Development of a novel BSL2 system for high-throughput analysis of hantavirus entry glycoproteins

开发用于汉坦病毒侵入糖蛋白高通量分析的新型 BSL2 系统

• 批准号: 10597757
• 负责人: Rohit K Jangra
• 金额: $ 20.86万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-28 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10597757
• 关键词: Address; Americas; Biology; Cardiopulmonary; Case Fatality Rates; Containment; Development; FDA approved; Glycoproteins; Hantavirus; Hantavirus Infections; Hemorrhagic Fever with Renal Syndrome; Human; Immune response; Immunology; Mediating; Molecular; Pathologic Processes; Persons; Plasmids; Public Health; Research; Rodent; Role; South American; Surface; Syndrome; System; Vaccines; Virion; Virus; base; genetic analysis; high throughput analysis; novel; reverse genetics; transmission process

Hantaviruses are the causative agents of hantavirus cardiopulmonary syndrome (HCPS) and hemorrhagic fever with renal syndrome (HFRS) in Americas and Eurasia, respectively. These viruses are usually transmitted from their rodent reservoirs to human. However, multiple incidents of person-to-person transmission of a South American Andes hantavirus raises significant public health concern about these deadly viruses with up to 40% case fatality rates. No FDA-approved hantavirus vaccines and therapies exist. Cellular entry and infection of hantaviruses is mediated by its virion surface Gn/Gc glycoproteins, which are also the main target of protective immune responses. However, our understanding of the molecular determinants of Gn/Gc in hantavirus entry and antigenicity remains limited, at least partly, due to the general requirement of Biosafety level-3 (BSL3) containment for hantavirus research and the lack of a reverse genetics system. To address these limitations, multiple BSL2 pseudovirus systems have been developed. However, current BSL2 systems are limited by poor scalability and limited diversity of the generated viruses due to their inefficient plasmid-based rescue, which makes them incompatible with a more comprehensive analysis of the biology and function of the entry glycoproteins. Here, we propose to fix this major shortcoming of the hantavirus field by developing a novel BSL2 system that allows comprehensive reverse as well as forward genetic analysis of Gn/Gc’s role in virus entry and antigenicity.

汉坦病毒分别是美洲和欧亚大陆汉坦病毒心肺综合征（HCPS）和肾综合征出血热（HFRS）的病原体。这些病毒通常从啮齿动物宿主传播给人类。然而，南美安第斯山脉汉坦病毒的多起人际传播事件引发了人们对这些致命病毒的严重公共卫生担忧，病死率高达 40%。目前还没有 FDA 批准的汉坦病毒疫苗和疗法。汉坦病毒的细胞进入和感染是由其病毒颗粒表面 Gn/Gc 糖蛋白介导的，这也是保护性免疫反应的主要目标。然而，我们对汉坦病毒进入和抗原性中 Gn/Gc 的分子决定因素的理解仍然有限，至少部分是由于汉坦病毒研究的生物安全 3 级（BSL3）遏制的一般要求以及反向遗传学系统的缺乏。为了解决这些限制，开发了多种 BSL2 假病毒系统。然而，当前的 BSL2 系统由于基于质粒的救援效率低下，受到可扩展性差和所生成病毒多样性有限的限制，这使得它们与对进入糖蛋白的生物学和功能进行更全面的分析不相容。在这里，我们建议通过开发一种新型 BSL2 系统来解决汉坦病毒领域的这一主要缺点，该系统允许对 Gn/Gc 在病毒进入和抗原性中的作用进行全面的反向和正向遗传分析。