#METABOLOMICS WORKBENCH dmukha_20240309_152224 DATATRACK_ID:4697 STUDY_ID:ST003152 ANALYSIS_ID:AN005172 PROJECT_ID:PR001941 VERSION 1 CREATED_ON March 27, 2024, 1:21 pm #PROJECT PR:PROJECT_TITLE The shutdown of NADH oxidation via Respiratory Complex I mimics fatty acid PR:PROJECT_TITLE biosynthesis inhibition PR:PROJECT_TYPE LC-MS Quantitative Analysis PR:PROJECT_SUMMARY Proliferating cancer cells actively utilize anabolic processes for biomass PR:PROJECT_SUMMARY production, including de novo biosynthesis of amino acids, nucleotides, and PR:PROJECT_SUMMARY fatty acids. The key enzyme of the fatty acid biosynthesis pathway, fatty acid PR:PROJECT_SUMMARY synthase (FASN), is widely recognized as a promising therapeutic target in PR:PROJECT_SUMMARY cancer and other health conditions. Here, we establish a metabolic signature of PR:PROJECT_SUMMARY FASN inhibition using a panel of pharmacological inhibitors (GSK2194069, PR:PROJECT_SUMMARY TVB-2640, TVB-3166, C75, cerulenin, and Fasnall). We find that the activity of PR:PROJECT_SUMMARY some commonly used FASN inhibitors is inconsistent with the metabolic signature PR:PROJECT_SUMMARY of FASN inhibition (accumulation of malonate, succinate, malonyl coenzyme A, PR:PROJECT_SUMMARY succinyl coenzyme A, and other metabolic perturbations). Moreover, we show that PR:PROJECT_SUMMARY one of these putative FASN inhibitors, Fasnall, is a respiratory Complex I PR:PROJECT_SUMMARY inhibitor that mimics FASN inhibition through NADH accumulation and consequent PR:PROJECT_SUMMARY depletion of the tricarboxylic acid cycle metabolites. We demonstrate that PR:PROJECT_SUMMARY Fasnall impairs tumor growth in several oxidative phosphorylation-dependent PR:PROJECT_SUMMARY cancer models, including combination therapy-resistant melanoma patient-derived PR:PROJECT_SUMMARY xenografts. Fasnall administration does not reproduce neurological side effects PR:PROJECT_SUMMARY in mice reported for other Complex I inhibitors. Our results have significant PR:PROJECT_SUMMARY implications for understanding the FASN role in human health and disease and PR:PROJECT_SUMMARY provide evidence of therapeutic potential for Complex I inhibitors with fast PR:PROJECT_SUMMARY systemic clearance. Our findings also highlight the continuing need for PR:PROJECT_SUMMARY validation of small molecule inhibitors to distinguish high-quality chemical PR:PROJECT_SUMMARY probes and to expand the understanding of their application. PR:INSTITUTE Wistar Institute PR:DEPARTMENT Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer PR:DEPARTMENT Center PR:LABORATORY Schug's Lab PR:LAST_NAME Mukha PR:FIRST_NAME Dzmitry PR:ADDRESS 3601 Spruce St., Philadelphia, Pennsylvania 19104, USA PR:EMAIL dmukha@wistar.org PR:PHONE +12154956903 PR:FUNDING_SOURCE This work was supported by grants from the National Institutes of Health (NIH) PR:FUNDING_SOURCE National Cancer Institute (NCI) DP2 CA249950-01 (Z.T.S.), NIH NCI P01 CA114046 PR:FUNDING_SOURCE (Z.T.S.), Melanoma Research Foundation 717173 (Z.T.S.), and Susan G. Komen PR:FUNDING_SOURCE CCR19608782 (Z.T.S.). PR:PUBLICATIONS Submission Pending PR:CONTRIBUTORS Dzmitry Mukha, Jena Dessain, Seamus O’Connor, Katherine Pniewski, Fabrizio PR:CONTRIBUTORS Bertolazzi, Jeet Patel, Mary Mullins, Zachary T. Schug #STUDY ST:STUDY_TITLE Metabolomics analysis of murine xenograft tumors derived from human breast ST:STUDY_TITLE cancer cell line MCF7 1 h after isotopic glucose bolus ST:STUDY_TYPE Intracellular metabolomics, [U-13C6] glucose tracing ST:STUDY_SUMMARY Metabolomics analysis of murine xenograft tumors derived from human breast ST:STUDY_SUMMARY cancer cell line MCF7 1 h after isotopic glucose bolus. The NSG female mice were ST:STUDY_SUMMARY ~55 days after grafting tumors. [U-13C6] D-Glucose bolus was delivered ST:STUDY_SUMMARY intraperitoneally with a 10 mg/kg dose of Fasnall. ST:INSTITUTE Wistar Institute ST:DEPARTMENT Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer ST:DEPARTMENT Center ST:LABORATORY Schug's Lab ST:LAST_NAME Mukha ST:FIRST_NAME Dzmitry ST:ADDRESS 3601 Spruce St, Philadelphia, PA 19104, USA ST:EMAIL dmukha@wistar.org ST:PHONE 2154956903 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 12 ST:NUM_FEMALES 12 ST:PUBLICATIONS Submission Pending #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN NOD Scid Gamma (NSG) SU:AGE_OR_AGE_RANGE ~4.5 months SU:WEIGHT_OR_WEIGHT_RANGE 24.3 +/- 1.9 g (mean +/- SD) SU:GENDER Female SU:ANIMAL_HOUSING Wistar Animal Facility SU:SPECIES_GROUP Rodents (order Rodentia) #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 01_Fasnall_MCF7_Tum_Gluc_1h_F_1066 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=01_Fasnall_MCF7_Tum_Gluc_1h_F_1066.raw; RAW_FILE_NAME(Raw File Name)=NEG_01_Fasnall_MCF7_Tum_Gluc_1h_F_1066.mzXML; RAW_FILE_NAME(Raw File Name )=POS_01_Fasnall_MCF7_Tum_Gluc_1h_F_1066.mzXML SUBJECT_SAMPLE_FACTORS - 02_Fasnall_MCF7_Tum_Gluc_1h_F_1068 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=02_Fasnall_MCF7_Tum_Gluc_1h_F_1068.raw; RAW_FILE_NAME(Raw File Name)=NEG_02_Fasnall_MCF7_Tum_Gluc_1h_F_1068.mzXML; RAW_FILE_NAME(Raw File Name )=POS_02_Fasnall_MCF7_Tum_Gluc_1h_F_1068.mzXML SUBJECT_SAMPLE_FACTORS - 03_Fasnall_MCF7_Tum_Gluc_1h_F_1069 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=03_Fasnall_MCF7_Tum_Gluc_1h_F_1069.raw; RAW_FILE_NAME(Raw File Name)=NEG_03_Fasnall_MCF7_Tum_Gluc_1h_F_1069.mzXML; RAW_FILE_NAME(Raw File Name )=POS_03_Fasnall_MCF7_Tum_Gluc_1h_F_1069.mzXML SUBJECT_SAMPLE_FACTORS - 04_Fasnall_MCF7_Tum_Gluc_1h_F_1070 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=04_Fasnall_MCF7_Tum_Gluc_1h_F_1070.raw; RAW_FILE_NAME(Raw File Name)=NEG_04_Fasnall_MCF7_Tum_Gluc_1h_F_1070.mzXML; RAW_FILE_NAME(Raw File Name )=POS_04_Fasnall_MCF7_Tum_Gluc_1h_F_1070.mzXML SUBJECT_SAMPLE_FACTORS - 05_Fasnall_MCF7_Tum_Gluc_1h_F_1071 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=05_Fasnall_MCF7_Tum_Gluc_1h_F_1071.raw; RAW_FILE_NAME(Raw File Name)=NEG_05_Fasnall_MCF7_Tum_Gluc_1h_F_1071.mzXML; RAW_FILE_NAME(Raw File Name )=POS_05_Fasnall_MCF7_Tum_Gluc_1h_F_1071.mzXML SUBJECT_SAMPLE_FACTORS - 06_Fasnall_MCF7_Tum_Gluc_1h_F_1072 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=06_Fasnall_MCF7_Tum_Gluc_1h_F_1072.raw; RAW_FILE_NAME(Raw File Name)=NEG_06_Fasnall_MCF7_Tum_Gluc_1h_F_1072.mzXML; RAW_FILE_NAME(Raw File Name )=POS_06_Fasnall_MCF7_Tum_Gluc_1h_F_1072.mzXML SUBJECT_SAMPLE_FACTORS - 07_Fasnall_MCF7_Tum_Gluc_1h_F_1073 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall RAW_FILE_NAME(Raw File Name)=07_Fasnall_MCF7_Tum_Gluc_1h_F_1073.raw; RAW_FILE_NAME(Raw File Name)=NEG_07_Fasnall_MCF7_Tum_Gluc_1h_F_1073.mzXML; RAW_FILE_NAME(Raw File Name )=POS_07_Fasnall_MCF7_Tum_Gluc_1h_F_1073.mzXML SUBJECT_SAMPLE_FACTORS - 08_Fasnall_MCF7_Tum_Gluc_1h_D_1074 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=08_Fasnall_MCF7_Tum_Gluc_1h_D_1074.raw; RAW_FILE_NAME(Raw File Name)=NEG_08_Fasnall_MCF7_Tum_Gluc_1h_D_1074.mzXML; RAW_FILE_NAME(Raw File Name )=POS_08_Fasnall_MCF7_Tum_Gluc_1h_D_1074.mzXML SUBJECT_SAMPLE_FACTORS - 09_Fasnall_MCF7_Tum_Gluc_1h_D_1075 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=09_Fasnall_MCF7_Tum_Gluc_1h_D_1075.raw; RAW_FILE_NAME(Raw File Name)=NEG_09_Fasnall_MCF7_Tum_Gluc_1h_D_1075.mzXML; RAW_FILE_NAME(Raw File Name )=POS_09_Fasnall_MCF7_Tum_Gluc_1h_D_1075.mzXML SUBJECT_SAMPLE_FACTORS - 10_Fasnall_MCF7_Tum_Gluc_1h_D_1077 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=10_Fasnall_MCF7_Tum_Gluc_1h_D_1077.raw; RAW_FILE_NAME(Raw File Name)=NEG_10_Fasnall_MCF7_Tum_Gluc_1h_D_1077.mzXML; RAW_FILE_NAME(Raw File Name )=POS_10_Fasnall_MCF7_Tum_Gluc_1h_D_1077.mzXML SUBJECT_SAMPLE_FACTORS - 11_Fasnall_MCF7_Tum_Gluc_1h_D_1079 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=11_Fasnall_MCF7_Tum_Gluc_1h_D_1079.raw; RAW_FILE_NAME(Raw File Name)=NEG_11_Fasnall_MCF7_Tum_Gluc_1h_D_1079.mzXML; RAW_FILE_NAME(Raw File Name )=POS_11_Fasnall_MCF7_Tum_Gluc_1h_D_1079.mzXML SUBJECT_SAMPLE_FACTORS - 12_Fasnall_MCF7_Tum_Gluc_1h_D_1080 Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=12_Fasnall_MCF7_Tum_Gluc_1h_D_1080.raw; RAW_FILE_NAME(Raw File Name)=NEG_12_Fasnall_MCF7_Tum_Gluc_1h_D_1080.mzXML; RAW_FILE_NAME(Raw File Name )=POS_12_Fasnall_MCF7_Tum_Gluc_1h_D_1080.mzXML SUBJECT_SAMPLE_FACTORS - 135standard_091823_1 Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=135standard_091823_1.raw; RAW_FILE_NAME(Raw File Name)=NEG_135standard_091823_1.mzXML; RAW_FILE_NAME(Raw File Name )=POS_135standard_091823_1.mzXML SUBJECT_SAMPLE_FACTORS - 135standard_091823_2 Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=135standard_091823_2.raw; RAW_FILE_NAME(Raw File Name)=NEG_135standard_091823_2.mzXML; RAW_FILE_NAME(Raw File Name )=POS_135standard_091823_2.mzXML SUBJECT_SAMPLE_FACTORS - 80MeOH_Blank Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=80MeOH_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_80MeOH_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_80MeOH_Blank.mzXML SUBJECT_SAMPLE_FACTORS - 80MeOH_Blank_20230918145946 Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=80MeOH_Blank_20230918145946.raw; RAW_FILE_NAME(Raw File Name)=NEG_80MeOH_Blank_20230918145946.mzXML; RAW_FILE_NAME(Raw File Name )=POS_80MeOH_Blank_20230918145946.mzXML SUBJECT_SAMPLE_FACTORS - 80MeOH_Blank_20230918155328 Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=80MeOH_Blank_20230918155328.raw; RAW_FILE_NAME(Raw File Name)=NEG_80MeOH_Blank_20230918155328.mzXML; RAW_FILE_NAME(Raw File Name )=POS_80MeOH_Blank_20230918155328.mzXML SUBJECT_SAMPLE_FACTORS - 80MeOH_Blank_20230918220912 Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=80MeOH_Blank_20230918220912.raw; RAW_FILE_NAME(Raw File Name)=NEG_80MeOH_Blank_20230918220912.mzXML; RAW_FILE_NAME(Raw File Name )=POS_80MeOH_Blank_20230918220912.mzXML SUBJECT_SAMPLE_FACTORS - 80MeOH_Blank_20230918223604 Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=80MeOH_Blank_20230918223604.raw; RAW_FILE_NAME(Raw File Name)=NEG_80MeOH_Blank_20230918223604.mzXML; RAW_FILE_NAME(Raw File Name )=POS_80MeOH_Blank_20230918223604.mzXML #COLLECTION CO:COLLECTION_SUMMARY Tissue metabolite extracts were prepared from ~50 mg snap-frozen tissue samples. CO:COLLECTION_SUMMARY Each sample was weighed while frozen. Frozen samples were ground at the CO:COLLECTION_SUMMARY temperature of liquid nitrogen by Retch Cryomill. Then, 80% methanol was added CO:COLLECTION_SUMMARY to each tube at a ratio of 1 ml per 50 mg of tissue. CO:COLLECTION_PROTOCOL_FILENAME DM_metabolomics_samples.txt CO:SAMPLE_TYPE Tumor cells CO:COLLECTION_METHOD 80% methanol extraction CO:VOLUMEORAMOUNT_COLLECTED ~1 ml CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS 1.5 ml plastic centrifuge tubes CO:STORAGE_VIALS 1.5 ml plastic centrifuge tubes #TREATMENT TR:TREATMENT_SUMMARY Animals received 1 g/kg [U-13C6] D-glucose bolus intraperitoneally with a 10 TR:TREATMENT_SUMMARY mg/kg dose of Fasnall. TR:TREATMENT_COMPOUND Fasnall TR:TREATMENT_ROUTE Intraperitoneal injection TR:TREATMENT_DOSE 10 mg/kg TR:TREATMENT_DOSEVOLUME 50 ul TR:TREATMENT_DOSEDURATION 1 h TR:TREATMENT_VEHICLE 1:1 DMSO/PBS TR:ANIMAL_ANESTHESIA None TR:ANIMAL_FASTING None TR:ANIMAL_ENDP_EUTHANASIA CO2 TR:ANIMAL_ENDP_TISSUE_COLL_LIST 50 mg tumor samples #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Tissue metabolite extracts were prepared from ~50 mg snap-frozen tissue samples. SP:SAMPLEPREP_SUMMARY Each sample was weighed while frozen. Frozen samples were ground at the SP:SAMPLEPREP_SUMMARY temperature of liquid nitrogen by Retch Cryomill. Then, 80% methanol was added SP:SAMPLEPREP_SUMMARY to each tube at a ratio of 1 ml per 50 mg of tissue. All samples were SP:SAMPLEPREP_SUMMARY centrifuged at 18,000 g and 4 °C for 20 min. The supernatant was then SP:SAMPLEPREP_SUMMARY transferred to new Eppendorf tubes and centrifuged again with the same SP:SAMPLEPREP_SUMMARY parameters. After centrifugation, the extracts were transferred to glass LC-MS SP:SAMPLEPREP_SUMMARY vials. SP:SAMPLEPREP_PROTOCOL_FILENAME DM_metabolomics_samples.txt SP:EXTRACTION_METHOD 80% methanol SP:EXTRACT_ENRICHMENT None SP:EXTRACT_CLEANUP None SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION None SP:SAMPLE_DERIVATIZATION None SP:SAMPLE_SPIKING None SP:SUBCELLULAR_LOCATION Intracellular metabolites #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 RS CH:COLUMN_NAME Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) CH:SOLVENT_A Water with 0.01% ammonium hydroxide and 20 mM ammonium bicarbonate CH:SOLVENT_B Acetonitrile CH:FLOW_GRADIENT A linear solvent gradient of a total duration 22.5 min was starting with 0.2 CH:FLOW_GRADIENT ml/min flow rate of 80% solvent B, 12.5 min – 30%, 15 min – 30%, 15.2 min CH:FLOW_GRADIENT – 80%, 20 min – 80%, 21 min – flow rate 0.3 ml/min, 22 min – flow rate CH:FLOW_GRADIENT 0.3 ml/min, 22.1 min – flow rate 0.2 ml/min. CH:FLOW_RATE 0.2 ml/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Wistar Institute Proteomics & Metabolomic Core AN:OPERATOR_NAME Nicole Gorman AN:DETECTOR_TYPE Orbitrap #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The HESI ion source voltage was set to the following parameters: 3,600 V in both MS:MS_COMMENTS polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 MS:MS_COMMENTS °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer MS:MS_COMMENTS was set to acquire data in the polarity-switching mode averaging two microscans MS:MS_COMMENTS with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range MS:MS_COMMENTS 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were MS:MS_COMMENTS converted into the mzXML format by ProteoWizard and analyzed in MAVEN. MS:ION_SOURCE_TEMPERATURE 200 MS:ION_SPRAY_VOLTAGE 3600 MS:IONIZATION Both MS:SOURCE_TEMPERATURE 200 MS:AUTOMATIC_GAIN_CONTROL 5e6 MS:DATAFORMAT RAW #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS AU MS_METABOLITE_DATA_START Samples 01_Fasnall_MCF7_Tum_Gluc_1h_F_1066 02_Fasnall_MCF7_Tum_Gluc_1h_F_1068 03_Fasnall_MCF7_Tum_Gluc_1h_F_1069 04_Fasnall_MCF7_Tum_Gluc_1h_F_1070 05_Fasnall_MCF7_Tum_Gluc_1h_F_1071 06_Fasnall_MCF7_Tum_Gluc_1h_F_1072 07_Fasnall_MCF7_Tum_Gluc_1h_F_1073 08_Fasnall_MCF7_Tum_Gluc_1h_D_1074 09_Fasnall_MCF7_Tum_Gluc_1h_D_1075 10_Fasnall_MCF7_Tum_Gluc_1h_D_1077 11_Fasnall_MCF7_Tum_Gluc_1h_D_1079 12_Fasnall_MCF7_Tum_Gluc_1h_D_1080 135standard_091823_1 135standard_091823_2 80MeOH_Blank 80MeOH_Blank_20230918145946 80MeOH_Blank_20230918155328 80MeOH_Blank_20230918220912 80MeOH_Blank_20230918223604 Factors Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:10 mg/kg Fasnall Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle Sample source:Xenograft tumors | Isotopic Tracer:1 g/kg [U-13C6] D-glucose in PBS | Drug Treatment:Vehicle Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Sample source:NA | Isotopic Tracer:NA | Drug Treatment:NA Acetyl-CoA C12 PARENT 209042.9 61946.19 93089.56 50019.67 177328.5 111109.8 203671.8 158513.2 237993.5 247305.2 122608.7 175351.4 0 0 0 0 0 2130231 2294503 Acetyl-CoA C13-label-1 68854.13 21979.43 32683.68 10244.43 62698.05 40916.6 63474.5 46759.3 61720.67 85098.74 33801.22 53502.52 0 0 0 0 0 635557.5 674353.7 Acetyl-CoA C13-label-2 29408.75 14642.22 23119.23 13081.69 26270.96 22267.64 50559.89 24184.75 43349.52 51334.3 9803.318 10208.91 0 0 0 0 0 18601.42 79896.95 Acetyl-CoA C13-label-3 3634.4 0 2967.131 0 3570.365 3440.157 8869.429 3317.896 3329.163 11442.55 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2098.417 0 0 0 0 Acetyl-CoA C13-label-7 0 0 0 0 0 0 0 4290.523 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-8 0 0 0 0 0 0 0 0 2795.058 2778.127 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-11 3417.838 0 0 0 0 0 3248.349 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-12 0 0 0 0 0 0 0 0 0 4781.052 3220.252 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-13 0 0 0 0 0 0 0 0 0 0 0 4487.317 0 0 0 0 0 0 0 Acetyl-CoA C13-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-18 0 0 3468.439 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-22 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-23 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Dimethyl sulfoxide (DMSO) C12 PARENT 1.42E+08 1.31E+08 1.44E+08 1.28E+08 1.61E+08 1.40E+08 1.51E+08 1.16E+08 1.76E+08 1.24E+08 1.61E+08 1.49E+08 94918.38 146967.9 110036.4 133384.7 122833.8 152391.4 121633.5 Dimethyl sulfoxide (DMSO) C13-label-1 3686443 3177516 3747818 3269921 4134047 3892390 3934929 2905766 4644828 3069655 4020344 3732068 0 0 0 0 0 0 0 Dimethyl sulfoxide (DMSO) C13-label-2 0 9947.674 10064.14 9759.642 10878.85 7025.199 12381.35 0 11654.64 0 0 0 0 0 0 0 0 0 0 Fasnall C12 PARENT 2.27E+07 9893311 1.85E+07 7087194 1.80E+07 1.18E+07 1.04E+07 456680.2 0 0 23454.69 0 47310.83 0 0 0 0 0 0 Fasnall C13-label-1 4296348 2072216 3704906 1354357 3568540 2203533 2023841 0 0 0 0 0 119345.8 0 0 0 0 0 0 Fasnall C13-label-2 357198.2 147113.3 324176 120647.8 283934.2 245433.7 118232 0 0 0 0 0 92950.48 0 0 0 0 0 0 Fasnall C13-label-3 0 0 36736.52 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-10 0 0 0 0 0 0 0 24220.54 0 0 0 0 0 52574.84 66572.48 0 0 0 0 Fasnall C13-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 21938.22 0 0 Fasnall C13-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7002.311 0 0 4901.964 7755.701 Fasnall C13-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C12 PARENT 1.25E+07 4763742 5944884 8672385 2.66E+07 1.49E+07 2.20E+07 1.39E+07 1.30E+07 2.52E+07 1.12E+07 2.21E+07 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-1 3504989 1127209 1460808 1872187 6761810 3366473 5394682 3450721 3114889 6376062 2570554 5368820 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-2 2435125 525670.2 871947.6 1134223 5843286 1515285 4495981 2790095 1711713 5502271 1513459 3463094 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-3 671174.4 156168.6 272681.1 331306.1 1760640 394362.7 1473670 841720.1 451029.6 1473794 413530.2 948905.6 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-4 830219 227073.7 379511.8 497267.3 1662877 520223.5 1713034 1045252 718153 2039276 522718.9 1069434 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-5 226426 65760.21 122786.2 158630.4 462617.5 146837.8 510416.7 279450.2 210769 516429.2 160626.4 335962.6 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-6 236518.5 69226.3 104469.1 198586.1 345260 160106.9 482111.3 191820.8 207035.7 507000.4 130881.2 232642.6 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-7 20420.48 14317.11 20121.94 38896.15 72709.81 0 126045.9 27245.61 20230.6 115619.5 20423.55 26827.65 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-8 28512.33 0 22623.91 18563.35 36621.67 24393.52 56028.26 28266.76 20650.62 62519 27490.33 22572.2 0 0 7710.009 0 0 0 0 Myristoyl-L-carnitine C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-15 0 0 0 17689.21 0 0 0 0 0 0 0 0 0 4484.366 4603.932 0 7627.178 0 0 Myristoyl-L-carnitine C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention time quantified m/z parent pubchem id kegg id regno comments Acetyl-CoA C12 PARENT 8.079143 810.1327 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-1 8.083757 811.1354 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-2 8.098002 812.1372 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-3 8.099983 813.1405 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-4 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-5 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-6 8.145617 816.1545 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-7 8.019383 817.1552 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-8 8.212826 818.1617 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-9 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-10 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-11 8.2184 821.1707 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-12 8.089258 822.1719 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-13 8.147017 823.176 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-14 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-15 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-16 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-17 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-18 8.005217 828.1924 810.1328 444493 C00024 50043 Acetyl-CoA C13-label-19 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-20 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-21 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-22 NA NA 810.1328 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-23 NA NA 810.1328 444493 C00024 50043 Peak was not detected Dimethyl sulfoxide (DMSO) C12 PARENT 2.540911 79.02119 79.0212 679 C11143 38047 Dimethyl sulfoxide (DMSO) C13-label-1 2.447631 80.02449 79.0212 679 C11143 38047 Dimethyl sulfoxide (DMSO) C13-label-2 2.471638 81.02775 79.0212 679 C11143 38047 Fasnall C12 PARENT 1.579835 339.1633 339.1633 16230362 NA NA Fasnall C13-label-1 1.577026 340.1667 339.1633 16230362 NA NA Fasnall C13-label-2 1.574856 341.17 339.1633 16230362 NA NA Fasnall C13-label-3 1.62751 342.1742 339.1633 16230362 NA NA Fasnall C13-label-4 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-5 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-6 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-7 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-8 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-9 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-10 1.616105 349.1982 339.1633 16230362 NA NA Fasnall C13-label-11 1.796783 350.2018 339.1633 16230362 NA NA Fasnall C13-label-12 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-13 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-14 1.644285 353.2097 339.1633 16230362 NA NA Fasnall C13-label-15 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-16 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-17 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-18 NA NA 339.1633 16230362 NA NA Peak was not detected Fasnall C13-label-19 NA NA 339.1633 16230362 NA NA Peak was not detected Myristoyl-L-carnitine C12 PARENT 1.741611 372.3103 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-1 1.73831 373.3138 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-2 1.738097 374.3169 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-3 1.746872 375.3205 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-4 1.739996 376.3236 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-5 1.753668 377.3272 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-6 1.725926 378.3304 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-7 1.751529 379.3342 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-8 1.730362 380.3371 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-9 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-10 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-11 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-12 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-13 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-14 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-15 1.774794 387.3603 372.3103 53477791 NA 38728 Myristoyl-L-carnitine C13-label-16 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-17 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-18 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-19 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-20 NA NA 372.3103 53477791 NA 38728 Peak was not detected Myristoyl-L-carnitine C13-label-21 NA NA 372.3103 53477791 NA 38728 Peak was not detected METABOLITES_END #END