Liu Xianzhi, Chen Zhiyuan, Lin Jun, Lian Yifan, Gan Wenxuan, Liu Huajie, Huang Xingxiang, Mei Jiaxin, Ma Tianrong, Lu Zhi, Zeng Wei, Gong Yihang, Chen Shuai, He Weiling
Department of Gastroenterology, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian, 361000, China.
Department of Pediatrics, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian, 361000, China.
Int J Biol Sci. 2025 Aug 30;21(12):5563-5585. doi: 10.7150/ijbs.118024. eCollection 2025.
Pyroptosis is a novel programmed cell death that exists in inflammatory diseases and methyltransferase-like 3 (METTL3) is a core N6-methyladenosine (m6A) modified methyltransferase that has been shown to regulate cell fate. However, the role of pyroptosis in acute liver injury (ALI) is still unknown and whether it is regulated by m6A modification needs to be elucidated. Here, mutant and knockout mouse were constructed, CCl- and TAA-induced ALI models were established and primary cells were isolated, and cell pyroptosis and m6A modification were evaluated. We found that hepatocyte pyroptosis is a key characteristic of ALI, and METTL3-mediated m6A modification was upregulated in hepatocytes during ALI. Inhibition of METTL3-mediated m6A modification alleviated hepatocyte pyroptosis and ALI. Through MeRIP-seq analysis and verification, was determined as the target of METTL3-mediated m6A modification, which was regulated in an IGF2BP2-dependent manner. Mechanistically, METTL3 can bind to DLGAP5, and then DLGAP5 promoted pyroptosis through NF-κB-dependent NLRP3 inflammasome activation and direct potentiation of inflammasome structure formation and assembly. mutation or AT9283-mediated DLGAP5 inhibition alleviated pyroptosis and ALI. The effects of hepatocyte pyroptosis on cell interaction were then explored and we revealed that NLRP3 inflammasome and interleukin releasing by the GSDMD-N-dependent membrane pores from pyroptotic hepatocytes activated macrophage metabolic reprogramming and M1 polarization, further exacerbating ALI. deficiency alleviated ALI by suppressing hepatocyte pyroptosis and blocking communication between macrophages and hepatocytes. Our findings indicate the potential mechanisms of ALI from an intercellular communication perspective, and targeted-inhibition of DLGAP5 and -blockade of hepatocyte-macrophage interaction provide promising strategies for ALI treatment.
细胞焦亡是一种存在于炎症性疾病中的新型程序性细胞死亡，而甲基转移酶样3（METTL3）是一种核心的N6-甲基腺苷（m6A）修饰甲基转移酶，已被证明可调节细胞命运。然而，细胞焦亡在急性肝损伤（ALI）中的作用仍不清楚，其是否受m6A修饰调控尚待阐明。在此，构建了突变和敲除小鼠，建立了CCl4和TAA诱导的ALI模型并分离了原代细胞，对细胞焦亡和m6A修饰进行了评估。我们发现肝细胞焦亡是ALI的关键特征，且ALI期间肝细胞中METTL3介导的m6A修饰上调。抑制METTL3介导的m6A修饰可减轻肝细胞焦亡和ALI。通过MeRIP-seq分析和验证，确定IGF2BP2为METTL3介导的m6A修饰的靶点，其以IGF2BP2依赖的方式受到调控。机制上，METTL3可与DLGAP5结合，然后DLGAP5通过NF-κB依赖的NLRP3炎性小体激活以及直接增强炎性小体结构形成和组装来促进细胞焦亡。IGF2BP2突变或AT9283介导的DLGAP5抑制可减轻细胞焦亡和ALI。随后探讨了肝细胞焦亡对细胞相互作用的影响，我们发现焦亡肝细胞中由GSDMD-N依赖性膜孔释放的NLRP3炎性小体和白细胞介素激活了巨噬细胞代谢重编程和M1极化，进一步加重了ALI。IGF2BP2缺乏通过抑制肝细胞焦亡和阻断巨噬细胞与肝细胞之间的通讯减轻了ALI。我们的研究结果从细胞间通讯角度揭示了ALI的潜在机制，靶向抑制DLGAP5和阻断肝细胞-巨噬细胞相互作用为ALI治疗提供了有前景的策略。
