Modeling HIV and methamphetamine-induced neuroinflammation in cerebral organoids

模拟 HIV 和甲基苯丙胺诱导的脑类器官神经炎症

• 批准号: 10528845
• 负责人: PAUL W. SPEARMAN
• 金额: $ 59.88万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10528845
• 关键词: 3-Dimensional; Acute; Adverse effects; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Astrocytes; Autopsy; Biological Models; Brain; Cell surface; Cells; Central Nervous System Diseases; Cerebrum; Chronic; Clinical Research; Development; Electrophysiology (science); Evaluation; Functional disorder; HIV; HIV Infections; HIV-associated neurocognitive disorder; Health; Human; Immunofluorescence Microscopy; Individual; Inflammation; Inflammation Mediators; Inflammatory; Investigation; Knock-out; Longevity; Measures; Mediating; Methamphetamine; Microelectrodes; Microglia; Modeling; Molecular; Morbidity - disease rate; Multiple Sclerosis; Myeloid Cells; NF-kappa B; Neurocognitive Deficit; Neuronal Dysfunction; Neurons; Organoids; Pathogenesis; Pathway interactions; Persons; Play; Process; Production; Role; SIV; SYK gene; Signal Pathway; Signal Transduction; Source; Stimulant; Study models; Testing; Time; Tyrosine Kinase Inhibitor; Viral; Viral Proteins; Viral reservoir; acute infection; antiretroviral therapy; brain cell; cell injury; cell type; chronic infection; comorbidity; crosslink; cytokine; defined contribution; experimental study; induced pluripotent stem cell; inhibitor; insight; knockout gene; methamphetamine effect; methamphetamine exposure; methamphetamine use; neurocognitive disorder; neuroinflammation; particle; recruit; relating to nervous system; single-cell RNA sequencing; src-Family Kinases; transcriptome sequencing

Combination antiretroviral therapy (cART) has led to dramatic increases in lifespan among HIV-infected individuals. Despite effective cART, however, HIV-associated morbidities exert a significant toll. HIV- associated neurocognitive disorders (HAND) occur in up to 50% of chronically infected individuals despite cART. The pathogenesis of HAND remains under investigation. Neuroinflammation is a hallmark of HAND, as established by clinical studies, autopsy studies, and animal models. Ongoing or intermittent replication of HIV in the CNS is likely to contribute to neuroinflammation through direct effects on the infected cells or through release of viral proteins and inflammatory mediators. Use of neural stimulants including methamphetamine can exacerbate the neurocognitive decline seen in HAND, but the mechanisms underlying this comorbidity are not understood. Microglia are the primary resident myeloid cells of the brain, are infected at early times following acute infection with HIV or SIV, can act as a CNS viral reservoir, and are thought to play a central role in the development of HAND. The pathways responsible for microglial activation and dysfunction following HIV infection remain incompletely defined. Microglia derived from induced pluripotent stem cells (iPSCs) provide a unique opportunity to examine the molecular mechanisms underlying microglial activation. iPSC-derived microglia will be introduced into cerebral organoids, providing the additional opportunity to define the effects of microglial activation on surrounding astrocytes, neurons, and other cells. Tetherin is a host restriction factor that captures HIV during the assembly process in infected cells and generates a proinflammatory signaling cascade within infected cells. Experiments in Aim 1 of this project will evaluate HIV-induced neuroinflammation both in an unbiased way and through a directed evaluation of the role of tetherin-mediated signaling as a trigger of microglial inflammation. RNAseq, cytokine production, and immunofluorescence microscopy will be employed to define microglial activation following HIV infection. In Aim 2, we will introduce HIV-infected microglia into cerebral organoids to define the molecular basis of HIV-induced neuroinflammation and neuronal dysfunction. Single-cell RNAseq and evaluation of neuronal health and electrophysiology will be performed in models representing acute infection and in ART-suppressed, chronic infection of the brain. The potential of methamphetamine to contribute to neuroinflammation and neuronal damage in the HIV-infected microglia/organoid model will then be defined, and the relevant pathways identified. Together, these studies will provide insights into the pathogenesis of HAND and the potential contribution of methamphetamine to neurocognitive decline.

联合抗逆转录病毒疗法 (cART) 显着延长了 HIV 感染者的寿命 个人。然而，尽管 cART 有效，但与 HIV 相关的发病率仍造成重大损失。艾滋病病毒- 高达 50% 的慢性感染者会出现相关神经认知障碍 (HAND)，尽管 大车。 HAND 的发病机制仍在研究中。神经炎症是 HAND 的一个标志， 由临床研究、尸检研究和动物模型确定。持续或间歇性复制 中枢神经系统中的艾滋病毒可能通过直接影响受感染的细胞或导致神经炎症 通过释放病毒蛋白和炎症介质。使用神经兴奋剂包括 甲基苯丙胺会加剧 HAND 中的神经认知能力下降，但其机制 这种合并症的背后尚不清楚。小胶质细胞是大脑中主要的髓样细胞， 在急性感染 HIV 或 SIV 后的早期被感染，可以充当 CNS 病毒库，并且 被认为在 HAND 的发展中发挥着核心作用。负责小胶质细胞的通路 HIV 感染后的激活和功能障碍仍未完全确定。小胶质细胞源自 诱导多能干细胞（iPSC）为检查分子机制提供了独特的机会 潜在的小胶质细胞激活。 iPSC 衍生的小胶质细胞将被引入大脑类器官中，提供 额外的机会来定义小胶质细胞激活对周围星形胶质细胞、神经元的影响， 和其他细胞。 Tetherin 是一种宿主限制因子，可在组装过程中捕获 HIV 感染细胞并在感染细胞内产生促炎信号级联。目标实验 该项目的第一部分将以公正的方式并通过 直接评估系链蛋白介导的信号传导作为小胶质细胞炎症触发因素的作用。 RNAseq、细胞因子产生和免疫荧光显微镜将用于定义小胶质细胞 HIV感染后激活。在目标 2 中，我们将把感染 HIV 的小胶质细胞引入大脑类器官中 定义艾滋病毒引起的神经炎症和神经元功能障碍的分子基础。单细胞 RNAseq 以及神经元健康和电生理学的评估将在代表的模型中进行 急性感染和 ART 抑制的脑部慢性感染。甲基苯丙胺的潜力 导致 HIV 感染的小胶质细胞/类器官模型中的神经炎症和神经元损伤 然后进行定义，并确定相关途径。总之，这些研究将提供有关以下方面的见解： HAND 的发病机制以及甲基苯丙胺对神经认知能力下降的潜在贡献。