Cerebral organoid and IPSC derived microglia: Modeling of HIV and methamphetamine co-morbidity

大脑类器官和 IPSC 衍生的小胶质细胞：HIV 和甲基苯丙胺共病模型

基本信息

批准号：
10687067
负责人：
Peng Jiang
金额：
$ 53.11万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10687067
关键词：
3-Dimensional Abuse Reporting Acquired Immunodeficiency Syndrome Address Adolescent Adult Anti-Inflammatory Agents Anti-Retroviral Agents Applications Grants Astrocytes Autopsy Biogenesis Biological Assay Biology Blood - brain barrier anatomy Brain CRISPR/Cas technology Cells Central Nervous System Cerebrum Chronic Clinical Coculture Techniques Data Dendritic Spines Development Diagnosis Disease Drug abuse End Point Assay Ensure Erythro Etiology Event Genes Goals HIV HIV Genome HIV Infections HIV-associated neurocognitive disorder High Prevalence Human Individual Inflammation Inflammatory Knowledge Macrophage Measures Mediating Methamphetamine MicroRNAs Microglia Mitochondria Modeling Molecular Molecular Profiling Monkeys Mus Myelogenous Nerve Degeneration Neuroglia Neuroimmune Neuronal Injury Neurons Neuropathogenesis Organoids Outcome Pathogenesis Pathology Pathway interactions Patients Pattern Penetration Persons Pharmaceutical Preparations Population Property Proteins Regimen Reproducibility Role SIV Sampling Series Signal Pathway Signal Transduction Societies Synapses Technology Toxic effect Treatment Efficacy Viral Virus Diseases Yolk Sac antiretroviral therapy brain dysfunction brain tissue cell type comorbidity design drug of abuse experimental study extracellular vesicles genome editing graft vs host disease human pluripotent stem cell humanized mouse in vivo induced pluripotent stem cell metabolic rate methamphetamine abuse neuroinflammation neurotoxicity non-invasive imaging novel novel therapeutics pharmacokinetics and pharmacodynamics postsynaptic presynaptic progenitor progenitor system psychostimulant receptor response self organization single-cell RNA sequencing success synaptic pruning therapeutic evaluation three-dimensional modeling tool

项目摘要

Abstract: An estimated ~1.3 million adults and adolescents are living with human immunodeficiency virus (HIV) infection in the USA of which 15% of them undiagnosed. The initiation of combination antiretroviral therapy (cART) has transformed AIDS from a fatal illness into a chronic and manageable disease. The presence of HIV infected microglia and macrophage reservoirs in the central nervous system (CNS) and variable penetration of anti- retroviral drugs across the blood brain barrier after cART are likely factors for the persistence of HIV associated neurocognitive disorder (HAND). Drugs of abuse such as the potent psychostimulant methamphetamine (Meth) that is abused by an estimated 30 million people in the world further minimize the efficacy of cART and in conjunction with HIV exacerbate CNS pathology. Thus, significant gap in knowledge in the field is to better understand the neuropathogenesis and the etiology of clinical outcomes observed in HIV+ patient’s dependent on Meth abuse. The current grant proposal focuses on developing a human induced pluripotent stem cell based cerebral organoid model to investigate underlying brain dysfunction during HIV/Meth interaction. Cerebral organoids are 3-dimensional “mini brains” that can self-organize and recapitulate many milestone events seen in in vivo brain development. Our recent study on generating a novel organoid model with the feasibility in controlling the microglia ratio and microenvironment of organoid formation to recapitulate changes in brain functions under various conditions (e.g. virus and viral infection-induced inflammation and synaptic pruning) makes this model ideal for our proposed studies. The central hypothesis of this proposal is that Meth in conjunction with HIV causes significant neuronal damage. The combination of microglia-containing cerebral organoid model, extracellular vesicle biology, single-cell RNA-sequencing, CRISPR/Cas9-mediated gene editing will decipher novel intercellular and molecular mechanisms and pathways that underlie abnormalities in neuronal functions and connectivity caused by HIV infection. We will investigate this hypothesis under three specific aims: Specific Aim 1 will examine neuronal function, synaptic and mitochondrial perturbations during HIV/meth interactions in 3D microglia-containing cerebral organoid model; Specific Aim 2 will investigate the role of extracellular vesicles in microglial organoids treated under HIV/meth conditions; and Specific Aim 3 will characterize the molecular mechanisms underlying HIV infection-induced neuronal injury and further evaluate our microglial organoid model as a reliable tool to identify molecular signatures of HAND. Our proposed experiments will decipher molecular mechanisms, novel signaling events and molecular partners underlying the neuronal injury in HAND. Through the series of the experiments we aim to present an easily scalable and reproducible model to study HAND pathogenesis. The data obtained from these studies can be used to design novel therapeutics to control HIV infection.

抽象的：估计约有130万成人和青少年患有人类免疫缺陷病毒（HIV）感染在美国，其中15％的未诊断。组合抗逆转录病毒疗法（CART）的倡议具有将艾滋病从致命疾病转变为一种慢性且易于控制的疾病。感染HIV的存在中枢神经系统（CNS）中的小胶质细胞和巨噬细胞储量以及抗侵蚀的可变渗透卡车后，横跨血液屏障的逆转录病毒药物可能是与HIV相关的持续性的因素神经认知障碍（手）。滥用药物，例如潜在的心理刺激甲基苯丙胺（METH）世界上估计有3000万人滥用这进一步最小化购物车的效率与HIV加剧的CNS病理学结合。那是在现场了解的显着差距是更好了解在HIV+患者的依赖性中观察到的临床结局的神经病生成和病因关于滥用甲基苯丙胺。当前的赠款提案着重于开发人类诱导的多能干细胞脑器官模型研究HIV/METH相互作用期间的潜在脑功能障碍。大脑器官是三维“迷你大脑”，可以自组织和概括许多里程碑事件体内脑发育。我们最近关于生成具有可行性的新型器官模型的研究控制器官形成的小胶质细胞比率和微环境，以概括大脑的变化在各种条件下的功能（例如病毒和病毒感染诱导的感染和突触修剪）使该模型非常适合我们提议的研究。该提议的核心假设是与HIV的结合会引起明显的神经元损害。含小胶质细胞的大脑的组合器官模型，细胞外囊泡生物学，单细胞RNA测序，CRISPR/CAS9介导的基因编辑将破译新型的细胞间和分子机制和途径，这些机制是神经元异常的基础 HIV感染引起的功能和连通性。我们将根据三个具体目的研究这一假设：特定的目标1将检查HIV/METH期间的神经元功能，突触和线粒体扰动含3D小胶质细胞的大脑器官模型中的相互作用；具体目标2将调查在艾滋病毒/甲基甲基条件下处理的小胶质细胞器中的细胞外蔬菜；和特定的目标3将表征HIV感染引起的神经元损伤的分子机制和进一步评估我们的小胶质器官模型是识别手分子特征的可靠工具。我们提出的实验将破译分子机制，新的信号事件和分子伙伴的基础手中的神经元损伤。通过一系列实验，我们旨在提出易于扩展的实验和可再现的模型研究手发病。从这些研究中获得的数据可用于设计控制HIV感染的新型疗法。