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：
10301839
负责人：
Schahram Akbarian
金额：
$ 74.44万
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/10301839
关键词：
3-Dimensional AIDS/HIV problem 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 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 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 Colony-Stimulating Factor Maps Measures Mediating Microglia Modeling Molecular Conformation Molecular Profiling Mus Neonatal Neurobiology Neuroglia Pattern Peripheral Pharmaceutical Preparations Primary Cell Cultures RNA Regimen Reporter Rewards Site Specimen Substance Use Disorder Switch Genes System Technology Testing Time Tissues Transcript Transplantation Treatment Protocols Umbilical Cord Blood Variant Viral Work addiction antiretroviral therapy base cannabinoid receptor cell type cognitive function epigenome epigenomics improved induced pluripotent stem cell integration site latent infection macrophage mouse model neonatal mice nervous system disorder 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.

HIV相关的神经认知障碍在联合抗逆转录病毒治疗时代持续存在 (cART)，而人类中枢神经系统中的 HIV 潜伏期和 HIV 转录物的细胞特异性表达仍然存在与艾滋病毒相关的神经系统疾病的患病率很高，而且还在不断增加。在 cART 稳定抑制的人群中识别中枢神经系统病毒逃逸，通常进一步复杂化 HIV 感染者/艾滋病患者 (PLWHA) 中共同登记的物质使用障碍 (SUD) 流行病，因为 SUD 也对中枢神经系统功能产生深远的影响，我们实验室正在进行的工作首先提供了这一点。评估细胞类型特异性艾滋病毒“分子特征”，包括基因组整合模式和奖赏和成瘾回路中转录组和表观基因组水平的改变正如“初步数据”部分详细描述的那样，我们发现了戏剧性的结果。死后标本的小胶质细胞子集中有高水平的 HIV 表达，并带有 HIV 转录本水平位居小胶质细胞中表达最高的 5 个 RNA 之列，即所有 RNA 的 99.9% 相应地，成瘾回路中的HIV基因组整合位点占主导地位。然而，小胶质细胞特异性基因对活性染色质区室具有强烈偏好。 cART 期间持续存在的潜伏感染的挥之不去的影响尚未得到很好的表征——部分由于确定小胶质细胞对细胞的贡献程度存在根本性挑战我们的初步研究还提供了一个模型系统，我们可以通过它来跟踪和隔离。持续感染的细胞，可以应用于小胶质细胞室，并使我们能够定义通过研究人类小胶质细胞中的 HIV 基因组，研究 cART 期间持续存在的基因组扰动。驻留在小鼠大脑中并将其与追踪持续感染的小胶质细胞的技术联系起来，我们将能够首次对实验疗法和干预措施进行建模，以补充我们的研究具体来说，我们基于 Cre-reporter 的 HIV 诱导谱系的描述性工作。追踪（HILT）标记系统将使我们能够量化和分离罕见的潜伏感染小胶质细胞，在 cART 期间持续存在，并分别绘制感染者和感染者的转录组和表观基因组改变图谱未感染的小胶质细胞，均来自同一小鼠大脑，我们将重点关注附加电路。研究接受标准 cART 方案治疗的小鼠的神经炎症、认知和奖赏行为以及一种涉及大麻素受体 2 激动剂药物的实验性疗法，根据我们的研究初步数据显示，其与限制组织中艾滋病毒感染程度的抗炎活性有关。