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

研究支持核心 B：原代细胞、仿生和 iPSC 衍生细胞模型

• 批准号: 10304584
• 负责人: JONATHAN KARN
• 金额: $ 73.12万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10304584
• 关键词: Behavior; Biological; Biological Assay; Biological Models; Biomimetics; Blood Cells; Blood Circulation; Brain; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cannabis; Cell Line; Cell model; Cells; Cerebrum; Clinical; Clinical Trials; Cocaine; Coculture Techniques; Collaborations; Color; Complex; DNA; Drug Targeting; Drug usage; Endothelial Cells; Endothelium; Epithelial Cells; Gap Junctions; Genes; Goals; Guide RNA; HIV; HIV Genome; HIV Infections; Immune system; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Inflammation; Innate Immune System; Institution; Intestines; Measures; Methamphetamine; Methods; Microbiology; Microglia; Migration Assay; Modeling; Modification; Molecular; Mucous Membrane; Neurodegenerative Disorders; Neurons; Nucleic Acids; Opioid; Organ; Organoids; Pathogenesis; Patients; Peripheral; Persons; Pharmaceutical Preparations; Primary Cell Cultures; Proteomics; RNA; Research; Research Personnel; Research Support; Ribonucleoproteins; Sampling; Services; Small Intestines; Stimulus; Study models; Substance Use Disorder; System; T memory cell; T-Lymphocyte; Techniques; Technology; Tissue Sample; Tissues; Training; Vascular Permeabilities; Viral reservoir; Virus; antiretroviral therapy; base; brain endothelial cell; cell transformation; cell type; digital; droplet sequencing; drug of abuse; genome editing; human model; immunoregulation; in vivo; induced pluripotent stem cell; innovation; intestinal epithelium; latent HIV reservoir; metabolomics; monolayer; nervous system disorder; new technology; novel; phenotypic biomarker; response; single-cell RNA sequencing; stem cell technology; substance use; tool; transcriptomics; viral DNA; viral RNA; viral detection

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.

项目总结-原代细胞、仿生学和iPSC衍生细胞模型核心B 了解可卡因、甲基苯丙胺、大麻和阿片类药物 扩大艾滋病毒潜伏期的损害是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 由于在循环中发现的潜伏感染细胞数量很少， 从患者获得全面的组织样本集，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 关于体内组织库的行为。核心B将提供两种离体原代细胞模型 (Aim 1）和高灵敏度的基于核酸的测定，以测量物质使用的影响 对临床样本中的HIV储存库（Aim 2）的影响。一个关键的悬而未决的问题是， 药物调节多种细胞类型和组织中潜伏的HIV库。主要细胞， 仿生和iPSC衍生细胞模型核心B团队将为中心提供和开发 研究人员为三个主要的创新和高度信息化的细胞和仿生模型 吸毒和艾滋病毒的器官靶点-大脑，肠道和免疫系统。类器官 诱导多能干细胞（iPSC）将用于模拟组织 微环境，并扩大了测定范围，超出了使用单细胞 离体模型。Core B提供的模型将在整个卓越中心使用 以评价药物使用对HIV潜伏期的影响，也将用样本离体建立 来自接受抗逆转录病毒治疗的物质使用障碍和艾滋病毒感染者。这些 每个模型都将反映出控制药物反应的分子机制的多样性， 来自外周的CD 4 T细胞、来自肠道的粘膜T细胞中的HIV使用和潜伏期，以及 小胶质细胞、神经元和微血管内皮细胞， 独特的机会，以对比物质使用在体内的影响与直接细胞生物学 离体分析。重要的是，核心B团队将继续开发新的检测和方法 研究药物使用对艾滋病毒持久性和潜伏机制的影响。在 具体而言，这一努力包括（a）开发单细胞RNA诱导测定，（B）模拟 TFH隔室，以及（c）推进iPSC衍生的脑类器官以研究药物使用和HIV CNS中的潜伏期。因此，核心B将与核心C中的分子服务协作， 提供新的技术，用于评估细胞，蛋白质组学，微生物， 与药物使用相关的代谢组学和转录组学变化及其对HIV的影响 核心D中可用的患者样本中的储库以及原代细胞培养物和工具， 操纵细胞，否则将无法提供给物质使用研究人员在我们的 机构和全国。