Reversal of HIV latency by METH and Inflammation

通过冰毒和炎症逆转 HIV 潜伏期

基本信息

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
项目类别：
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

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

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项目摘要

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HIV感染者发展出某种形式的HIV相关神经认知障碍（手）。消费诸如甲基苯丙胺（甲基苯丙胺）等滥用药物加剧手的症状，但是这些药物会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。在这项研究中，我们将彻底检验以下假设，即HAND是HIV蛋白的神经毒性作用的结果当潜在感染的小胶质细胞对神经退行性的迹象响应时，合成。我们将还测试了暴露于滥用药物的假设会增强艾滋病毒的复制和加剧手。这项高度多学科的调查是实验室之间的密切合作分子机制艾滋病毒潜伏期专家乔纳森·卡恩（CWRU）博士和库尔特博士 Hauser，神经生物学家和药物滥用专家（VCU）。我们最近建立了一个可靠且健壮的方法来发展从源自原代胶质细胞的永生小胶质细胞新鲜的CNS组织，并使用它们创建小胶质细胞/HIV细胞模型。该提案资本化关于无偏的shrna库屏幕的发现，该屏幕表明Nurr1/corest Trans- 阻遏物复合物在沉默的小胶质细胞中起关键作用，这种机制是不同于记忆中的沉默。基于这些观察，我们将研究表观遗传机制导致HIV启动子的沉默，包括Nurr1/Corest，由染色质免疫沉淀实验（CHIP-SEQ），研究急性和用甲基甲基苯丙胺治疗逆转艾滋病毒潜伏期。我们的实验也将发现炎症信号与甲基相同，诱导HIV表达。最后，使用两本小说共培养系统，我们将演示甲基化的小胶质细胞/HIV细胞如何恶化神经元损伤。这些研究的成功完成将提供详细的理解响应于甲基的HIV感染小胶质细胞的基本生物学。通过建立艾滋病毒潜伏期，炎症和神经元损害之间的关系我们将提供新的洞悉HIV患者的手发展以及如何通过甲基甲基苯丙胺增加虐待。