Efficacy of the antiviral state versus Sindbis and Venezuelan equine encephalitis

抗病毒状态对辛德比斯和委内瑞拉马脑炎的疗效

基本信息

批准号：
8312719
负责人：
WILLIAM B KLIMSTRA
金额：
$ 33.75万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8312719
关键词：
5&apos -exoribonuclease Ablation Accounting Alphavirus Alphavirus Infections Animal Model Antiviral Agents Attenuated Live Virus Vaccine Benign Candidate Disease Gene Cell physiology Cells Cellular Structures Collaborations Cytoplasm Data Dendritic Cells Disease Drug Delivery Systems Evaluation Exhibits Exoribonucleases Gene Proteins Generations Genes Genetic Transcription Genome Guanosine Triphosphate Phosphohydrolases Host Defense Human Immune response In Vitro Individual Interferon Type I Interferons Letters Mediating Membrane Metabolism Modeling Molecular Mus Mutagenesis Nucleocapsid Process Production Property Proteins RNA RNA Viruses Relative (related person)Resistance Ribonucleases Signal Transduction Sindbis Virus Specificity Structure System Techniques Therapeutic Up-Regulation Vaccine Design Vaccines Venezuelan Equine Encephalitis Virus Venezuelan Equine Encephalomyelitis Viral Viral Drug Resistance Viral Proteins Virulence Virulent Virus Virus Diseases Virus Replication Zinc Fingers base cell type design exonuclease II in vivo insight model development pathogen prevent protein Mx protein protein interaction public health relevance research study resistance mechanism response

项目摘要

DESCRIPTION (provided by applicant): The type I interferon (IFN-1/2) system, if activated in the absence of virus antagonism, is very effective at blocking the replication of multiple viruses, either preventing or substantially reducing disease in animal models. Furthermore, some components of this system have evolved such that they can effectively distinguish between host and viral structural and metabolic processes, suppressing the viral processes with minimal non- target effects upon the host. These attributes provide an excellent conceptual model for development of new antiviral drugs: a virus suppression system that distinguishes "self" (host) from "nonself" (virus) and effectively suppresses nonself, in some cases with broad spectrum activity versus multiple viruses. While this system is generally effective, we have found that the human-virulent alphavirus, Venezuelan equine encephalitis virus (VEEV), is much more resistant to the activity of the antiviral state than the human-avirulent Sindbis virus (SINV) and that the virulence of the viruses in animal models is largely reflective of these differences. Using global transcription profiling of dendritic cells, a cell-type highly relevant to alphavirus dissemination, amplification and disease in vivo, we have identified two IFN-1/2-inducible proteins with potent antiviral activity versus SINV, the ISG20 exoribonuclease and the Mx2 GTPase. To discover the mechanisms of antiviral activity employed by these proteins and the means by which VEEV resists these effectors of the antiviral state, we propose to characterize the relative efficacy of inhibition and mechanisms of antiviral activity exerted by each protein versus SINV and VEEV. To achieve this, we will use over-expression and interfering-RNA knockdown techniques to identify the point in the virus replication cycle at which each effector acts followed by mutagenesis to identify the responsible functional domains in the individual effectors and subcellular localization and protein-protein interaction studies to identify the viral structures and/or processes targeted for inhibition. The results of these studies will: i) determine the mechanism(s) of action versus alphaviruses of Mx2 and ISG20, ii) identify points of vulnerability of each virus as a first step in design of antiviral therapeutics, and iii) provide insights into molecular mechanisms underlying the resistance of VEEV to the activities of the antiviral state that be utilized in design of vaccines. PUBLIC HEALTH RELEVANCE: The results of these studies will identify the mechanisms of action of two highly active IFN-induced antiviral effector proteins that are involved in interferon-mediated suppression of alphavirus replication and the relative sensitivity of benign and highly virulent viruses to the effectors. This information can be used in the design of antiviral drugs that artificially mimic the activity or artificially stimulate the induction of these effectors. Furthermore, these studies will provide information regarding the contribution of resistance to particular effectors to the virulence of alphaviruses in mice and humans that can be used in design of alphavirus vaccines.

描述（由申请人提供）：I型干扰素（IFN-1/2）系统如果在没有病毒拮抗作用的情况下被激活，则可以非常有效地阻断多种病毒的复制，从而预防或显着减少动物模型中的疾病。此外，该系统的一些组件已经进化，使得它们能够有效地区分宿主和病毒的结构和代谢过程，抑制病毒过程，同时对宿主的非目标影响最小。这些属性为开发新型抗病毒药物提供了一个极好的概念模型：一种病毒抑制系统，可以区分“自身”（宿主）和“非自身”（病毒）并有效抑制非自身，在某些情况下与多种病毒相比具有广谱活性。虽然该系统总体上是有效的，但我们发现对人类无毒力的甲病毒、委内瑞拉马脑炎病毒 (VEEV) 比对人类无毒力的辛德比斯病毒 (SINV) 更能抵抗抗病毒状态的活性，并且其毒力动物模型中病毒的变化很大程度上反映了这些差异。利用树突状细胞（一种与甲病毒体内传播、扩增和疾病高度相关的细胞类型）的全局转录分析，我们鉴定了两种 IFN-1/2 诱导蛋白，它们比 SINV 具有更强的抗病毒活性，即 ISG20 核糖核酸外切酶和 Mx2 GTPase 。为了发现这些蛋白的抗病毒活性机制以及 VEEV 抵抗这些抗病毒状态效应子的方式，我们建议表征每种蛋白相对于 SINV 和 VEEV 的抑制相对功效和抗病毒活性机制。为了实现这一目标，我们将使用过表达和干扰 RNA 敲除技术来确定病毒复制周期中每个效应子发挥作用的点，然后进行诱变来确定各个效应子中负责的功能域以及亚细胞定位和蛋白质-蛋白质相互作用研究以确定针对抑制的病毒结构和/或过程。这些研究的结果将：i) 确定 Mx2 和 ISG20 相对于甲病毒的作用机制，ii) 确定每种病毒的脆弱点，作为抗病毒治疗设计的第一步，以及 iii) 提供对分子分子的见解。 VEEV 对用于疫苗设计的抗病毒状态活性产生抵抗的机制。公共健康相关性：这些研究的结果将确定两种高活性干扰素诱导的抗病毒效应蛋白的作用机制，这些效应蛋白参与干扰素介导的甲病毒复制抑制，以及良性和高毒力病毒对效应蛋白的相对敏感性。该信息可用于设计抗病毒药物，人工模拟这些效应物的活性或人工刺激这些效应物的诱导。此外，这些研究将提供有关特定效应物的抗性对小鼠和人类甲病毒毒力的影响的信息，这些信息可用于甲病毒疫苗的设计。