肠道病毒D68(EV-D68),导致严重的呼吸系统疾病和不可逆的中枢神经系统损伤,已成为世界性的严重公共卫生问题。然而,EV-D68发挥神经毒性的机制尚不清楚.因此,我们旨在分析EV-D68感染对卵裂的影响,亚细胞易位,和TARDNA结合蛋白43kDa(TDP-43)在呼吸或神经细胞中的致病性聚集。结果表明,EV-D68编码的蛋白酶2A和3C诱导了TDP-43的易位和切割,分别。具体来说,在图3C中,TDP-43的裂解残基327Q。与野生型TDP-43相比,3C介导的切割的TDP-43片段具有显著降低的蛋白质溶解度。因此,3C活动促进了TDP-43的聚集,对不同的人类细胞产生细胞毒性,包括胶质母细胞瘤T98G细胞。筛选了市售抗病毒药物对3C介导的TDP-43裂解的影响,结果表明,洛匹那韦是EV-D683C蛋白酶的有效抑制剂。总的来说,这些结果表明TDP-43是EV-D683C的保守宿主靶标。这项研究首次提供了TDP-43失调参与EV-D68发病机制的证据。重要性在过去的十年里,肠道病毒D68(EV-D68)感染的发病率在全球范围内有所增加.EV-D68感染可引起不同的呼吸道症状和严重的神经系统并发症,包括急性弛缓性脊髓炎.因此,阐明EV-D68毒性的潜在机制对于开发预防EV-D68感染相关疾病的新方法很重要。这项研究表明,EV-D68感染引发了转位,乳沟,和TDP-43的聚集,TDP-43是一种与退行性神经系统疾病密切相关的细胞内蛋白。病毒蛋白酶3C降低了TDP-43的溶解度,从而对宿主细胞施加细胞毒性,包括人类胶质母细胞瘤细胞.因此,抵消3C活性是缓解EV-D68触发的细胞死亡的有效策略。TDP-43的细胞质聚集是退行性疾病的标志,有助于神经细胞损伤和中枢神经系统(CNS)疾病。这项关于EV-D68诱导的TDP-43形成的研究结果扩展了我们对病毒介导的细胞毒性以及感染患者中TDP-43功能障碍相关的认知障碍和神经症状的潜在风险的理解。
Enterovirus D68 (EV-D68), which causes severe respiratory diseases and irreversible central nervous system damage, has become a serious public health problem worldwide. However, the mechanisms by which EV-D68 exerts neurotoxicity remain unclear. Thus, we aimed to analyze the effects of EV-D68 infection on the cleavage, subcellular translocation, and pathogenic aggregation of TAR DNA-binding protein 43 kDa (TDP-43) in respiratory or neural cells. The results showed that EV-D68-encoded proteases 2A and 3C induced TDP-43 translocation and cleavage, respectively. Specifically, 3C cleaved residue 327Q of TDP-43. The 3C-mediated cleaved TDP-43 fragments had substantially decreased protein solubility compared with the wild-type TDP-43. Hence, 3C activity promoted TDP-43 aggregation, which exerted cytotoxicity to diverse human cells, including glioblastoma T98G cells. The effects of commercially available antiviral drugs on 3C-mediated TDP-43 cleavage were screened, and the results revealed lopinavir as a potent inhibitor of EV-D68 3C protease. Overall, these results suggested TDP-43 as a conserved host target of EV-D68 3C. This study is the first to provide evidence on the involvement of TDP-43 dysregulation in EV-D68 pathogenesis. IMPORTANCE Over the past decade, the incidence of enterovirus D68 (EV-D68) infection has increased worldwide. EV-D68 infection can cause different respiratory symptoms and severe neurological complications, including acute flaccid myelitis. Thus, elucidating the mechanisms underlying EV-D68 toxicity is important to develop novel methods to prevent EV-D68 infection-associated diseases. This study shows that EV-D68 infection triggers the translocalization, cleavage, and aggregation of TDP-43, an intracellular protein closely related to degenerative neurological disorders. The viral protease 3C decreased TDP-43 solubility, thereby exerting cytotoxicity to host cells, including human glioblastoma cells. Thus, counteracting 3C activity is an effective strategy to relieve EV-D68-triggered cell death. Cytoplasmic aggregation of TDP-43 is a hallmark of degenerative diseases, contributing to neural cell damage and central nervous system (CNS) disorders. The findings of this study on EV-D68-induced TDP-43 formation extend our understanding of virus-mediated cytotoxicity and the potential risks of TDP-43 dysfunction-related cognitive impairment and neurological symptoms in infected patients.