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

基本信息

批准号：
10674037
负责人：
JONATHAN KARN
金额：
$ 78.49万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
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/10674037
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 Hand 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 Production Productivity 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 glial activation induced pluripotent stem cell inhibitor knock-down methamphetamine effect methamphetamine exposure migration mortality multidisciplinary neurocognitive disorder neurotoxic neurotransmission novel pharmacologic protective effect receptor reconstitution response small hairpin RNA substance use transcriptome sequencing

项目摘要

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% 的艾滋病毒阳性者有药物滥用行为。不仅是发病率和死亡率增加的主要原因，而且还与神经认知障碍的风险增加，例如手部神经元。通过传播“关”和“开”来与神经胶质细胞保持持续通信的系统在永生化人类中使用 HIV 潜伏期模型控制小胶质细胞激活状态。小胶质细胞（hμglia/HIV），我们之前已经证明细胞激活和炎症值得注意的是，有效感染的小胶质细胞与艾滋病毒的共培养。我们还发现，hμglia/HIV 细胞过多会导致病毒沉默。可以迁移到大脑类器官中，在那里它们会变得沉默，但会损害神经元。各种药物，包括甲基苯丙胺 (METH)，一种经常使用的滥用物质在艾滋病毒感染者中，产生艾滋病毒重新激活信号，这启动了一个循环小胶质细胞激活和进一步的神经损伤似乎是这种关闭和重新激活的循环。与小胶质细胞的 M1 到 M2 转变模型相似，就像 T 细胞中的 HIV 潜伏期一样在这个提议中，我们寻求效应细胞自然过渡到静息记忆细胞的产物。定义介导小胶质细胞病毒沉默和重新激活周期的关键信号在 iPSC 衍生的大脑类器官的背景下，由神经元进行这项多学科研究。是 Jonathan Karn 博士（CWRU、HIV 分子生物学）、Anthony Wynshaw-Boris 博士（CWRU、iPSC 细胞、脑类器官）、Kurt 博士 Hauser（VCU，神经生物学和药物滥用）和 Pamela Knapp 博士（VCU，脑类器官）。为了避免使用转化细胞的局限性，我们最近开始了实验使用 iPSC 衍生的脑类器官和小胶质细胞之间的共培养。在 iPSC 衍生的脑类器官和小胶质细胞之间，我们将彻底检验这一假设 HIV感染的小胶质细胞对神经元损伤的过度反应导致增强鉴于此，我们还将检验接触冰毒的假设。有缺陷的小胶质细胞-神经元串扰的背景，利用基因组编辑增强了 HIV 复制。方法，我们将确定“开”和“关”受体系统的具体贡献控制小胶质细胞中的 HIV 潜伏期，并研究 METH 如何影响神经元-小胶质细胞信号传导为了增加艾滋病毒的产生，在进行基因研究的同时，我们还将评估针对小胶质细胞受体、HIV 转录抑制剂的药物数量，以及炎症介质以确定可能预期的治疗方法减缓手部疾病的发展，尤其是滥用药物的患者。