HBGA receptors in host cell entry and infection of norovirus

HBGA受体在诺如病毒进入宿主细胞和感染中的作用

• 批准号: 9182813
• 负责人: Wen Jiang
• 金额: $ 48.98万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9182813
• 关键词: Alpha Cell; Antibodies; Antigen Receptors; Antiviral Agents; Binding; Biological Assay; Biological Models; Blood Group Antigens; Caliciviridae; Calicivirus; Capsid; Capsid Proteins; Carbohydrates; Cell Culture Techniques; Cell Line; Cells; Collaborations; Confined Spaces; Cryoelectron Microscopy; Disease Outbreaks; Economics; Evaluation; Family; Food Contamination; Gastroenteritis; Genetic; Genome; Goals; Health; Health care facility; Hospitals; Hot Spot; Human; Infection; Kinetics; Knowledge; Laboratories; Life Cycle Stages; Link; Mediating; Membrane; Modeling; Molecular; Molecular Conformation; Molecular Virology; Monitor; Mucins; Mutagenesis; Mutation; Norovirus; Pathogenesis; Pathway interactions; Penetration; Persons; Play; Population; Predisposition; Primates; Procedures; Process; Public Health; RNA; Reagent; Research; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; Saliva; Schools; Site; Societies; Structure; System; Testing; Time; United States; Vertebral column; Viral; Viral Genome; Virion; Virus; Virus Diseases; Virus Replication; Water; antigen binding; base; experimental study; flexibility; genomic RNA; improved; novel; operation; pandemic disease; particle; pathogen; permissiveness; public health relevance; receptor; reverse genetics; screening; structural biology; viral RNA; virus culture; virus host interaction

DESCRIPTION (provided by applicant): We seek to understand the interactions between Human noroviruses (HuNoVs) and their Histo-blood group antigen (HBGA) receptors in the attachment/penetration into host cells and potentially triggering conformational dynamics of the viral capsid and genome release in the early stage of viral infection. HuNoVs in the Caliciviridae family are the major cause of nonbacterial gastroenteritis worldwide. The highly contagious HuNoVs can rapidly spread among the population through contaminated food, water and person-to-person contact in confined spaces, such as cruises, hotels, schools, hospitals and other long-term health care facilities. There are approximately 21 million cases of infection in ~1,500 HuNoVs outbreaks in the United States annually, which causes severe economic loss to society and threatens human health. Despite this serious public health concern, knowledge about the infection mechanism and pathogenesis of HuNoVs is limited due to the lack of a permissive cell line. Our past decade studies showed a strong association of HBGAs as a susceptibility factor in NoV infection, and therefore further studies to seek direct evidence on HBGAs as a receptor of NoVs are necessary. In this R01 application, we will use the recently discovered primate Tulane virus (TV) as the model system to fulfill our goals because of its close genetic and structural similarities to HuNoVs, the availability of an established cultivation system, and the fact that TV also utilizes HBGAs as receptors. We will first test the HBGA requirement in TV infection in cell cultures by performing blocking or inhibition experiments using HBGA-specific blocking reagents. We also will study the roles of HBGAs in the attachment and/or penetration of TV entry into host cells. Furthermore, we will test a novel hypothesis of HBGA-triggered dynamic change of TV capsid on viral genome release to initiate infection. This hypothesis is based on our recent observation of conformational changes of TV following interaction with HBGAs. Finally, we will validate these findings using a reverse genetic system of TV by mutagenesis studies in attempt to determine the hot spots in the capsid protein responsible for these interactions. We also aim to develop the TV culture system into a useful surrogate for antiviral screening/evaluation against HuNoVs. The proposed studies in this application will be performed by collaboration between two research teams on structural biology (Wen Jiang, PI) and molecular virology (Xi Jiang, co-PI) with an excellent collaboration track record in the past. We are confident that we will make a rapid progress in fulfilling our aims.

描述（由申请人提供）：我们寻求了解人类诺如病毒 (HuNoV) 与其组织血型抗原 (HBGA) 受体在附着/渗透到宿主细胞中的相互作用，并可能触发病毒衣壳和基因组释放的构象动力学在病毒感染的早期阶段。杯状病毒科的 HuNoV 是全世界非细菌性胃肠炎的主要原因。具有高度传染性的 HuNoV 可以通过受污染的食物、水以及在邮轮、酒店、学校、医院和其他长期医疗机构等密闭空间内的人与人接触而在人群中迅速传播。美国每年约1,500起HuNoV疫情中约有2100万例感染病例，给社会造成严重经济损失并威胁人类健康。尽管存在严重的公共卫生问题，但由于缺乏允许的细胞系，对 HuNoV 的感染机制和发病机制的了解仍然有限。我们过去十年的研究表明，HBGA 作为 NoV 感染的易感因素有很强的相关性，因此有必要进一步研究以寻求 HBGA 作为 NoV 受体的直接证据。在这个 R01 应用中，我们将使用最近发现的灵长类杜兰病毒 (TV) 作为模型系统来实现我们的目标，因为它与 HuNoV 具有密切的遗传和结构相似性，并且可以建立培养 系统，而且电视也利用 HBGA 作为受体。我们将首先通过使用 HBGA 特异性阻断试剂进行阻断或抑制实验来测试细胞培养物中 TV 感染的 HBGA 要求。我们还将研究 HBGA 在 TV 进入宿主细胞的附着和/或渗透中的作用。此外，我们将测试 HBGA 触发 TV 衣壳对病毒基因组释放的动态变化以引发感染的新假设。这一假设基于我们最近对 TV 与 HBGA 相互作用后构象变化的观察。最后，我们将通过诱变研究使用 TV 的反向遗传系统来验证这些发现，试图确定衣壳蛋白中负责这些相互作用的热点。我们还旨在将电视文化系统发展成为针对 HuNoV 的抗病毒筛查/评估的有用替代品。本申请中拟议的研究将由过去具有良好合作记录的结构生物学（PI，文江）和分子病毒学（Co-PI）两个研究团队合作进行。我们有信心在实现我们的目标方面取得快速进展。