#METABOLOMICS WORKBENCH ptth222_20180124_071451_mwtab.txt DATATRACK_ID:1305 STUDY_ID:ST000928 ANALYSIS_ID:AN001521 PROJECT_ID:PR000642 VERSION 1 CREATED_ON January 24, 2018, 6:44 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 III) 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_U13CGlc_CTL_med24h Cell Line:HMEC Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Not Cancer; Extraction Time (hours)=24 SUBJECT_SAMPLE_FACTORS - C13-U_Ctr_1_t24_PRESAT01 Cell Line:None Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Not Cancer; Extraction Time (hours)=24 SUBJECT_SAMPLE_FACTORS - ZR751_Media_Ctl_24h_13_PRESAT01 Cell Line:ZR-75-1 Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Cancer; Extraction Time (hours)=24 SUBJECT_SAMPLE_FACTORS - MDAMB231_13C_U_Ctr_24h_PRESAT01 Cell Line:MDA-MB-231 Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Cancer; Extraction Time (hours)=24 SUBJECT_SAMPLE_FACTORS - HMEC_U13CGlc_MSA_med0h Cell Line:HMEC Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Not Cancer; Extraction Time (hours)=- SUBJECT_SAMPLE_FACTORS - C13-U_Ctr_1_t0_PRESAT01 Cell Line:None Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Not Cancer; Extraction Time (hours)=- SUBJECT_SAMPLE_FACTORS - ZR751_Media_Ctl_0h_9_PRESAT01 Cell Line:ZR-75-1 Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Cancer; Extraction Time (hours)=- SUBJECT_SAMPLE_FACTORS - MDAMB231_13C_U_Ctr_0h_PRESAT01 Cell Line:MDA-MB-231 Tracer=Uniformly Labelled Glucose; Tracer_Abbreviated=U-C13-Glc; Cancer=Cancer; Extraction Time (hours)=- #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 C13-U_Ctr_1_t0_PRESAT01 C13-U_Ctr_1_t24_PRESAT01 HMEC_U13CGlc_CTL_med24h HMEC_U13CGlc_MSA_med0h MDAMB231_13C_U_Ctr_0h_PRESAT01 MDAMB231_13C_U_Ctr_24h_PRESAT01 ZR751_Media_Ctl_0h_9_PRESAT01 ZR751_Media_Ctl_24h_13_PRESAT01 Factors Cell Line:None Cell Line:None Cell Line:HMEC Cell Line:HMEC Cell Line:MDA-MB-231 Cell Line:MDA-MB-231 Cell Line:ZR-75-1 Cell Line:ZR-75-1 aGlc 13C-A 17378.15303 14818.53582 18242.41806 24938.86377 5379.08966 4759.404956 1664.175292 aGlc 13C-B 18700.17112 15550.41949 184.21 19230.7089 26473.46382 6174.153267 5613.86219 1838.700809 DSS 18222.56847 19242.60919 16906.48 16710.92822 20038.83914 18490.75061 17212.42347 14520.63211 Lac-3 15538.79 13082.62006 23003.05 3666.58 6557.39 12130.23624 1389.06 3252.019162 Lac13C-A 3476.901273 19364.88174 117000.26 11994.30115 4589.062897 99978.69827 17673.16071 Lac13C-B 4063.751829 21934.84855 124753.09 13277.52384 3859.167198 98854.99649 444.0870333 17475.85354 Thr-4 20445.52933 22131.75 22237.98 25666.94206 27699.81521 20342.54 2219.526891 1483.28 Val-Me 19992.65589 20590.35333 16364.09 20806.33091 23690.30993 16792.25673 2327.347805 1432.670026 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 aGlc 13C-A alpha-glucose 1 13C 5.0601840406437 - 5.089217417 aGlc 13C-B alpha-glucose 1 13C 5.34227566658487 - 5.370053034 DSS 2,2-Dimethyl-2-silapentane-5-sulfonate 2 12C -0.005872257 Lac-3 lactate 3 12C 1.4097129589625 - 1.422250453 Lac13C-A lactate 3 13C 1.29146759828232 - 1.323241202 Lac13C-B lactate 3 13C 1.50692207285929 - 1.538418541 Thr-4 threonine 4 12C 1.3442860987363 - 1.362396707 Val-Me valine 5 12C 1.06006730095028 - 1.080016776 METABOLITES_END #END