Reversal of HIV latency by METH and Inflammation

通过冰毒和炎症逆转 HIV 潜伏期

• 批准号: 9236600
• 负责人: JONATHAN KARN
• 金额: $ 75.15万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9236600
• 关键词: Acute; Affect; Agonist; Astrocytes; Biology; Blood Circulation; Brain; Cell model; Cells; ChIP-seq; Chronic; Coculture Techniques; Collaborations; Complex; Computer Retrieval of Information on Scientific Projects Database; Consumption; DNA; Data; Development; Disease Progression; Drug abuse; Epigenetic Process; Etiology; Event; Exposure to; Genes; Genetic; Grant; HIV; HIV Infections; HIV-associated neurocognitive disorder; Highly Active Antiretroviral Therapy; Human; IL8 gene; Imaging technology; Immune system; Individual; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-1 beta; Interleukin-6; Investigation; Knock-out; Laboratories; Latent Virus; Lead; Libraries; Life; Measures; Mediating; Methamphetamine; Methods; Microglia; Molecular; Mus; Nerve Degeneration; Neuraxis; Neurocognitive Deficit; Neuroglia; Neuronal Injury; Neurons; Pathway interactions; Patients; Peripheral; Pharmaceutical Preparations; Play; Proteins; Reaction; Regimen; Residual state; Rest; Signal Transduction; Symptoms; Synapses; System; T memory cell; TNF gene; Testing; Tissues; Variant; Ventral Tegmental Area; Viral; Viral Load result; Virus; Virus Diseases; Work; chemokine; chromatin immunoprecipitation; cytokine; drug of abuse; epigenetic marker; immortalized cell; insight; macrophage; memory CD4 T lymphocyte; methamphetamine abuse; methamphetamine use; multidisciplinary; neurotoxic; neurotoxicity; novel; promoter; receptor; research study; response; sigma receptors; signal processing; small hairpin RNA; tool; transcriptome sequencing

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.

概括 三分之一的艾滋病毒感染者会出现某种形式的艾滋病毒相关神经认知功能 紊乱（手）。吸食甲基苯丙胺 (METH) 等滥用药物会加重病情 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 疾病进展仍不明确。在这项研究中我们 将彻底检验 HAND 是 HIV 蛋白神经毒性作用结果的假设 当潜伏感染的小胶质细胞对神经退行性变的迹象做出反应时合成。我们将 还检验了以下假设：接触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 HIV 潜伏期分子机制专家 Jonathan Karn 博士 (CWRU) 和 Kurt 博士 豪瑟（Hauser），神经生物学家和药物滥用（VCU）专家。我们最近成立了一个 可靠而稳健的方法从源自原代胶质细胞中开发永生化小胶质细胞 新鲜的中枢神经系统组织，并用它们创建小胶质细胞/艾滋病毒细胞模型。该提案大写 一项公正的 shRNA 文库筛选结果显示，Nurr1/CoREST 反式 阻遏物复合物在小胶质细胞中沉默 HIV 方面发挥着关键作用，这一机制是 与记忆 T 细胞的沉默不同。基于这些观察，我们将研究 表观遗传机制导致 HIV 启动子沉默，包括 Nurr1/CoREST，通过 染色质免疫沉淀实验（Chip-Seq），并研究急性和 长期使用冰毒治疗可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 HIV 表达。最后用两本小说 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感染的小胶质细胞响应METH的基础生物学研究。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 患者中 HAND 的发展以及冰毒如何增强这一发展 虐待。