Reversal of HIV latency by METH and Inflammation

摘要 三分之一的HIV感染者会发展成某种形式的HIV相关神经认知 无序(手)。滥用甲基苯丙胺(冰毒)等药物的消费会加剧 手部症状，但这些药物影响的细胞和分子机制 HIV疾病在中枢神经系统(CNS)的进展仍然不明确。在这项研究中，我们 将彻底检验这一假设，即手是艾滋病毒蛋白质神经毒性作用的结果 当潜伏感染的小胶质细胞对神经变性的迹象做出反应时合成的。我们会 也要检验这样一种假设，即接触滥用药物会增加艾滋病毒的复制并加剧 手。这项高度多学科的调查是两个实验室之间的密切合作 乔纳森·卡恩博士(CWRU)，艾滋病毒潜伏期分子机制专家，以及库尔特博士 豪泽是一位神经生物学家和药物滥用(VCU)专家。我们最近建立了一个 从原代小胶质细胞培养永生化小胶质细胞的可靠方法 新鲜的中枢神经系统组织，并用它们建立小胶质细胞/HIV细胞模型。这项提议充分利用了 根据无偏向shRNA文库筛选的结果，发现Nurr1/corest反式- 抑制子复合体在抑制小胶质细胞中的艾滋病毒方面发挥着关键作用，这一机制是 有别于记忆T细胞中的沉默。在这些观察的基础上，我们将研究 导致HIV启动子沉默的表观遗传机制，包括Nurr1/corest，通过 染色质免疫沉淀实验(ChIP-Seq)，并研究急性和 冰毒慢性治疗逆转艾滋病毒潜伏期。我们的实验还将揭示 炎症信号，与冰毒一起，诱导艾滋病毒表达。最后，用两部小说 共培养系统，我们将演示冰毒引发的小胶质细胞/HIV细胞如何加剧 神经元损伤。成功完成这些研究将提供详细的了解 HIV感染的小胶质细胞对冰毒的基本生物学反应。通过建立 我们将提供新的HIV潜伏期、炎症和神经元损伤之间的关系 对HIV患者手发育的洞察以及冰毒如何增强手的发育 虐待。

Summary One in three HIV-infected individuals develops some form of HIV-associated neurocognitive disorder (HAND). Consumption of drugs of abuse such as methamphetamine (METH) aggravates the symptoms of HAND, but the cellular and molecular mechanisms by which these drugs impact HIV disease progression in the central nervous system (CNS) remain ill-defined. In this study we will thoroughly test the hypothesis that HAND is the result of the neurotoxic effects of HIV proteins synthesized when latently infected microglial cells respond to signs of neurodegeneration. We will also test the hypothesis that exposure to drugs of abuse enhance HIV replication and exacerbate HAND. This highly multidisciplinary investigation is a close collaboration between the laboratories of Dr. Jonathan Karn (CWRU), an expert in the molecular mechanisms HIV latency, and Dr. Kurt Hauser, a neurobiologist and expert on drug abuse (VCU). We have recently established a reliable and robust method to develop immortalized microglial cells from primary glia derived from fresh CNS tissue, and used them to create microglia/HIV cellular models. The proposal capitalizes on findings of an unbiased shRNA library screen, which revealed that the Nurr1/CoREST trans- repressor complex plays a key role in silencing HIV in microglial cells, a mechanism which is distinct from silencing in memory T-cells. Building on these observations, we will study the epigenetic machinery leading to silencing of the HIV promoter, including Nurr1/CoREST, by chromatin immunoprecipitation experiments (Chip-Seq), and study the impact of both acute and chronic treatment with METH on reversing HIV latency. Our experiments will also uncover the inflammatory signals that, together with METH, induce HIV expression. Finally, using two novel co-culture systems, we will demonstrate how METH-primed microglia/HIV cells exacerbate neuronal damage. Successful completion of these studies will provide a detailed understanding of fundamental biology of HIV-infected microglia in response to METH. By establishing the relationship between HIV latency, inflammation, and neuronal damage we will provide new insights into the development of HAND in HIV patients and how this is augmented by METH abuse.