Elimination of persistently HIV-infected cells by targeting host factors

描述：HIV-1的治愈现在似乎是可以实现的，这项研究的主要目标是通过确定新的策略来消除持续感染的细胞或预防持续/潜伏的细胞感染，从而为治愈做出贡献。艾滋病毒在宿主细胞(CD4+T细胞和巨噬细胞)中的持久性是治愈的主要障碍，因为目前的治疗方法只针对病毒复制，留下了持续感染的细胞的储备库，只要这些细胞存活下来，这些细胞就携带艾滋病毒基因组。我们将重点关注感染艾滋病毒的巨噬细胞作为持续艾滋病毒感染的储存库和导致潜伏感染CD4+T细胞的关键角色。为了实现我们的目标，我们将追求三个相互关联的目标。每个目标都将侧重于消除感染艾滋病毒的巨噬细胞或防止感染艾滋病毒的巨噬细胞在CD4+T细胞中潜伏感染艾滋病毒的独特战略。目的1研究USP18和ISG15两个基因/蛋白在HIV萌发中的作用。萌芽是艾滋病毒离开受感染细胞的机制，由宿主和病毒蛋白控制。我们建议USP18在允许艾滋病毒萌芽发生方面发挥新的作用。我们预测，USP18活性的破坏将阻止HIV萌芽导致细胞内促凋亡病毒蛋白的积累，这一点单独或与其他策略结合将导致受感染的巨噬细胞死亡。我们将使用shRNAs的过度表达或基因沉默来确定USP18在HIV萌芽和HIV感染的巨噬细胞存活中的作用。巨噬细胞通过蛋白酶体或溶酶体途径有效地降解和回收细胞内的蛋白质。在破坏HIV萌芽的同时，我们将抑制蛋白质循环，以促进HIV蛋白质的积累，从而增加细胞毒细胞死亡。目的2将研究细胞基因/蛋白质如何调节巨噬细胞中艾滋病毒的复制速度。通过识别抑制或增强HIV基因表达、蛋白质翻译或萌芽的因素，我们将识别可被利用来杀死HV感染的巨噬细胞的其他漏洞。当与防止艾滋病毒萌芽和/或艾滋病毒蛋白质循环的战略相结合时，增强艾滋病毒复制将增加导致艾滋病毒感染的巨噬细胞死亡的细胞压力。目的3研究HIV感染的巨噬细胞在驱动CD4+T细胞潜伏感染中的作用。我们已经确定了HIV感染巨噬细胞的独特表型变化，包括可能支持潜伏感染CD4+T细胞的特定结果。潜伏期的两条途径将被研究，包括导致HIV感染的CD4+T细胞生长停滞的机制和增强静止的CD4+T细胞的HIV感染的机制。这三个目标共同提供了一项综合战略，以确定/利用艾滋病毒感染巨噬细胞的独特特性，以破坏持续/潜伏感染的生物途径。

DESCRIPTION: A cure for HIV-1 now seems achievable and the primary objective of this study is to contribute to a cure by identifying novel strategies to eliminate persistently-infected cellsor prevent persistent/latent infection of cells. HIV persistence in host cells (CD4+ T cells and macrophages) is a major obstacle to a cure because current therapies only target viral replication, leaving a reservoir of persistently-infected cells that carry the HIV genome for as long as those cells survive. We will focus on the role of HIV-infected macrophages as a reservoir for persistent HIV infection and as a key player in causing latent infection of CD4+ T cells. To achieve our goal, we will pursue three interrelated aims. Each aim will focus on unique strategies to either eliminate HIV-infected macrophages or prevent HIV-infected macrophages from driving HIV latency in CD4+ T cells. Aim 1 will investigate the role of two genes/proteins, USP18 and ISG15, in HIV budding. Budding, the mechanism by which HIV leaves an infected cell, is controlled by host and viral proteins. We propose a novel role for USP18 in allowing HIV budding to occur. We predict that disruption of USP18 activity will prevent HIV budding leading to accumulation of intracellular pro-apoptotic viral proteins, which alone or in combination with other strategies will cause death of infected macrophages. We will use over-expression or gene silencing with shRNAs to determine the role of USP18 in HIV budding and survival of HIV-infected macrophages. Macrophages efficiently degrade and recycle intracellular proteins through proteasome or lysosome pathways. In conjunction with disrupted HIV budding, we will inhibit protein recycling to enhance accumulation of HIV proteins to increase cytotoxic cell death. Aim 2 will investigate how cellular genes/proteins regulate the rate of HIV replication in macrophages. By identifying factors that inhibit or enhance HIV gene expression, protein translation or budding, we will identify additional vulnerabilities that can be exploited to kill HV-infected macrophages. When paired with a strategy to prevent HIV budding and/or HIV protein recycling, enhanced HIV replication will augment cell stress leading to death of HIV-infected macrophages. Aim 3 will study the role of HIV-infected macrophages in driving latent infection of CD4+ T cells. We have identified unique phenotypic changes to HIV-infected macrophages that include specific outcomes likely to support latent infection of CD4+ T cells. Two pathways of latency will be studied including a mechanism to cause growth arrest of HIV-infected CD4+ T cells and a mechanism to enhance HIV infection of resting CD4+ T cells. Together these three aims provide a comprehensive strategy to identify/exploit unique properties of HIV- infected macrophages to disrupt biological pathways of persistent/latent infection.