#METABOLOMICS WORKBENCH hormel101_20161207_151358_mwtab.txt DATATRACK_ID:791 STUDY_ID:ST000519 ANALYSIS_ID:AN000793 PROJECT_ID:PR000383 VERSION 1 CREATED_ON December 13, 2016, 12:30 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 Investigating ceremide concentrations in mice muscle tissue lacking insulin ST:STUDY_TITLE receptors and IGF-1 receptors ST:STUDY_SUMMARY Quantitative measures of ceramide metabolite concentrations in muscle from ST:STUDY_SUMMARY control, M-IR-/-, M-IGF1R-/- , and MIGIRKO mice. Also compare mice on a chow ST:STUDY_SUMMARY diet to mice on a high fat diet (HFD). 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 719 ms5689-1 Group:Control | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 721 ms5689-2 Group:Control | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 813 ms5689-3 Group:Control | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 546 ms5689-4 Group:Control | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 722 ms5689-5 Group:IRKO | Genotype:IR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 723 ms5689-6 Group:IRKO | Genotype:IR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 814 ms5689-7 Group:IRKO | Genotype:IR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 817 ms5689-8 Group:IRKO | Genotype:IR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 549 ms5689-9 Group:IRKO | Genotype:IR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 551 ms5689-10 Group:IRKO | Genotype:IR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 743 ms5689-11 Group:Control | Genotype:IGFR lox | Diet:Chow SUBJECT_SAMPLE_FACTORS 747 ms5689-12 Group:Control | Genotype:IGFR lox | Diet:Chow SUBJECT_SAMPLE_FACTORS 783 ms5689-13 Group:Control | Genotype:IGFR lox | Diet:Chow SUBJECT_SAMPLE_FACTORS 740 ms5689-14 Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 741 ms5689-15 Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 742 ms5689-16 Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 780 ms5689-17 Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 781 ms5689-18 Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow SUBJECT_SAMPLE_FACTORS 1191 ms5689-19 Group:CD | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 1192 ms5689-20 Group:CD | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 1193 ms5689-21 Group:CD | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 1195 ms5689-22 Group:CD | Genotype:Irlox | Diet:Chow SUBJECT_SAMPLE_FACTORS 850 ms5689-23 Group:HFD | Genotype:Irlox | Diet:High Fat Diet SUBJECT_SAMPLE_FACTORS 864 ms5689-24 Group:HFD | Genotype:Irlox | Diet:High Fat Diet SUBJECT_SAMPLE_FACTORS 865 ms5689-25 Group:HFD | Genotype:Irlox | Diet:High Fat Diet SUBJECT_SAMPLE_FACTORS 867 ms5689-26 Group:HFD | Genotype:Irlox | Diet:High Fat Diet SUBJECT_SAMPLE_FACTORS 869 ms5689-27 Group:HFD | Genotype:Irlox | Diet:High Fat Diet #COLLECTION CO:COLLECTION_SUMMARY mouse 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 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 ms5689-1 ms5689-2 ms5689-3 ms5689-4 ms5689-5 ms5689-6 ms5689-7 ms5689-8 ms5689-9 ms5689-10 ms5689-11 ms5689-12 ms5689-13 ms5689-14 ms5689-15 ms5689-16 ms5689-17 ms5689-18 ms5689-19 ms5689-20 ms5689-21 ms5689-22 ms5689-23 ms5689-24 ms5689-25 ms5689-26 ms5689-27 Factors Group:Control | Genotype:Irlox | Diet:Chow Group:Control | Genotype:Irlox | Diet:Chow Group:Control | Genotype:Irlox | Diet:Chow Group:Control | Genotype:Irlox | Diet:Chow Group:IRKO | Genotype:IR -/- | Diet:Chow Group:IRKO | Genotype:IR -/- | Diet:Chow Group:IRKO | Genotype:IR -/- | Diet:Chow Group:IRKO | Genotype:IR -/- | Diet:Chow Group:IRKO | Genotype:IR -/- | Diet:Chow Group:IRKO | Genotype:IR -/- | Diet:Chow Group:Control | Genotype:IGFR lox | Diet:Chow Group:Control | Genotype:IGFR lox | Diet:Chow Group:Control | Genotype:IGFR lox | Diet:Chow Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow Group:IGFRKO | Genotype:IGFR -/- | Diet:Chow Group:CD | Genotype:Irlox | Diet:Chow Group:CD | Genotype:Irlox | Diet:Chow Group:CD | Genotype:Irlox | Diet:Chow Group:CD | Genotype:Irlox | Diet:Chow Group:HFD | Genotype:Irlox | Diet:High Fat Diet Group:HFD | Genotype:Irlox | Diet:High Fat Diet Group:HFD | Genotype:Irlox | Diet:High Fat Diet Group:HFD | Genotype:Irlox | Diet:High Fat Diet Group:HFD | Genotype:Irlox | Diet:High Fat Diet Sph 0.010 0.004 0.002 0.010 0.004 0.002 0.004 0.004 0.004 0.005 0.001 0.005 0.002 0.002 0.000 0.003 0.002 0.002 0.003 0.003 0.004 0.004 0.004 0.002 0.003 0.004 0.004 SPA 0.005 0.002 0.001 0.007 0.001 0.002 0.003 0.002 0.003 0.002 0.003 0.002 0.000 0.001 0.001 0.001 0.001 0.002 0.001 0.003 0.003 0.002 0.001 0.001 0.001 0.002 0.001 S1P 0.005 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.000 0.001 0.001 0.001 0.001 0.001 C8-cer 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 C14-cer 0.005 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.002 0.001 0.002 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.002 0.001 0.001 0.002 0.001 C16-cer 0.399 0.084 0.106 0.122 0.146 0.130 0.135 0.100 0.064 0.139 0.090 0.119 0.091 0.054 0.109 0.121 0.112 0.135 0.133 0.150 0.098 0.145 0.141 0.117 0.099 0.139 0.123 C18:1-cer 0.045 0.006 0.006 0.010 0.010 0.007 0.008 0.005 0.006 0.009 0.006 0.006 0.005 0.005 0.005 0.005 0.005 0.006 0.006 0.008 0.010 0.009 0.014 0.011 0.010 0.015 0.014 C18-cer 2.617 0.169 0.209 0.280 0.347 0.337 0.337 0.151 0.215 0.299 0.235 0.262 0.155 0.101 0.139 0.156 0.190 0.247 0.206 0.336 0.390 0.303 0.514 0.341 0.300 0.377 0.456 C20-cer 0.085 0.007 0.011 0.012 0.016 0.016 0.015 0.007 0.008 0.015 0.006 0.010 0.002 0.001 0.004 0.004 0.005 0.006 0.011 0.018 0.014 0.015 0.022 0.019 0.016 0.021 0.019 C22-cer 0.110 0.014 0.020 0.017 0.023 0.022 0.019 0.015 0.014 0.020 0.013 0.017 0.011 0.010 0.014 0.013 0.014 0.015 0.016 0.021 0.018 0.021 0.027 0.024 0.021 0.026 0.025 C24-1-cer 0.815 0.164 0.187 0.197 0.239 0.229 0.223 0.165 0.121 0.210 0.135 0.186 0.136 0.105 0.174 0.189 0.176 0.210 0.226 0.239 0.181 0.226 0.191 0.161 0.129 0.187 0.181 C24-cer 0.236 0.034 0.045 0.042 0.054 0.051 0.048 0.036 0.030 0.045 0.039 0.045 0.026 0.024 0.035 0.031 0.032 0.040 0.038 0.049 0.039 0.049 0.047 0.039 0.034 0.050 0.040 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Sph SPA S1P C8-cer C14-cer C16-cer C18:1-cer C18-cer C20-cer C22-cer C24-1-cer C24-cer METABOLITES_END #END