METTL3-dependent m 6 A modification of SNAP29 induces “autophagy-mitochondrial crisis” in the ischemic microenvironment after soft tissue transplantation

作者：Ningning Yang, Yingying Lai, Gaoxiang Yu, Xuzi Zhang, Jingwei Shi, Linyi Xiang, Jiacheng Zhang, Yuzhe Wu, Xiaoqiong Jiang, Xuanlong Zhang, Liangliang Yang, Weiyang Gao, Jian Ding, Xiangyang Wang, Jian Xiao, Kailiang Zhou · 发表于：Autophagy · 年份：2025 · DOI：10.1080/15548627.2025.2493455 · 被引用次数：37 · 研究领域：RNA modifications and cancer、Cancer-related gene regulation、HVDC Systems and Fault Protection

Necrosis at the ischemic distal end of flap transplants increases patients’ pain and economic burden. Reactive oxygen species (ROS) and mitochondrial damage are crucial in regulating parthanatos, but the mechanisms linking disrupted macroautophagic/autophagic flux to parthanatos in ischemic flaps remain unclear. The results of western blotting, immunofluorescence staining, and a proteomic analysis revealed that the autophagic protein SNAP29 was deficient in ischemic flaps, resulting in disrupted autophagic flux, increased ROS-induced parthanatos, and aggravated ischemic flap necrosis. The use of AAV vector to restore SNAP29 in vivo mitigated the disruption of autophagic flux and parthanatos. Additionally, quantification of the total m6A level and RIP-qPCR, MeRIP-qPCR, and RNA stability assessments were performed to determine differential Snap29 mRNA m6A methylation levels and mRNA stability in ischemic flaps. Various in vitro and in vivo tests were conducted to verify the ability of METTL3-mediated m6A methylation to promote SNAP29 depletion and disrupt autophagic flux. Finally, we concluded that restoring SNAP29 by inhibiting METTL3 and YTHDF2 reversed the “autophagy-mitochondrial crisis”, defined for the first time as disrupted autophagic flux, mitochondrial damage, mitochondrial protein leakage, and the occurrence of parthanatos. The reversal of this crisis ultimately promoted the survival of ischemic flaps.Abbreviations: AAV = adeno-associated virus; ACTA2/α-SMA = actin a...