#METABOLOMICS WORKBENCH hormel101_20161207_150111_mwtab.txt DATATRACK_ID:790 STUDY_ID:ST000518 ANALYSIS_ID:AN000792 PROJECT_ID:PR000383 VERSION 1 CREATED_ON December 13, 2016, 12:24 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 ceramide 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 ceramide concentration 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 ms5632-1 group:control age=7.9; tissue weight (mg)=32.51 SUBJECT_SAMPLE_FACTORS F471 ms5632-2 group:control age=7.0; tissue weight (mg)=38.4 SUBJECT_SAMPLE_FACTORS 2095 ms5632-3 group:control age=9.6; tissue weight (mg)=49.05 SUBJECT_SAMPLE_FACTORS 2103 ms5632-4 group:control age=8.3; tissue weight (mg)=53.96 SUBJECT_SAMPLE_FACTORS MF1092 ms5632-5 group:control age=9.7; tissue weight (mg)=47.92 SUBJECT_SAMPLE_FACTORS MF1103 ms5632-6 group:control age=8.7; tissue weight (mg)=54.03 SUBJECT_SAMPLE_FACTORS 2096 ms5632-7 group:MIGIRKO age=9.6; tissue weight (mg)=46.6 SUBJECT_SAMPLE_FACTORS 2102 ms5632-8 group:MIGIRKO age=8.3; tissue weight (mg)=48.1 SUBJECT_SAMPLE_FACTORS 2109 ms5632-9 group:MIGIRKO age=8.6; tissue weight (mg)=40.8 SUBJECT_SAMPLE_FACTORS 2111 ms5632-10 group:MIGIRKO age=8.6; tissue weight (mg)=45.7 SUBJECT_SAMPLE_FACTORS 2064 ms5632-11 group:MIGIRKO age=10.1; tissue weight (mg)=29.4 SUBJECT_SAMPLE_FACTORS MF991 ms5632-12 group:MIGIRKO age=9.4; tissue weight (mg)=37.1 SUBJECT_SAMPLE_FACTORS F453 ms5632-13 group:Fox0 TKO age=9.7; tissue weight (mg)=44.14 SUBJECT_SAMPLE_FACTORS F455 ms5632-14 group:Fox0 TKO age=9.7; tissue weight (mg)=57.93 SUBJECT_SAMPLE_FACTORS F472 ms5632-15 group:Fox0 TKO age=7.0; tissue weight (mg)=47.52 SUBJECT_SAMPLE_FACTORS F394 ms5632-16 group:Fox0 TKO age=13.1; tissue weight (mg)=51.34 SUBJECT_SAMPLE_FACTORS F323 ms5632-17 group:Fox0 TKO age=9.9; tissue weight (mg)=41.06 SUBJECT_SAMPLE_FACTORS MF496 ms5632-18 group:Fox0 TKO age=12.9; tissue weight (mg)=52.07 SUBJECT_SAMPLE_FACTORS MF1091 ms5632-19 group:QKO age=9.7; tissue weight (mg)=47.77 SUBJECT_SAMPLE_FACTORS MF1104 ms5632-20 group:QKO age=8.7; tissue weight (mg)=60.11 SUBJECT_SAMPLE_FACTORS MF1105 ms5632-21 group:QKO age=8.7; tissue weight (mg)=58.32 SUBJECT_SAMPLE_FACTORS MF1107 ms5632-22 group:QKO age=8.7; tissue weight (mg)=56.07 SUBJECT_SAMPLE_FACTORS MF1119 ms5632-23 group:QKO age=8.5; tissue weight (mg)=66.63 SUBJECT_SAMPLE_FACTORS MF1064 ms5632-24 group:QKO age=9.0; tissue weight (mg)=65.87 SUBJECT_SAMPLE_FACTORS F444 ms5632-25 group:STZ age=10.0; tissue weight (mg)=54.4 SUBJECT_SAMPLE_FACTORS F463 ms5632-26 group:STZ age=8.3; tissue weight (mg)=39.02 SUBJECT_SAMPLE_FACTORS F304 ms5632-27 group:STZ age=11.9; tissue weight (mg)=41.38 SUBJECT_SAMPLE_FACTORS F322 ms5632-28 group:STZ age=9.9; tissue weight (mg)=43.23 SUBJECT_SAMPLE_FACTORS MF569 ms5632-29 group:STZ age=11.4; tissue weight (mg)=43.99 SUBJECT_SAMPLE_FACTORS MF1046 ms5632-30 group:STZ age=8.9; tissue weight (mg)=26.92 SUBJECT_SAMPLE_FACTORS F454 ms5632-31 group:STZ FoxO TKO age=9.7; tissue weight (mg)=48.86 SUBJECT_SAMPLE_FACTORS F456 ms5632-32 group:STZ FoxO TKO age=9.7; tissue weight (mg)=41.49 SUBJECT_SAMPLE_FACTORS F462 ms5632-33 group:STZ FoxO TKO age=8.3; tissue weight (mg)=32.12 SUBJECT_SAMPLE_FACTORS F325 ms5632-34 group:STZ FoxO TKO age=9.9; tissue weight (mg)=44.06 SUBJECT_SAMPLE_FACTORS MF512 ms5632-35 group:STZ FoxO TKO age=12.1; tissue weight (mg)=44.26 SUBJECT_SAMPLE_FACTORS MF568 ms5632-36 group:STZ FoxO TKO age=11.4; tissue weight (mg)=57.64 #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 ceramide concentrations 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 ceramide concentrations in muscle tissue #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C8 (150 x 2mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Quantiva QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ng/mg MS_METABOLITE_DATA_START Samples ms5632-1 ms5632-2 ms5632-3 ms5632-4 ms5632-5 ms5632-6 ms5632-7 ms5632-8 ms5632-9 ms5632-10 ms5632-11 ms5632-12 ms5632-13 ms5632-14 ms5632-15 ms5632-16 ms5632-17 ms5632-18 ms5632-19 ms5632-20 ms5632-21 ms5632-22 ms5632-23 ms5632-24 ms5632-25 ms5632-26 ms5632-27 ms5632-28 ms5632-29 ms5632-30 ms5632-31 ms5632-32 ms5632-33 ms5632-34 ms5632-35 ms5632-36 Factors group:control group:control group:control group:control group:control group:control group:MIGIRKO group:MIGIRKO group:MIGIRKO group:MIGIRKO group:MIGIRKO group:MIGIRKO 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 FoxO TKO group:STZ FoxO TKO group:STZ FoxO TKO group:STZ FoxO TKO group:STZ FoxO TKO group:STZ FoxO TKO Sph 0.03 0.04 0.03 0.03 0.03 0.04 0.07 0.09 0.08 0.06 0.11 0.09 0.03 0.08 0.08 0.03 0.04 0.04 0.05 0.04 0.04 0.08 0.04 0.06 0.05 0.10 0.05 0.03 0.05 0.10 0.08 0.10 0.08 0.07 0.05 0.04 Spa 0.01 0.01 0.01 0.01 0.01 0.01 0.03 0.04 0.03 0.03 0.05 0.04 0.01 0.02 0.04 0.01 0.02 0.01 0.01 0.01 0.01 0.03 0.01 0.02 0.01 0.03 0.01 0.01 0.01 0.03 0.03 0.02 0.02 0.02 0.01 0.01 S1P 0.01 0.01 0.01 0.01 0.01 0.01 0.02 0.02 0.02 0.01 0.04 0.04 0.01 0.01 0.01 0.01 0.01 0.01 0.00 0.00 0.00 0.01 0.00 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.00 C8 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 C14 0.01 0.00 0.00 0.01 0.00 0.00 0.01 0.01 0.02 0.01 0.02 0.02 0.01 0.07 0.01 0.00 0.01 0.00 0.01 0.01 0.00 0.02 0.00 0.01 0.00 0.01 0.00 0.00 0.01 0.01 0.01 0.01 0.01 0.00 0.00 0.00 C16 0.43 0.50 0.35 0.71 0.44 0.43 1.15 1.32 1.54 1.90 2.25 2.04 0.38 1.51 1.13 0.42 0.49 0.47 0.75 0.55 0.41 0.95 0.48 0.79 0.53 0.87 0.55 0.89 0.64 1.13 0.71 0.51 0.69 0.86 0.50 0.41 C18:1 0.04 0.02 0.02 0.02 0.03 0.04 0.05 0.06 0.06 0.05 0.07 0.08 0.02 0.07 0.05 0.04 0.04 0.03 0.04 0.02 0.04 0.03 0.03 0.01 0.10 0.12 0.05 0.08 0.13 0.12 0.10 0.08 0.22 0.04 0.04 0.06 C18 7.93 5.63 4.37 8.82 5.59 4.99 2.51 4.82 5.10 2.38 5.07 4.31 5.47 6.69 6.45 4.17 5.88 4.73 6.53 6.29 5.11 6.78 4.96 5.07 9.18 15.17 12.97 8.20 8.96 19.83 9.54 13.31 12.53 7.76 9.19 6.48 C20 0.09 0.08 0.07 0.15 0.07 0.07 0.18 0.40 0.35 0.24 0.36 0.51 0.07 0.18 0.15 0.05 0.10 0.06 0.21 0.14 0.10 0.34 0.10 0.15 0.13 0.32 0.24 0.14 0.31 0.70 0.41 0.41 0.41 0.24 0.15 0.12 C22 0.20 0.18 0.15 0.30 0.16 0.14 0.27 0.43 0.39 0.35 0.60 0.55 0.16 0.29 0.30 0.14 0.21 0.15 0.25 0.19 0.16 0.31 0.16 0.22 0.33 0.52 0.41 0.28 0.38 0.83 0.44 0.41 0.50 0.32 0.27 0.23 C24:1 1.27 1.35 0.98 1.85 1.41 1.37 2.68 3.74 4.29 2.95 6.07 5.67 1.01 1.57 2.07 1.19 1.35 1.07 1.67 1.36 1.15 2.69 1.21 1.49 1.32 2.32 1.58 1.45 1.63 3.36 2.12 1.84 2.56 1.45 1.31 0.92 C24 0.43 0.43 0.29 0.71 0.38 0.35 0.65 1.25 1.22 0.87 1.81 1.69 0.33 1.09 0.99 0.34 0.54 0.33 0.60 0.50 0.34 1.09 0.39 0.70 0.51 1.25 0.84 0.58 0.61 1.44 1.10 0.85 1.04 0.85 0.66 0.48 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Sph Spa S1P C8 C14 C16 C18:1 C18 C20 C22 C24:1 C24 METABOLITES_END #END