Single Cell Analysis of Epigenetic Mechanisms that Regulate HIV-1 CNS Latency and Neuropathogenesis

调节 HIV-1 CNS 潜伏期和神经发病机制的表观遗传机制的单细胞分析

• 批准号: 10748578
• 负责人: CRISTIAN COARFA
• 金额: $ 88.56万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-06 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748578
• 关键词: Antiviral Therapy; Autopsy; Bioinformatics; Blood; Brain; CD4 Positive T Lymphocytes; Cell Line; Cell Nucleus; Cells; Cessation of life; Chromatin; Cognition; Cognitive; Cognitive deficits; Collection; Data; Dementia; Diagnosis; Effectiveness; Epigenetic Process; Gene Expression; Genetic Transcription; Goals; HIV-1; HIV-associated neurocognitive disorder; High Prevalence; Hippocampus; Human; In Vitro; Individual; Infection; Knock-out; Lymphoid Tissue; Messenger RNA; MicroRNAs; Microglia; Minor; Neurocognitive Deficit; Neuropathogenesis; Persons; Prefrontal Cortex; Prior Therapy; Research; Role; Sample Size; Sampling; Site; Specimen; Technology; Therapeutic; Toxic effect; United States; Untranslated RNA; Validation; Viral; Viral reservoir; Virus; Virus Latency; Virus Replication; Work; antiretroviral therapy; cell type; experimental study; gene network; induced pluripotent stem cell; insight; latent virus activation; motor disorder; neuroAIDS; neurocognitive disorder; neurotropic; novel; novel therapeutic intervention; overexpression; reactivation from latency; single cell analysis; single-cell RNA sequencing; transcriptome sequencing

Current antiretroviral therapies (ART) are successful in suppressing HIV-1 replication in most individuals. However, cessation of ART results in emergence of HIV-1 from a latent viral reservoir, thereby requiring a lifetime ART regime. The best-described HIV-1 reservoir is that of CD4+ T lymphocytes in circulating blood and lymphoid tissues that contain a transcriptionally silent but replication-competent virus. The brain is also an important viral reservoir site, but it is poorly characterized due to the challenges in obtaining specimens for analyses. Despite its effectiveness, multiple toxicities are associated with ART, especially the high prevalence of HIV-associated neurocognitive deficits. The research described in this application will use human autopsy specimens to investigate two key issues regarding HIV-1 infection of the brain: 1) epigenetic mechanisms that regulate active and latent HIV-1 infection in the brain; 2) mechanisms underlying neuropathogenesis and neurocognitive deficits in HIV-1-infected individuals on suppressive ART. Brain specimens from prefrontal cortex and hippocampus from HIV-1 infected individuals treated with ART death will be obtained from the National NeuroAIDS Consortium. Matched HIV-1-negative control specimens will be obtained from the UTHealth Brain Collection. Single cell RNA sequencing technology will be utilized to identify long noncoding RNAs (lncRNAs), mRNAs, and microRNAs (miRNAs) that are dysregulated in HIV-1-infected individuals. Validation experiments will be performed in HIV-1-infected microglia cells to identify dysregulated lncRNAs and miRNAs that contribute to HIV-1 replication, latency, and neuropathogenesis. Completion of the proposed work is likely to identify therapeutic approaches to treat HIV-1 infection of the brain.

目前的抗逆转录病毒疗法 (ART) 可以成功抑制大多数个体中的 HIV-1 复制。 然而，停止 ART 会导致 HIV-1 从潜伏病毒库中出现，因此需要 终身 ART 制度。最清楚描述的 HIV-1 储存库是循环血液中的 CD4+ T 淋巴细胞 以及含有转录沉默但具有复制能力的病毒的淋巴组织。大脑也是一个 重要的病毒储存库位点，但由于获取样本的挑战，对其特征了解甚少。 分析。尽管 ART 有效，但多种毒性与 ART 相关，尤其是高患病率 HIV相关的神经认知缺陷。本申请中描述的研究将使用人体尸检 样本来研究有关大脑 HIV-1 感染的两个关键问题：1）表观遗传机制 调节大脑中活跃和潜伏的 HIV-1 感染； 2）神经发病机制和 接受抑制性 ART 治疗的 HIV-1 感染者的神经认知缺陷。前额叶的大脑标本 接受 ART 死亡治疗的 HIV-1 感染者的皮质和海马体将从 国家神经艾滋病联盟。匹配的 HIV-1 阴性对照标本将从 UTHealth 大脑集合。单细胞RNA测序技术将用于识别长非编码 HIV-1 感染者体内失调的 RNA (lncRNA)、mRNA 和 microRNA (miRNA)。 将在 HIV-1 感染的小胶质细胞中进行验证实验，以识别失调的 lncRNA 和 有助于 HIV-1 复制、潜伏期和神经发病机制的 miRNA。完成拟议的 这项工作可能会确定治疗大脑 HIV-1 感染的治疗方法。