Abstract 7075: Global epitranscriptomic and transcriptomic footprint revealed upregulation of methylated single minded 2-small variant (meSIM2-SV) in colorectal cancer (CRC) by NTMT1(METTL11A) via m6A-YTHDF1-dependent manner

Abstract Establishing global footprint of altered m6A mRNA is a unique method to identify the new prognostics biomarkers. Altered N6-methyladenosine(m6A) RNA methylation regulates cancer progression with uncharacterized mechanism. Here, we present novel link between m6A modification on 5’-UTR SIM2-SV contribution in colorectal cancer progression. MeRIP and RNA-seq data from 68 fresh frozen CRC samples revealed 13,843 altered m6A methylated peaks and found highest in chromosomes 1, 2, and 3. Analysis uncovered 119 overlapping genes and clusters into 32 up-regulated genes with 56 up-hyper peaks, 8 up-regulated genes with 9 up-hypo peaks, 12 down-regulated genes with 14 down-hyper peaks, and 28 down-regulated genes with 40 down-hypo peaks. A significant increase in the SIM2 m6A and RNA level led us to perform correlation analysis between the SIM2SV and m6A RNA regulators. SIM2SV was significantly correlated with YTHDF1 m6A-readers and m6A-NTMT1 writer, suggesting important roles of SIM2SV in CRC oncogenicity. m6A-sequencing identified m6A enrichment in 5’-UTR of SIM2SV gene which further verified. siNTMT1 and siYTHDF1 manipulation in HCT116 and RKO cell lines verified a decrease in SIM2SV gene/protein expression, and m6A level. Knockdown effects also showed decrease in cell proliferation, clonogenicity and induced cell cycle arrest in the G2/M phase for NTMT1 and YTHDF1 versus SIM2 Knockdown showed cell cycle arrest in S/G2 phase. Our investigation revealed the role of new writer NTMT1 for methylated SIM2SV gene expression in colorectal cancer proliferation via m6A-YTHDF1(as “decoder”) dependent manner. Further investigation on m6A modification specifically on SIM2 mRNA revealed the essential roles of its writer NTMT1 and its reader YTHDF1 in CRC development, evidently supporting the importance of the unique landscape of m6A epigenetic modification in colorectal neoplasm, which not only advance our understanding of CRC pathobiology but also provides a new set of molecular targets for building up effective tools to fight CRC. Citation Format: Sudhir Kumar Rai, Ting Gong, Zitong Gao, Yujia Qin, Matthew Huo, Ken Nakastu, Hua Yang, Yuanyuan Fu, Lang Wu, Mayumi Jijiwa, Masaki Nasu, Shaoqiu Chen, Gang Huang, Peiwen Fei, Youping Deng. Global epitranscriptomic and transcriptomic footprint revealed upregulation of methylated single minded 2-small variant (meSIM2-SV) in colorectal cancer (CRC) by NTMT1(METTL11A) via m6A-YTHDF1-dependent manner [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7075.