Effects of Interferon on primate lentiviruses

干扰素对灵长类慢病毒的影响

基本信息

批准号：
10619797
负责人：
Paul D. Bieniasz
金额：
$ 71.01万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-21 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10619797
关键词：
Acquired Immunodeficiency Syndrome Acute Affect Amino Acid Substitution Animal Model Attenuated Bar Codes Binding Binding Sites Biological Models CD8-Positive T-Lymphocytes CRISPR screen Cell Culture Techniques Cells Chronic Clinical Development Drug Kinetics Exhibits Genes Goals Grant HIV resistance HIV-1 Host Defense Human Image In Vitro Infection Innate Immune Response Interferon Type I Interferon-alpha Interferons Knock-out Knowledge Laboratories Lentivirus Infections Link Macaca Macaca nemestrina Mediating Methods Modeling Molecular Molecular Cloning Pathogenesis Phase Predisposition Primate Lentiviruses Property Proteins Role SIV Testing Time Variant Viral Virus Virus Inhibitors Virus Replication Work acute infection base cell type cross-species transmission env Gene Products experimental study gene product in vivo nonhuman primate novel preservation response simian human immunodeficiency virus tool viron

项目摘要

ABSTRACT This proposal is a multi PI proposal that aims to understand how antiviral proteins limit host range. Prior work has demonstrated that type I interferon (IFN) induced proteins limit primate lentivirus replication in natural and non-natural hosts. Indeed, our previous studies have led to the discovery of antiviral interferon stimulated genes (ISGs), elucidated their mechanism of action and uncovered ways in which primate lentiviruses evolve evade or counteract antiviral proteins. Additionally, we have exploited this knowledge to generate novel chimeric viruses that better represent the HIV-1 strains circulating in humans for use in non-human primate models, including a minimally modified HIV-1 strain (stHIV) that can cause AIDS in pigtail macaques when CD8+ cells are transiently depleted during the acute infection. In this new proposal, we will explore the role of type I IFN and novel antiviral ISGs in limiting primate lentivirus replication in vitro and in vivo. Specific Aim 1 will explore the mechanism of action of antiviral ISGs affecting viral entry. Using a CRISPR screen, we have found novel ISGs whose expression appears to inhibit HIV-1 infection, specifically at the virus entry step. Knockout of one of these genes enhances HIV-1 infection in a manner that varies dramatically according to the particular strain from which the Env protein is derived. Importantly, the magnitude of effect of the ISG on HIV-1 infection mediated by a particular Env variant correlates with the sensitivity of a virus carrying that Env variant to inhibition by type I IFN. We will determine the molecular details of the how IFN/ISGs inhibits HIV-1 entry by elucidating viral and host determinants of inhibition, across cell types and species, and use imaging and other approaches to determine how inhibition affects incoming viron fate. We will also determine how novel ISGs contribute to the differential IFN sensitivity of primary transmitted founder and chronic HIV-1 strains and adapted/unadapted SHIVs. In Specific Aim 2, we will use newly developed, more effective methods to perturb type I IFN in macaques to determine the effect of IFN on primate lentivirus replication therein. In particular, we will determine the effect of type I IFN blockade on stHIV replication and clinical course in pigtail macaques. We will also use SHIV to determine whether HIV-1 Env variants that exhibit differential type I IFN sensitivity in vitro, also do so in vivo. Finally, we will determine how type I IFN affects SHIV dissemination and competitiveness use barcoded SHIVs, bearing IFN/ISG-sensitive and resistant HIV-1 Env proteins, defined in Aim 1, by determining how efficiently each SHIV disseminates in macaques in the presence and absence of type I IFN blockade.

抽象的该提案是一项多 PI 提案，旨在了解抗病毒蛋白如何限制宿主范围。之前的工作已证明 I 型干扰素 (IFN) 诱导蛋白限制灵长类慢病毒在自然和环境中的复制非自然宿主。事实上，我们之前的研究已经发现了抗病毒干扰素刺激基因（ISG），阐明了它们的作用机制并揭示了灵长类慢病毒进化的方式逃避或抵消抗病毒蛋白。此外，我们还利用这些知识来生成新颖的更好地代表人类中传播的 HIV-1 病毒株的嵌合病毒，用于非人类灵长类动物模型，包括经过最低限度改造的 HIV-1 毒株（stHIV），该毒株在以下情况下可导致猪尾猕猴感染艾滋病： CD8+ 细胞在急性感染期间短暂耗尽。在这个新提案中，我们将探讨 I 型 IFN 和新型抗病毒 ISG 限制灵长类慢病毒体外和体内复制。具体目标 1 将探讨抗病毒 ISG 影响病毒进入的作用机制。使用 CRISPR 筛选，我们有发现了新的 ISG，其表达似乎可以抑制 HIV-1 感染，特别是在病毒进入步骤。敲除这些基因之一会增强 HIV-1 感染，其增强方式根据病毒的不同而有显着差异。 Env 蛋白源自的特定菌株。重要的是，ISG 对 HIV-1 的影响程度由特定 Env 变体介导的感染与携带该 Env 变体的病毒的敏感性相关 I 型 IFN 的抑制作用。我们将通过以下方式确定 IFN/ISG 如何抑制 HIV-1 进入的分子细节：阐明跨细胞类型和物种的病毒和宿主抑制决定因素，并使用成像和其他方法确定抑制如何影响传入病毒命运的方法。我们还将确定 ISG 的新颖程度导致原发性HIV-1病毒株和慢性HIV-1病毒株对干扰素的敏感性不同，适应/未适应的SHIV。在具体目标2中，我们将使用新开发的、更有效的方法来扰动在猕猴体内进行 I 型 IFN，以确定 IFN 对灵长类动物慢病毒复制的影响。特别是，我们将确定 I 型 IFN 阻断对猪尾猕猴中 STHIV 复制和临床病程的影响。我们还将使用 SHIV 来确定 HIV-1 Env 变体是否在体外表现出不同的 I 型 IFN 敏感性，在体内也这样做。最后，我们将确定 I 型干扰素如何影响 SHIV 传播和竞争力使用条形码 SHIV，带有目标 1 中定义的 IFN/ISG 敏感且耐药的 HIV-1 Env 蛋白，通过确定每种 SHIV 在存在和不存在 I 型 IFN 的情况下在猕猴中传播的效率封锁。