State Key Laboratory of Common Mechanism Research for Major Diseases, and Key Laboratory of Synthetic Biology Regulatory Elements, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Suzhou, China.
Cyrus Tang Hematology Center and Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
J Cell Biol. 2023 Dec 4;222(12). doi: 10.1083/jcb.202303108. Epub 2023 Oct 31.
Enterovirus 71 (EV71) and Coxsackie A16 (CVA16) are two major causative agents of hand, foot, and mouth disease (HFMD) in young children. However, the mechanisms regulating the replication and pathogenesis of EV71/CVA16 remain incompletely understood. We performed a genome-wide CRISPR-Cas9 knockout screen and identified Ragulator as a mediator of EV71-induced apoptosis and pyroptosis. The Ragulator-Rag complex is required for EV71 and CVA16 replication. Upon infection, the Ragulator-Rag complex recruits viral 3D protein to the lysosomal surface through the interaction between 3D and RagB. Disruption of the lysosome-tethered Ragulator-Rag-3D complex significantly impairs the replication of EV71/CVA16. We discovered a novel EV71 inhibitor, ZHSI-1, which interacts with 3D and significantly reduces the lysosomal tethering of 3D. ZHSI-1 treatment significantly represses replication of EV71/CVA16 as well as virus-induced pyroptosis associated with viral pathogenesis. Importantly, ZHSI-1 treatment effectively protects against EV71 infection in neonatal and young mice. Thus, our study indicates that targeting lysosome-tethered Ragulator-Rag-3D may be an effective therapeutic strategy for HFMD.
肠道病毒 71 型（EV71）和柯萨奇病毒 A16 型（CVA16）是导致婴幼儿手足口病（HFMD）的两种主要病原体。然而，调节 EV71/CVA16 复制和发病机制的机制仍不完全清楚。我们进行了全基因组 CRISPR-Cas9 敲除筛选，发现 Ragulator 是 EV71 诱导细胞凋亡和细胞焦亡的介质。Ragulator-Rag 复合物是 EV71 和 CVA16 复制所必需的。感染后，Ragulator-Rag 复合物通过 3D 和 RagB 之间的相互作用将病毒 3D 蛋白募集到溶酶体表面。溶酶体 tethered Ragulator-Rag-3D 复合物的破坏显著抑制 EV71/CVA16 的复制。我们发现了一种新型 EV71 抑制剂 ZHSI-1，它与 3D 相互作用，并显著减少 3D 的溶酶体 tethered。ZHSI-1 处理显著抑制 EV71/CVA16 的复制以及与病毒发病机制相关的病毒诱导的细胞焦亡。重要的是，ZHSI-1 处理可有效预防新生和幼鼠的 EV71 感染。因此，我们的研究表明，靶向溶酶体 tethered Ragulator-Rag-3D 可能是治疗 HFMD 的有效策略。
