RNA modification has emerged as an important epigenetic mechanism that controls abnormal metabolism and growth in acute myeloid leukaemia (AML). However, the roles of RNA N⁴-acetylcytidine (ac4C) modification in AML remain elusive. Here, we report that ac4C and its catalytic enzyme NAT10 drive leukaemogenesis and sustain self-renewal of leukaemic stem cells/leukaemia-initiating cells through reprogramming serine metabolism. Mechanistically, NAT10 facilitates exogenous serine uptake and de novo biosynthesis through ac4C-mediated translation enhancement of the serine transporter SLC1A4 and the transcription regulators HOXA9 and MENIN that activate transcription of serine synthesis pathway genes. We further characterize fludarabine as an inhibitor of NAT10 and demonstrate that pharmacological inhibition of NAT10 targets serine metabolic vulnerability, triggering substantial anti-leukaemia effects both in vitro and in vivo. Collectively, our study demonstrates the functional importance of ac4C and NAT10 in metabolism control and leukaemogenesis, providing insights into the potential of targeting NAT10 for AML therapy.

RNA 修饰已成为控制急性髓系白血病 （AML） 异常代谢和生长的重要表观遗传机制。然而，RNA N⁴-乙酰胞苷 （ac4C） 修饰在 AML 中的作用仍然难以捉摸。在这里，我们报道了 ac4C 及其催化酶 NAT10 通过重编程丝氨酸代谢驱动白血病发生并维持白血病干细胞/白血病起始细胞的自我更新。从机制上讲，NAT10 通过 ac4C 介导的丝氨酸转运蛋白 SLC1A4 和转录调节因子 HOXA9 和 MENIN 的翻译增强促进外源丝氨酸摄取和从头生物合成，这些转录调节因子激活丝氨酸合成途径基因的转录。我们进一步将氟达拉滨表征为 NAT10 的抑制剂，并证明 NAT10 的药理学抑制针对丝氨酸代谢脆弱性，在体外和体内触发显着的抗白血病作用。总的来说，我们的研究证明了 ac4C 和 NAT10 在代谢控制和白血病发生中的功能重要性，为靶向 NAT10 进行 AML 治疗的潜力提供了见解。