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

• 批准号: 8520159
• 负责人: WILLIAM B KLIMSTRA
• 金额: $ 31.72万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 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/8520159
• 关键词: 5' -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 Targeting; 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

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激活，在阻断多种病毒的复制方面非常有效，可以预防动物模型中的疾病。此外，该系统的某些组成部分已经进化，因此它们可以有效地区分宿主和病毒结构和代谢过程，从而抑制病毒过程，对宿主的非目标影响最小。这些属性为开发新抗病毒药的开发提供了出色的概念模型：一种将“自我”（宿主）与“非自然”（病毒）区分开的病毒抑制系统，并在某些情况下有效地抑制了非妇女，在具有广泛频谱活动的情况下，与多种病毒相比。虽然该系统通常是有效的，但我们发现人性化的α病毒，委内瑞拉马脑炎病毒（VEEV），对抗病毒状态的活性比人类狂欢性的sindbis病毒（SINV）更具抵抗力，并且在动物模型中的毒力在动物模型中具有很大的反映。利用树突状细胞的全球转录分析，一种与体内α病毒传播，扩增和疾病高度相关的细胞类型，我们已经确定了两个具有有效抗病毒活性的IFN-1/2-诱导蛋白，与SINV相比，ISG20 Exoribibonclease和MX2 GTPASE。为了发现这些蛋白质使用的抗病毒活性的机制以及VEEV抵抗抗病毒状态的这些效应因子的手段，我们建议表征抑制的相对疗效和每种蛋白质与SINV和VEEV施加的抗病活性的机制。为了实现这一目标，我们将使用过表达和干扰RNA敲低技术来识别病毒复制周期中每个效应子的作用，然后进行诱变，以识别单个效应子中的负责任功能域以及亚细胞定位和蛋白质 - 蛋白质相互作用研究，以识别靶向抑制的病毒结构和/或过程。这些研究的结果将：i）确定MX2和ISG20的作用与α20，ii）确定每个病毒的脆弱性点作为抗病毒疗法设计的第一步，以及III）提供了对Veev对抗病毒状态的抵抗力的分子机制的洞察力。 公共卫生相关性：这些研究的结果将确定两个高度活跃的IFN诱导的抗病毒效应子蛋白的作用机制，这些蛋白参与干扰素介导的降低α病毒复制的抑制以及对效应子的良性和高毒性病毒的相对敏感性。这些信息可用于设计抗病毒药物，这些抗病毒药物可以人为地模仿活性或人为地刺激这些效应子的诱导。此外，这些研究将提供有关特定效应子对小鼠和人类α杀伤力的贡献的信息，这些抗性可用于α病毒疫苗的设计。