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

目前的抗逆转录病毒疗法(ART)在抑制大多数人的HIV-1复制方面是成功的。 然而，停止抗逆转录病毒疗法会导致HIV-1从潜伏的病毒储备库中出现，因此需要 终身艺术制度。被描述得最好的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复制能力强的病毒的淋巴组织。大脑也是一种 重要的病毒储存点，但由于在获取样本方面的挑战，它的特征很差 分析。尽管ART有效，但它的多种毒性与ART有关，特别是高患病率 与艾滋病毒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 研究与HIV-1脑感染有关的两个关键问题的标本：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 HIV-1感染者在抑制性ART方面的神经认知缺陷。前额叶脑组织标本 接受抗逆转录病毒治疗的HIV-1感染者的大脑皮质和海马体将从 国家神经艾滋病联盟。匹配的HIV-1阴性对照样本将从 UTHealth大脑收藏。单细胞RNA测序技术将被用来识别长非编码 在HIV-1感染者中不受调控的RNA(LncRNAs)、mRNAs和microRNAs(MiRNAs)。 验证实验将在感染HIV-1的小胶质细胞中进行，以识别失调的lncRNA和 有助于HIV-1复制、潜伏和神经发病的miRNAs。完成拟议的 这项工作可能会找到治疗脑内HIV-1感染的治疗方法。

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.