Cell Biology and Functional Analysis Core for Institution # 269291

摘要 细胞生物学和功能分析(CBFA)核心的总体目标是为基础科学家提供 随着临床研究人员在细胞和分子水平上研究HIV-1导致的神经缺陷， 指导和培训。此外，将为过渡到以下领域的研究人员提供资源 神经艾滋病毒。尽管目前使用的抗逆转录病毒疗法(ART)在控制病毒载量方面取得了成功 几乎所有的HIV-1患者，这种病毒抑制剂的混合物都不能潜伏地从病毒中清除 感染的细胞包括T细胞、巨噬细胞、脑小胶质细胞和其他潜伏期的细胞 身体。因此，一旦停止ART方案，患者仍面临病毒反弹的风险。在最近 几年来，已经实施了几种策略来删除/编辑潜伏感染的病毒序列 水库。同样重要的是，能够识别这些细胞，以打击和摧毁它们。CBFA的核心是 旨在为调查人员提供培训和技术专长，以利用细胞和分子工具 进行与破译HIV-1诱导的中枢神经系统疾病的分子机制相关的研究，并最终 设计分子疗法来对抗大脑中的艾滋病毒-1感染。这个核心将为调查人员提供 具有良好特性的高纯度脑细胞，包括神经元、星形胶质细胞、少突胶质细胞、小胶质细胞 内皮细胞、神经前体细胞以及培养的人外周血细胞 分子、细胞和病毒学研究。此外，人诱导多能干细胞(HiPSC)系，以及 来自IPSCs的皮质球体和脑器官将被生成、表征并提供给 神经艾滋病毒领域的核心用户。此外，还将为隔离提供专业知识， HIV-1、HIV-2和SIV各种分支的特征、繁殖和维护。在本意见书中， 我们采用新的方法来研究大脑不同区域的神经元功能。 微电极阵列(MEA)技术。此外，显微镜方面的专业知识和培训， 免疫组织化学分析被提供给研究人员。我们的核心将与其他核心密切合作，以 推进综合性多学科协同中心计划。这种协同方式将确保 CNHC发展奖获得者和CNHC用户在进行生产性高影响力方面的成功 神经艾滋病毒的研究。我们项目的主要优势是我们的专家团队具有互补的专业知识 让研究人员在与病毒载体设施密切合作的同时进行神经艾滋病毒研究，以及 最终在高纯度的中枢神经系统细胞培养中进行实验，以评估 体外和体外培养的神经细胞。

SUMMARY The overall goal of the Cell Biology and Functional Analyses (CBFA) core is to provide basic scientists as well as clinical researchers studying HIV-1 induced neurological deficits at cell and molecular levels, resources, mentorship, and training. In addition, resources would be available to researchers transitioning to the area of NeuroHIV. Despite the success of the currently used antiretroviral therapy (ART) in controlling viral load in virtually all HIV-1 patients, this cocktail of viral inhibitors has not been able to eliminate the virus from latently infected cells including T-cells, macrophages, brain microglial cells, and cells in other sites of latency throughout the body. Therefore, the individuals remain at risk of viral rebound once they stop ART regimen. In the recent years, several strategies have been implemented to remove/edit viral sequences from the latently infected reservoirs. Equally as important is to be able to identify such cells to target and destroy them. The CBFA core is designed to provide investigators with training and technical expertise to utilize cellular and molecular tools to conduct research related to deciphering molecular mechanisms of HIV-1-induced CNS disease and ultimately design of molecular therapeutics to combat HIV-1 infection in the brain. This core would provide investigators with well characterized highly purified brain cells including neurons, astrocytes, oligodendrocytes, microglia, endothelial cells, and neural progenitors as well as cultured human peripheral blood cells for performing molecular, cellular, and virological studies. Moreover, human induced pluripotent stem cell (hiPSC) lines, and cortical spheroids and cerebral organoids from iPSCs, will be generated, characterized and made available for the core users in the field of NeuroHIV. In addition, expertise will be also provided for the isolation, characterization, propagation and maintenance of a variety of HIV-1 clades, HIV-2 and SIV. In this submission, we have included novel methods to study neuronal function in different regions of the brain using our Microelectrode Array (MEA) technology. In addition, expertise and training in microscopical, immunohistochemical analyses are offered to investigators. Our core will work closely with the other cores to promote a comprehensive multidisciplinary collaborative center program. This synergistic approach will ensure the success of CNHC developmental award recipients and CNHC users in conducting productive high impact research in neuroHIV. The main strength of our program is our team of experts with complementary expertise for investigators to conduct their neuroHIV research while working closely with the viral vector facility, and ultimately performing experiments in highly purified CNS cell cultures to assess the functional aspects of neuronal cells both in vitro and ex vivo.