Reversal of HIV latency by METH and Inflammation

总结 三分之一的HIV感染者会出现某种形式的HIV相关神经认知障碍。 手（Hand）滥用药物的消费，如甲基苯丙胺（METH）兴奋剂 HAND的症状，但这些药物影响的细胞和分子机制 中枢神经系统（CNS）中的HIV疾病进展仍然不明确。本研究 将彻底检验HAND是HIV蛋白质神经毒性作用的结果的假设 当潜伏感染的小胶质细胞对神经变性的迹象作出反应时合成。我们将 同时也检验了暴露于滥用药物会增强艾滋病毒复制并加剧 手这项高度多学科的研究是实验室之间的密切合作 乔纳森·卡恩博士（CWRU），艾滋病毒潜伏期的分子机制专家，和库尔特博士 豪瑟是一位神经生物学家和药物滥用专家。我们最近成立了一个 一种可靠和稳健的方法，从来源于以下的原代神经胶质细胞发育永生化小神经胶质细胞： 新鲜的CNS组织，并使用它们来创建小胶质细胞/HIV细胞模型。提案资本化 无偏shRNA文库筛选的结果显示，Nurr 1/CoREST反式- 阻遏物复合物在小胶质细胞中沉默HIV中起着关键作用，这是一种 与记忆T细胞中的沉默不同。在这些观察的基础上，我们将研究 表观遗传机制导致HIV启动子沉默，包括Nurr 1/CoREST， 染色质免疫沉淀实验（Chip-Seq），并研究急性和 METH慢性治疗逆转HIV潜伏期。我们的实验还将揭示 炎症信号，与METH一起诱导HIV表达。最后，使用两个新的 共培养系统，我们将展示如何METH引发的小胶质细胞/艾滋病毒细胞加剧 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METH的反应的基础生物学。通过建立 HIV潜伏期，炎症和神经元损伤之间的关系，我们将提供新的 深入了解HIV患者HAND的发展以及METH如何增强HAND 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.