HIV Infection and Latency In Astrocytes

HIV 感染和星形胶质细胞潜伏期

• 批准号: 10129115
• 负责人: Johnny J He
• 金额: $ 57.57万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10129115
• 关键词: HIV; Infection; Latency; Astrocytes

ABSTRACT Combination antiretroviral therapy (cART) has considerably prolonged the lifespan of HIV-infected individuals and changed the landscape of the HIV/AIDS disease. However, latent HIV in the cells/tissues has prevented from achieving a functional cure or complete eradication. Limited access to cART and the ability of HIV to establish latent infection have made the central nervous system (CNS) unique HIV reservoirs. Microglia/macrophages are the main target cells for HIV infection in the CNS and all can be productively, latently or persistently infected. In contrast, astrocytes are infected but in a restricted manner; our understanding of these cells as HIV latent reservoirs and their roles of HIV latency in HIV/neuroAIDS is quite limited. We have recently found that cell-cell contact leads to successful HIV infection of astrocytes and establishment of HIV latency in these cells with an extremely low level of ongoing HIV replication. In addition, we have found that expression of HIV early gene Tat in astrocytes induces miR-132 and down-modulates methyl CpG-binding protein 2 (MeCP2), a chromatin-remodeling epigenetic factor and causes neurotoxicity through miRNA- containing exosomes. Moreover, we have shown that cocaine activates HIV replication in HIV latently infected astrocytes through miR-132 expression and that Tat expression is linked to establishment of HIV latency in astrocytes. Lastly, we have obtained the preliminary data that miR-132 expression in the cerebrospinal fluid (CSF) is elevated in HIV-infected subjects with minor cognitive and motor disorder. As a logical extension of our published and preliminary studies, we propose to characterize HIV infection and latency in astrocytes and their contribution to astrocyte function and HIV/neuroAIDS in the era of cART. The underlying hypothesis of this proposal is that HIV-infected astrocytes constitute latent HIV reservoirs in the CNS and directly contribute to HIV/neuroAIDS. To test this hypothesis, we propose to address the following four interrelated specific aims: (1) To characterize cell-cell contact-mediated HIV infection of astrocytes; (2) To elucidate molecular mechanisms of HIV latency in astrocytes; (3) To determine effects of latent HIV infection on astrocytes and neurons; and (4) To identify CSF biomarkers for HIV latency in the brain. We will use a combined molecular, cellular, biochemical, and genetic approach including use of primary mouse cortical astrocyte cultures and neuron cultures, doxycycline-inducible brain-specific HIV Tat transgenic mice (iTat), primary human fetal brain/astrocyte cultures, brain tissues and CSF samples of HIV-1 cohorts in our studies. We anticipate that this proposal will allow us to determine the regulatory mechanisms of HIV latency in astrocytes and the significance of these cells as HIV reservoirs in the ear of cART and in the context of cocaine abuse. The findings will likely inform control and eradication strategies for the HIV reservoirs in the CNS. The enormous amount of information available on HIV infection and pathogenesis in the CNS/astrocytes and HIV latency in the periphery and the results obtained from our preliminary studies make accomplishment of these aims practical.

抽象的 联合抗逆转录病毒疗法（CART）已大大延长了HIV感染者的寿命， 改变了艾滋病毒/艾滋病疾病的景观。但是，细胞/组织中的潜在艾滋病毒已阻止 功能治愈或完全消除。获得购物车的机会有限，艾滋病毒建立潜在感染的能力已有 使中枢神经系统（CNS）独特的艾滋病毒水库。小胶质细胞/巨噬细胞是HIV的主要靶细胞 中枢神经系统的感染和所有人都可以有效，潜在或持续感染。相反，星形胶质细胞被感染，但 以有限的方式；我们对这些细胞作为艾滋病毒潜伏储藏的理解及其在艾滋病毒潜伏期中的作用 艾滋病毒/神经助剂非常有限。我们最近发现细胞 - 细胞接触导致成功的HIV感染 这些细胞中持续的HIV复制水平极低的细胞中星形胶质细胞和HIV潜伏期的建立。在 此外，我们发现，艾滋病毒早期基因在星形胶质细胞中的表达会诱导miR-132并下调甲基 CpG结合蛋白2（MECP2），一种染色质复制的表观遗传因子，并通过miRNA-引起神经毒性 含有外泌体。此外，我们已经表明可卡因激活HIV中的HIV复制 通过miR-132的表达，TAT表达与星形胶质细胞中HIV潜伏期的建立有关。最后，我们 已经获得了脑脊液（CSF）中的miR-132表达在HIV感染中升高的初步数据 患有较小认知和运动障碍的受试者。作为我们发表和初步研究的逻辑扩展，我们 建议表征星形胶质细胞中的HIV感染和潜伏期及其对星形胶质细胞功能的贡献 购物车时代的艾滋病毒/神经辅助。该提议的基本假设是，感染HIV的星形胶质细胞构成 中枢神经系统中的潜在艾滋病毒水库，直接促成艾滋病毒/神经助剂。为了检验这一假设，我们建议 解决以下四个相互关联的特定目的：（1）表征细胞 - 细胞接触介导的HIV感染 星形胶质细胞； （2）阐明星形胶质细胞中艾滋病毒潜伏期的分子机制； （3）确定潜在艾滋病毒的影响 星形胶质细胞和神经元的感染； （4）识别脑脊液生物标志物，以用于大脑中的艾滋病毒潜伏期。我们将使用一个 结合分子，细胞，生化和遗传方法，包括使用原代小鼠星形胶质细胞 培养和神经元培养物，强力霉素 - 可诱导的脑特异性HIV TAT转基因小鼠（ITAT），原代人胎儿 在我们的研究中，脑/星形胶质细胞培养物，脑组织和HIV-1队列的CSF样品。我们预计这个建议 将使我们能够确定星形胶质细胞中艾滋病毒潜伏期的调节机制，以及这些细胞的重要性 在购物车耳朵和可卡因滥用的背景下，艾滋病毒水库。调查结果可能会为控制和 中枢神经系统中的艾滋病毒水库的根除策略。有关艾滋病毒感染的大量信息 中枢神经系统/星形胶质细胞和外围的HIV潜伏期的发病机理以及从我们的初步获得的结果 研究使这些目的实现了实用。