Immune Network Dysregulation of the Central Nervous System with HIV Persistence and Opioid Abuse

项目摘要/摘要 尽管用抗逆转录病毒疗法(ART)成功地抑制了病毒复制，但艾滋病毒仍在体内存在， 阻止了病毒的根除。即使大脑中持续存在的艾滋病毒水平很低，也可能导致神经疾病 损害，因为30-50%接受抗逆转录病毒治疗的艾滋病毒抑制良好的人会发展出与艾滋病毒相关的神经认知 精神障碍(手)。中枢神经系统的这些异常因阿片类药物滥用而进一步复杂化，如阿片类药物暴露 会导致过度的神经炎症。HIV和OUD诱导的中枢神经系统免疫激活不完全 在细胞水平上被理解。了解中枢神经系统持续免疫激活的细胞基础至关重要 用于减少日益增长的HIV和OUD成人人口的神经系统疾病。的最新进展 大规模并行的单细胞RNA测序已经广泛地发现了大量的神经元和神经胶质细胞群体 在健康和疾病中贯穿中枢神经系统。这样的方法提供了一种强大而公正的方式 为了了解基于转录的中枢神经系统细胞和免疫网络的组织 单个单元级别的配置文件。在这里，我们建议使用最先进和新颖的方法来 神经科学，免疫生物学和计算生物学，解剖失控的免疫网络 艾滋病毒患者和OUD患者的中枢神经系统以及在艾滋病毒背景下对阿片类药物的细胞类型特异性反应。我们会 应用最先进的单细胞转录组分析揭示分子结构和免疫 HIV和OUD患者脑脊液的活化我们将利用我们的人脑单个大型数据集 NIDA支持的单细胞阿片类药物中枢数据生成中心的核RNA测序 耶鲁大学艾滋病毒背景下的反应(Y-SCOCH)以表征神经免疫的变化 OUD和HIV中的通信。我们将开发和应用现代针对细胞类型的新方法，如 作为体外培养系统，测定脑免疫细胞类型的阿片类反应，包括小胶质细胞和 在艾滋病毒的背景下，大脑驻留的T细胞。确定调控网络和分子机制 艾滋病毒对阿片类药物暴露的持续反应将极大地促进我们对艾滋病毒潜伏期的理解，并可能 为治疗手部和根除HIV病毒提供新的见解和众多的药物靶点 患有阿片类药物使用障碍的持久者。

PROJECT SUMMARY/ABSTRACT HIV persists within the body despite successful suppression of viral replication with antiretroviral therapy (ART), preventing eradication of the virus. Even a low level of persistent HIV in the brain may cause neurological damage, as 30-50% of well-suppressed HIV individuals under ART develop HIV-associated neurocognitive disorders (HAND). These abnormalities in the CNS are further complicated by opioid abuse, as opioid exposure results in exaggerated neuroinflammation. CNS immune activation induced by HIV and OUD is incompletely understood at the cellular level. Understanding the cellular basis for persistent CNS immune activation is critical for reducing neurological morbidities in the growing population of adults with HIV and OUD. Recent advances in massively parallel single cell RNA sequencing have uncovered numerous neuronal and glial populations widely across the central nervous system in health and disease. Such methods provide a powerful and unbiased way for understanding the organization of the cellular and immune network of the CNS based on transcriptional profiles at the single cell level. Here, we propose to employ state-of-the-art and novel methodologies in neuroscience, immunobiology, and computational biology to dissect the dysregulated immune network in the CNS of patients with HIV and OUD and the cell-type-specific response to opioid in the context of HIV. We will apply state-of-the-art single cell transcriptome analysis to uncover the molecular architecture and immune activation of the CSF in the patients with HIV and OUD. We will leverage our large dataset of human brain single nucleus RNA sequencing generated at the NIDA-supported CNS data generation center of Single Cell Opioid Responses in the Context of HIV at Yale (Y-SCORCH) to characterize the alterations of neuroimmune communications in OUD and HIV. We will develop and employ novel modern cell-type-specific approaches, such as ex vivo culture system, to determine the opioid responses of brain immune cell types, including microglia and brain-resident T cells, in the context of HIV. Determining the regulatory networks and molecular mechanisms of HIV persistence in response to opioid exposure will greatly advance our understanding of HIV latency and may provide novel insights and numerous pharmaceutical targets for treating HAND and eradicating HIV virus in HIV persistent individuals with opioid use disorder.