#METABOLOMICS WORKBENCH hormel101_20161021_150158 DATATRACK_ID:764 STUDY_ID:ST000505 ANALYSIS_ID:AN000774 PROJECT_ID:PR000371 VERSION 1 CREATED_ON November 29, 2016, 3:06 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_TYPE MS targeted analysis 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:LABORATORY Mayo Metabolomics Core 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 Amino Acid Metabolites in LNCaP Cell pellets ST:STUDY_TYPE timecourse ST:STUDY_SUMMARY Timecourse of Amino Acid Metabolites in LNCaP Cell pellets ST:INSTITUTE Mayo Clinic ST:LABORATORY Mayo Metabolomics Core 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 - ms5459-1 Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-2 Treatment Factor:treated-100 pM R1881 | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-3 Treatment Factor:treated-1 nM R1881 | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-4 Treatment Factor:treated-10 nM R1881 | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-5 Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-6 Treatment Factor:treated-100 pM R1881 | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-7 Treatment Factor:treated-10 nM R1881 | 13C media incubation time:24 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-8 Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-9 Treatment Factor:treated-100 pM R1881 | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-10 Treatment Factor:treated-1 nM R1881 | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-11 Treatment Factor:treated-10 nM R1881 | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-12 Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-13 Treatment Factor:treated-100 pM R1881 | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media SUBJECT_SAMPLE_FACTORS - ms5459-14 Treatment Factor:treated-10 nM R1881 | 13C media incubation time:72 hours assay=Concentration of AA + metabolites in cell media #COLLECTION CO:COLLECTION_SUMMARY LNCaP cell 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 Concentration of Amino Acids metabolites in cell media #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Quantum Ultra MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS micromolar MS_METABOLITE_DATA_START Samples ms5459-1 ms5459-2 ms5459-3 ms5459-4 ms5459-5 ms5459-6 ms5459-7 ms5459-8 ms5459-9 ms5459-10 ms5459-11 ms5459-12 ms5459-13 ms5459-14 Factors Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:24 hours Treatment Factor:treated-100 pM R1881 | 13C media incubation time:24 hours Treatment Factor:treated-1 nM R1881 | 13C media incubation time:24 hours Treatment Factor:treated-10 nM R1881 | 13C media incubation time:24 hours Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:24 hours Treatment Factor:treated-100 pM R1881 | 13C media incubation time:24 hours Treatment Factor:treated-10 nM R1881 | 13C media incubation time:24 hours Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:72 hours Treatment Factor:treated-100 pM R1881 | 13C media incubation time:72 hours Treatment Factor:treated-1 nM R1881 | 13C media incubation time:72 hours Treatment Factor:treated-10 nM R1881 | 13C media incubation time:72 hours Treatment Factor:treated-vehicle (ethanol) | 13C media incubation time:72 hours Treatment Factor:treated-100 pM R1881 | 13C media incubation time:72 hours Treatment Factor:treated-10 nM R1881 | 13C media incubation time:72 hours Histidine 36.0 31.6 32.2 25.2 31.2 27.3 28.8 33.8 29.4 20.7 23.0 26.5 22.4 22.5 Hydroxyproline 192.6 177.4 205.6 194.1 191.5 190.7 182.8 189.7 180.0 193.4 188.0 185.8 180.6 181.2 1-Methylhistidine 0.0 3.2 1.1 11.0 2.5 3.9 1.1 2.3 5.0 2.8 0.0 11.1 3.8 3.3 3-Methylhistidine 10.8 12.5 11.2 11.2 12.9 12.8 14.5 11.1 9.2 12.7 13.0 15.6 7.2 9.4 Phosphoethanolamine 0.2 0.0 0.1 0.0 0.1 0.0 0.0 0.7 0.3 0.2 0.1 0.6 0.3 0.2 Carnosine 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Anserine 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Ethanolamine 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.4 0.0 0.0 0.0 0.4 0.0 0.0 Aspartic Acid 132.2 126.1 129.7 132.6 126.2 129.7 137.7 127.0 122.1 119.6 114.7 127.5 118.5 126.0 Sarcosine 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.2 0.3 0.4 0.1 0.2 0.4 beta-Alanine 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.0 0.0 0.1 gamma-Amino-N-butyric-acid 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 alpha-Aminoadipic-acid 0.1 0.1 0.1 0.1 0.1 0.1 0.0 0.4 0.3 0.3 0.4 0.5 0.3 0.5 beta-Aminoisobutyric-acid 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Hydroxylysine 1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Hydroxylysine 2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 alpha-Amino-N-butyric-acid 0.0 0.1 0.0 0.1 0.0 0.0 0.0 0.0 0.1 0.0 0.0 0.1 0.0 0.0 Cystathionine 1 8.4 9.2 9.1 9.8 10.0 9.2 9.0 33.6 42.6 46.7 44.5 30.9 43.4 44.3 Cystathionine 2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Methionine 35.5 34.7 36.1 33.2 38.5 35.5 36.0 36.0 18.3 16.6 14.4 33.9 19.6 15.4 allo-Isoleucine 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Homocystine 2.2 2.7 2.5 2.5 1.9 2.4 2.2 2.0 1.9 1.7 1.7 0.7 2.1 1.8 Asparagine 494.1 515.9 458.5 493.5 496.4 480.2 467.3 508.0 449.4 437.6 438.0 469.1 437.4 425.9 Arginine 946.0 888.1 1004.3 970.2 954.3 935.4 907.4 619.1 387.6 461.8 379.8 617.5 387.6 363.3 Taurine 1.8 1.4 1.7 1.4 2.0 1.4 1.4 1.2 0.5 0.5 0.8 1.3 0.6 0.7 Serine 289.7 294.2 294.1 300.5 303.8 291.2 278.4 240.1 117.0 99.1 81.8 197.5 105.0 80.8 Glutamine 15.4 14.3 16.8 15.4 16.4 16.3 14.5 53.3 37.3 59.2 57.9 33.2 36.2 55.3 Glycine 184.4 187.5 191.2 192.2 191.1 193.7 193.5 359.2 352.8 323.7 327.9 377.6 349.8 364.4 Citrulline 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Glutamic Acid 35.6 34.9 36.2 34.6 37.2 35.8 34.3 54.3 64.9 56.8 65.1 46.6 65.4 63.3 Threonine 164.6 161.0 162.8 163.7 176.8 159.2 159.9 176.9 135.8 120.1 119.0 166.1 131.1 120.7 Alanine 95.6 91.9 87.7 90.7 92.7 93.2 88.9 250.4 192.7 169.5 175.5 193.4 197.3 162.4 Proline 176.7 164.6 170.0 161.2 178.4 172.7 162.9 192.6 174.4 155.6 156.1 193.4 174.7 163.3 Ornithine 116.4 154.6 132.6 150.6 125.8 135.8 129.1 526.2 740.8 595.9 742.7 438.5 717.3 739.0 Lysine 218.9 192.4 184.3 189.0 198.5 193.5 189.1 198.3 153.8 151.4 141.5 191.9 149.5 146.6 Cystine 164.6 171.9 182.2 166.0 190.1 163.9 198.7 131.0 115.1 114.0 107.5 145.6 116.4 118.5 Tyrosine 112.8 109.2 116.8 111.9 114.0 107.2 117.8 112.5 97.6 89.7 86.6 111.1 94.2 91.1 Valine 163.7 157.6 159.4 158.9 159.7 155.8 156.1 159.9 118.6 103.3 102.8 158.5 113.5 108.2 Isoleucine 372.0 383.5 383.9 379.4 392.1 368.8 380.3 368.4 337.1 312.9 306.1 358.4 327.5 326.7 Leucine 346.4 356.9 354.8 347.5 358.9 340.5 350.2 336.3 283.7 261.2 252.5 327.6 278.3 262.1 Phenylalanine 89.8 86.5 88.1 86.0 92.9 83.5 84.3 93.2 67.8 62.5 60.0 91.3 68.2 62.4 Tryptophan 8.9 7.5 7.9 7.1 9.0 7.2 7.4 8.0 1.6 0.3 0.0 8.0 1.6 0.0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Histidine Hydroxyproline 1-Methylhistidine 3-Methylhistidine Phosphoethanolamine Carnosine Anserine Ethanolamine Aspartic Acid Sarcosine beta-Alanine gamma-Amino-N-butyric-acid alpha-Aminoadipic-acid beta-Aminoisobutyric-acid Hydroxylysine 1 Hydroxylysine 2 alpha-Amino-N-butyric-acid Cystathionine 1 Cystathionine 2 Methionine allo-Isoleucine Homocystine Asparagine Arginine Taurine Serine Glutamine Glycine Citrulline Glutamic Acid Threonine Alanine Proline Ornithine Lysine Cystine Tyrosine Valine Isoleucine Leucine Phenylalanine Tryptophan METABOLITES_END #END