The CVB3-induced P62 cleavage promotes ferroptosis via NRF2/GPX4 and facilitates viral replication

Hailan Yao^*Feng HeZhuo LiuMiao FengDingding CaoSen LiXuelai LiuYiting JiangXiaoyu YiZhewei Liu

• Capital Institute of Pediatrics, Beijing, China

Coxsackie B group type III virus (CVB3) is a serious pathogen of viral myocarditis and its replication in the heart and other organs could cause severe damage to the hosts. The mechanisms underlying CVB3-induced pathological effects, particularly concerning ferroptosis-a regulated cell death driven by iron-dependent lipid peroxidation-remain incompletely understood. In our study, we demonstrate that CVB3 infection triggers ferroptosis by cleaving the selective autophagy receptor P62 and this cleavage leads to the degradation of NRF2 and the subsequent downregulation of GPX4, a central ferroptosis inhibitor. We identified that CVB3 replication results in the proteolytic cleavage of P62 into Cand N-terminal fragments. Exogenous non-cleavable P62 mutant significantly attenuated ferroptosis both in vitro and in vivo by stabilizing the KEAP1/NRF2/GPX4 axis. Although P62 typically functions in ubiquitin-dependent selective autophagy, we found that GPX4 expression were not regulated by ubiquitination during CVB3 infection. Additionally, a high diet of selenium which is also an essential element in the human antioxidant system and is necessary for the synthesis of GPX4, supressed ferroptosis and improved survival rate in CVB3 infected mice. Collectively, our findings establish a novel mechanism that CVB3 cleaves P62 to downregulate the KEAP1/NRF2/GPX4 axis, thereby driving ferroptosis and disease progression, and identify restoration of P62 or selenium supplementation as a potential therapeutic strategy.