Regulation of HIV latency by microglial-neuronal interactions

小胶质细胞-神经元相互作用对 HIV 潜伏期的调节

• 批准号: 10450662
• 负责人: JONATHAN KARN
• 金额: $ 78.81万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10450662
• 关键词: 3-Dimensional; Biological Models; Brain; CRISPR/Cas technology; CX3CL1 gene; Cells; Cerebrum; Coculture Techniques; Collaborations; Communication; Defect; Development; Drug abuse; Drug usage; Effector Cell; Fractalkine; GABA Receptor; Genetic; Genetic Transcription; Glutamate Receptor; HIV; HIV Infections; HIV Seropositivity; Human; Imaging technology; Individual; Infection; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Injury; Investigation; Laboratories; Lead; Libraries; Mediating; Memory; Methamphetamine; Microglia; Modeling; Molecular Biology; Morbidity - disease rate; Mutation; Natural Products; Nerve Degeneration; Neurobiology; Neurocognitive; Neuroglia; Neurons; Organoids; Pathway interactions; Patients; Pattern recognition receptor; Pharmaceutical Preparations; Pharmacology; Production; Purinoceptor; Rattus; Receptor Gene; Receptor Signaling; Recovery; Regulation; Rest; Risk; Role; Signal Pathway; Signal Transduction; Stress; Symptoms; System; T-Lymphocyte; Testing; Therapeutic; United States; Viral; Viral Proteins; Virus; antiretroviral therapy; cell transformation; design; dopaminergic neuron; drug of abuse; experimental study; gamma-Aminobutyric Acid; genome editing; induced pluripotent stem cell; inhibitor; knock-down; methamphetamine effect; methamphetamine exposure; mortality; multidisciplinary; neurocognitive disorder; neurotoxic; neurotransmission; novel; protective effect; receptor; reconstitution; response; small hairpin RNA; substance use; transcriptome sequencing

Summary Over 40% of HIV-positive individuals in the United States engage in substance use. This not only represents a major cause of enhanced morbidity and mortality, but also is associated with increased risks of neurocognitive disorders, such as HAND. Neurons possess refined systems for maintaining constant communication with glia through propagation of “Off” and “On” signals controlling microglial activation states. Using HIV latency models in immortalized human microglial cells (hµglia/HIV), we have shown previously that cellular activation and inflammatory responses induce HIV production. Remarkably, co-culture of productively infected microglia with an excess of healthy neurons leads to viral silencing. We have also shown that hµglia/HIV cells can migrate into brain organoids where they become silenced. However, damaging neurons with a variety of agents, including methamphetamine (METH), a frequently-used abuse substance among HIV-infected individuals, produce reactivation signals for HIV, and this initiates a cycle of microglial activation and further neuronal damage. This cycle of shutdown and reactivation seems to parallel the M1 to M2 transition model of microglial cells, much as HIV latency in T cells is a product of the natural transition of effector cells to resting memory cells. In this proposal, we seek to define the key signals mediating the cycle of viral silencing and reactivation in microglial cells by neurons in the context of iPSC-derived cerebral organoids. This multidisciplinary investigation is designed as a close collaboration between the laboratories of Dr. Jonathan Karn (CWRU, HIV molecular biology), Dr. Anthony Wynshaw-Boris (CWRU, iPSC cells, brain organoids), Dr. Kurt Hauser (VCU, neurobiology and drug abuse), and Dr. Pamela Knapp (VCU, brain organoids). To avoid the limitations of working with transformed cells, we have recently initiated experiments using co-cultures between iPSC-derived cerebral organoids and microglia. Using co-cultures between iPSC-derived cerebral organoids and microglia, we will thoroughly test the hypothesis that the exaggerated responses of HIV-infected microglia to neuronal damage leads to enhanced neurodegeneration. We will also test the hypothesis that exposure to METH, given this background of faulty microglia-neuron crosstalk, enhances HIV replication. Using genome editing approaches, we will identify the specific contribution of “On” and “Off” receptor systems in controlling HIV latency in microglia, and study how METH impacts neuronal-microglial signaling to augment HIV production. In parallel with our genetic investigations, we will also evaluate a number of pharmacological agents against microglial receptors, HIV transcription inhibitors, and mediators of inflammation in order to define therapeutic approaches that might be expected to slow the development of HAND, especially in patients who abuse drugs.

概括 在美国，超过40％的HIV阳性个体从事药物使用。这 不仅代表了增强发病率和死亡率的主要原因，而且还相关 随着神经认知障碍（例如手）的风险增加。神经元具有精致 通过传播“ OFF”和“ ON”来维持与神经胶质的持续沟通的系统 控制小胶质激活状态的信号。在永生的人类中使用艾滋病毒潜伏期模型 小胶质细胞（HµGLIA/HIV），我们先前已经证明了细胞活化和炎症性 备注诱导艾滋病毒产生。值得注意的是，与 过多的健康神经元会导致病毒沉默。我们还显示了HµGlia/HIV细胞 可以迁移到脑器官中，在那里它们被沉默。但是，用 各种特工，包括甲基苯丙胺（METH），一种常用的滥用物质 在感染HIV的个体中，为HIV产生重新激活信号，这引发了 小胶质激活和进一步的神经元损伤。这个关闭和重新激活的周期似乎 要与小胶质细胞的M1和M2过渡模型平行，与T细胞中的HIV潜伏期一样 效应细胞向静止记忆细胞的自然过渡的产物。在此提案中，我们寻求 定义介导小胶质细胞中病毒沉默和重新激活周期的键信号 在IPSC衍生的大脑器官的背景下神经元。这项多学科调查 被设计为乔纳森·卡恩（Jonathan Karn）博士实验室之间的密切合作（CWRU，HIV 分子生物学），Anthony Wynshaw-Boris博士（CWRU，IPSC细胞，脑官体），Kurt博士 Hauser（VCU，神经生物学和药物滥用），以及Pamela Knapp博士（VCU，脑器官）。到 避免使用转化的细胞的局限性，我们最近开始了实验 使用IPSC衍生的大脑器官和小胶质细胞之间的共培养。使用共培养 在IPSC衍生的大脑器官和小胶质细胞之间，我们将彻底检验假设 艾滋病毒感染的小胶质细胞对神经元损害的夸大反应导致 神经变性。鉴于此，我们还将测试暴露于甲基苯丙胺的假设 小胶质细胞 - 神经元串扰的背景，增强了HIV复制。使用基因组编辑 方法，我们将确定“ ON”和“ OFF”受体系统在 控制小胶质细胞中的艾滋病毒潜伏期，并研究甲基甲基元如何影响神经元 - 神经信号传导 增强艾滋病毒的产生。与我们的遗传研究并行，我们还将评估 针对小胶质细胞受体，HIV转录抑制剂和 炎症的介体是为了定义可能期望的理论方法 减慢手的发展，尤其是在滥用药物的患者中。