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

HIV、甲基苯丙胺和人类 iPSC 衍生的含有小胶质细胞的大脑类器官

• 批准号: 10055449
• 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/10055449
• 关键词: AIDS prevention; Acute; Astrocytes; Autopsy; Biological Assay; Biological Models; Biopsy; Brain; Cell Line; Cells; Cellular Structures; 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（METH）是一种强效成瘾性精神兴奋剂，是最常滥用的药物之一。 在美国，冰毒滥用在艾滋病毒感染者中非常普遍，这具有独特性。 鉴于冰毒的使用和艾滋病毒对神经元的重叠影响，艾滋病毒预防和治疗面临的挑战。 中枢神经系统（CNS）损伤，因此迫切需要了解之间相互作用的作用 然而，METH 和 HIV 在 HIV 相关神经认知障碍 (HAND) 发病机制中的研究。 HAND 因从 HIV 患者尸检或活检中收集活脑细胞存在困难而受到阻碍。 最近成功利用人类诱导多能干细胞生成小胶质细胞和脑类器官 （iPSC）现在提供了一个很好的机会来研究冰毒和/或艾滋病毒对中枢神经系统的影响。 该项目的目标是验证并建立临床相关的体外脑模型 该项目将由两名拥有药物滥用背景的 P.I. 进行。 何博士一直致力于研究所提出的研究的互补专业知识和经验。 滥用（冰毒和阿片类药物）对外周/中枢神经系统先天免疫和艾滋病毒感染的影响 自1999年以来一直致力于巨噬细胞/小胶质细胞。胡博士的研究重点是神经发生、iPSC生成、 他们最近的合作工作在炎症小体、CRISPR/Cas HIV 根除和 HAND 方面取得了成功。 生成并表征了人类 iPSC 衍生的含有小胶质细胞的大脑类器官 (MCO)， 表达主要的中枢神经系统细胞类型：神经干/祖细胞、神经元、星形胶质细胞和小胶质细胞。 重要的是，这些 MCO 可能会感染 HIV。根据这些重要发现，我们提出了三种建议。 具体目标： 目标 1. 研究人类 iPSC 衍生的 MCO 的动态 HIV 感染以及 METH 的影响 MCO 的 HIV 感染；目标 2. 确定 HIV 感染细胞类型和主要靶细胞/储存细胞 MCO 中的 HIV；目标 3. 研究 HIV 感染对 MCO 发育和神经元回路的影响。 该项目的成功完成将为研究艾滋病毒感染的作用提供体外脑模型 以及神经艾滋病发病机制中的滥用。