#METABOLOMICS WORKBENCH araskind_20150413_9550621_mwtab.txt DATATRACK_ID:233 STUDY_ID:ST000170 ANALYSIS_ID:AN000447 PROJECT_ID:PR000223 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Leukemia stem cells studies_dup PR:PROJECT_TYPE Glycolysis/TCA/Nucleotide analysis (tissue/cells) PR:PROJECT_SUMMARY Define the function of AMPK metabolic sensor in leukemia stem cells PR:INSTITUTE Baylor College of Medicine PR:DEPARTMENT Molecular and Human genetics PR:LABORATORY Nakada Lab PR:LAST_NAME Nakada PR:FIRST_NAME Daisuke PR:ADDRESS Houston, TX PR:EMAIL nakada@bcm.edu PR:PHONE 713-798-1175 #STUDY ST:STUDY_TITLE 13C mass isotopomer analysis (LCMS flux studies) MLL-AF9 (part III) ST:STUDY_TYPE Glycolysis/TCA/Nucleotide analysis (tissue/cells) ST:STUDY_SUMMARY How cancer cells adapt to metabolically adverse conditions in patients and ST:STUDY_SUMMARY to proliferate is a fundamental question in cancer biology. Here we show that ST:STUDY_SUMMARY protein kinase (AMPK), a metabolic checkpoint kinase, confers metabolic stress ST:STUDY_SUMMARY to leukemia-initiating cells (LICs) and promotes leukemogenesis. Upon dietary ST:STUDY_SUMMARY MLL-AF9-induced murine acute myeloid leukemia (AML) activated AMPK and ST:STUDY_SUMMARY leukemogenic potential. AMPK deletion significantly delayed leukemogenesis and ST:STUDY_SUMMARY LICs by reducing the expression of glucose transporter 1 (Glut1), compromising ST:STUDY_SUMMARY flux, and increasing oxidative stress and DNA damage. LICs were particularly ST:STUDY_SUMMARY on AMPK to suppress oxidative stress in the hypoglycemic bone marrow ST:STUDY_SUMMARY Strikingly, AMPK inhibition synergized with physiological metabolic stress ST:STUDY_SUMMARY by dietary restriction and profoundly suppressed leukemogenesis. Our results ST:STUDY_SUMMARY that AMPK protects LICs from metabolic stress and that combining AMPK ST:STUDY_SUMMARY with physiological metabolic stress potently suppresses AML by inducing ST:STUDY_SUMMARY stress and DNA damage. ST:INSTITUTE University of Michigan ST:DEPARTMENT Molecular and Human genetics (Baylor College of Medicine) ST:LABORATORY Nakada Lab ST:LAST_NAME Saitoh ST:FIRST_NAME Yusuke ST:ADDRESS Houston, TX ST:EMAIL araskind@med.umich.edu> ST:PHONE 713-798-1175 ST:NUM_GROUPS 1 ST:TOTAL_SUBJECTS 9 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:SPECIES_GROUP Mammal #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - S00017008 GENOTYPE:AMPKαfl/fl | GROUP:Bone marrow SUBJECT_SAMPLE_FACTORS - S00017010 GENOTYPE:AMPKαfl/fl | GROUP:Bone marrow SUBJECT_SAMPLE_FACTORS - S00017012 GENOTYPE:AMPKαfl/fl | GROUP:Bone marrow SUBJECT_SAMPLE_FACTORS - S00017009 GENOTYPE:AMPKαfl/fl | GROUP:Spleen SUBJECT_SAMPLE_FACTORS - S00017011 GENOTYPE:AMPKαfl/fl | GROUP:Spleen SUBJECT_SAMPLE_FACTORS - S00017013 GENOTYPE:AMPKαfl/fl | GROUP:Spleen SUBJECT_SAMPLE_FACTORS - S00017014 GENOTYPE:AMPKαΔ/Δ | GROUP:Bone marrow SUBJECT_SAMPLE_FACTORS - S00017015 GENOTYPE:AMPKαΔ/Δ | GROUP:Bone marrow SUBJECT_SAMPLE_FACTORS - S00017016 GENOTYPE:AMPKαΔ/Δ | GROUP:Bone marrow #COLLECTION CO:COLLECTION_SUMMARY - CO:SAMPLE_TYPE Cells #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME Gly-TCA-nucleotides_analysis_protocol-2015-03-09.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 1260 CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 1mm, 3um) CH:METHODS_ID AQM020 CH:METHODS_FILENAME QTOF-002-HILIC-35min-1mm.m #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME MRC2 (University of Michigan) AN:ACQUISITION_PARAMETERS_FILE QTOF-002-HILIC-35min-1mm.m AN:PROCESSING_PARAMETERS_FILE EX00389-Flux-Quant-method.m #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6520 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS mM MS_METABOLITE_DATA_START Samples S00017008 S00017010 S00017012 S00017009 S00017011 S00017013 S00017014 S00017015 S00017016 Factors GENOTYPE:AMPKαfl/fl | GROUP:Bone marrow GENOTYPE:AMPKαfl/fl | GROUP:Bone marrow GENOTYPE:AMPKαfl/fl | GROUP:Bone marrow GENOTYPE:AMPKαfl/fl | GROUP:Spleen GENOTYPE:AMPKαfl/fl | GROUP:Spleen GENOTYPE:AMPKαfl/fl | GROUP:Spleen GENOTYPE:AMPKαΔ/Δ | GROUP:Bone marrow GENOTYPE:AMPKαΔ/Δ | GROUP:Bone marrow GENOTYPE:AMPKαΔ/Δ | GROUP:Bone marrow 2 or 3-phosphoglycerate 0.2749 0.2178 0.1060 0.0784 0.0791 0.0442 6-Phosphogluconate 0.0355 0.0303 0.0504 0.0209 0.0412 0.0522 0.0721 0.0286 0.0356 Acetyl-CoA 0.0540 0.0442 0.1001 0.0116 0.0125 Adenosine diphosphate 3.2166 2.9132 3.3073 4.8869 2.6028 3.9940 1.9633 0.6554 1.0599 Adenosine monophosphate 0.9395 0.5836 1.2922 4.9193 5.0201 10.4192 2.8176 1.4996 0.9127 Adenosine triphosphate 1.7826 1.6929 1.6539 2.0781 1.2834 1.3987 1.7462 1.1473 1.2682 Citrate or Isocitrate 1.1254 0.9106 2.5543 0.7856 1.6035 2.7287 1.6892 1.0634 2.4373 Erythrose 4-phosphate 1.2759 0.6095 1.9125 0.5430 1.1120 3.9477 1.6767 1.6723 1.4242 Fructose 1_6-bisphosphate 0.8975 0.7937 6.2013 1.6072 1.5003 10.4554 1.6042 0.5498 0.5502 Malate 4.2773 4.6436 6.8832 3.4428 3.0444 3.1205 2.8250 0.3788 1.9067 Nicotinamide Adenine Dinucleotide oxidized 0.1260 0.0450 0.0191 0.0429 0.0146 0.0113 0.0224 0.0101 0.1229 Nicotinamide Adenine Dinucleotide reduced 9.7775 10.8438 8.3556 9.1727 3.4163 2.8091 2.0664 Phosphoenolpyruvate 0.1169 0.1560 Ribose-5-phosphate or xylulose 5-phosphate 1.8613 0.9631 1.5465 0.9984 2.3023 2.8097 1.5578 1.2378 2.0549 Sedoheptulose 7-phosphate 0.2601 0.1196 0.4332 0.1314 0.2188 0.5454 0.2105 0.1866 0.1462 Sucrose 0.0019 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type 2 or 3-phosphoglycerate 724 C00597 2PG/3PG Nakada_Lab_ID 6-Phosphogluconate 91493 C00345 6PG Nakada_Lab_ID Acetyl-CoA 444493 C00024 aCoA Nakada_Lab_ID Adenosine diphosphate 6022 C00008 ADP Nakada_Lab_ID Adenosine monophosphate 6083 C00020 AMP Nakada_Lab_ID Adenosine triphosphate 5957 C00002 ATP Nakada_Lab_ID Citrate or Isocitrate 311 C00158 CIT/ICIT Nakada_Lab_ID Erythrose 4-phosphate 122357 C00279 E4P Nakada_Lab_ID Fructose 1,6-bisphosphate 172313 C00354 FBP Nakada_Lab_ID Malate 222656 C00149 MAL Nakada_Lab_ID Nicotinamide Adenine Dinucleotide oxidized 10897651 C00003 NAD Nakada_Lab_ID Nicotinamide Adenine Dinucleotide reduced 439153 C00004 NADH Nakada_Lab_ID Phosphoenolpyruvate 1005 C00074 PEP Nakada_Lab_ID Ribose-5-phosphate or xylulose 5-phosphate 77982 C00117 R5P/X5P Nakada_Lab_ID Sedoheptulose 7-phosphate 165007 C05382 S7P Nakada_Lab_ID Sucrose 5988 C00089 SUC Nakada_Lab_ID METABOLITES_END #END