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

基本信息

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
项目类别：
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

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

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项目摘要

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（HCP）和肾脏综合征（HFRS）的出血热，病例死亡率分别高达40％和14.5％。不存在FDA-批准的汉塔病毒药物和疫苗。 RNA测序技术的最新进展导致发现了40多种啮齿动物，昆虫动物（痣和sh）和蝙蝠携带的遗传上不同的汉坦病毒。因此，随着森林砍伐，栖息地破坏和气候变化的机会可能会使更多的汉塔病毒储量宿主更加接近人类人口，将来，人畜共患溢出的机会可能会更高。尽管它们具有巨大的遗传多样性和公共健康的重要性，但我们对汉坦病毒易感性（病毒入口的能力）的分子决定因素和细胞水平上的允许性（病毒复制能力）的理解仅从几个汉坦病毒的研究中得出，并且仍然是原始的。这在很大程度上是由于缺乏分子工具和汉坦病毒分离株，并且通常需要研究生物安全3级（BSL3）遏制中的汉坦病毒。病毒体表面GN/GC糖蛋白是汉塔病毒易感性的唯一病毒决定因素。尽管对于某些汉坦病毒而存在，但在我们以外的研究人员中仅对一种汉塔病毒（Andes病毒）生成了自我复制的替代病毒，但自我复制的替代病毒仅生成了自我复制的替代病毒。自我复制，荧光标记的RVSV提供了可靠的可扩展系统，这些系统非常适合使用BSL-2上的生存和/或基于基于FACS的全基因组基因筛选的新型宿主因子发现。鉴于GN（GN-NTD）的N末端结构域形成了GN/GC在病毒体表面上尖峰的大多数表面暴露部分，并且是GN/GC最不同的区域，我们假设（i）GNTD多样性是Hantavirus的关键病毒性，不适合（II）的（ii）的（II）的（II）均不及格（II），并且是-II（II）的ntress ersiatient n n n n n n n n n sendy（II）。汉坦病毒的子集。我们的主要目标是生成良好的分子工具来定义汉塔病毒易感性和允许性的病毒和宿主决定因素。 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正宗的非隆起汉坦病毒。这些工具将有助于我们实现长期目标，即对汉塔病毒易感性和分子水平上的汉塔病毒敏感性和允许性的宿主决定因素进行更全面的了解。