY  	Profiling of WT, IDH1 R132H, WT + 2HG, Mut + AG!5198
PR:INSTITUTE        	University of Michigan
PR:DEPARTMENT       	Neursurgery
PR:LABORATORY       	Orringer Lab
PR:LAST_NAME        	Orringer
PR:FIRST_NAME       	Daniel
PR:ADDRESS          	Ann Arbor, MI
PR:EMAIL            	dorringe@umich.edu
PR:PHONE            	734-647-9222
PR:FUNDING_SOURCE   	
PR:PROJECT_COMMENTS 	
PR:PUBLICATIONS     	
#STUDY
ST:STUDY_TITLE      	Metabolomic profiling of IDH1 mutation 2
ST:STUDY_TYPE       	MS analysis
ST: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.
ST:INSTITUTE        	University of Michigan
ST:DEPARTMENT       	Biomedical Research Core Facilities
ST:LABORATORY       	Metabolomics core
ST:LAST_NAME        	Kachman
ST:FIRST_NAME       	Maureen
ST:ADDRESS          	6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
ST:EMAIL            	mkachman@med.umich.edu
ST:PHONE            	(734) 232-8175
ST:SUBMIT_DATE      	03/22/2017 09:48:08 AM
ST:NUM_GROUPS       	4
ST:TOTAL_SUBJECTS   	9
ST:STUDY_COMMENTS   	
#SUBJECT_SAMPLE_FACTORS:          	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS            	SU0018656	S00028825	Cell Line:IDH1 WILD TYPE | Treatment:NORMAL CULTURE MEDIA	
SUBJECT_SAMPLE_FACTORS            	SU0018657	S00028826	Cell Line:IDH1 WILD TYPE | Treatment:NORMAL CULTURE MEDIA	
SUBJECT_SAMPLE_FACTORS            	SU0018658	S00028827	Cell Line:IDH1 WILD TYPE | Treatment:NORMAL CULTURE MEDIA	
SUBJECT_SAMPLE_FACTORS            	SU0018659	S00028828	Cell Line:IDH1 R132H | Treatment:NORMAL CULTURE MEDIA	
SUBJECT_SAMPLE_FACTORS            	SU0018660	S00028829	Cell Line:IDH1 R132H | Treatment:NORMAL CULTURE MEDIA	
SUBJECT_SAMPLE_FACTORS            	SU0018661	S00028830	Cell Line:IDH1 R132H | Treatment:NORMAL CULTURE MEDIA	
SUBJECT_SAMPLE_FACTORS            	SU0018662	S00028831	Cell Line:IDH1 WILD TYPE | Treatment:CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF	
SUBJECT_SAMPLE_FACTORS            	SU0018663	S00028832	Cell Line:IDH1 WILD TYPE | Treatment:CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF	
SUBJECT_SAMPLE_FACTORS            	SU0018664	S00028833	Cell Line:IDH1 WILD TYPE | Treatment:CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF	
#SUBJECT
SU:SUBJECT_TYPE                   	HUMAN
SU:SUBJECT_SPECIES                	Homo sapiens
SU:TAXONOMY_ID                    	9606
#COLLECTION
CO:COLLECTION_SUMMARY             	-
#TREATMENT
TR:TREATMENT_SUMMARY              	-
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY             	-
SP:SAMPLEPREP_PROTOCOL_FILENAME   	A037-2HG cells updated-20160721.pdf
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE            	C18
CH:INSTRUMENT_NAME                	6490A QQQ LC/MS (Agilent)
CH:COLUMN_NAME                    	ACQUITY UPLC HSS T3, 100A, 1.8mkm, 2.1mm X 50mm (Waters Corporation)
CH:METHODS_FILENAME               	2016-03-16-2HG-6490
#ANALYSIS
AN:ANALYSIS_TYPE                  	MRM
AN:INSTRUMENT_NAME                	6490A QQQ LC/MS (Agilent)
AN:ANALYSIS_PROTOCOL_FILE         	A037-2HG cells updated-20160721.pdf
AN:ACQUISITION_PARAMETERS_FILE    	2016-03-16-2HG-6490
AN:PROCESSING_PARAMETERS_FILE     	-
#MS
MS:INSTRUMENT_TYPE                	QQQ
MS:MS_TYPE                        	
MS:ION_MODE                       	NEGATIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	uM/ng protein
MS_METABOLITE_DATA_START
Sample	S00028825	S00028826	S00028827	S00028828	S00028829	S00028830	S00028831	S00028832	S00028833
Factors	Cell Line:IDH1 WILD TYPE | Treatment:NORMAL CULTURE MEDIA	Cell Line:IDH1 WILD TYPE | Treatment:NORMAL CULTURE MEDIA	Cell Line:IDH1 WILD TYPE | Treatment:NORMAL CULTURE MEDIA	Cell Line:IDH1 R132H | Treatment:NORMAL CULTURE MEDIA	Cell Line:IDH1 R132H | Treatment:NORMAL CULTURE MEDIA	Cell Line:IDH1 R132H | Treatment:NORMAL CULTURE MEDIA	Cell Line:IDH1 WILD TYPE | Treatment:CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF	Cell Line:IDH1 WILD TYPE | Treatment:CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF	Cell Line:IDH1 WILD TYPE | Treatment:CULTURE MEDIA + OCTYL-2HG (0.2mM) IN DMF
(2R)-2-HYDROXYPENTANEDIOIC ACID	2.274904019	2.626153718	.16017799	47.05407046	49.21195136	32.08372226	28.16127607	13.87789563	18.60918156
(2S)-2-HYDROXYPENTANEDIOIC ACID	2.827622943	3.395280567	.71648629	0	0	0	0	0	0
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	moverz_quant	ri	ri_type	pubchem_id	inchi_key	kegg_id	other_id	other_id_type
(2R)-2-HYDROXYPENTANEDIOIC ACID				439391				
(2S)-2-HYDROXYPENTANEDIOIC ACID				439939				
METABOLITES_END
