Research Support Core B: Primary Cell, Biomimetic, and iPSC-derived Cell Models

项目摘要-原代细胞、仿生学和IPSC衍生细胞模型Core B 了解可卡因、甲基苯丙胺、大麻和阿片类药物的作用机制(S) 放大HIV潜伏期的损害是CWRU卓越中心的中心目标 物质使用对艾滋病毒的影响。我们对艾滋病毒潜伏期和持久性的理解是 由于在循环中发现了少量潜伏感染细胞，这一困难 在从患者那里获得全面的组织样本方面，缺乏已知的表型 可以区分潜伏感染细胞和未感染细胞的标记，以及有限的信息 关于组织储存库在体内的行为。Core B将提供两种体外原代细胞模型 (目标1)和高灵敏的核酸分析，以衡量物质使用的影响 关于临床样本中的艾滋病毒储存库(目标2)。一个关键的悬而未决的问题是每个人如何 药物调节多种细胞类型和组织中的潜伏的艾滋病毒储存库。初级细胞， 仿生和iPSC衍生的细胞模型Core B团队将为Center提供和开发 研究人员创新和高度信息量的细胞和仿生模型的三个主要 毒品使用和艾滋病毒的器官靶标--大脑、肠道和免疫系统。有机化合物衍生 来自组织外植体和诱导的多能干细胞(IPSC)将被用来模拟组织 微环境，并将分析范围扩大到使用单个细胞无法实现的范围 体外模型。Core B提供的模型将在整个卓越中心使用 评估药物使用对艾滋病毒潜伏期的影响，并将在体外用样本建立 来自接受抗逆转录病毒治疗的物质使用障碍患者和艾滋病毒患者。这些 每个模型都将反映控制药物反应的分子机制的多样性。 外周CD4T细胞、肠道粘膜T细胞的使用和HIV潜伏期 来自中枢神经系统的小胶质细胞、神经元和微血管内皮细胞，也将提供 对比体内物质使用和直接细胞生物学影响的独特机会 体外分析。重要的是，核心B团队将继续开发新的分析和方法 研究物质使用对HIV持续和潜伏机制的影响。在……里面 特别是，这项工作包括(A)开发单细胞RNA诱导分析，(B)模拟 TFH分室，以及(C)推进IPSC衍生的脑有机体研究药物使用和艾滋病毒 中枢神经系统的潜伏期。因此，核心B与核心C中的分子服务合作，将 提供获取新技术的途径，以评估细胞、蛋白质组、微生物、 与物质使用相关的代谢学、转录学和转录变化及其对艾滋病毒的影响 在Core D和原代细胞培养中提供的患者样本中的储存库和工具 我们的物质使用研究人员无法操纵细胞 机构和全国范围内。

Project Summary - Primary Cell, Biomimetics, and iPSC-derived Cellular Models Core B Understanding the mechanism(s) by which cocaine, methamphetamine, cannabis, and opioids amplify the damage of HIV latency is a central goal of the CWRU Center for Excellence on the Impact of Substance Use on HIV. Our understanding of HIV latency and persistence has been complicated by the small numbers of latently infected cells found in the circulation, the difficulty of obtaining comprehensive sets of tissue samples from patients, the lack of known phenotypic markers that can distinguish latently infected cells from uninfected ones, and limited information about the behavior of tissue reservoirs in vivo. Core B will provide both ex vivo primary cell models (Aim 1) and highly sensitive nucleic acid-based assays to measure the impact of substance use on the HIV reservoir (Aim 2) in clinical samples. A key unresolved issue is how each individual drug modulates the latent HIV reservoir in multiple cell types and tissues. The Primary Cell, Biomimetic, and iPSC-derived Cell Models Core B team will provide and develop for Center investigators innovative and highly informative cellular and biomimetic models for the three main organ targets of drug use and HIV – the brain, gut, and the immune system. Organoids derived from tissue explants and induced pluripotent stem cells (iPSC) will be used to mimic the tissue microenvironment and extend the range of assays beyond what can be achieved using single cell ex vivo models. The models offered by Core B will be used throughout the Center of Excellence to evaluate the impact of drug use on HIV latency and will also be established ex vivo with samples derived from persons with Substance Use Disorder and HIV on anti-retroviral therapy. These models will each reflect the diversity of molecular mechanisms governing the response to drug use and HIV latency in CD4 T cells from the periphery, mucosal T cells from the gut, and microglial, neuronal, and microvascular endothelial cells from the CNS and will also provide unique opportunities to contrast the effects of substance use in vivo with direct cell biological analyses ex vivo. Importantly, the Core B team will continue to develop new assays and methods to study the impact of substance use on the mechanisms of HIV persistence and latency. In particular, this effort includes (a) developing single cell RNA induction assays, (b) modeling the TFH compartment, and (c) advancing iPSC-derived cerebral organoids to study drug use and HIV latency in the CNS. Thus, Core B, in collaboration with the molecular services in Core C, will provide access to novel technologies for evaluating the cellular, proteomic, microbiological, metabolomic, and transcriptomic changes associated with substance use and their impact on HIV reservoirs in patient samples available in Core D and to primary cell cultures and tools to manipulate cells that would not otherwise be available to Substance Use researchers at our institutions and nationwide.