#METABOLOMICS WORKBENCH hormel101_20161021_150630 DATATRACK_ID:765 STUDY_ID:ST000506 ANALYSIS_ID:AN000775 PROJECT_ID:PR000371 VERSION 1 CREATED_ON November 29, 2016, 3:15 pm #PROJECT PR:PROJECT_TITLE Mayo Metabolomics Pilot and Feasibility Award: Metabolic Dissection of Aberrant PR:PROJECT_TITLE Glutamine Metabolism in Prostate Cancer PR:PROJECT_SUMMARY In this project, we propose to define the specific role(s) of three crucial PR:PROJECT_SUMMARY oncogenic signaling cascades (androgen receptor, Myc and AMPK) in regulating PR:PROJECT_SUMMARY glutamine metabolism in prostate cancer using 13C-labeled glutamine with mass PR:PROJECT_SUMMARY spectrometry-based metabolic flux analyses. PR:INSTITUTE Mayo Clinic PR:LAST_NAME Frigo PR:FIRST_NAME Daniel PR:ADDRESS Department of Biology and Biochemistry, University of Houston, Houston, Texas PR:ADDRESS 77204-5001 PR:EMAIL frigo@uh.edu PR:PHONE 832-842-8824 #STUDY ST:STUDY_TITLE Timecourse of TCA panel in LNCaP Cells ST:STUDY_SUMMARY Timecourse of TCA panel in LNCaP Cells ST:INSTITUTE Mayo Clinic ST:LAST_NAME Frigo ST:FIRST_NAME Daniel ST:ADDRESS Department of Biology and Biochemistry, University of Houston, Houston, Texas ST:ADDRESS 77204-5001 ST:EMAIL frigo@uh.edu ST:PHONE 832-842-8824 #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 - sample # 1 Treatment:vehicle (ethanol) 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 2 Treatment:100 pM R1881 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 3 Treatment:1 nM R1881 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 4 Treatment:10 nM R1881 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 5 Treatment:vehicle (ethanol) 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 6 Treatment:100 pM R1881 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 7 Treatment:10 nM R1881 13C media incubation time=24 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 8 Treatment:vehicle (ethanol) 13C media incubation time=72 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 9 Treatment:100 pM R1881 13C media incubation time=72 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 10 Treatment:1 nM R1881 13C media incubation time=72 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 11 Treatment:10 nM R1881 13C media incubation time=72 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 12 Treatment:vehicle (ethanol) 13C media incubation time=72 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 13 Treatment:100 pM R1881 13C media incubation time=72 hours; assay=TCA media concentrations SUBJECT_SAMPLE_FACTORS - sample # 14 Treatment:10 nM R1881 13C media incubation time=72 hours; assay=TCA media concentrations #COLLECTION CO:COLLECTION_SUMMARY LNCaP media #TREATMENT TR:TREATMENT_SUMMARY LNCaP cells that were serum starved and incubated with 2mM [U-13C]glutamine and TR:TREATMENT_SUMMARY treated with dose response of a synthetic androgen (R1881) for 24 or 72 hours. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY TCA media concentrations #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 5975 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/ml 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 Factors Treatment:vehicle (ethanol) Treatment:100 pM R1881 Treatment:1 nM R1881 Treatment:10 nM R1881 Treatment:vehicle (ethanol) Treatment:100 pM R1881 Treatment:10 nM R1881 Treatment:vehicle (ethanol) Treatment:100 pM R1881 Treatment:1 nM R1881 Treatment:10 nM R1881 Treatment:vehicle (ethanol) Treatment:100 pM R1881 Treatment:10 nM R1881 Lactate 683.4 714.3 710.6 768.9 668.1 727.3 759.0 1075.7 1891.0 2333.7 2349.2 1182.0 1986.3 2413.2 Succinate 1.470 1.069 0.938 1.022 0.946 1.001 0.890 3.634 4.450 4.133 4.252 3.787 4.621 4.492 Fumarate 1.453 0.907 0.627 0.646 0.612 0.558 0.443 0.710 0.815 0.826 0.791 0.637 0.810 0.850 Oxaloacetate 1.980 1.947 1.916 1.935 2.011 2.044 1.985 1.994 2.098 2.499 2.144 3.074 2.661 4.237 Ketoglutarate 0.826 0.871 0.793 0.925 0.811 0.943 0.872 1.582 2.152 1.780 1.829 1.878 2.129 2.152 Malate 1.215 0.728 0.687 0.657 0.679 0.644 0.597 2.033 2.425 2.415 2.526 1.813 2.497 2.521 Glutamate 8.165 8.042 7.931 7.672 8.092 7.906 7.751 11.933 14.600 13.059 14.831 9.537 14.450 14.049 cis-Aconitic Acid 1.369 0.848 0.677 0.630 0.666 0.593 0.549 0.712 0.635 0.537 0.568 0.674 0.601 0.485 Citrate 5.650 5.389 6.286 6.899 4.778 5.694 6.613 18.315 27.089 30.453 31.496 17.766 27.150 31.203 Isocitrate 2.947 0.917 0.501 0.309 0.288 0.219 0.151 0.205 0.157 0.135 0.133 0.171 0.152 0.136 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Lactate Succinate Fumarate Oxaloacetate Ketoglutarate Malate Glutamate cis-Aconitic Acid Citrate Isocitrate METABOLITES_END #END