Viral and host determinants of susceptibility of diverse hantaviruses

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- 经批准的汉坦病毒药物和疫苗不存在。RNA测序的最新进展 科技导致发现了40多种不同基因的汉坦病毒，这些病毒是由 啮齿动物、食虫动物(鼹鼠和地鼠)和蝙蝠。因此，人畜共患病溢出的可能性 未来可能会更高，因为森林砍伐、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群体。 尽管它们具有巨大的遗传多样性和对公共健康的重要性，但我们对 汉坦病毒易感性(病毒进入能力)和通透性的分子决定因素 (病毒复制能力)在细胞水平上是根据仅有的几项研究得出的 汉坦病毒，目前仍处于萌芽状态。这在很大程度上是由于缺乏分子工具和 汉坦病毒分离株及生物安全3级(BSL3)汉坦病毒研究的一般需要 遏制。 代用病毒，如重组水疱性口炎病毒(RVSV)，假型为 汉坦病毒进入Gn/GC糖蛋白为研究病毒进入和 BSL-2控制下的病毒-宿主相互作用。给定G_n(G_n-)的N-末端整环 NTD)形成了病毒粒子表面Gn/GC尖峰的大部分表面暴露部分，并且是 Gn/GC的最大分歧区域，我们假设：(I)Gn-NTD多样性是一种关键的病毒 汉坦病毒易感性的决定因素，以及(Ii)我们最近发现的新的汉坦病毒受体 PCDH1是钙粘附素超家族成员，至少需要一个新的子集才能进入 汉坦病毒。我们的主要目标是生成具有良好特性的分子工具来定义 汉坦病毒易感性和易感性的病毒和宿主决定因素。与 夏威夷大学的Richard Yanagihara说，我们将(I)产生携带汉坦病毒的rVSV 代表哺乳动物汉坦病毒多样性的gn/gc糖蛋白，(Ii)定义候选 新型汉坦病毒进入细胞所需的汉坦病毒受体，以及(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化细胞系 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 在分子水平上的汉坦病毒易感性和通透性。

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.