#METABOLOMICS WORKBENCH krystinemansfield_20230113_130754 DATATRACK_ID:3690 STUDY_ID:ST002446 ANALYSIS_ID:AN003984 VERSION 1 CREATED_ON 09-01-2023 #PROJECT PR:PROJECT_TITLE Reprogramming of miR-142-dependent Metabolism Drives Blast Transformation of PR:PROJECT_TITLE Chronic Myelogenous Leukemia PR:PROJECT_TYPE MS quantitative analysis PR:PROJECT_SUMMARY Metabolic profiling of miR-142+/+BCR-ABL Leukemic stem cell (LSC)-enriched PR:PROJECT_SUMMARY Lineage-Sca-1+c-Kit+ cells (LSKs) compared to miR-142-/-BCR-ABL controls PR:INSTITUTE Translational Genomics Research Institute PR:LAST_NAME Mansfield PR:FIRST_NAME Krystine PR:ADDRESS 445 N 5th St, Phoenix, AZ, 85004, USA PR:EMAIL kgarcia@tgen.org PR:PHONE 602-343-8832 PR:PUBLICATIONS https://www.nature.com/articles/s41467-023-41167-z PR:DOI http://dx.doi.org/10.21228/M8VD8T #STUDY ST:STUDY_TITLE Untargeted metabolomics of miR-142 WT vs KO CML cells ST:STUDY_SUMMARY MiR-142 is dynamically expressed and plays a regulatory role in hematopoiesis. ST:STUDY_SUMMARY Based on the simple observation that miR-142 levels are significantly lower in ST:STUDY_SUMMARY CD34+CD38- cells from blast crisis (BC) chronic myeloid leukemia (CML). CML ST:STUDY_SUMMARY patients compared with chronic phase (CP) CML patients (p=0.002), we ST:STUDY_SUMMARY hypothesized that miR-142 deficit plays a role in BC transformation. To test ST:STUDY_SUMMARY this hypothesis, we generated a miR-142 KO BCR-ABL (i.e., miR-142−/−BCR-ABL) ST:STUDY_SUMMARY mouse by crossing a miR-142−/− mouse with a miR-142+/+BCR-ABL mouse. While ST:STUDY_SUMMARY the miR-142+/+BCR-ABL mice developed and died of CP CML, the ST:STUDY_SUMMARY miR-142−/−BCR-ABL mice developed a BC-like phenotype in the absence of any ST:STUDY_SUMMARY other acquired gene mutations and died significantly sooner than ST:STUDY_SUMMARY miR-142+/+BCR-ABL CP controls (p=0.001). Leukemic stem cell (LSC)-enriched ST:STUDY_SUMMARY Lineage-Sca-1+c-Kit+ cells (LSKs) from diseased miR-142−/−BCR-ABL mice ST:STUDY_SUMMARY transplanted into congenic recipients, recapitulated the BC features thereby ST:STUDY_SUMMARY suggesting stable transformation of CP-LSCs into BC-LSCs in the miR-142 KO CML ST:STUDY_SUMMARY mouse. Single cell (sc) RNA-seq profiling showed that miR-142 deficit changed ST:STUDY_SUMMARY the cellular landscape of the miR-142−/−BCR-ABL LSKs compared with ST:STUDY_SUMMARY miR-142+/+BCR-ABL LSKs with expansion of myeloid-primed and loss of ST:STUDY_SUMMARY lymphoid-primed factions. Bulk RNA-seq analyses along with unbiased metabolomic ST:STUDY_SUMMARY profiling and functional metabolic assays demonstrated enhanced fatty acid ST:STUDY_SUMMARY β-oxidation (FAO) and oxidative phosphorylation (OxPhos) in ST:STUDY_SUMMARY miR-142−/−BCR-ABL LSKs vs miR-142+/+BCR-ABL LSKs. MiR-142 deficit enhanced ST:STUDY_SUMMARY FAO in miR-142−/−BCR-ABL LSKs by increasing the expression of CPT1A and ST:STUDY_SUMMARY CPT1B, that controls the cytosol-to-mitochondrial acyl-carnitine transport, a ST:STUDY_SUMMARY critical step in FAO. MiR-142 deficit also enhanced OxPhos in ST:STUDY_SUMMARY miR-142−/−BCR-ABL LSKs by increasing mitochondrial fusion and activity. As ST:STUDY_SUMMARY the homeostasis and activity of LSCs depend on higher levels of these oxidative ST:STUDY_SUMMARY metabolism processes, we then postulate that miR-142 deficit is a potentially ST:STUDY_SUMMARY druggable target for BC-LSCs. To this end, we developed a novel CpG-miR-142 ST:STUDY_SUMMARY mimic oligonucleotide (ODN; i.e., CpG-M-miR-142) that corrected the miR-142 ST:STUDY_SUMMARY deficit and alone or in combination with a tyrosine kinase inhibitor (TKI) ST:STUDY_SUMMARY significantly reduced LSC burden and prolonged survival of miR-142−/−BCR-ABL ST:STUDY_SUMMARY mice. The results from murine models were validated in BC CD34+CD38- primary ST:STUDY_SUMMARY blasts and patient-derived xenografts (PDXs). In conclusion, an acquired miR-142 ST:STUDY_SUMMARY deficit sufficed in transforming CP-LSCs into BC-LSCs, via enhancement of ST:STUDY_SUMMARY bioenergetic oxidative metabolism in absence of any additional gene mutations, ST:STUDY_SUMMARY and likely represent a novel therapeutic target in BC CML. ST:INSTITUTE Translational Genomics Research Institute ST:LAST_NAME Mansfield ST:FIRST_NAME Krystine ST:ADDRESS 445 N 5th St, Phoenix, AZ, 85004, USA ST:EMAIL kgarcia@tgen.org ST:PHONE 602-343-8832 ST:SUBMIT_DATE 2023-01-13 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Male and female #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS A09xKOxFx01 Norm. Area: 088_0003_02B_R_A09xKOxFx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F9) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A09xKOxFx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A10xKOxFx02 Norm. Area: 088_0003_02B_R_A10xKOxFx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F10) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A10xKOxFx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A11xKOxFx03 Norm. Area: 088_0003_02B_R_A11xKOxFx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F11) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A11xKOxFx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A12xKOxFx04 Norm. Area: 088_0003_02B_R_A12xKOxFx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F12) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A12xKOxFx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A13xKOxFx05 Norm. Area: 088_0003_02B_R_A13xKOxFx05xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F13) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A13xKOxFx05xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A14xKOxMx01 Norm. Area: 088_0003_02B_R_A14xKOxMx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F14) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A14xKOxMx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A15xKOxMx02 Norm. Area: 088_0003_02B_R_A15xKOxMx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F15) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A15xKOxMx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A16xKOxMx03 Norm. Area: 088_0003_02B_R_A16xKOxMx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F16) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A16xKOxMx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A17xKOxMx04 Norm. Area: 088_0003_02B_R_A17xKOxMx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F17) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A17xKOxMx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A18xKOxMx05 Norm. Area: 088_0003_02B_R_A18xKOxMx05xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F18) Genotype:KO RAW_FILE_NAME=088_0003_02B_R_A18xKOxMx05xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A09xKOxFx01 Norm. Area: 088_0003_02D_R_A09xKOxFx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F175) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A09xKOxFx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A10xKOxFx02 Norm. Area: 088_0003_02D_R_A10xKOxFx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F176) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A10xKOxFx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A11xKOxFx03 Norm. Area: 088_0003_02D_R_A11xKOxFx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F177) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A11xKOxFx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A12xKOxFx04 Norm. Area: 088_0003_02D_R_A12xKOxFx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F178) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A12xKOxFx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A13xKOxFx05 Norm. Area: 088_0003_02D_R_A13xKOxFx05xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F179) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A13xKOxFx05xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A14xKOxMx01 Norm. Area: 088_0003_02D_R_A14xKOxMx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F180) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A14xKOxMx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A15xKOxMx02 Norm. Area: 088_0003_02D_R_A15xKOxMx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F181) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A15xKOxMx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A16xKOxMx03 Norm. Area: 088_0003_02D_R_A16xKOxMx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F182) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A16xKOxMx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A17xKOxMx04 Norm. Area: 088_0003_02D_R_A17xKOxMx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F183) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A17xKOxMx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A18xKOxMx05 Norm. Area: 088_0003_02D_R_A18xKOxMx05xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F184) Genotype:KO RAW_FILE_NAME=088_0003_02D_R_A18xKOxMx05xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A09xKOxFx01 Norm. Area: 088_0003_02E_R_A09xKOxFx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F206) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A09xKOxFx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A10xKOxFx02 Norm. Area: 088_0003_02E_R_A10xKOxFx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F207) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A10xKOxFx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A11xKOxFx03 Norm. Area: 088_0003_02E_R_A11xKOxFx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F208) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A11xKOxFx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A12xKOxFx04 Norm. Area: 088_0003_02E_R_A12xKOxFx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F209) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A12xKOxFx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A13xKOxFx05 Norm. Area: 088_0003_02E_R_A13xKOxFx05xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F210) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A13xKOxFx05xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A14xKOxMx01 Norm. Area: 088_0003_02E_R_A14xKOxMx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F211) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A14xKOxMx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A15xKOxMx02 Norm. Area: 088_0003_02E_R_A15xKOxMx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F212) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A15xKOxMx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A16xKOxMx03 Norm. Area: 088_0003_02E_R_A16xKOxMx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F213) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A16xKOxMx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A17xKOxMx04 Norm. Area: 088_0003_02E_R_A17xKOxMx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F214) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A17xKOxMx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A18xKOxMx05 Norm. Area: 088_0003_02E_R_A18xKOxMx05xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F215) Genotype:KO RAW_FILE_NAME=088_0003_02E_R_A18xKOxMx05xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A09xKOxFx01 Norm. Area: 088_0003_02G_R_A09xKOxFx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F145) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A09xKOxFx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A10xKOxFx02 Norm. Area: 088_0003_02G_R_A10xKOxFx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F146) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A10xKOxFx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A11xKOxFx03 Norm. Area: 088_0003_02G_R_A11xKOxFx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F147) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A11xKOxFx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A12xKOxFx04 Norm. Area: 088_0003_02G_R_A12xKOxFx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F148) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A12xKOxFx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A13xKOxFx05 Norm. Area: 088_0003_02G_R_A13xKOxFx05conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F149) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A13xKOxFx05conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A14xKOxMx01 Norm. Area: 088_0003_02G_R_A14xKOxMx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F150) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A14xKOxMx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A15xKOxMx02 Norm. Area: 088_0003_02G_R_A15xKOxMx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F151) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A15xKOxMx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A16xKOxMx03 Norm. Area: 088_0003_02G_R_A16xKOxMx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F152) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A16xKOxMx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A17xKOxMx04 Norm. Area: 088_0003_02G_R_A17xKOxMx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F153) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A17xKOxMx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A18xKOxMx05 Norm. Area: 088_0003_02G_R_A18xKOxMx05conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F154) Genotype:KO RAW_FILE_NAME=088_0003_02G_R_A18xKOxMx05conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A01xWTxFx01 Norm. Area: 088_0003_02B_R_A01xWTxFx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F1) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A01xWTxFx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A02xWTxFx02 Norm. Area: 088_0003_02B_R_A02xWTxFx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F2) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A02xWTxFx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A03xWTxFx03 Norm. Area: 088_0003_02B_R_A03xWTxFx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F3) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A03xWTxFx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A04xWTxFx04 Norm. Area: 088_0003_02B_R_A04xWTxFx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F4) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A04xWTxFx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A05xWTxMx01 Norm. Area: 088_0003_02B_R_A05xWTxMx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F5) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A05xWTxMx01xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A06xWTxMx02 Norm. Area: 088_0003_02B_R_A06xWTxMx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F6) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A06xWTxMx02xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A07xWTxMx03 Norm. Area: 088_0003_02B_R_A07xWTxMx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F7) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A07xWTxMx03xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A08xWTxMx04 Norm. Area: 088_0003_02B_R_A08xWTxMx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F8) Genotype:WT RAW_FILE_NAME=088_0003_02B_R_A08xWTxMx04xHILICpos_09SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A01xWTxFx01 Norm. Area: 088_0003_02D_R_A01xWTxFx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F167) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A01xWTxFx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A02xWTxFx02 Norm. Area: 088_0003_02D_R_A02xWTxFx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F168) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A02xWTxFx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A03xWTxFx03 Norm. Area: 088_0003_02D_R_A03xWTxFx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F169) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A03xWTxFx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A04xWTxFx04 Norm. Area: 088_0003_02D_R_A04xWTxFx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F170) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A04xWTxFx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A05xWTxMx01 Norm. Area: 088_0003_02D_R_A05xWTxMx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F171) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A05xWTxMx01xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A06xWTxMx02 Norm. Area: 088_0003_02D_R_A06xWTxMx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F172) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A06xWTxMx02xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A07xWTxMx03 Norm. Area: 088_0003_02D_R_A07xWTxMx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F173) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A07xWTxMx03xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A08xWTxMx04 Norm. Area: 088_0003_02D_R_A08xWTxMx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F174) Genotype:WT RAW_FILE_NAME=088_0003_02D_R_A08xWTxMx04xconcRPpos_21SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A01xWTxFx01 Norm. Area: 088_0003_02E_R_A01xWTxFx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F198) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A01xWTxFx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A02xWTxFx02 Norm. Area: 088_0003_02E_R_A02xWTxFx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F199) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A02xWTxFx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A03xWTxFx03 Norm. Area: 088_0003_02E_R_A03xWTxFx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F200) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A03xWTxFx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A04xWTxFx04 Norm. Area: 088_0003_02E_R_A04xWTxFx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F201) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A04xWTxFx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A05xWTxMx01 Norm. Area: 088_0003_02E_R_A05xWTxMx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F202) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A05xWTxMx01xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A06xWTxMx02 Norm. Area: 088_0003_02E_R_A06xWTxMx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F203) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A06xWTxMx02xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A07xWTxMx03 Norm. Area: 088_0003_02E_R_A07xWTxMx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F204) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A07xWTxMx03xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A08xWTxMx04 Norm. Area: 088_0003_02E_R_A08xWTxMx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.raw (F205) Genotype:WT RAW_FILE_NAME=088_0003_02E_R_A08xWTxMx04xconcRPneg_28SEP21_CEL_MOU_LUM_0000_04AC_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A01xWTxFx01 Norm. Area: 088_0003_02G_R_A01xWTxFx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F137) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A01xWTxFx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A02xWTxFx02 Norm. Area: 088_0003_02G_R_A02xWTxFx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F138) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A02xWTxFx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A03xWTxFx03 Norm. Area: 088_0003_02G_R_A03xWTxFx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F139) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A03xWTxFx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A04xWTxFx04 Norm. Area: 088_0003_02G_R_A04xWTxFx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F140) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A04xWTxFx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A05xWTxMx01 Norm. Area: 088_0003_02G_R_A05xWTxMx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F141) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A05xWTxMx01conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A06xWTxMx02 Norm. Area: 088_0003_02G_R_A06xWTxMx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F142) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A06xWTxMx02conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A07xWTxMx03 Norm. Area: 088_0003_02G_R_A07xWTxMx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F143) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A07xWTxMx03conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML SUBJECT_SAMPLE_FACTORS A08xWTxMx04 Norm. Area: 088_0003_02G_R_A08xWTxMx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.raw (F144) Genotype:WT RAW_FILE_NAME=088_0003_02G_R_A08xWTxMx04conxHILICneg_29SEP21_CEL_MOU_LUM_0000_04AB_15MIN_KP_KP_01.mzML #COLLECTION CO:COLLECTION_SUMMARY Lin-c-Kit+ cells, a fraction where LSKs reside, were selected four weeks after CO:COLLECTION_SUMMARY tet-off and BCR-ABL induction from the BM of miR-142−/−BCR-ABL and CO:COLLECTION_SUMMARY miR-142+/+BCR-ABL mice. Lin- cells were firstly selected using Lineage depletion CO:COLLECTION_SUMMARY microbeads and c-kit+ cells were then selected using anti-mouse CD117 microbeads CO:COLLECTION_SUMMARY (both from Miltenyi Biotec, San Diego, CA). CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY NA #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites extraction was performed by adding chilled methanol: acetonitrile: SP:SAMPLEPREP_SUMMARY water (2:1:1, v/v/v)) to the cell pellet of pooled Lin-c-kit+ cells followed by SP:SAMPLEPREP_SUMMARY three freeze-thaw cycles. The lysed cells were precipitated by centrifugation at SP:SAMPLEPREP_SUMMARY 15,000 rpm for 10 min at 4 °C and metabolites in the supernatant were vacuum SP:SAMPLEPREP_SUMMARY concentrated and subjected to LC-MS analysis. #CHROMATOGRAPHY CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 RS CH:COLUMN_NAME Thermo Hypersil GOLD C18 (150 x 2.1mm, 1.9um) CH:COLUMN_TEMPERATURE 40 CH:FLOW_GRADIENT 10 min linear gradient from 100% solvent A and 0% solvent B to 2% solvent A and CH:FLOW_GRADIENT 98% solvent B, and 5 min of equilibration time CH:FLOW_RATE 0.35 mL/min CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile; 0.1% formic acid CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Fusion Tribrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS MS1 data were acquired over a mass range of 70 to 1500 m/z in the orbitrap MS:MS_COMMENTS operated at a resolution of 60,000. MS/MS data were acquired in AcquireX mode MS:MS_COMMENTS using the iterative precursor exclusion workflow, with a stepped HCD collision MS:MS_COMMENTS energy at 20, 35 and 50, at a resolution of 30,000 in the orbitrap MS:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST002446_AN003984_Results.txt UNITS:Peak Area Has RT:Yes RT units:Minutes #END