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: 10303812
• 负责人: Rohit K Jangra
• 金额: $ 10万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 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 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10303812
• 关键词: Ablation; Andes Virus; Biochemical; Biological Assay; Biology; CRISPR/Cas technology; Cadherins; Cardiopulmonary; Case Fatality Rates; Cell Culture Techniques; Cell Line; Cells; Chiroptera; Codon Nucleotides; Collaborations; Collection; Containment; Data; Deforestation; Disease; Endothelial Cells; Engineering; Epidemiology; FDA approved; Family; Fatality rate; Future; GC gene; Generations; Genetic; Genetic Screening; Genetic Variation; Glycoproteins; Goals; Habitats; Hantavirus; Hantavirus Infections; Haploid Cells; Hawaii; Head; Hemorrhagic Fever with Renal Syndrome; Human; ITGB3 gene; Immune response; In Vitro; Incidence; Infection; Integration Host Factors; Knowledge; Label; Libraries; Light; Mediating; Mole the mammal; Molecular; Molecular Virology; Mutation; N-terminal; Pharmaceutical Preparations; Phylogenetic Analysis; Plasmids; Population; Population Growth; Predisposition; Primary carcinoma of the liver cells; Prospect Hill virus; Public Health; Reagent; Reporter; Research; Research Personnel; Retroviridae; Rodent; Sequence Analysis; Serology; Shrews; Sin Nombre virus; Surface; Syndrome; System; TNFSF5 gene; Technology; Testing; Therapeutic; Therapeutic antibodies; Universities; Vaccines; Viral; Virion; Virulence; Virus; Virus Replication; Zoonoses; antibody inhibitor; base; climate change; experience; genome-wide; in vivo; insight; member; novel; pathogen; permissiveness; receptor; response; tool; transcriptome sequencing; vaccine candidate; virus host interaction; zoonotic spillover

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 permissiveness (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. The virion surface Gn/Gc glycoproteins are the sole viral determinants of hantavirus susceptibility. Although single-cycle pseudotyped viruses carrying Gn/Gc exist for some hantaviruses, self-replicating surrogate viruses have only been generated for one hantavirus (Andes virus) by researchers other than us. Self-replicating, fluorescent-labeled rVSVs provide robust, scalable systems that are uniquely suited for novel host factor discovery using survival and/or FACS-based genome-wide genetic screens at BSL-2. 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) unidentified non- PCDH1 receptor(s) mediate entry of at least a subset of 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 host factor 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 permissiveness at the molecular level.

汉坦病毒引起汉坦病毒心肺综合征（HCPS）和肾综合征出血热（HFRS），病死率分别高达40%和14.5%。FDA批准的汉他病毒药物和疫苗并不存在。RNA测序技术的最新进展已经发现了40多种基因上不同的汉坦病毒，这些病毒由啮齿动物、食虫动物（鼹鼠和地鼠）和蝙蝠携带。因此，随着森林砍伐、栖息地破坏和气候变化将使更多的汉坦病毒宿主更接近人类，人畜共患疾病蔓延的可能性将来可能更高。尽管汉他病毒具有巨大的遗传多样性和公共卫生重要性，但我们对细胞水平上汉他病毒易感性（病毒进入的能力）和容许性（病毒复制的能力）的分子决定因素的理解仅来自对少数汉他病毒的研究，并且仍然是初步的。这在很大程度上是由于缺乏分子工具和汉他病毒分离株，以及普遍需要研究生物安全3级（BSL 3）遏制汉他病毒。病毒粒子表面的Gn/Gc糖蛋白是汉坦病毒易感性的唯一决定因素。虽然携带Gn/Gc的单循环假型病毒存在于一些汉他病毒中，但除我们之外的研究人员仅为一种汉他病毒（安第斯山脉病毒）产生了自我复制的替代病毒。自我复制、荧光标记的rVSV提供了稳健的、可扩展的系统，其独特地适合于在BSL-2处使用存活和/或基于FACS的全基因组遗传筛选来发现新的宿主因子。考虑到Gn的N-末端结构域（Gn-NTD）形成了病毒体表面上Gn/Gc刺突的大部分表面暴露部分，并且是Gn/Gc最趋异的区域，我们假设（i）Gn-NTD多样性是汉坦病毒易感性的关键病毒决定因素，（ii）未鉴定的非PCDH 1受体介导了至少一个汉坦病毒亚群的进入。我们的主要目标是产生良好的特征分子工具，以确定病毒和宿主决定因素的汉坦病毒的易感性和持久性。与夏威夷大学的Richard Yanagihara合作，我们将（i）产生代表哺乳动物汉他病毒多样性的携带汉他病毒Gn/Gc糖蛋白的rVSV，（ii）确定新型汉他病毒进入细胞的宿主因子要求，（iii）最后，我们将应用这些知识产生过表达相关宿主因子的工程细胞系，用于分离真实的非啮齿动物传播的汉他病毒。这些工具将帮助我们实现我们的长期目标，即在分子水平上更全面地了解汉坦病毒易感性和宽容性的病毒和宿主决定因素。