#METABOLOMICS WORKBENCH ptth222_20180124_070246_mwtab.txt DATATRACK_ID:1304 STUDY_ID:ST000927 ANALYSIS_ID:AN001520 PROJECT_ID:PR000642 VERSION 1 CREATED_ON January 24, 2018, 6:41 pm #PROJECT PR:PROJECT_TITLE Breast Cancer Metabolism PR:PROJECT_SUMMARY The program comprises three project areas utilizing stable isotope resolved PR:PROJECT_SUMMARY metabolomics to gain a mechanistic understanding of NSCLC in situ. The projects PR:PROJECT_SUMMARY combine cell culture, animal models and human subjects to define the influence PR:PROJECT_SUMMARY of the tumor microenvironment on cancer progression. PR:INSTITUTE University of Kentucky PR:LAST_NAME Lane PR:FIRST_NAME Andrew PR:ADDRESS Rm 516 Biopharm Complex, 789 S. Limestone St.,Univ. of Kentucky, Lexington, KY PR:ADDRESS 40536 PR:EMAIL andrewlane@gmail.com PR:PHONE 8592182868 #STUDY ST:STUDY_TITLE Probing the metabolic phenotype of breast cancer cells by multiple tracer stable ST:STUDY_TITLE isotope resolved metabolomics (part II) ST:STUDY_SUMMARY Breast cancers vary by their origin and specific set of genetic lesions, which ST:STUDY_SUMMARY gives rise to distinct phenotypes and differential response to targeted and ST:STUDY_SUMMARY untargeted chemotherapies. To explore the functional differences of different ST:STUDY_SUMMARY breast cell types, we performed Stable Isotope Resolved Metabolomics (SIRM) ST:STUDY_SUMMARY studies of one primary breast (HMEC) and three breast cancer cells (MCF-7, ST:STUDY_SUMMARY MDAMB-231, and ZR75-1) having distinct genotypes and growth characteristics, ST:STUDY_SUMMARY using 13C6-glucose, 13C-1+2-glucose, 13C5,15N2-Gln, 13C3-glycerol, and ST:STUDY_SUMMARY 13C8-octanoate as tracers. These tracers were designed to probe the central ST:STUDY_SUMMARY energy producing and anabolic pathways (glycolysis, pentose phosphate pathway, ST:STUDY_SUMMARY Krebs Cycle, glutaminolysis, nucleotide synthesis and lipid turnover). We found ST:STUDY_SUMMARY that glycolysis was not associated with the rate of breast cancer cell ST:STUDY_SUMMARY proliferation, glutaminolysis did not support lipid synthesis in primary breast ST:STUDY_SUMMARY or breast cancer cells, but was a major contributor to pyrimidine ring synthesis ST:STUDY_SUMMARY in all cell types; anaplerotic pyruvate carboxylation was activated in breast ST:STUDY_SUMMARY cancer versus primary cells. We also found that glucose metabolism in individual ST:STUDY_SUMMARY breast cancer cell lines differed between in vitro cultures and tumor ST:STUDY_SUMMARY xenografts, but not the metabolic distinctions between cell lines, which may ST:STUDY_SUMMARY reflect the influence of tumor architecture/microenvironment. ST:INSTITUTE University of Kentucky ST:LAST_NAME Lane ST:FIRST_NAME Andrew ST:ADDRESS Rm 516 Biopharm Complex, 789 S. Limestone St.,Univ. of Kentucky, Lexington, KY ST:ADDRESS 40536 ST:EMAIL andrewlane@gmail.com ST:PHONE 8592182868 #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 - HMEC_UC13Glc_Ctl_toc50 Cell Line:HMEC | Tracer:Uniformly Labelled Glucose Tracer_Abbreviated=U-C13-Glc; Cancer=Not Cancer SUBJECT_SAMPLE_FACTORS - MCF7_UC13Glc_Ctl_toc50 Cell Line:MCF-7 | Tracer:Uniformly Labelled Glucose Tracer_Abbreviated=U-C13-Glc; Cancer=Cancer SUBJECT_SAMPLE_FACTORS - ZR75_1_cells_24h_c13glcSep09_toc50 Cell Line:ZR-75-1 | Tracer:Uniformly Labelled Glucose Tracer_Abbreviated=U-C13-Glc; Cancer=Cancer SUBJECT_SAMPLE_FACTORS - HMEC_C13Gln_Ctl_toc50 Cell Line:HMEC | Tracer:Uniformly Labelled Glutamine Tracer_Abbreviated=U-C13_N15-Gln; Cancer=Not Cancer SUBJECT_SAMPLE_FACTORS - MCF7_C13Gln_Ctl_toc50 Cell Line:MCF-7 | Tracer:Uniformly Labelled Glutamine Tracer_Abbreviated=U-C13_N15-Gln; Cancer=Cancer SUBJECT_SAMPLE_FACTORS - ZR751_Nor_HiGlc_U13C15NGln_TOCSY Cell Line:ZR-75-1 | Tracer:Uniformly Labelled Glutamine Tracer_Abbreviated=U-C13_N15-Gln; Cancer=Cancer #COLLECTION CO:COLLECTION_SUMMARY Cells were cultured in various media then harvested after 24 hours. CO:SAMPLE_TYPE Cells #TREATMENT TR:TREATMENT_SUMMARY None #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cell lines are seperated into polar, non-polar, protein, and media. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME INOVA NM:INSTRUMENT_TYPE CW-NMR NM:NMR_EXPERIMENT_TYPE Other NM:SPECTROMETER_FREQUENCY 800 MHz #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS area NMR_METABOLITE_DATA_START Samples HMEC_C13Gln_Ctl_toc50 HMEC_UC13Glc_Ctl_toc50 MCF7_C13Gln_Ctl_toc50 MCF7_UC13Glc_Ctl_toc50 ZR75_1_cells_24h_c13glcSep09_toc50 ZR751_Nor_HiGlc_U13C15NGln_TOCSY Factors Cell Line:HMEC | Tracer:Uniformly Labelled Glutamine Cell Line:HMEC | Tracer:Uniformly Labelled Glucose Cell Line:MCF-7 | Tracer:Uniformly Labelled Glutamine Cell Line:MCF-7 | Tracer:Uniformly Labelled Glucose Cell Line:ZR-75-1 | Tracer:Uniformly Labelled Glucose Cell Line:ZR-75-1 | Tracer:Uniformly Labelled Glutamine UXP-5 13C-A 50.74038416 57.01434766 315.6179647 415.98389 318.7578252 157.6554549 UXP-5 13C-B 86.42896764 157.3195258 340.7416454 404.7737169 225.5226737 425.8632207 UXP-5,6 13C-A 500.507461 103.9207387 922.8158171 627.2578756 1415.874749 622.1238667 UXP-5,6 13C-B 538.2189425 89.07780904 867.864989 447.1612448 1107.10075 1303.257013 UXP-6 456.3852955 1082.995872 2721.184463 2194.277233 3932.602853 617.0999188 UXP-6 13C-A 82.42272487 80.22959021 402.6092283 690.9317071 347.3962358 267.7202832 UXP-6 13C-B 238.1921457 111.4472882 435.3887976 508.4034492 332.1889378 150.29 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Common Name Hydrogen Number Isotope PPM Range (1) PPM Range (2) UXP-5 13C-A Uridine Phosphates 5 13C 7.93186439797907 - 7.967447801 5.79290009505425 - 5.834791062 UXP-5 13C-B Uridine Phosphates 5 13C 7.93610290815661 - 7.972723056 6.08994864749875 - 6.10982575 UXP-5,6 13C-A Uridine Phosphates 5 13C 7.77607998762907 - 8.106954655 5.78768499084674 - 6.110864608 UXP-5,6 13C-B Uridine Phosphates 5 13C 7.77890697522207 - 8.111280041 5.80200553239918 - 6.115338917 UXP-6 Uridine Phosphates 6 13C 7.93377342544742 - 7.983361653 5.942904357 - 5.96340861810778 UXP-6 13C-A Uridine Phosphates 6 13C 7.77851358447656 - 8.101883111 5.94440660086254 - 6.106839021 UXP-6 13C-B Uridine Phosphates 6 13C 8.08405619244877 - 8.111280042 5.94501676112817 - 5.967740509 METABOLITES_END #END