Viral and host determinants of susceptibility of diverse hantaviruses

不同汉坦病毒易感性的病毒和宿主决定因素

• 批准号: 10538154
• 负责人: Rohit K Jangra
• 金额: $ 16.6万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10538154
• 关键词: Viral; host; determinants; susceptibility; diverse

Hantaviruses cause hantavirus cardiopulmonary syndrome (HCPS) and hemorrhagic fever with renal syndrome (HFRS) with case fatality rates of up to 40% and 14.5%, respectively. FDA- approved hantavirus drugs and vaccine do not exist. Recent advances in RNA sequencing technology has led to the discovery of more than 40 genetically distinct hantaviruses carried by rodents, insectivores (moles and shrews) and bats. Consequently, chances of zoonotic spillover are likely to be higher in future as deforestation, habitat destruction and climate change will bring more of the hantavirus reservoir hosts in closer proximity to the human populations. Despite their huge genetic diversity and public health importance, our understanding of the molecular determinants of hantavirus susceptibility (capacity for virus entry) and permissivity (capacity for virus replication) at the cellular level is derived from studies of only a few hantaviruses and remains rudimentary. This is largely due to the lack of molecular tools and hantavirus isolates and the general need to study hantaviruses in biosafety level-3 (BSL3) containment. Surrogate viruses such as recombinant vesicular stomatitis viruses (rVSVs) pseudotyped with the hantavirus entry Gn/Gc glycoproteins provide excellent tools for investigating virus entry and virus-host interactions under BSL-2 containment. Given that the N-terminal domain of Gn (Gn- NTD) forms most of the surface-exposed part of the Gn/Gc spikes on the virion surface and is the most divergent region of Gn/Gc, we hypothesize that (i) Gn-NTD diversity is a key viral determinant of hantavirus susceptibility, and (ii) our recently identified novel hantavirus receptor PCDH1, a cadherin superfamily member, is required for the entry of at least a subset of novel hantaviruses. Our primary objective is to generate well-characterized molecular tools to define viral and host determinants of hantavirus susceptibility and permissivity. In collaboration with Richard Yanagihara at the University of Hawaii, we will (i) generate rVSVs bearing hantavirus Gn/Gc glycoproteins representing mammalian hantavirus diversity, (ii) define the candidate hantavirus receptor requirements for cellular entry of novel hantaviruses, and (iii) finally, we will apply this knowledge to generate engineered cell lines over-expressing the relevant host factors for the isolation of authentic non-rodent-borne hantaviruses. These tools will help us achieve our long-term goals of generating a more comprehensive picture of the viral and host determinants of hantavirus susceptibility and permissivity at the molecular level.

汉坦病毒引起汉坦病毒心肺综合征（HCPS）和出血热， 肾综合征（HFRS），病死率分别高达40%和14.5%。FDA- 目前还没有批准的汉他病毒药物和疫苗。RNA测序的最新进展 这项技术已经发现了40多种基因上不同的汉他病毒， 啮齿动物、食虫动物（鼹鼠和鼩 鼱）和蝙蝠。因此，人畜共患传染病的机会 由于森林砍伐、栖息地破坏和气候变化， 使更多的汉坦病毒宿主更接近人类。 尽管它们具有巨大的遗传多样性和公共卫生的重要性，但我们对它们的理解是， 汉坦病毒易感性的分子决定因素（病毒进入的能力）和持久性 在细胞水平上对病毒复制能力的研究， 汉他病毒和仍然是基本的。这主要是由于缺乏分子工具， 汉坦病毒分离株和研究生物安全等级3（BSL 3）汉坦病毒的一般需要 安全壳 替代病毒，如重组水泡性口炎病毒（rVSV），用 汉坦病毒进入Gn/Gc糖蛋白为研究病毒进入提供了极好的工具， BSL-2控制下的病毒-宿主相互作用。鉴于Gn的N-末端结构域（Gn- NTD）在病毒体表面上形成Gn/Gc尖峰的大部分表面暴露部分， Gn/Gc最趋异的区域，我们假设（i）Gn-NTD多样性是一个关键的病毒 汉坦病毒易感性的决定因素，以及（ii）我们最近鉴定的新型汉坦病毒受体 PCDH 1是钙粘蛋白超家族成员，是进入至少一个新的肿瘤细胞亚群所必需的。 汉他病毒我们的主要目标是产生良好表征的分子工具来定义 汉坦病毒易感性和持久性的病毒和宿主决定因素。协同 夏威夷大学的Richard Yanagihara博士说，我们将（i）生产携带汉坦病毒的rVSV Gn/Gc糖蛋白代表哺乳动物汉坦病毒多样性，（ii）定义候选 新型汉他病毒进入细胞的汉他病毒受体要求，和（iii）最后，我们将 应用这些知识来产生过表达相关宿主因子的工程细胞系 用于分离真正的非啮齿动物传播的汉坦病毒。这些工具将帮助我们实现 更全面地了解病毒和宿主决定因素的长期目标 在分子水平上对汉坦病毒的易感性和持久性进行研究。