#METABOLOMICS WORKBENCH hormel101_20161202_124855_mwtab.txt DATATRACK_ID:784 STUDY_ID:ST000513 ANALYSIS_ID:AN000786 PROJECT_ID:PR000383 VERSION 1 CREATED_ON December 8, 2016, 3:16 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 Inhibition of autophagy/proteasome degradation or inhibition of protein ST:STUDY_TITLE synthesis in models of muscle insulin resistance affect TCA cycle ST:STUDY_SUMMARY To determine which protein degradation pathways downstream of IR and IGF1R ST:STUDY_SUMMARY contribute to changes in amino acid and mitochondrial metabolite pools, we will ST:STUDY_SUMMARY treat control, M-IR-/-, MIGIRKO, and HFD obese mice with inhibitors of autophagy ST:STUDY_SUMMARY or proteasome. We will treat 5 animals each of control, M-IR-/-, MIGIRKO, and ST:STUDY_SUMMARY HFD mice with vehicle, colchicine to inhibit autophagy, or MG132 to inhibit ST:STUDY_SUMMARY proteasome activity, then measure TCA cycle in muscle tissue. 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 613 Sample # 1 group:Control/Saline | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 595 Sample # 2 group:Control/Saline | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1837 Sample # 3 group:Control/Saline | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1816 Sample # 4 group:Control/Saline | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1942 Sample # 5 group:Control/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1956 Sample # 6 group:Control/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1989 Sample # 7 group:Control/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 615 Sample # 8 group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 597 Sample # 9 group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1838 Sample # 10 group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1815 Sample # 11 group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1970 Sample # 12 group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1972 Sample # 13 group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1943 Sample # 14 group:IRKO/Saline | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1957 Sample # 15 group:IRKO/Saline | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1948 Sample # 16 group:IRKO/Saline | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1990 Sample # 17 group:IRKO/Saline | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1992 Sample # 18 group:IRKO/Saline | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1947 Sample # 19 group:HFD/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1950 Sample # 20 group:HFD/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1960 Sample # 21 group:HFD/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1964 Sample # 22 group:HFD/Saline | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1988 Sample # 23 group:HFD/Saline | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1849 Sample # 24 group:Control/Colch | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 606 Sample # 25 group:Control/Colch | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1826 Sample # 26 group:Control/Colch | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1847 Sample # 27 group:Control/Colch | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1855 Sample # 28 group:Control/Colch | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1944 Sample # 29 group:Control/Colch | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1958 Sample # 30 group:Control/Colch | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1852 Sample # 31 group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1853 Sample # 32 group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1856 Sample # 33 group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1817 Sample # 34 group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 607 Sample # 35 group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1824 Sample # 36 group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1945 Sample # 37 group:IRKO/Colch | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1954 Sample # 38 group:IRKO/Colch | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1955 Sample # 39 group:IRKO/Colch | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1959 Sample # 40 group:IRKO/Colch | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1993 Sample # 41 group:IRKO/Colch | genotype:IR -/- array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1951 Sample # 42 group:HFD+Colch | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1952 Sample # 43 group:HFD+Colch | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1965 Sample # 44 group:HFD+Colch | genotype:Irlox IGFRlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1991 Sample # 45 group:HFD+Colch | genotype:Irlox array=TCA concentrations SUBJECT_SAMPLE_FACTORS 1994 Sample # 46 group:HFD+Colch | genotype:Irlox array=TCA concentrations #COLLECTION CO:COLLECTION_SUMMARY mouse gastrocnemius muscle tissue #TREATMENT TR:TREATMENT_SUMMARY Mice lacking insulin receptors (IR -/- genotype), or IGF-1 receptors (ICF-1 -/- TR:TREATMENT_SUMMARY genotype), or both were generated using Cre lox recombination. Controls were IR TR:TREATMENT_SUMMARY lox/lox, IGF-1 lox/lox, or both. Additional, 10 mice were included that were fed TR:TREATMENT_SUMMARY different diets for 8 weeks, chow or high fat diet. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY TCA concentrations in muscle tissue #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A 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 5975 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/mg tissue 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 Sample # 40 Sample # 41 Sample # 42 Sample # 43 Sample # 44 Sample # 45 Sample # 46 Factors group:Control/Saline | genotype:Irlox IGFRlox group:Control/Saline | genotype:Irlox IGFRlox group:Control/Saline | genotype:Irlox IGFRlox group:Control/Saline | genotype:Irlox IGFRlox group:Control/Saline | genotype:Irlox group:Control/Saline | genotype:Irlox group:Control/Saline | genotype:Irlox group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- group:MIGIRKO/Saline | genotype:IR -/- IGFR -/- group:IRKO/Saline | genotype:IR -/- group:IRKO/Saline | genotype:IR -/- group:IRKO/Saline | genotype:IR -/- group:IRKO/Saline | genotype:IR -/- group:IRKO/Saline | genotype:IR -/- group:HFD/Saline | genotype:Irlox group:HFD/Saline | genotype:Irlox group:HFD/Saline | genotype:Irlox group:HFD/Saline | genotype:Irlox IGFRlox group:HFD/Saline | genotype:Irlox group:Control/Colch | genotype:Irlox IGFRlox group:Control/Colch | genotype:Irlox IGFRlox group:Control/Colch | genotype:Irlox IGFRlox group:Control/Colch | genotype:Irlox IGFRlox group:Control/Colch | genotype:Irlox IGFRlox group:Control/Colch | genotype:Irlox group:Control/Colch | genotype:Irlox group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- group:MIGIRKO/Colch | genotype:IR -/- IGFR -/- group:IRKO/Colch | genotype:IR -/- group:IRKO/Colch | genotype:IR -/- group:IRKO/Colch | genotype:IR -/- group:IRKO/Colch | genotype:IR -/- group:IRKO/Colch | genotype:IR -/- group:HFD+Colch | genotype:Irlox group:HFD+Colch | genotype:Irlox group:HFD+Colch | genotype:Irlox IGFRlox group:HFD+Colch | genotype:Irlox group:HFD+Colch | genotype:Irlox Lactate 28.252 31.567 27.085 28.506 30.447 26.637 28.718 25.292 22.233 15.958 18.839 16.045 12.504 24.745 25.896 25.374 22.545 25.158 27.864 30.908 29.657 25.066 27.280 30.118 29.378 32.528 32.185 30.772 29.199 33.448 20.513 13.282 19.610 22.856 17.554 25.329 27.252 31.725 26.296 25.279 26.483 29.822 29.524 13.279 32.375 29.144 Succinate 0.095 0.155 0.102 0.071 0.069 0.062 0.069 0.093 0.090 0.062 0.051 0.072 0.041 0.053 0.046 0.066 0.066 0.076 0.120 0.106 0.140 0.098 0.138 0.062 0.104 0.084 0.050 0.066 0.079 0.066 0.057 0.036 0.031 0.060 0.057 0.053 0.039 0.050 0.036 0.090 0.043 0.095 0.102 0.087 0.070 0.099 Fumarate 0.176 0.205 0.233 0.145 0.155 0.162 0.161 0.436 0.417 0.432 0.286 0.446 0.325 0.151 0.263 0.240 0.171 0.326 0.156 0.151 0.212 0.141 0.165 0.165 0.218 0.261 0.216 0.208 0.311 0.223 0.430 0.383 0.448 0.451 0.373 0.402 0.197 0.276 0.232 0.303 0.246 0.185 0.234 0.165 0.202 0.231 Oxaloacetate no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk no good pk Ketoglutarate 0.012 0.010 0.010 0.012 0.011 0.016 0.013 0.006 0.007 0.008 0.009 0.007 0.007 0.010 0.008 0.008 0.008 0.008 0.008 0.006 0.006 0.007 0.006 0.010 0.008 0.006 0.007 0.006 0.005 0.006 0.005 0.005 0.005 0.004 0.005 0.005 0.007 0.006 0.005 0.005 0.006 0.005 0.005 0.005 0.008 0.005 Malate 0.147 0.182 0.220 0.127 0.139 0.147 0.157 0.428 0.478 0.424 0.327 0.437 0.335 0.177 0.266 0.232 0.173 0.323 0.150 0.130 0.201 0.128 0.152 0.193 0.183 0.243 0.189 0.196 0.285 0.214 0.386 0.355 0.424 0.428 0.379 0.363 0.210 0.277 0.222 0.281 0.237 0.156 0.208 0.138 0.169 0.200 Glutamate 0.435 0.376 0.300 0.369 0.362 0.257 0.487 1.131 1.327 1.225 0.958 1.411 0.877 0.343 0.706 0.400 0.407 0.709 0.204 0.117 0.131 0.223 0.093 0.350 0.398 0.227 0.360 0.361 0.205 0.168 1.053 1.071 1.110 1.058 1.267 0.830 0.547 0.464 0.314 0.440 0.694 0.150 0.183 0.305 0.282 0.360 cis-Aconitic Acid 0.038 0.022 0.012 0.012 0.005 0.001 0.002 0.006 0.005 0.003 0.005 0.002 0.001 0.001 0.006 0.004 0.003 0.002 0.006 0.004 0.002 0.003 0.004 0.003 0.002 0.002 0.001 0.002 0.002 0.002 0.003 0.001 0.002 0.002 0.002 0.003 0.001 0.003 0.001 0.003 0.002 0.001 0.002 0.002 0.002 0.002 Citrate 0.088 0.118 0.061 0.059 0.084 0.062 0.073 0.141 0.161 0.166 0.129 0.146 0.140 0.056 0.097 0.080 0.073 0.114 0.048 0.079 0.059 0.082 0.103 0.087 0.091 0.119 0.081 0.110 0.114 0.090 0.165 0.160 0.164 0.155 0.163 0.135 0.098 0.089 0.057 0.090 0.105 0.084 0.074 0.090 0.075 0.093 Isocitrate 0.059 0.062 0.023 0.054 0.039 0.019 0.054 0.039 0.032 0.079 0.075 0.063 0.029 0.076 0.149 0.050 0.065 0.135 0.014 0.003 0.002 0.028 0.003 0.077 0.079 0.046 0.075 0.032 0.054 0.106 0.016 0.009 0.005 0.003 0.006 0.003 0.158 0.122 0.210 0.166 0.289 0.002 0.003 0.278 0.002 0.050 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Lactate Succinate Fumarate Oxaloacetate Ketoglutarate Malate Glutamate cis-Aconitic Acid Citrate Isocitrate METABOLITES_END #END