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衰变机制常规地管理细胞并有效地去除不需要的RNA。我们 最近发现，包括VEE在内的甲病毒具有特异性RNA稳定元件， 它们的3' UTR，其募集细胞因子并阻断病毒转录物的去腺苷化/衰变，从而 促进有效和富有成效的感染。基于这些数据，我们假设， 编码加帽和多腺苷酸化转录物的RNA病毒已经进化出选择性地 抑制作为有效病毒基因表达的先决条件的细胞mRNA衰变机制。目标 本提案的目的一是通过评估我们是否可以将我们的观察扩展到以下方面来检验这一假设： 细胞RNA去腺苷化/衰变机制对另外的甲病毒的病毒抑制（WEE和 病毒）以及负义RNA病毒（埃博拉病毒、马尔堡病毒和尼帕病毒）。除了扩大我们的 了解分子宿主-病毒相互作用后，这些研究还将测试这些药物是否采用了 与甲病毒类似的策略用于介导病毒RNA稳定性。如果这些RNA病毒使用单个（如 初步数据表明，甲病毒的情况是如此）或有限数量的策略来抑制细胞凋亡， mRNA衰变机制，这代表了抗病毒治疗的新的和有吸引力的靶标。为此目的， 目标2的目标是优化我们建立的用于测量病毒mRNA稳定性的体外测定， 快速有效地筛选化合物库。该提案的最终目的是执行 化学库筛选，以识别和验证克服病毒抑制的候选先导化合物 细胞RNA衰变机制。这些化合物将代表进一步研究的有吸引力的候选物。 作为抗病毒治疗剂的开发，其可以非常好地具有针对多种 具有生物防御意义的病毒该研究项目符合RMRCE综合研究重点 将直接与RP 3.1、3.2、3.5、3.7和3.8相互作用，并利用 核心E该项目的目标应增加重要的专业知识，并与RMRCE协同工作 病毒治疗焦点小组，特别是由于创新的基本病毒学问题正在提出， 开发抗病毒药物的新方法可以靶向多个感兴趣的RNA病毒家族， 组