Role of non-coding RNA in establishing and maintaining HIV latency

 描述(由申请人提供)：尽管HIV潜伏期和重新激活具有关键的临床重要性，但我们对建立潜伏期和随后的重新激活所涉及的机制的了解非常不完整。在这项应用中，我们将研究非编码RNA，包括miRNA和lncRNA，在控制HIV转录和建立HIV前病毒的表观遗传沉默方面的作用。在许多系统中的研究表明，不同染色质修饰因子之间的复杂相互作用，以及lncRNAs和miRNA在调节特定位点的表观遗传变化中的广泛参与。虽然类似的RNA-蛋白质网络可能控制着表观遗传的变化，从而控制HIV前病毒群体的潜伏期和重新激活，但具体的机制和RNA尚未确定。我们将检验这一假设，即潜伏期的建立、维持和退出是由染色质修饰复合体施加的表观遗传变化控制的，染色质修饰复合体通过与一个或多个lncRNAs和miRNAs结合被引导到前病毒位点。这项工作将利用Karn实验室开发的新的体外系统，该系统使我们能够研究在统一的初级T效应记忆细胞群中潜伏和重新激活的诱导，Valadkhan实验室在lncRNAs方面的丰富经验，以及张实验室对miRNA和lncRNA在HIV TAT和转录调控中的作用的新见解。这项工作将围绕以下具体目标展开：目标1(美国)。鉴定与诱发潜伏期相关的表观遗传学改变有关的染色质修饰因子。目标2(美国)。明确长非编码RNA(LncRNAs)在HIV相关表观遗传学变化中的作用。目的3(中国)：确定miRNAs是否导致HIV-1潜伏期，并研究其作用机制。目的4(中国)：研究NRON lncRNA诱导的Tat蛋白降解及其在HIV-1潜伏时间中的作用。成功完成这项申请的四个协同目标将需要美国和中国团体之间交换信息和材料。我们将利用美国首席研究员Jonathan Karn博士及其同事Saba Valadkhan博士在HIV潜伏期的分子研究和lncRNA分析方面的经验，以及中国首席研究员张辉博士在microRNAs(MiRNAs)功能分析方面的补充专业知识。

 DESCRIPTION (provided by applicant): Despite the crucial clinical importance of HIV latency and reactivation, our knowledge of the mechanisms involved in establishment of latency and the subsequent reactivation is very incomplete. In this application we will examine the role of non-coding RNAs, both miRNA and lncRNA, on the control of HIV transcription and establishment of epigenetic silencing of HIV proviruses. Studies in numerous systems have pointed to complex interactions among different chromatin modifying factors and the extensive involvement of lncRNAs and miRNA in regulation of locus-specific epigenetic changes. While it is likely that similar RNA-protein networks govern the epigenetic changes and thus latency and reactivation of HIV proviral population, the specific mechanisms and RNAs have yet to be identified. We will test the hypothesis that the establishment, maintenance and exit from the latent state is governed by epigenetic changes imposed by chromatin modifying complexes which are guided to the proviral locus through association with one or more lncRNAs and miRNAs. This work will take advantage of a novel ex vivo system developed in the Karn laboratory which enables us to investigate the induction of both latency and reactivation in uniform primary T-effector memory cell populations, extensive experience with lncRNAs in the Valadkhan laboratory and new insights into the role of miRNA and lncRNA in the regulation of HIV Tat and transcription from the Zhang laboratory. The work will focus around the following Specific Aims: Aim 1 (US). Identification of chromatin modifying factors involved in inducing latency-related epigenetic changes. Aim 2 (US). To define the role of long non-coding RNAs (lncRNAs) in HIV-related epigenetic changes. Aim 3 (China): To determine whether miRNAs contribute to HIV-1 latency and to study their mechanisms of action. Aim 4 (China): To study degradation of Tat protein induced by NRON lncRNA and its contribution to HIV- 1 latency. Successful completion of the four synergistic aims of this application will require the exchange of information and materials between the US and Chinese groups. We will leverage the experience of the US principal investigator, Dr. Jonathan Karn and his colleague Dr. Saba Valadkhan, on molecular studies of HIV latency and the analysis of lncRNA and the complementary expertise of the Chinese principal investigator, Dr. Hui Zhang, on analysis of the function of microRNAs (miRNAs).