HIV, Methamphetamine and Human iPSC-derived Microglia-containing Cerebral Organoids

HIV、甲基苯丙胺和人 iPSC 衍生的含有小胶质细胞的大脑类器官

• 批准号: 10398189
• 负责人: WENZHE HO
• 金额: $ 61.76万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10398189
• 关键词: AIDS prevention; Acute; Astrocytes; Autopsy; Biological Assay; Biological Models; Biopsy; Brain; Cell Line; Cells; Cerebrum; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; DLG4 gene; DNA; Development; Drug abuse; Enzyme-Linked Immunosorbent Assay; Feasibility Studies; Flow Cytometry; GAG Gene; Generations; Glial Fibrillary Acidic Protein; Growth; HIV; HIV Infections; HIV-associated neurocognitive disorder; Human; ITGAM gene; Immunohistochemistry; In Vitro; Individual; Inflammasome; MCM2 gene; Methamphetamine; Methods; Microglia; Modeling; Molecular; National Institute of Drug Abuse; Natural Immunity; Nature; Neuraxis; Neurons; Opioid; Organ; Organoids; Pathogenesis; Patients; Peripheral; Physiological; Protocols documentation; RNA; Reproducibility; Research; Role; Synaptophysin; Time; United States; Virus; Western Blotting; Work; acute infection; base; brain cell; cell type; chronic infection; clinically relevant; confocal imaging; drug of abuse; experience; human model; induced pluripotent stem cell; interest; macrophage; methamphetamine abuse; methamphetamine effect; methamphetamine use; methamphetamine user; nervous system development; nestin protein; neuroAIDS; neurogenesis; neuron apoptosis; neuronal circuitry; neuronal survival; neurotropic; psychostimulant; relating to nervous system; stem; stem cells; success; synaptogenesis; synaptotagmin

Abstract Methamphetamine (METH), a potent addictive psychostimulant, is one of the most commonly abused drugs in the United States. METH abuse is highly prevalent in HIV-infected individuals, which presents unique challenges for HIV prevention and treatment. Given the overlap impact of METH use and HIV on neuronal damage in the central nervous system (CNS), it becomes urgent to understand the role of interplays between METH and HIV in the pathogenesis of HIV-associated neurocognitive disorders (HAND). However, studies of HAND have been hampered by difficulties in collecting live brain cells from autopsy or biopsy of HIV patients. Recent success in generating microglia and cerebral organoids from human induced pluripotent stem cells (iPSCs) now offers a great opportunity to investigate the impact of METH and/or HIV on the CNS. The overall objective of this project is to validate and establish a clinically relevant in vitro brain model for NeuroAIDS research in the context of drug abuse. The project will be carried out by two P.I.s who have the complementary expertise and experience in the proposed studies. Dr. Ho has been working on the impact of abused drugs (METH and opioids) on peripheral/CNS innate immunity and HIV infection of macrophages/microglia since 1999. Dr. Hu’s research focuses on neurogenesis, iPSC generation, inflammasomes, CRISPR/Cas HIV eradication and HAND. Their recent collaborative work has successfully generated and characterized the human iPSC-derived microglia-containing cerebral organoids (MCOs) which express the major CNS cell types: neural stem/progenitor cells, neurons, astrocytes, and microglia. More importantly, these MCOs could be infected by HIV. Based on these important findings, we propose three specific aims: Aim 1. To study dynamic HIV infection of human iPSC-derived MCOs and the impact of METH on HIV infection of MCOs; Aim 2. To determine HIV-infected cell types and the primary target/reservoir cells of HIV in MCOs; Aim 3. To study the impact of HIV infection on the development and neuronal circuitry in MCOs. Successful completion of this project should provide an in vitro brain model to study the roles of HIV infection and drugs of abuse in the pathogenesis of NeuroAIDS.

抽象的 甲基苯丙胺（甲基苯丙胺）是一种潜在的加性心理刺激剂，是最常见的滥用药物之一 美国。在感染艾滋病毒的人中滥用甲基苯丙胺很普遍，这表现出独特的 艾滋病毒预防和治疗面临的挑战。鉴于甲基苯甲酸和HIV对神经元的重叠影响 中枢神经系统（CNS）的损害，迫切需要了解之间的相互作用 艾滋病和艾滋病毒在与HIV相关的神经认知疾病的发病机理中（手）。但是，研究 从尸检或HIV患者的活检中收集活脑细胞的困难使手受到阻碍。 最近从人类诱导的多能干细胞产生小胶质细胞和脑器官方面的成功 （IPSC）现在提供了一个很好的机会，可以调查甲基苯丙胺和/或艾滋病毒对中枢神经系统的影响。总体 该项目的目的是验证和建立与临床相关的体外大脑模型 神经辅助研究在药物滥用的背景下进行研究。该项目将由两个拥有的P.I.进行 在拟议的研究中完全专业知识和经验。 Ho博士一直在研究 滥用药物（甲基苯甲酸属和阿片类药物）在外围/中枢神经系统的先天免疫和HIV感染上 自1999年以来巨噬细胞/小胶质细胞。HU博士的研究重点是神经发生，IPSC的产生， 炎症，CRISPR/CAS HIV消除和手。他们最近的合作工作已成功 生成并表征了人类IPSC衍生的含小胶质细胞的大脑器官（MCOS） 表达主要的CNS细胞类型：神经元/祖细胞，神经元，星形胶质细胞和小胶质细胞。更多的 重要的是，这些MCO可以被HIV感染。基于这些重要发现，我们提出了三个 具体目的：目的1。研究人IPSC衍生的MCO的动态HIV感染和甲基苯丙胺的影响 关于MCO的HIV感染；目标2。确定感染HIV的细胞类型和主要目标/储层细胞 MCOS的艾滋病毒；目的3。研究艾滋病毒感染对MCOS中发育和神经元电路的影响。 该项目的成功完成应提供一个体外大脑模型来研究HIV感染的作用 和神经助剂的发病机理中的滥用药物。