A DUAL CHIMERIC HUMAN ASTROGLIAL-MICROGLIAL MODEL OF HIV AND HAND

HIV和手的双嵌合人类星形胶质细胞-小胶质细胞模型

基本信息

批准号：
10625341
负责人：
STEVEN Alan GOLDMAN
金额：
$ 64.51万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10625341
关键词：
Address Adult Affect Amphetamine Abuse Anatomy Animal Experiments Animal Model Animals Architecture Astrocytes Autopsy Behavior Behavior assessment Behavioral Blood Vessels Brain CSF1R gene Categories Cells Central Nervous System Chimera organism Chronic Cognition Cognitive Cognitive deficits Communicable Diseases Corpus striatum structure Dendrites Deterioration Development Engraftment Environment Event Fiber Gene Expression Gene Expression Profile Genetic Transcription Goals HIV HIV Infections HIV-1 HIV-associated neurocognitive disorder Hominidae Human Immune Immunodeficient Mouse Infection Knockout Mice Light Lymphocyte Macrophage Maps Methods Microglia Modeling Mus Nature Neonatal Neostriatum Neurodegenerative Disorders Neuroglia Neurons Neurophysiology - biologic function Oligodendroglia Output Pathology Phenotype Pleomorphism Process Rabies Rewards Role Secondary to Structure Synapses Systemic infection T-Lymphocyte Techniques Testing Time Toxic effect Transplantation Vertebral column Viral Virulence Virus Xenograft procedure addiction brain cell cognitive enhancement comorbidity defined contribution density drug of abuse embryonic stem cell functional disability glial activation human embryonic stem cell humoral immunity deficiency implantation in vivo in vivo Model methamphetamine exposure methamphetamine use monocyte mouse model neonatal mice neural network neurocognitive disorder neurophysiology progenitor response single-cell RNA sequencing stem cells synaptic failure synaptic function trafficking

项目摘要

Abstract Synaptic failure is an important feature of HIV infection of the brain, and a likely key contributor to HIV- associated neurocognitive disorders (HAND). Yet current animal models have proven of limited utility in defining the mechanisms of this process, in part because of the species-specific nature of HIV infection, but also because of the greater complexity of human astrocytes relative to those of mice. To address this issue, we will utilize mice chimeric for both human microglia and human astrocytes, to assess the effects of HIV infection on central neurons. To that end, we will engraft mice with both human glial progenitor cells (hGPCs) and microglia, each derived from embryonic stem cells (hESCs). We have established the methods of generating these human glial chimeras, by the neonatal implantation of hGPCs, which outcompete and ultimately replace the host mouse GPCs, yielding adult chimeras broadly colonized with human astroglia1-5. This process is especially robust in the neostriatum, allowing the glial humanization of regions critically involved in striatal reward and addiction circuits. We have recently extended this approach to include chimerization with hESC-derived microglia, paired with the use of CSF1R null mice lacking host microglia, crossed to NSG SGM3 mice to allow the stable xenograft of hGPCs. The mice are thus chimeric for hGPC-derived astrocytes as well as microglia, in a T- and B-cell deficient background that allows the effects of glial HIV infection on neurons to be isolated, following intracerebral inoculation with HIV-infected microglia. These chimeric human astroglial-microglial (CHAM) mice are especially attractive, since they incorporate the hominid-specific features of human astroglia, which are themselves key components of central synapses. Using this model, we will test the postulate that astrocytes become both structurally and functionally impaired by microglial HIV infection, resulting in the loss of synaptic engagement by affected astrocytes, with consequent dendritic involution and network disruption. By infecting CHAM mice with HIV, and using rabies viral-EGFP to trace striatal dendrites, we will assess the effects of astrocytic HIV infection on the dendritic architecture and synaptic structure of resident medium spiny neurons. In parallel, we will study the effects in CHAM mice of HIV infection complicated by methamphetamine use – a common and disabling comorbidity that suppresses dopaminergic input to the striatum – focusing on the structural and transcriptional responses of human glia to the combination of infection and addiction, as well as on the behavioral effects of that combination. To that end, we will use single cell RNA-Seq to assess the changes in gene expression by human astrocytes and their partnered mouse neurons caused by HIV infection, both alone and together with chronic methamphetamine use, to identify those changes that contribute to the striatal synaptic disruption and behavioral pathology of these mice. Our goal is to test the hypothesis that the HIV-infected striatum, by virtue of astrocytic fiber disengagement from dopaminergic synapses in particular, is especially vulnerable to the effects of amphetamine abuse, while defining the transcriptional basis for the glial pathology underlying that vulnerability.

抽象的突触衰竭是大脑HIV感染的重要特征，也是艾滋病毒的关键因素相关的神经认知障碍（手）。然而，当前的动物模型已证明有限的效用此过程的机制，部分原因是艾滋病毒感染的物种特定性质，也是因为人类星形胶质细胞相对于小鼠的复杂性较高。为了解决这个问题，我们将利用老鼠人类小胶质细胞和人星形胶质细胞的嵌合，以评估HIV感染对中央的影响神经元。为此，我们将与人神经胶质祖细胞（HGPC）和小胶质细胞一起植入小鼠源自胚胎干细胞（HESC）。我们已经建立了产生这些人神经胶质的方法嵌合体，通过HGPC的新生儿植入，它超过并最终替换了宿主鼠标 GPC产生的成年嵌合体通过人类星形胶质素1-5广泛定植。这个过程在 Neostriatum，允许与纹状体奖励和成瘾电路的区域进行神经素化。最近，我们扩展了这种方法，以与hESC衍生的小胶质细胞一起使用，并与使用缺乏宿主小胶质细胞的CSF1R无效小鼠，交叉到NSG SGM3小鼠以允许稳定的异种移植物 HGPC。因此，小鼠是HGPC衍生的星形胶质细胞和小胶质细胞的嵌合后背景允许神经胶质HIV感染对神经元的影响，后脑内脑内用HIV感染的小胶质细胞接种。这些嵌合人星形胶质细胞膜（CHAM）小鼠尤其是有吸引力，因为它们包含了人类星形胶质细胞的人类特定特征，这本身就是关键中央突触的组成部分。使用此模型，我们将测试星形胶质细胞成为两者的假设小胶质细胞HIV感染在结构和功能上受损，导致突触接合的丧失影响星形胶质细胞，因此树突状相关和网络破坏。通过感染的Cham小鼠 HIV，并使用狂犬病病毒-EGFP追踪纹状体树突，我们将评估星形胶质细胞HIV感染的影响关于居民中刺神经元的树突状结构和突触结构。同时，我们将学习甲基苯丙胺使用复杂的艾滋病毒感染的CHAM小鼠的作用 - 一种常见和残疾抑制纹状体的多巴胺能输入的合并症 - 重点是结构和转录人神经胶质对感染和成瘾的组合以及对行为影响的反应该组合。为此，我们将使用单细胞RNA-seq评估基因表达的变化人类星形胶质细胞及其伴侣的小鼠神经元，由HIV感染引起慢性甲基苯丙胺的使用，以识别导致纹状体突触中断和这些小鼠的行为病理。我们的目标是检验以下假设：艾滋病毒感染的纹状体尤其是多巴胺能突触的星形胶质细胞纤维脱离接触，特别容易受到影响苯丙胺滥用，同时定义了该脆弱性的神经胶质病理的转录基础。