Effect of cellular microRNAs on HIV-1 replication

Pi：Zhang，Huong授权号：1RO1AI078812-01A2资助题目：细胞microRNAs对HIV-1复制的影响 修订后的摘要部分 MicroRNAs(MiRNAs)是一种具有抑制蛋白质翻译能力的短~21个核苷酸的调节性RNA。它们参与了包括植物、昆虫、脊椎动物和哺乳动物在内的许多真核生物谱系的各种生物学功能的调节。越来越多的证据表明，人类免疫缺陷病毒1型(HIV-1)的复制受细胞或病毒来源的miRNAs调控。最近，我们发现一些细胞内的miRNAs参与了HIV-1潜伏期的建立和维持。各种HIV-1mRNAs的3‘末端是一组细胞miRNAs的结合位点，包括mir-28、mir-125b、mir-150、mir-223和mir-382，这些miRNAs富含在静止的CD4+T细胞中，而不是激活的CD4+细胞中。这些miRNAs的反义抑制剂可以显著地抵消其相应的miRNAs对HIV-1感染克隆的静息CD4T细胞中HIV-1蛋白翻译的抑制作用，或对接受抑制性高效抗逆转录病毒治疗(HAART)的静息CD4+T细胞产生HIV-1的抑制作用。此外，我们还发现病毒粒子相关的细胞miRNA可能在HIV-1的复制中发挥作用。由于miRNAs影响HIV-1复制的证据是确凿的，我们想提出以下项目来系统地研究细胞miRNAs对HIV-1复制的影响：(1)进一步阐明细胞miRNAs如何在静止的CD4T淋巴细胞中导致HIV-1整合后潜伏期的机制研究(2)。进一步研究细胞miRNAs与静息的CD4T淋巴细胞中TAT或REV的低效表达之间的相互作用以及它们对HIV-1潜伏期的可能协同贡献。这项提议中的实验旨在详细了解细胞miRNAs如何影响HIV-1复制的机制。这些信息对于进一步利用miRNA操纵在控制HIV-1复制和传播方面的相关性至关重要。以下修订的目标是为两年的ARRA奖提出的，随后将是两年的NIAID奖。

PI: Zhang, Hui Grant Number: 1RO1AI078812-01A2 Grant title: Effect of cellular microRNAs on HIV-1 replication Revised Abstract Section MicroRNAs (miRNAs) are short ~21-nt-long regulatory RNAs with the ability to repress protein translation. They are involved in the regulation of various biological functions in numerous eukaryotic lineages, including plants, insects, vertebrate, and mammals. Accumulating evidence has indicated that human immunodeficiency virus type 1 (HIV-1) replication is regulated by cellor virus-derived miRNAs. Recently, we have found that several cellular miRNAs are involved in establishing and maintaining HIV-1 latency in resting primary CD4 T-lymphocytes. The 3'termini of various HIV-1 mRNAs are the binding site of a cluster of cellular miRNAs including mir-28, mir-125b, mir-150, mir-223, and mir-382, which is enriched in the resting CD4+ T cells rather than in activated CD4+ cells. The antisense inhibitors of these miRNAs can significantly counteract the inhibitory effects of their corresponding miRNAs upon either HIV-1 protein translation in the resting CD4 T-cells transfected with HIV-1 infectious clone, or HIV-1 production from the resting CD4+ T-cells isolated from HIV-1-infected individuals receiving suppressive highly active antiretroviral treatment (HAART). Besides, we have also found that virion associated cellular miRNA could play a role in HIV-1 replication. As the evidence that miRNAs affect HIV-1 replication is solid, we would like to propose the following projects to systematically examine the effect of cellular miRNAs on HIV-1 replication: (1). Mechanistic studies to further clarify how cellular miRNAs contribute to HIV-1 postintegration latency in resting CD4 T-lymphocytes (2). Further investigate the interaction between cellular miRNAs and the inefficient expression of Tat or Rev and their possible synergetic contribution to HIV-1 latency in resting CD4 T-lymphocytes. Experiments in this proposal are designed to provide a detail understanding of the mechanism how cellular miRNAs affect HIV-1 replication. Such information is crucial for further exploiting the relevance of miRNA manipulation in controlling HIV-1 replication and transmission. Following revised aims are proposed for the 2-year ARRA award, which will be followed by a 2-year NIAID award.