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.

项目摘要 - 原代细胞，仿生和IPSC衍生的细胞模型核心B 了解可卡因，甲基苯丙胺，大麻和阿片类药物的机制 放大艾滋病毒潜伏期的损害是CWRU卓越中心的核心目标 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已经 在循环中发现的少量潜在感染细胞变得复杂，困难 从患者那里获得全面的组织样品集，缺乏已知的表型 可以将潜在感染细胞与未感染细胞区分开的标记和有限的信息 关于体内组织储层的行为。核心B将提供两个离体主细胞模型 （AIM 1）和高度敏感的基于核酸的测定法以测量物质使用的影响 在临床样品中的HIV储层（AIM 2）上。一个未解决的关键问题是每个人如何 药物在多种细胞类型和组织中调节潜在的HIV储量。主要细胞， 仿生和IPSC衍生的细胞模型核心B团队将为中心提供和开发 研究人员的创新性和信息丰富的细胞和仿生模型，用于三个主要的 源自药物使用和艾滋病毒的器官 - 大脑，肠道和免疫系统。 来自组织外植体和诱导的多能干细胞（IPSC）将用于模仿组织 微环境并将测定的范围扩展到使用单个单元的实现的范围 离体模型。核心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，并将在体内建立样品 源自抗逆转录病毒疗法的药物使用障碍和HIV的人。这些 模型将每个模型反映有关对药物反应的分子机制的多样性 来自外周，粘膜T细胞的CD4 T细胞中的使用和HIV潜伏期，来自肠道的粘膜T细胞， 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 与直接细胞生物学对比的独特机会将物质使用的影响与物质使用的影响 分析离体。重要的是，核心B团队将继续开发新的测定法和方法 研究物质使用对HIV持久性和潜伏期机制的影响。在 特别是，这项工作包括（a）开发单细胞RNA诱导分析，（b）建模 TFH隔室，（c）推进IPSC衍生的大脑器官来研究药物使用和HIV 中枢神经系统的延迟。核心B与Core C中的分子服务合作，将 提供对新技术的访问，以评估细胞，蛋白质组学，微生物学， 代谢组学和与药物使用相关的转录组变化及其对HIV的影响 核心D中可用的患者样品中的水库以及原代细胞培养物和工具 操纵细胞，否则在我们的 机构和全国。