Direct RNA sequencing (m6A analysis) and RNA sequencing on DMG cultures after FTO inhibition via FB23-2 treatment
Diffuse Midline Gliomas (DMG) are deadly paediatric brain cancers with limited therapeutic options. These tumors are thought to arise from oligodendrocyte precursor cells (OPC), which acquire a driver H3K27M histone mutation that leads to an aberrant histone modification landscape. RNA-N6-methyladenosine (m6A) is a crucial epi-transcriptomic modification that regulates RNA stability, splicing, translation, and degradation. It plays a significant role in OPC development through its cross-talking with histone modifications. Given the pivotal role of m6A in normal OPC biology, investigating its dysregulation in DMG could uncover new therapeutic options and improve our understanding of the disease's underlying mechanisms. Here, for the first time, we perform direct RNA-sequencing on two patient-derived DMG cultures to generate transcript and base-resolution m6A landscapes of DMG. Furthermore, we interrogate the therapeutic benefits of the Methyltransferase 13 (METTL13) inhibitor STM2457 and the Fat mass and obesity associated (FTO) player inhibitor FB23-2 in DMG using direct RNA sequencing, RNA sequencing, Mass spectrometry and Functional assays.