#METABOLOMICS WORKBENCH araskind_20170714_124505 DATATRACK_ID:1138 STUDY_ID:ST000822 ANALYSIS_ID:AN001305 PROJECT_ID:PR000585
VERSION             	1
CREATED_ON             	August 2, 2017, 10:51 am
#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
#STUDY
ST:STUDY_TITLE                   	Metabolomic profiling of IDH1 mutation (part II)
ST:STUDY_TYPE                    	MS analysis
ST:STUDY_SUMMARY                 	We are interested in using both spontaneous and stimulated Raman microscopy to
ST:STUDY_SUMMARY                 	visualize these metabolomic changes as spectral alterations. We have two
ST:STUDY_SUMMARY                 	isogneic cell lines of normal human astrocytes differing only by a point
ST:STUDY_SUMMARY                 	mutation in the IDH-1 gene. We will work with the metabolomics core to elucidate
ST:STUDY_SUMMARY                 	the changes in central metabolism and lipid synthesys in an effort to determine
ST:STUDY_SUMMARY                 	the precise biochemical alterations underlying observed spectral differences. We
ST:STUDY_SUMMARY                 	wil then use a selective inhibitor of the IDH1 R132H to demonstrate to attempt
ST:STUDY_SUMMARY                 	to return TCA metabolome and lipidome to WT phenotype. Lastly, we will use a
ST:STUDY_SUMMARY                 	cell-permeablized variant of 2HG (2R-octyl-alpha-hydroxyglutarate) to
ST:STUDY_SUMMARY                 	recaptitulate the R132H mutant phenotype in wild-type cells, providing strong
ST:STUDY_SUMMARY                 	evidence that 2HG accumulation uderlies the metabolomic (and thus, spectral)
ST:STUDY_SUMMARY                 	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:NUM_GROUPS                    	4
ST:TOTAL_SUBJECTS                	9
#SUBJECT
SU:SUBJECT_TYPE                  	HUMAN
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#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	
#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           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent
CH:COLUMN_NAME                   	Waters Acquity HSS T3 (50 x 2.1mm, 1.8um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	A037-2HG cells updated-20160721.pdf
#MS
MS:MS_COMMENTS                   	-
MS:INSTRUMENT_NAME               	Agilent 6490A QQQ
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	uM/ng protein
MS_METABOLITE_DATA_START
Samples	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
#END