Modeling HIV Microglia-Associated Infection and Inflammation in a Chimeric Mouse Brain

在嵌合小鼠大脑中模拟 HIV 小胶质细胞相关的感染和炎症

基本信息

批准号：
10632139
负责人：
Schahram Akbarian
金额：
$ 74.35万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
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/10632139
关键词：
3-Dimensional Affect Agonist Anti-Inflammatory Agents Antiinflammatory Effect Autopsy Award Behavior Biological Models Brain CD34 gene CD8B1 gene CNR1 gene CNR2 gene Cells Chromatin Cocaine Abuse Cognition Combined Modality Therapy Complement Data Diagnosis Disease DsRed Encephalitis Enterobacteria phage P1 Cre recombinase Epidemic Funding Gene Expression Genes Genetic Recombination Genome Genomics HIV HIV Genome HIV Infections HIV encephalitis HIV-associated neurocognitive disorder HIV/AIDS Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hi-C High Prevalence Human Immune system Immunodeficient Mouse Infection Inflammation Injections Intervention Investigational Therapies Knock-in Laboratories Link Liver Macrophage Macrophage Colony-Stimulating Factor Maps Measures Mediating Microglia Modeling Molecular Conformation Molecular Profiling Mus Neonatal Neuroanatomy Neurobiology Neuroglia Pattern Peripheral Persons Pharmaceutical Preparations Predisposition Primary Cell Cultures RNA Regimen Reporter Rewards Site Sorting Specimen Substance Use Disorder Switch Genes System Technology Testing Time Tissues Transcript Transplantation Treatment Protocols Umbilical Cord Blood Variant Viral Work addiction antiretroviral therapy cannabinoid receptor cell type cognitive function epigenome epigenomics improved induced pluripotent stem cell integration site latent infection mouse model neonatal mice neuroAIDS neurogenomics neuroinflammation novel opioid abuse preference progenitor reconstitution transcriptome transcriptome sequencing transcriptomics

项目摘要

HIV-associated neurocognitive disorders persist in the era of combination antiretroviral therapy (cART) while HIV latency, and cell-specific expression of HIV transcript in human CNS remains incompletely understood. There is high prevalence of HIV-associated neurologic disease and increasing recognition of CNS viral escape in people stably suppressed with cART, often further complicated by the co-registered epidemic of substance use disorders (SUD) in people living with HIV/AIDS (PLWHA), as SUD also have profound impact on CNS function. Ongoing work in our laboratory is providing first assessments of cell-type specific HIV 'molecular signatures', including genome integration patterns and alterations on the level of the transcriptome and epigenome in reward- and addiction circuitry of the human postmortem brain. As described in detail in the Preliminary Data section, we found dramatically high levels of HIV expression in a subset of microglia from postmortem specimens, with HIV transcript levels ranking among the top 5 highest expressed RNAs in microglia, or the 99.9% percentile of all microglial transcript. Correspondingly, HIV genome integration sites in addiction circuitry are dominated by microglia-specific genes, with strong preference for active chromatin compartments. However, lingering effects of latent infection that persist during cART have not been well characterized—in part because of fundamental challenges in identifying the extent to which microglial cells contribute to the latent reservoir. Our preliminary studies also provide a model system whereby we can track and isolate persistently infected cells which can be applied to the microglial compartment and will allow us to define the genomic perturbations that persist during cART. By studying HIV genomics in human microglia residing in the mouse brain and linking this with technology to track persistently infected microglia, we will be able to model, for the first time, experimental therapies and interventions to complement our descriptive work in human postmortem brain. Specifically, our Cre-reporter based HIV-induced lineage tracing (HILT) marking system will allow us to quantify and isolate the rare latently infected microglia that persist during cART, and map transcriptomic and epigenomic alterations separately both for infected, and non-infected microglia, both collected from the same mouse brain. With focus on addition circuitry, we will study neuroinflammation, cognition and reward behavior in mice treated with standard cART regimens and an experimental therapy involving Cannabinoid receptor 2 agonist drugs that, according to our preliminary data, are linked to anti-inflammatory activity limiting the extent of HIV infection in tissues.

在抗逆转录病毒疗法的时代，与HIV相关的神经认知疾病持续存在（CART）而HIV潜伏期和HIV转录本在人CNS中的细胞特异性表达仍然存在不完全理解。与HIV相关的神经系统疾病的患病率很高认识到CNS病毒逃生的人稳定地被推车抑制，通常会更加复杂艾滋病毒/艾滋病患者（PLWHA）的物质使用障碍（SUD）共同注册的流行病，由于SUD对中枢神经系统功能也有深远的影响。我们实验室正在进行的工作首先提供评估细胞类型的特异性HIV“分子特征”，包括基因组整合模式和奖励和成瘾电路中转录组和表观基因组水平的改变人类后大脑。如“初步数据”部分中详细描述的，我们发现了巨大的 hiv转录本的小胶质细胞中的小胶质细胞中高水平的HIV表达在小胶质细胞中排名最高的5个最高表达RNA，或所有分数中排名最高的99.9％小胶质成绩单。相应地，成瘾电路中的HIV基因组整合位点主导通过小胶质细胞特异性基因，对活性染色质区室具有强烈的偏爱。然而，在购物车期间持续存在的潜在感染的挥之不去的影响尚未得到很好的特征 - 部分由于基本挑战在确定小胶质细胞在多大程度上有助于潜在水库。我们的初步研究还提供了一个模型系统，我们可以跟踪和隔离持续感染的细胞，可应用于小胶质细胞室，并允许我们定义在购物车期间持续存在的基因组扰动。通过研究人类小胶质细胞中的HIV基因组学居住在鼠标大脑中，并将其与技术联系在一起以跟踪持续感染的小胶质细胞，我们将能够首次建模实验疗法和干预措施以完成我们的人类验尸大脑中的描述性工作。具体而言，我们基于CRE-Reporter的HIV诱导谱系跟踪（HILT）标记系统将使我们能够量化和隔离很少受感染的小胶质细胞在卡车期间持续存在，并分别映射转录组和表观基因组改变，既有感染，又未感染的小胶质细胞，都是从同一小鼠脑收集的。专注于加法电路，我们将研究用标准推车治疗的小鼠中的神经炎症，认知和奖励行为并且一种实验疗法涉及大麻素受体2激动剂药物，根据我们的初步数据与抗炎活性有关，限制了组织中HIV感染的程度。