#METABOLOMICS WORKBENCH hormel101_20161207_152448_mwtab.txt DATATRACK_ID:792 STUDY_ID:ST000520 ANALYSIS_ID:AN000794 PROJECT_ID:PR000383
VERSION             	1
CREATED_ON             	December 13, 2016, 4:46 pm
#PROJECT
PR:PROJECT_TITLE                 	Mayo Metabolomics Pilot and Feasibility Award: Role of muscle insulin and IGF-1
PR:PROJECT_TITLE                 	signaling on serum and muscle metabolite profiles
PR:PROJECT_SUMMARY               	Skeletal muscle insulin resistance is a cardinal feature of the pathogenesis of
PR:PROJECT_SUMMARY               	type 2 diabetes. Insulin and IGF-1 signal through their highly related receptors
PR:PROJECT_SUMMARY               	to impact on many aspects of muscle physiology including glucose homeostasis,
PR:PROJECT_SUMMARY               	protein metabolism, and mitochondrial function. Early physiological studies, as
PR:PROJECT_SUMMARY               	well as recent large scale metabolomic studies, have shown that changes in
PR:PROJECT_SUMMARY               	specific pools of circulating amino acid metabolites, such as branched chain
PR:PROJECT_SUMMARY               	amino acids (BCAAs), are associated with insulin resistance and can predict
PR:PROJECT_SUMMARY               	future diabetes, but the source and impact of these changes in amino acids are
PR:PROJECT_SUMMARY               	not fully understood. We have recently generated mice which lack insulin
PR:PROJECT_SUMMARY               	receptors (IR) or IGF-1 receptors (IGF1R) or both in muscle using Cre lox
PR:PROJECT_SUMMARY               	recombination. We find that mice which lack only IR or only IGF1R in muscle show
PR:PROJECT_SUMMARY               	minimal changes in muscle mass, but do display increases in proteasomal activity
PR:PROJECT_SUMMARY               	and autophagy in muscle. On the other hand, mice with combined loss of both IR
PR:PROJECT_SUMMARY               	and IGF1R display markedly decreased muscle mass and enhanced degradation
PR:PROJECT_SUMMARY               	pathways, associated with increased protein synthesis, and display changes in
PR:PROJECT_SUMMARY               	mitochondrial gene regulation, indicating that both receptors can compensate to
PR:PROJECT_SUMMARY               	some extent for loss of the other. We hypothesize that IR and IGF1R signaling in
PR:PROJECT_SUMMARY               	muscle coordinate amino acid metabolite turnover and fuel
PR:PROJECT_SUMMARY               	substrate/mitochondrial metabolism, and that in insulin resistant states,
PR:PROJECT_SUMMARY               	changes in protein metabolism and mitochondrial function disrupt relative
PR:PROJECT_SUMMARY               	proportions of amino acid metabolites, which in turn contribute to diabetes risk
PR:PROJECT_SUMMARY               	and/or muscle pathology. We propose to test this hypothesis by performing large
PR:PROJECT_SUMMARY               	scale metabolomics on serum and muscle from mice lacking IR, IGF1R or both in
PR:PROJECT_SUMMARY               	muscle, and we will compare these changes to both insulin deficient
PR:PROJECT_SUMMARY               	streptozotocin-treated and insulin resistant diet-induced obese mouse models. To
PR:PROJECT_SUMMARY               	gain insight into which pathways are critical for metabolite changes, we will
PR:PROJECT_SUMMARY               	also treat mice with specific inhibitors of mTOR, a common protein synthesis
PR:PROJECT_SUMMARY               	pathway, as well as inhibitors of autophagy or proteasomal degradation and
PR:PROJECT_SUMMARY               	determine metabolite concentrations in muscle and serum. These studies will
PR:PROJECT_SUMMARY               	identify specific pathways that impact amino acid and mitochondrial metabolite
PR:PROJECT_SUMMARY               	flux which are perturbed in insulin resistant states, and potentially provide
PR:PROJECT_SUMMARY               	insights into how changes in amino acid metabolites contribute to diabetes risk.
PR:INSTITUTE                     	Mayo Clinic
PR:LAST_NAME                     	O'Neill
PR:FIRST_NAME                    	Brian
PR:ADDRESS                       	One Joslin Place, Boston, MA 02215
PR:EMAIL                         	brian.o'neill@joslin.harvard.edu
PR:PHONE                         	617-309-2400
#STUDY
ST:STUDY_TITLE                   	Measuring TCA cycle concentrations in insulin resistant and insulin deficient
ST:STUDY_TITLE                   	mouse tissue models
ST:STUDY_SUMMARY                 	To compare models of insulin resistance to a model of loss of insulin signaling,
ST:STUDY_SUMMARY                 	we will also determine TCA cycle metabolites in muscle, using control and
ST:STUDY_SUMMARY                 	streptozotocin (STZ) treated mice as a model of insulin deficient diabetes.
ST:INSTITUTE                     	Mayo Clinic
ST:LAST_NAME                     	O'Neill
ST:FIRST_NAME                    	Brian
ST:ADDRESS                       	One Joslin Place, Boston, MA 02215
ST:EMAIL                         	brian.o'neill@joslin.harvard.edu
ST:PHONE                         	617-309-2400
#SUBJECT
SU:SUBJECT_TYPE                  	Mouse
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	F467	Sample # 1	group:control	Age=7.9; Group label=Cont 2/24 sac 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	F471	Sample # 2	group:control	Age=7.0; Group label=Cont 2/24 sac 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	2095	Sample # 3	group:control	Age=9.6; Group label=WT 8-10 wks 3/9
SUBJECT_SAMPLE_FACTORS           	2103	Sample # 4	group:control	Age=8.3; Group label=WT 8-10 wks 3/9
SUBJECT_SAMPLE_FACTORS           	MF1092	Sample # 5	group:control	Age=9.7; Group label=8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	MF1103	Sample # 6	group:control	Age=8.7; Group label=8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	F453	Sample # 7	group:Fox0 TKO	Age=9.7; Group label=FoxO TKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	F455	Sample # 8	group:Fox0 TKO	Age=9.7; Group label=FoxO TKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	F472	Sample # 9	group:Fox0 TKO	Age=7.0; Group label=FoxO TKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	F394	Sample # 10	group:Fox0 TKO	Age=13.1; Group label=FoxO TKO 6/23/15
SUBJECT_SAMPLE_FACTORS           	F323	Sample # 11	group:Fox0 TKO	Age=9.9; Group label=FoxO TKO 6/23/15
SUBJECT_SAMPLE_FACTORS           	MF496	Sample # 12	group:Fox0 TKO	Age=12.9; Group label=FoxO TKO 6/23/15
SUBJECT_SAMPLE_FACTORS           	MF1091	Sample # 13	group:QKO	Age=9.7; Group label=QKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	MF1104	Sample # 14	group:QKO	Age=8.7; Group label=QKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	MF1105	Sample # 15	group:QKO	Age=8.7; Group label=QKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	MF1107	Sample # 16	group:QKO	Age=8.7; Group label=QKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	MF1119	Sample # 17	group:QKO	Age=8.5; Group label=QKO 8-10 wk on 3/9
SUBJECT_SAMPLE_FACTORS           	MF1064	Sample # 18	group:QKO	Age=9.0; Group label=QKO
SUBJECT_SAMPLE_FACTORS           	F444	Sample # 19	group:STZ	Age=10.0; Group label=STZ 2/24 sac 8-10wk 3/9
SUBJECT_SAMPLE_FACTORS           	F463	Sample # 20	group:STZ	Age=8.3; Group label=STZ 2/24 sac 8-10wk 3/9
SUBJECT_SAMPLE_FACTORS           	F304	Sample # 21	group:STZ	Age=11.9; Group label=STZ
SUBJECT_SAMPLE_FACTORS           	F322	Sample # 22	group:STZ	Age=9.9; Group label=STZ
SUBJECT_SAMPLE_FACTORS           	MF569	Sample # 23	group:STZ	Age=11.4; Group label=STZ
SUBJECT_SAMPLE_FACTORS           	MF1046	Sample # 24	group:STZ	Age=8.9; Group label=STZ Sal
SUBJECT_SAMPLE_FACTORS           	F454	Sample # 25	group:STZ	Age=9.7; Group label=STZ 2/24 sac 8-10wk 3/9
SUBJECT_SAMPLE_FACTORS           	F456	Sample # 26	group:STZ	Age=9.7; Group label=STZ 2/24 sac 8-10wk 3/9
SUBJECT_SAMPLE_FACTORS           	F462	Sample # 27	group:STZ	Age=8.3; Group label=STZ 2/24 sac 8-10wk 3/9
SUBJECT_SAMPLE_FACTORS           	F325	Sample # 28	group:STZ FoxO TKO	Age=9.9; Group label=STZ FoxO TKO
SUBJECT_SAMPLE_FACTORS           	MF512	Sample # 29	group:STZ FoxO TKO	Age=12.1; Group label=STZ FoxO TKO
SUBJECT_SAMPLE_FACTORS           	MF568	Sample # 30	group:STZ FoxO TKO	Age=11.4; Group label=STZ FoxO TKO
SUBJECT_SAMPLE_FACTORS           	2125	Sample # 31	group:control	Age=13.4; Group label=WT
SUBJECT_SAMPLE_FACTORS           	2126	Sample # 32	group:MIGIRKO	Age=13.4; Group label=MIGIRKO
SUBJECT_SAMPLE_FACTORS           	2176	Sample # 33	group:MIGIRKO	Age=9.9; Group label=MIGIRKO
SUBJECT_SAMPLE_FACTORS           	2185	Sample # 34	group:control	Age=8.1; Group label=WT
SUBJECT_SAMPLE_FACTORS           	2186	Sample # 35	group:MIGIRKO	Age=8.1; Group label=MIGIRKO
SUBJECT_SAMPLE_FACTORS           	1142	Sample # 36	group:control	Age=14.7; Group label=WT
SUBJECT_SAMPLE_FACTORS           	1143	Sample # 37	group:MIGIRKO	Age=14.7; Group label=MIGIRKO
SUBJECT_SAMPLE_FACTORS           	1185	Sample # 38	group:control	Age=11.3; Group label=WT
SUBJECT_SAMPLE_FACTORS           	1187	Sample # 39	group:MIGIRKO	Age=11.3; Group label=MIGIRKO
#COLLECTION
CO:COLLECTION_SUMMARY            	To determine the role of FoxO transcription factors in muscle atrophy and
CO:COLLECTION_SUMMARY            	increased autophagy in MIGIRKO mice, we crossed MIGIRKO (lacking IR and IGF1R)
CO:COLLECTION_SUMMARY            	mice with mice in which FoxO1, FoxO3, and FoxO4 genes were floxed to delete all
CO:COLLECTION_SUMMARY            	the major isoforms of FoxO expressed in muscle. Mice in which 5 separate genes
CO:COLLECTION_SUMMARY            	— IR, Igf1r, FoxO1, FoxO3, and FoxO4 — were specifically deleted in muscle
CO:COLLECTION_SUMMARY            	(muscle quintuple-knockout mice, hereafter referred to as QKO mice); were born
CO:COLLECTION_SUMMARY            	in normal Mendelian ratios, and appeared normal both on external inspection and
CO:COLLECTION_SUMMARY            	following dissection compared with littermate controls and with muscle FoxO1/3/4
CO:COLLECTION_SUMMARY            	triple-knockout mice (FoxO TKO). Streptozotocin (STZ) treated mice were used as
CO:COLLECTION_SUMMARY            	a model of insulin deficient diabetes.
#TREATMENT
TR:TREATMENT_SUMMARY             	To determine the relevance of the changes in M-IR-/-, M-IGF1R-/- , and MIGIRKO
TR:TREATMENT_SUMMARY             	mice to insulin resistant states, we will perform large scale metabolomics and
TR:TREATMENT_SUMMARY             	determine amino acid and TCA cycle metabolites in muscle and serum from 5 mice
TR:TREATMENT_SUMMARY             	fed chow diet or 5 fed a high fat diet (HFD) for 8 weeks. Lastly, to compare
TR:TREATMENT_SUMMARY             	models of insulin resistance to a model of loss of insulin signaling, we will
TR:TREATMENT_SUMMARY             	also determine muscle and serum metabolites in 5 control and 5 streptozotocin
TR:TREATMENT_SUMMARY             	(STZ) treated mice as a model of insulin deficient diabetes.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	TCA concentrations in muscle tissue
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Agilent 7890B
CH:COLUMN_NAME                   	Agilent HP5-MS (30m × 0.25mm, 0.25 um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:MS_COMMENTS                   	-
MS:INSTRUMENT_NAME               	Agilent 5977
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	nmol/mg
MS_METABOLITE_DATA_START
Samples	Sample # 1	Sample # 2	Sample # 3	Sample # 4	Sample # 5	Sample # 6	Sample # 7	Sample # 8	Sample # 9	Sample # 10	Sample # 11	Sample # 12	Sample # 13	Sample # 14	Sample # 15	Sample # 16	Sample # 17	Sample # 18	Sample # 19	Sample # 20	Sample # 21	Sample # 22	Sample # 23	Sample # 24	Sample # 25	Sample # 26	Sample # 27	Sample # 28	Sample # 29	Sample # 30	Sample # 31	Sample # 32	Sample # 33	Sample # 34	Sample # 35	Sample # 36	Sample # 37	Sample # 38	Sample # 39
Factors	group:control	group:control	group:control	group:control	group:control	group:control	group:Fox0 TKO	group:Fox0 TKO	group:Fox0 TKO	group:Fox0 TKO	group:Fox0 TKO	group:Fox0 TKO	group:QKO	group:QKO	group:QKO	group:QKO	group:QKO	group:QKO	group:STZ	group:STZ	group:STZ	group:STZ	group:STZ	group:STZ	group:STZ	group:STZ	group:STZ	group:STZ FoxO TKO	group:STZ FoxO TKO	group:STZ FoxO TKO	group:control	group:MIGIRKO	group:MIGIRKO	group:control	group:MIGIRKO	group:control	group:MIGIRKO	group:control	group:MIGIRKO
Lactate	2.74	71.98	60.90	104.59	63.73	64.20	70.04	110.35	75.69	79.89	104.18	53.88	98.23	61.13	55.82	58.46	64.17	123.78	281.04	69.82	47.59	41.64	126.80	167.34	72.26	91.09	165.37	60.43	81.80	53.95	44.73	96.14	63.85	31.38	120.99	22.74	166.86	21.43	181.27
Succinate	0.27	0.28	0.36	0.12	0.26	0.21	0.21	0.13	0.36	0.26	0.12	0.18	0.20	0.23	0.29	0.29	0.36	0.41	0.46	1.14	0.13	0.27	0.26	0.47	0.12	0.12	0.11	0.19	0.29	0.30	0.28	0.21	0.31	0.31	0.59	0.33	0.58	0.25	0.82
Fumarate	0.28	0.26	0.34	0.21	0.27	0.24	0.26	0.16	0.46	0.37	0.28	0.28	0.28	0.29	0.32	0.31	0.33	0.28	0.19	0.24	0.20	0.34	0.35	0.17	0.22	0.20	0.23	0.39	0.34	0.34	0.19	0.33	0.38	0.19	0.37	0.19	0.37	0.19	0.36
Oxaloacetate	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.00	0.00	0.00	0.00	0.01	0.03	0.02	0.02	0.01	0.01	0.02	0.01	0.01	0.02	0.01	0.01	0.01	0.00	0.01	0.01	0.00	0.01	0.00	0.01	0.00	0.01
Ketoglutarate	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
Malate	0.26	0.24	0.31	0.20	0.25	0.22	0.24	0.15	0.43	0.34	0.26	0.25	0.26	0.26	0.28	0.28	0.30	0.26	0.18	0.23	0.19	0.31	0.32	0.16	0.21	0.20	0.22	0.36	0.31	0.30	0.17	0.31	0.35	0.17	0.35	0.17	0.35	0.17	0.34
Glutamate	0.39	0.54	0.67	0.35	0.58	0.33	0.67	0.52	1.98	0.36	0.73	0.65	0.90	0.57	0.61	0.88	0.70	1.00	1.23	0.90	0.31	0.45	0.85	1.19	1.04	0.50	0.73	1.05	0.61	0.88	0.46	0.88	0.84	0.42	1.24	0.33	0.75	0.48	1.06
cis-Aconitic Acid	0.00	0.00	0.00	0.01	0.00	0.00	0.00	0.00	0.01	0.01	0.01	0.00	0.00	0.00	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.00	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.00	0.00	0.00	0.01	0.00	0.01	0.00	0.01	0.00	0.01
Citrate	0.59	0.17	0.14	0.12	0.17	0.14	0.20	0.08	0.29	0.20	0.15	0.16	0.19	0.15	0.15	0.18	0.17	0.21	0.09	0.19	0.09	0.12	0.13	0.16	0.40	0.21	0.09	0.23	0.16	0.14	0.14	0.24	0.30	0.13	0.27	0.19	0.22	0.20	0.26
Isocitrate	0.00	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.02	0.01	0.01	0.01	0.01	0.01	0.01	0.02	0.02	0.02	0.01	0.01	0.01	0.01	0.01	0.01	0.03	0.02	0.01	0.02	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01	0.01
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
Lactate
Succinate
Fumarate
Oxaloacetate
Ketoglutarate
Malate
Glutamate
cis-Aconitic Acid
Citrate
Isocitrate
METABOLITES_END
#END