甲型流感病毒SUMO化修饰的条件性敲除小鼠感染模型和致病性研究

The pathogenesis study on influenza A virus is the basis for disease treatments and epidemic controls. We and others previously reported that influenza A virus exploits cellular SUMOylation system for their own benefits. However, how sumoylation contributes to disease outcomes and the virus pathogenicity is yet unknown. In this study, we will take advantage of the reverse genetic technology to generate a recombinant virus stably expressing Cre recombinase. This recombinant Cre-expressing virus will be used to infect loxP-floxed sumoylation enzymes conditional KO mice, so that sumoylation enzymes will be knocked off particularly in the infected-tissues but not in the healthy tissues. By using this infection model, we will be able to study the role of sumoylation in influenza A virus infected animals, to demonstrate the location and the pathogenesis of viral sumoylation in vivo, and to validate the contribution of sumoylation to disease outcomes. This study will help to demonstrate the role of sumoylation in virus pathogenicity; will generate a conditional-KO mouse model in infected tissues; will validate the relevance of the key virus-host interaction to disease development; and will provide theoretic supports for developing anti-viral drugs based on inhibiting the virus-host interactions.

甲型流感病毒的致病机制研究是疾病治疗和疫情防控的基础，近年来相继报道了甲型流感病毒的复制强烈依赖于宿主细胞的SUMO化修饰系统，但SUMO化在体内感染中如何影响了病毒的致病性以及作用机制仍不清楚。本研究拟通过反向遗传学技术构建表达Cre重组酶的甲型流感病毒，用来感染“LoxP-floxed”转基因小鼠，从而实现在病毒感染组织中特异性敲除宿主SUMO化修饰酶，研究体内感染状态下病毒蛋白SUMO化发生的部位及其致病机制，验证SUMO化修饰对甲型流感病毒致病性的重要贡献。该研究对于揭示SUMO化修饰如何影响感染性疾病的发生发展有重要意义；感染部位条件性敲除技术有助于评价病毒宿主蛋白相互作用的疾病相关性；为研发以阻断病毒蛋白SUMO化修饰为靶标的抗病毒药物提供小鼠感染模型和科学依据。

甲型流感病毒的致病机制研究是疾病治疗和疫情防控的基础，近年来相继报道了甲型流感病毒的复制强烈依赖于宿主细胞的SUMO化修饰系统，但SUMO化在体内感染中如何影响了病毒的致病性以及作用机制仍不清楚。本研究拟通过反向遗传学技术构建表达Cre重组酶的甲型流感病毒，用来感染“LoxP-floxed”转基因小鼠，从而实现在病毒感染组织中特异性敲除宿主SUMO化修饰酶，研究体内感染状态下病毒蛋白SUMO化发生的部位及其致病机制，验证SUMO化修饰对甲型流感病毒致病性的重要贡献。结果表明SUMO化修饰促进了甲型流感病毒的致病性，条件性敲除SUMO修饰的E2酶可以减轻流感病毒感染引起的死亡，揭示了蛋白修饰是有效的抗病毒靶点。项目相关成果还发现了多种蛋白修饰对甲型流感病毒的促进作用。以通讯作者（含共同）在Nature Communications, PLoS Pathogens，Vaccine发表3篇研究论文。以第一发明人申请专利2项，其中1项靶向宿主酶分子的抗病毒候选新药专利实现专利成果转让，合同额2000万人民币。项目负责人获得2019年国家自然科学基金优秀青年基金，项目培养的博士后获得包括博士后面上基金、国家自然科学基金青年基金等项目支持。