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• W4361280293 abstract "N6-methyladenosine (m6A) modification contributes to the pathogenesis and development of various cancers, including bladder cancer (BCa). In particular, integrin α6 (ITGA6) promotes BCa progression by cooperatively regulating multisite m6A modification. However, the therapeutic effect of targeting ITGA6 multisite m6A modifications in BCa remains unknown. We aim to develop a multisite dCasRx– m6A editor for assessing the effects of the multisite dCasRx–m6A editor targeted m6A demethylation of ITGA6 mRNA in BC growth and progression. The multisite dCasRx– m6A editor was generated by cloning. m6A-methylated RNA immunoprecipitation (meRIP), luciferase reporter, a single-base T3 ligase-based qPCR-amplification, Polysome profiling and meRIP-seq experiments were performed to determine the targeting specificity of the multisite dCasRx–m6A editor. We performed cell phenotype analysis and used in vivo mouse xenograft models to assess the effects of the multisite dCasRx–m6A editor in BC growth and progression. We designed a targeted ITGA6 multi-locus guide (g)RNA and established a bidirectional deactivated RfxCas13d (dCasRx)-based m6A-editing platform, comprising a nucleus-localized dCasRx fused with the catalytic domains of methyltransferase-like 3 (METTL3-CD) or α-ketoglutarate-dependent dioxygenase alkB homolog 5 (ALKBH5-CD), to simultaneously manipulate the methylation of ITGA6 mRNA at four m6A sites. The results confirmed the dCasRx–m6A editor modified m6A at multiple sites in ITGA6 mRNA, with low off-target effects. Moreover, targeted m6A demethylation of ITGA6 mRNA by the multisite dCasRx–m6A editor significantly reduced BCa cell proliferation and migration in vitro and in vivo. Furthermore, the dCasRx–ALKBH5-CD and ITGA6 multi-site gRNA delivered to 5-week-old BALB/cJNju-Foxn1nu/Nju nude mice via adeno-associated viral vectors significantly inhibited BCa cell growth. Our study proposes a novel therapeutic tool for the treatment of BC by applying the multisite dCasRx–m6A editor while highlighting its potential efficacy for treating other diseases associated with abnormal m6A modifications." @default.
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• W4361280293 date "2023-03-01" @default.
• W4361280293 modified "2023-10-16" @default.
• W4361280293 title "Targeted m6A demethylation of ITGA6 mRNA by a multisite dCasRx–m6A editor inhibits bladder cancer development" @default.
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• W4361280293 doi "https://doi.org/10.1016/j.jare.2023.03.010" @default.
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