Modeling HIV-1 Neuropathogenesis and neuronal dysregulation using 3D-organoids containing multiple CNS cell lineages

项目摘要 HIV相关的神经认知障碍（HAND）的范围从轻度到严重的认知缺陷， 超过60%的HIV-1受试者接受联合抗逆转录病毒治疗（cART）。病理学评价 HIV-1阳性受试者的额叶皮质显示存在HIV-1感染的神经炎症 小胶质细胞/巨噬细胞、星形胶质细胞增生、树突损失和神经元中的突触修剪。的降解 伴随这些神经元变化的树突导致突触丧失、神经元和认知能力下降， 这导致艾滋病毒感染者发生HAND。此外，这些影响可能会因以下因素而加剧： 非法药物使用滥用，如阿片类药物，这是这一人群经常滥用的药物，因此， 越来越需要研究HAND和了解HIV相关的神经发病机制，以开发更好的治疗方法 选择和改善疾病管理。然而，这些努力由于缺乏一个 生理学相关的脑/CNS模型。为了满足这一需求，我们开发了一种尖端的三文化人类 包含初级神经元、星形胶质细胞和小胶质细胞的3D脑类器官（hBORG）（HIV-1感染和 未感染）来精确地模拟大脑环境、慢性病毒复制和神经炎症环境 在HIV-1感染者中观察到。使用这个3D-BORG模型，我们建议测试我们的假设， 由HIV-1感染的小胶质细胞引起的神经炎症，诱导神经元变性，突触修剪和 树突的丧失、星形细胞增多症以及这些作用因阿片类药物滥用而恶化。我们建议 为实现这些目标，我们采取了以下措施：目标1。描述受感染的小胶质细胞如何有助于修剪和缩放 突触树突、突触功能丧失和神经元丧失;目的2.确定如何使用非法药物，例如 吗啡驱动神经元功能障碍和病理学的发展和/或恶化;以及目的3. 确定来自HIV-1感染的小胶质细胞的病毒和细胞因子如何引起HAND相关的病理性 功能.我们已经组建了一个研究团队，他们在HIV-1病毒学，3D类器官和 因此，药物滥用和突触功能将决定受感染的小胶质细胞如何与神经元相互作用， 星形胶质细胞，并进一步描绘参与树突状细胞损伤，突触可塑性， 存在和不存在药物滥用时的神经元存活。这将帮助我们开发新的疗法， 可以预防神经元损伤和突触功能丧失以及在PLWH中观察到的认知障碍。

Project Summary HIV-associated neurocognitive disorder (HAND) range from mild to severe deficits in cognition and occurs in more than 60% of HIV-1 subjects receiving combined antiretroviral therapy (cART). Pathological evaluation of the frontal cortex of HIV-1 positive subjects revealed neuroinflammation with presence of HIV-1 infected microglia/macrophages, astrogliosis, dendrites loss and synaptic pruning in neurons. The degradation of dendrites that accompany these neuronal changes leads to synaptic loss, neuronal and cognitive decline, and this causes development of HAND in people living with HIV. Moreover, these effects may be exacerbated by illicit drug use abuse such as opioids, a drug commonly abused by this population, thus, there is a great and increasing need to study HAND and understand HIV-associated neuropathogenesis to develop better treatment options and improve disease management. However, these efforts are significantly hampered by the lack of a physiologically relevant brain/CNS model. To address this need, we developed a cutting-edge tri-culture human 3D-brain organoids (hBORGs) that contains primary neurons, astrocytes and microglia (HIV-1-infected and uninfected) to accurately model the brain environment, chronic virus replication, and neuroinflammatory milieu observed in HIV-1 infected individuals. Using this 3D-BORG model, we propose to test our hypothesis that neuroinflammation caused by HIV-1 infected microglia, induces degeneration of neurons, synaptic pruning and loss of dendrites, astrocytosis and these effects worsen by the addition of oipoid drug abuse. We propose the following aims to achieve the goals: Aim 1. Delineate how infected microglia contribute to pruning and scaling of synaptodendrites, loss of synaptic functions and neuronal loss; Aim 2. Determine how use of illicit drugs such as Morphine drives the development and/or worsening of the neuronal dysfunction and pathology; and Aim 3. Identify the how viral and cellular factors from HIV-1 infected microglia cause the HAND-associated pathological features. We have assembled a team of investigators who have expertise in HIV-1 Virology, 3D-Organoids and drug abuse and synaptic function, thus, are poised to define how infected microglia interact with neurons and astrocytes and further delineate the molecular mechanisms involved in dendritic damage, synaptic plasticity and neuronal survival in the presence and absence of drug abuse. This will help us develop novel therapeutics that can prevent neuronal damage and loss of synaptic function and cognitive impairment observed in PLWH.