表观转录组RNA修饰构成了影响癌症进展的关键基因调控成分。在这些修饰中，RNA N4-乙酰基胞苷（ac4C）修饰，由ac4C writer N-乙酰转移酶10（NAT10）介导，调控了mRNA的稳定性。在此，我们确定ac4C修饰在顺铂治疗中被诱导，并且与膀胱癌（BCa）的化疗抗性相关。在体内外，NAT10通过增强DNA损伤修复，促进了BCa细胞的顺铂化疗抗性。机械上，NAT10通过保护AHNAK mRNA免受外切酶攻击来结合并稳定AHNAK mRNA，AHNAK介导的DNA损伤修复对NAT10诱导的顺铂抗性至关重要。临床上，NAT10过表达与顺铂抗性、复发和BCa患者的不良临床结果相关。顺铂诱导NFκB信号激活对NAT10表达的上调是必需的，NFκB p65直接结合NAT10启动子来激活转录。此外，Remodelin对NAT10的药理抑制能提高BCa器官样体和小鼠异种移植物对顺铂的敏感性。总的来说，本研究揭示了NAT10介导的mRNA稳定机制在BCa中的作用，为NAT10作为治疗靶点克服BCa的顺铂抗性奠定了基础。

Epitranscriptomic RNA modifications constitute a critical gene regulatory component that can affect cancer progression. Among these, the RNA N4-acetylcytidine (ac4C) modification, which is mediated by the ac4C writer N-acetyltransferase 10 (NAT10), regulates the stabilization of mRNA. Here, we identified that the ac4C modification is induced upon cisplatin treatment and correlates with chemoresistance in bladder cancer (BCa). Both in vitro and in vivo, NAT10 promoted cisplatin chemoresistance in BCa cells by enhancing DNA damage repair. Mechanistically, NAT10 bound and stabilized AHNAK mRNA by protecting it from exonucleases, and AHNAK-mediated DNA damage repair was required for NAT10-induced cisplatin resistance. Clinically, NAT10 overexpression was associated with chemoresistance, recurrence, and worse clinical outcome in BCa patients. Cisplatin-induced NFκB signaling activation was required for the upregulation of NAT10 expression, and NFκB p65 directly bound to the NAT10 promoter to activate transcription. Moreover, pharmacological inhibition of NAT10 with Remodelin sensitized BCa organoids and mouse xenografts to cisplatin. Overall, the present study uncovered a mechanism of NAT10-mediated mRNA stabilization in BCa, laying the foundation for NAT10 as a therapeutic target to overcome cisplatin resistance in BCa.