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肠道病毒D68(Enterovirus D68, EV-D68)与儿童严重呼吸道感染和急性弛缓性脊髓炎（Acute flaccid myelitis, AFM）相关，但其导致瘫痪的分子机制尚未阐明。为深入探究这一机制，本研究以2018年从儿童患者体内分离的一株EV-D68(D2基因亚型)为亲本株，通过构建ICR小鼠适应性株，成功获得了P0代至P9代的系列传代毒株。经全基因组测序分析，显示VP1-N90K是唯一位于病毒与受体结合区的位点，也是P2代与P4代毒株之间的唯一氨基酸差异位点。体内体外实验显示VP1-N90K突变可导致小鼠出现更强的致病性和细胞病变效应（Cytopathic effect, CPE）。为揭示VP1-N90K突变增强病毒致病性的分子基础，通过分子模拟技术分析发现VP1-N90K突变显著增强与宿主受体的结合亲和力。综上，本研究得出核心结论：VP1-N90K是增强EV-D68神经毒力的关键突变。该突变通过优化VP1与宿主受体的结合能力，显著促进病毒在宿主细胞表面的吸附、内化过程，以及在神经细胞内的复制效率，最终导致AFM瘫痪表型的发生。本研究结果为EV-D68致病机制的深入解析提供了关键实验依据，也为针对性抗病毒策略（如靶向VP1受体结合域的药物研发）的制定奠定了重要基础。
Abstract：Enterovirus D68(EV-D68) is associated with severe respiratory illness and acute flaccid myelitis(AFM) in children, yet the molecular mechanism driving paralysis remains poorly defined. In this study, an EV-D68 D2-subclade clinical isolate obtained from a pediatric patient in 2018 was used as the parental strain to generate a mouse-adapted virus through serial passage in ICR neonatal mice, yielding viral populations from passages P0 to P9. Whole-genome sequencing revealed that VP1-N90K was the only amino acid substitution within the receptor-binding region and the sole difference between P2 and P4 viral populations. In vivo and in vitro experiments demonstrated that the VP1-N90K mutation significantly increased viral pathogenicity in neonatal mice and induced stronger cytopathic effects in SH-SY5Y neuroblastoma and RD rhabdomyosarcoma cells. Molecular simulation analysis further showed that VP1-N90K markedly enhanced the binding affinity of VP1 to its host receptors, including sialic acid and MFSD6. Collectively, these findings identify VP1-N90K as a critical determinant of EV-D68 neurovirulence. By strengthening VP1– receptor interactions, this mutation promotes viral attachment, internalization, and replication within neuronal cells, ultimately contributing to AFM-like paralysis. This work provides mechanistic insight into EV-D68-induced neuropathogenesis and informs the development of targeted antiviral interventions, including agents directed against the VP1 receptor-binding interface.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250320
中图分类号:R373.2
引用信息:
[1]段伟,李冀琛,李慧洁,等.肠道病毒D68型VP1-N90K突变增强其在乳鼠感染致病力研究[J].病毒学报,2025,41(06):1710-1722.DOI:10.13242/j.cnki.bingduxuebao.250320.
基金信息:
国家疾病预防控制局公共卫生人才培养支持项目（项目号：GJJKJ-2024-ZY）; 传染病溯源预警与智能决策全国重点实验室（NITFID）资助项目（项目号：ZDGWNLJS25-36）,题目：未知及新发病原体监测及预测预警技术研究; 国家重点研发计划（项目号：2021YFC2302003）,题目：病毒监测网络数据标准及数据平台建设~~