A Genetic Analysis of Pneumotropism and Pneumovirulence in an MHV-1 Model of Sars

描述（由申请人提供）：本提案的目标是利用一种新的SARS啮齿动物模型来描述导致严重急性呼吸系统综合征（SARS）的病毒基因。SARS感染了全球8000多人，其中700多人死亡。我们建议使用我们小组开发的MHV-1感染的SARS小鼠模型来鉴定导致这种严重肺部疾病的冠状病毒基因。本研究的第一个目的是利用反向遗传学来检测结构蛋白刺突蛋白(S)、基质蛋白(E)、核衣壳蛋白(N)和非结构辅助蛋白在我们模型中sars样肺炎发病机制中的作用。这些研究将采用有针对性的重组方法。嵌合MHV-1/非嗜肺性重组病毒将用于确定靶向肺部和SARS样肺部疾病发展所需的MHV-1基因。在第二个目的中，我们将研究SARS- cov刺突蛋白（Srbd）的ACE2受体结合域除了在病毒附着细胞中起作用外，还通过取消ACE2的肺炎保护作用来参与SARS发病机制的假设。将分离出MHV-1/SARS-Srbd重组体，并确定它们在其他MHV-1抗性小鼠中增强毒力和引起致命的sars样肺部疾病的能力。这些研究将确定MHV-1基因对该病毒引起的独特的SARS样病理是必要和/或充分的，并评估SARS- cov Srbd在SARS小鼠模型中独立于其作为病毒附着蛋白的作用的发病机制中的作用。公共卫生相关性：本提案的目标是利用一种新的SARS啮齿动物模型来描述导致严重急性呼吸系统综合征（SARS）的病毒基因。SARS感染了全球8000多人，其中700多人死亡。我们将使用一种相关的冠状病毒，即小鼠肝炎病毒1型（MHV-1），它会在小鼠体内产生一种模仿SARS的疾病。我们使用这种新开发的SARS小鼠模型来识别导致这种严重肺部疾病的冠状病毒基因。这些信息应该有助于提高我们对SARS和其他冠状病毒肺部感染的理解，并可能为开发未来治疗这些疾病的方法提供基础。

DESCRIPTION (provided by applicant): The goal of this proposal is to delineate the viral genes that lead to severe acute respiratory syndrome (SARS) utilizing a novel rodent model of SARS. SARS infected over 8,000 persons globally with over 700 deaths. We propose to use a MHV-1 infected mouse model of SARS developed by our group to identify coronavirus genes that contribute to this severe pulmonary disease. The first aim of the research will utilize reverse genetics to examine the roles of structural proteins spike (S), matrix (E) and nucleocapsid (N) proteins and non-structural accessory proteins in the pathogenesis of SARS-like pneumonitis in our model. These studies will employ a targeted recombination approach. Chimeric MHV-1/non-pneumotropic recombinant viruses will be used to determine the MHV-1 genes necessary for targeting to the lung and for the development of SARS like pulmonary disease. In the second aim we will investigate the hypothesis that the ACE2 receptor binding domain of the SARS-CoV spike protein (Srbd) contributes to SARS pathogenesis by abrogating the pneumoprotective effect of ACE2 in addition to its role in viral attachment to cells. MHV-1/SARS-Srbd recombinants will be isolated and their ability to enhance virulence and cause lethal SARS-like pulmonary disease in otherwise MHV-1 resistant mice will be determined. These studies will identify MHV-1 genes that are necessary and/or sufficient for the unique SARS-like pathology induced by this virus, and assess the role of the SARS-CoV Srbd in pathogenesis independent of its role as a viral attachment protein in a mouse model of SARS. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to delineate the viral genes that lead to severe acute respiratory syndrome (SARS) utilizing a novel rodent model of SARS. SARS infected over 8,000 persons globally with over 700 deaths. We will use a related coronavirus, mouse hepatitis virus type 1 (MHV-1), that produces a disease in mice that mimics SARS. We use this newly developed mouse model of SARS to identify coronavirus genes that contribute to this severe pulmonary disease. This information should be useful in advancing our understanding of SARS and other coronavirus pulmonary infections and may provide a basis for developing future therapeutic approaches to these diseases.