Escobedo, A. K. | bioRxiv Science

bioRxiv2025

m6ASeqTools: An R toolkit for post-processing m6A sites detected by m6Anet

m6ASeqTools is an R package designed to streamline the post-processing and interpretation of site-level m6A predictions from m6Anet. It provides descriptive summaries of m6A distribution across transcripts, genes, biotypes, and transcript regions, and enables condition comparisons through a calculated weighted modification ratio. By integrating differential gene expression data, the package links methylation changes with expression differences, providing biotype-specific and region-specific insights into how m6A localization patterns relate to transcriptional regulation. Availability and implementationm6ASeqTools is freely available at https://github.com/hannalee809/m6ASeqTools Supplementary informationSupplementary data are available at Bioinformatics online.

bioinformatics↗

bioRxiv2024

Exploring N6-Methyladenosine and long transcripts in glioma cells and extracellular vesicles.

N6-methyladenosine (m6A) is the most prevalent internal mRNA modification, enriched in the CNS yet poorly characterized in glioma. Using long-read RNA sequencing, we mapped m6A in an in vitro glioma model following knockdown (KD) of the reader IGF2BP2, writer METTL3, and eraser ALKBH5, with naive glioma cells and astrocytes as controls. Glioma cells exhibited a two-fold reduction in global m6A, suggesting progressive loss from healthy to malignant states. Integrated analysis revealed that m6A mediated control of gene expression is influenced by modification topology (CDS:3'UTR), transcript biotype, and length. Regulator KD, particularly ALKBH5 induced redistribution of m6A toward 3'UTR with consequent gene upregulation. We also identified m6A-mediated isoform switching, with a higher usage of retained intron and nonsense-mediated decay isoforms. Structural and splicing alterations at the isoform level were identified unique to each KD condition indicating m6A driven aberrant alternative splicing. At the functional level, KD specific remodeling of oncogenic signaling was also observed. ALKBH5 KD suppressed MYC targets and pro-apoptotic signaling while METTL3 KD enhanced mTOR and PI3K-AKT signaling. Collectively, these results demonstrate that m6A mediated regulation in glioma is highly context-dependent, defining distinct clinically relevant phenotypes. This has implications for future biomarker discovery and development of targeted therapeutics.

cancer biology↗