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

基本信息

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
项目类别：
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&apos 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

项目摘要

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 降解机制定期监管细胞并有效去除不需要的 RNA。我们最近发现，包括 VEE 在内的甲病毒在他们的 3' UTR 招募细胞因子并阻止病毒转录本的去腺苷化/衰变，从而促进有效且富有成效的感染。根据这些数据，我们假设许多（如果不是全部）编码加帽和多聚腺苷酸转录物的 RNA 病毒已经进化出选择性的机制抑制细胞 mRNA 降解机制是有效病毒基因表达的先决条件。目标该提案的目标一是通过评估我们是否可以扩展我们的观察来检验这一假设病毒抑制细胞 RNA 去腺苷酸化/衰变机制以产生其他甲病毒（WEE 和 EEE）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-病毒相互作用的知识，这些研究还将测试这些药物是否采用与甲病毒类似的介导病毒RNA稳定性的策略。如果这些 RNA 病毒使用单个（如初步数据表明甲病毒就是这种情况）或抑制细胞的有限策略 mRNA 降解机制，这代表了抗病毒治疗的一个新颖且有吸引力的靶标。为此，目标 2 的目标是优化我们已建立的用于测量病毒 mRNA 稳定性的体外测定法，以允许快速有效地筛选化合物库。该提案的最终目标是执行化学库筛选，用于识别和验证克服病毒抑制的候选先导化合物细胞RNA衰变机制的研究。这些化合物将成为进一步研究的有吸引力的候选化合物开发作为抗病毒疗法，很可能对多种病毒具有广谱活性具有生物防御意义的病毒。该研究项目符合 RMRCE 综合研究重点病毒治疗学，并将与 RP 3.1、3.2、3.5、3.7 和 3.8 直接互动，并利用资源核心 E 的目标。该项目的目标应为 RMRCE 增加重要的专业知识并与其良好协同病毒治疗焦点小组，特别是由于提出了创新的基本病毒学问题开发抗病毒药物的新方法可以针对多个感兴趣的RNA病毒家族团体。