A novel antiviral approach using the cellular RNA decay machinery

一种利用细胞 RNA 衰变机制的新型抗病毒方法

• 批准号: 8261431
• 负责人: Jeffrey Wilusz
• 金额: $ 26.44万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261431
• 关键词: 3' Untranslated Regions; Alphavirus; Antiviral Agents; Biological Assay; Cells; Chemicals; Data; Development; Drops; Elements; Exonuclease; Family; Filoviridae; Focus Groups; Gene Expression; Goals; In Vitro; Indium; Infection; Knowledge; Laboratories; Lead; Libraries; Measures; Mediating; Messenger RNA; Modeling; Molecular; Noise; Nonsense Codon; Paramyxoviridae; Pharmaceutical Preparations; Phase; Police; Poly A; Quality Control; RNA; RNA Decay; RNA Degradation; RNA Stability; RNA Viruses; Reagent; Recruitment Activity; Research; Research Project Grants; Resources; Screening procedure; Sindbis Virus; System; Technology; Testing; Therapeutic; Togaviridae; Transcript; Untranslated Regions; Venezuelan Equine Encephalitis Virus; Viral; Virus; Virus Diseases; Virus Replication; base; biodefense; blocking factor; decapping enzyme; design; endonuclease; in vitro Assay; innovation; interest; mRNA Decay; mRNA Stability; mRNA Transcript Degradation; novel; novel strategies; polyadenylated messenger RNA; small molecule; small molecule libraries; tissue culture; viral RNA; virology; virus host interaction

The cellular RNA decay machinery routinely polices the cell and effectively removes unwanted RNAs. We have recently discovered that alphaviruses, including VEE, possess specific RNA stabilizing elements in their 3' UTR that recruit a cellular factor and block the deadenylation/decay of viral transcripts, thereby promoting an efficient and productive infection. Based on these data, we hypothesize that many if not all RNA viruses that encode capped and polyadenylated transcripts have evolved mechanisms to selectively suppress the cellular mRNA decay machinery as a prerequisite for efficient virus gene expression. The goal of Aim I of this proposal is to test this hypothesis by assessing whether we can extend our observations on viral suppression of the cellular RNA deadenylation/decay machinery to additional alphaviruses (WEE and EEE) as well as negative sense RNA viruses (Ebola, Marburg and Nipah). In addition to expanding our knowledge of molecular host-virus interactions, these studies will also test whether these agents employ a similar strategy as the alphaviruses for mediating viral RNA stability. If these RNA viruses use a single (as preliminary data indicate is the case for alphaviruses) or limited number of strategies to suppress the cellular mRNA decay machinery, this represents a novel and attractive target for antiviral therapeutics. To this end, the goal of Aim 2 is to optimize our established in vitro assays for measuring viral mRNA stability to allow for the rapid and effective screening of chemical compound libraries. The final aim of this proposal is to perform a chemical library screen to identify and validate candidate lead compounds that overcome viral suppression of the cellular RNA decay machinery. These compounds would represent attractive candidates for further development as antiviral therapeutics that may very well have a broad spectrum activity against a variety of viruses of biodefense significance. This research project fits within the RMRCE Integrated Research Focus on Viral Therapeutics, and will interact directly with RPs 3.1, 3.2, 3.5, 3.7 and 3.8, and utilize the resources of Core E. The goals of this project should add significant expertise to and synergize well with the RMRCE Viral Therapeutics Focus Group, particularly due to the innovative basic virology questions being asked and the novel approach to develop antivirals that could target multiple families of RNA viruses of interest to the Group.

细胞RNA衰减机械通常会保管细胞并有效去除不需要的RNA。我们 最近发现，包括VEE在内的α病毒具有特定的RNA稳定元件 他们的3'UTR募集细胞因子并阻止病毒转录本的脱甲基化/衰减，从而 促进有效和生产性感染。基于这些数据，我们假设许多（如果不是全部） 编码限制和聚腺苷酸化转录本的RNA病毒具有进化的机制，以选择性地 抑制细胞mRNA衰变机制，作为有效病毒基因表达的先决条件。目标 该提议的目的是通过评估我们是否可以扩展观察结果来检验该假设 病毒抑制细胞RNA降苯甲基化/衰减机械，以其他α病毒（Wee和Wee和 EEE）以及负性RNA病毒（Ebola，Marburg和Nipah）。除了扩大我们的 对分子宿主病毒相互作用的了解，这些研究还将测试这些药物是否采用 与介导病毒RNA稳定性的α病毒相似的策略。如果这些RNA病毒使用单个病毒（AS 初步数据表明α病毒是这种情况）或抑制细胞的策略数量有限 mRNA衰变机械，这代表了抗病毒疗法的新颖而有吸引力的靶标。为此， 目标2的目标是优化我们已建立的体外测定法，以测量病毒mRNA稳定性以允许 化合物库的快速有效筛选。该提议的最终目的是执行 化学库筛选以识别和验证克服病毒抑制的候选铅化合物 细胞RNA衰减机械的。这些化合物将代表有吸引力的候选人 作为抗病毒治疗剂的发展很可能具有广泛的活性 生物浓度意义的病毒。该研究项目符合RMRCE综合研究重点 在病毒疗法上，并将直接与RPS 3.1、3.2、3.5、3.7和3.8互动，并利用资源 核心E的目标应该为RMRCE增添重要的专业知识并协同作用 病毒疗法焦点小组，特别是由于提出了创新的基本病毒学问题和 开发抗病毒药的新方法，可以针对多种感兴趣的RNA病毒家族 团体。