#METABOLOMICS WORKBENCH ptth222_20180122_065609 DATATRACK_ID:1303 STUDY_ID:ST000926 ANALYSIS_ID:AN001519 PROJECT_ID:PR000642 VERSION 1 CREATED_ON January 25, 2018, 10:23 am #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 I) 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_13C1_2Glc_Ctl_toc50 Cell Line:HMEC Tracer_Abbreviated=1,2-C13-Glc; Tracer=C-1 and C-2 Labelled Glucose; Cancer=Not Cancer SUBJECT_SAMPLE_FACTORS - MDAMB231C12ctl_toc Cell Line:MDA-MB-231 Tracer_Abbreviated=1,2-C13-Glc; Tracer=C-1 and C-2 Labelled Glucose; Cancer=Cancer SUBJECT_SAMPLE_FACTORS - MCF7_13C1_2Glc_MSA_toc50 Cell Line:MCF-7 Tracer_Abbreviated=1,2-C13-Glc; Tracer=C-1 and C-2 Labelled Glucose; 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_13C1_2Glc_Ctl_toc50 MCF7_13C1_2Glc_MSA_toc50 MDAMB231C12ctl_toc Factors Cell Line:HMEC Cell Line:MCF-7 Cell Line:MDA-MB-231 AXP-1' 884.3108564 1943.991574 1055.396932 AXP-1' 13C-A 211.1878055 518.9944154 378.4871942 AXP-1' 13C-B 207.6454335 557.2099936 340.4890171 AXP-2' 13C-A 405.3317205 386.0306703 297.9435694 AXP-2' 13C-B 261.1339004 342.8770157 262.1082613 UXP-1' 1395.836977 3326.978039 2110.302819 UXP-1' 13C-A 273.391216 1261.528519 960.1687215 UXP-1' 13C-B 223.1022592 1273.602447 893.9571077 UXP-2' 13C-A 377.1720631 507.1312266 357.0022393 UXP-2' 13C-B 351.1434548 559.3359393 403.0921132 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) AXP-1' Adenosine Phosphates 1 12C 4.79170253811575 - 4.798012381 6.19048454074072 - 6.206438861 AXP-1' 13C-A Adenosine Phosphates 1 13C 4.79243083416649 - 4.799781414 6.04710978333285 - 6.065293797 AXP-1' 13C-B Adenosine Phosphates 1 13C 4.78860889596561 - 4.796593304 6.32930483403642 - 6.34593885 AXP-2' 13C-A Adenosine Phosphates 2 13C 4.66496300459883 - 4.683456738 6.19311305682752 - 6.208786156 AXP-2' 13C-B Adenosine Phosphates 2 13C 4.91438746129748 - 4.936932334 6.18735561411309 - 6.203440261 UXP-1' Uridine Phosphates 1 12C 4.33496306906378 - 4.363180841 5.95309996705757 - 5.982801171 UXP-1' 13C-A Uridine Phosphates 1 13C 4.3357775558663 - 4.365019251 5.80865161546149 - 5.840008113 UXP-1' 13C-B Uridine Phosphates 1 13C 4.33220162843475 - 4.366091787 6.09292618319445 - 6.123122931 UXP-2' 13C-A Uridine Phosphates 2 13C 4.20672169741188 - 4.255310679 5.95295381524286 - 5.985094849 UXP-2' 13C-B Uridine Phosphates 2 13C 4.45427821629896 - 4.486756248 5.94769401298603 - 5.979512045 METABOLITES_END #END