Wan Xinxing, Wang Linghao, Khan Md Asaduzzaman, Peng Lin, Sun Xiaoying, Yi Xuan, Wang Zhouqi, Chen Ke
Department of Endocrinology, The Third Xiangya Hospital of Central South University, Changsha, Hunan, PR China.
Department of Biochemistry and Microbiology, School of Health & Life Sciences, North South University, Dhaka, Bangladesh.
Cell Death Differ. 2025 Mar 23. doi: 10.1038/s41418-025-01483-x.
Dysfunctional adipogenesis is a major contributor of obesity. N-acetyltransferase 10 (NAT10) plays a crucial role in regulating N4-acetylcysteine (ac4C) modification in tRNA, 18SrRNA, and mRNA. As the sole "writer" in the ac4C modification process, NAT10 enhances mRNA stability and translation efficiency. There are few reports on the relationship between NAT10 and adipogenesis, as well as obesity. Our study revealed a significant upregulation of NAT10 in adipose tissues of obese individuals and high-fat diet-fed mice. Furthermore, our findings revealed that the overexpression of NAT10 promotes adipogenesis, while its silencing inhibits adipogenesis in both human adipose tissue-derived stem cells (hADSCs) and 3T3-L1 cells. These results indicate the intimate relationship between NAT10 and obesity. After silencing mouse NAT10 (mNAT10), we identified 30 genes that exhibited both hypo-ac4C modification and downregulation in their expression, utilizing a combined approach of acRIP-sequencing (acRIP-seq) and RNA-sequencing (RNA-seq). Among these genes, we validated KLF9 as a target of NAT10 through acRIP-PCR. KLF9, a pivotal transcription factor that positively regulates adipogenesis. Our findings showed that NAT10 enhances the stability of KLF9 mRNA and further activates the CEBPA/B-PPARG pathway. Furthermore, a dual-luciferase reporter assay demonstrated that NAT10 can bind to three motifs of mouse KLF9 and one motif of human KLF9. In vivo studies revealed that adipose tissue-targeted mouse AAV-NAT10 (AAV-shRNA-mNAT10) inhibits adipose tissue expansion in mice. Additionally, Remodelin, a specific NAT10 inhibitor, significantly reduced body weight, adipocyte size, and adipose tissue expansion in high-fat diet-fed mice by inhibiting KLF9 mRNA ac4C modification. These findings provide novel insights and experimental evidence of the prevention and treatment of obesity, highlighting NAT10 and its downstream targets as potential therapeutic targets.
脂肪生成功能障碍是肥胖的主要原因。N-乙酰转移酶10（NAT10）在调节tRNA、18SrRNA和mRNA中的N4-乙酰半胱氨酸（ac4C）修饰方面起着关键作用。作为ac4C修饰过程中的唯一“书写者”，NAT10可增强mRNA稳定性和翻译效率。关于NAT10与脂肪生成以及肥胖之间的关系，报道较少。我们的研究显示，肥胖个体和高脂饮食喂养小鼠的脂肪组织中NAT10显著上调。此外，我们的研究结果表明，NAT10的过表达促进脂肪生成，而其沉默则抑制人脂肪组织来源干细胞（hADSCs）和3T3-L1细胞中的脂肪生成。这些结果表明NAT10与肥胖之间存在密切关系。沉默小鼠NAT10（mNAT10）后，我们采用acRIP测序（acRIP-seq）和RNA测序（RNA-seq）相结合的方法，鉴定出30个表现出ac4C修饰降低和表达下调的基因。在这些基因中，我们通过acRIP-PCR验证KLF9是NAT10的一个靶点。KLF9是一种对脂肪生成起正向调节作用的关键转录因子。我们的研究结果表明，NAT10增强KLF9 mRNA的稳定性，并进一步激活CEBPA/B-PPARG通路。此外，双荧光素酶报告基因检测表明，NAT10可与小鼠KLF9的三个基序和人KLF9的一个基序结合。体内研究表明，脂肪组织靶向的小鼠AAV-NAT10（AAV-shRNA-mNAT10）可抑制小鼠脂肪组织扩张。此外，特异性NAT10抑制剂Remodelin通过抑制KLF9 mRNA的ac4C修饰，显著降低高脂饮食喂养小鼠的体重、脂肪细胞大小和脂肪组织扩张。这些发现为肥胖的预防和治疗提供了新的见解和实验证据，突出了NAT10及其下游靶点作为潜在治疗靶点的作用。
