The m6A Reader IGF2BP2 Regulates Glutamine Metabolism and Represents a Therapeutic Target in Acute Myeloid Leukemia [RNA-seq]

We report that m6A reader IGF2BP2 participates in the regulation of glutamine metabolism in AML. Targeting IGF2BP2 with our developed inhibitor CWI1-2 shows promising therapeutic efficacy in AML.To explore the mechanism of the inhibitor, we performed RNA-seq in AML cells upon treatment with CWI1-2 or DMSO. On the other hand, to distinguish the mechanism between IGF2BP2 and YTHDF2, we compare RNA-seq data of IGF2BP2 KD with YTHDF2 KD. Overall design: Total RNA from MonoMac6 cells with different treatments (i.e., CWI1-2 vs DMSO, YTHDF2 KD vs shNS) was isolated using the TRIzol reagent (Thermo fisher Scientific). Library construction of 1 µg RNA per sample was made using the NEBNext® Ultra RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations. PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumina) according to the manufacturer's instructions. The library preparations were sequenced on an Illumina NovaSeq platform and 150 bp paired-end reads were generated.