Discovery and Characterization of Capsid-Targeting Lentiviral Restriction Factors

项目摘要： 作为一种专性细胞内寄生虫，HIV像所有病毒一样，依赖于宿主编码的因子， 在宿主细胞内完成生命周期。然而，病毒也必须通过以下方式逃避识别： 专门的宿主因子已经进化到专门保护细胞免受病毒入侵。 一些抗病毒基因是干扰素诱导的转录程序的一部分， 宿主在感知到病毒入侵时就会释放出来。对宿主基因的全面描述 阻止灵长类慢病毒成功感染人类细胞的方法 使用当前方法进行表征。在目标1中，我们将描述 抑制HIV-1和相关灵长类猴免疫缺陷病毒（SIV）变体 TRIM 34是我们发现的一种TRIM 5蛋白，特异性靶向HIV-1衣壳变体， SIV在目标2中，我们将探索TRIM 34作为灵长类动物中广泛作用的抗病毒基因的作用 以及TRIM 34在限制HIV-1感染演变方面的潜在作用 在主机内。在目标3中，我们将进行全基因组HIV-CRISPR筛选方法， 找到抑制HIV-1衣壳突变体的干扰素刺激基因（ISG）。除了 在人类细胞中通过TRIM 34定义了一种新的HIV限制，这项工作有可能 发现新的宿主抗病毒基因，限制SIV和HIV病毒株在人类细胞中复制。 后续研究将集中在进一步了解的作用机制， 感兴趣的基因以及病毒拮抗或逃逸的机制。最终，操纵 这些因素对于旨在实现功能性HIV治愈的方法可能是重要的。

Project Summary: As an obligate intracellular parasite HIV, like all viruses, relies on host-encoded factors to complete its life cycle inside the host cell. However, the virus must also evade recognition by specialized host factors that have evolved specifically to defend cells against viral invasion. Some of the antiviral genes are part of an Interferon-induced transcriptional program that is deployed by the host on sensing of a viral invader. A comprehensive description of host genes that block primate lentiviruses from successfully infecting human cells has evaded characterization with current methodologies. In Aim 1 we will describe the mechanism of inhibition of HIV-1 and related primate Simian Immunodeficiency Virus (SIV) variants by TRIM34, a TRIM5 paralog we have discovered to specifically target HIV-1 capsid variants and SIVs. In Aim 2 we will explore the role of TRIM34 as a broadly acting antiviral gene in primates as well as the potential role of TRIM34 in contributing to limiting the evolution of HIV-1 infections within the host. In Aim 3 we will perform a whole-genome HIV-CRISPR screening approach to find the Interferon-Stimulated Genes (ISGs) that inhibit HIV-1 capsid mutants. In addition to defining a novel HIV restriction by TRIM34 in human cells, this work has the potential to discover novel host antiviral genes that limit SIV and HIV strains from replicating in human cells. Follow-up studies will be focused on furthering our understanding the mechanism of action of genes of interest as well as mechanisms of viral antagonism or escape. Ultimately, manipulation of these factors could be important for approaches aimed at achieving a functional HIV cure.