Macaque Model and Gene Expression Profiling of SARS

SARS 的猕猴模型和基因表达谱

• 批准号: 7163778
• 负责人: MICHAEL G KATZE
• 金额: $ 61.81万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7163778
• 关键词: A549; Adrenal Cortex Hormones; Adult Respiratory Distress Syndrome; Affect; Age; Alveolar; Animal Model; Animals; Antiviral Response; Area; Arts; Biological; Biological Assay; Cell Culture System; Cell Line; Cells; Cessation of life; Clinical Data; Coronavirus; Coronavirus Infections; Cultured Cells; Data; Dendritic Cells; Development; Diagnostic; Diffuse; Disease; Disease Outbreaks; Disease susceptibility; Engineering; Epithelial Cells; Gene Expression; Gene Expression Profiling; Genes; Genomics; Goals; Growth; Hamman-Rich syndrome; Human; Immune response; Immunosuppression; In Vitro; Infection; Information Technology; Integration Host Factors; Interferons; Knowledge; Lesion; Lung; Lung diseases; Macaca; Macaca fascicularis; Modeling; Molecular; Molecular Cloning; Morbidity - disease rate; Mutation; Numbers; Oligonucleotide Microarrays; Pathogenesis; Pathway interactions; Peripheral Blood Mononuclear Cell; Preventive; Primary Lesion; Pulmonary Pathology; Pulmonary alveolar structure; Pulmonology; RNA; Recombinants; Research Infrastructure; Research Personnel; Reserve Cell; Respiratory System; Role; SARS coronavirus; Severe Acute Respiratory Syndrome; Severities; Staging; Supporting Cell; Symptoms; System; Technology; Therapeutic Intervention; Therapeutic immunosuppression; Time; Tissues; Universities; Virulence; Virus; Virus Diseases; Washington; Week; base; body system; experience; functional genomics; insight; macrophage; mortality; novel; pathogen; pneumocyte; prognostic; programs; research study; response; surfactant; virus host interaction

DESCRIPTION (PROVIDED BY APPLICANT): Severe acute respiratory syndrome coronavirus (SARS-CoV) is an emerging pathogen responsible for considerable morbidity and mortality worldwide, yet little is known regarding the molecular basis for its pathogenesis. The most significant organ system affected in fatal SARS cases is the respiratory tract. The cynomolgus macaque model best replicates the pulmonary pathogenesis associated with human SARS and was used to prove that SARS-CoV is the primary etiologic agent of SARS, and that type-1 and -2 pneumocytes may be important target cells for SARS-CoV. Our overall goal is to increase our knowledge of SARS-CoV pulmonary pathogenesis. In Specific Aim 1 we will evaluate the pathogenesis of SARS lung disease expanding upon the macaque model. We intend to refine identification of 'target cells permissive for SARS-CoV and possible changes in their distribution at different times after infection. The role of age and immunosuppression, important host factors that increase the severity SARS, will be examined by comparing the development of SARS in macaques of different ages, or treated with different levels of corticosteroids. We will build on the macaque model to include the use of infectious molecularly cloned wild-type SARS-CoV and SARS-CoV containing mutations in specific accessory genes that may contribute to virulence. Specific Aim 2 will focus on the establishment of an appropriate cell culture system in which to evaluate the cellular response to wild-type and engineered SARS-CoV. This will include studies with human pneumocytes and small airway epithelial cells. We also will attempt to infect lung A549 cells and macaques primary macrophages and dendritic cells. Finally, Specific Aim 3 will determine molecular mechanisms and pathogenic signatures of SARS using global gene expression analysis and functional genomics. We will use oligonucleotide microarrays and sophisticated information technologies to examine gene expression changes in response to SARS-CoV infection. Experiments will be performed using RNA isolated from in vitro infections and from cells and tissues from experimentally infected macaques. Collectively, data obtained from the macaque model, cell culture systems, and gene expression analyses should provide new insights into the pathogenesis of SARS, aid in the development of diagnostic and prognostic assays, and identify novel targets for preventive or therapeutic intervention.

描述（由申请人提供）：严重急性呼吸综合征冠状病毒（SARS-CoV）是一种新兴病原体，在全球范围内造成相当大的发病率和死亡率，但对其发病机制的分子基础知之甚少。 SARS 致命病例中受影响最重要的器官系统是呼吸道。食蟹猴模型最好地复制了与人类SARS相关的肺部发病机制，并被用来证明SARS-CoV是SARS的主要病原体，并且1型和-2型肺细胞可能是SARS-CoV的重要靶细胞。我们的总体目标是增加我们对 SARS-CoV 肺部发病机制的了解。在具体目标 1 中，我们将评估在猕猴模型上扩展的 SARS 肺病的发病机制。我们打算完善对 SARS-CoV 允许的靶细胞的识别，以及它们在感染后不同时间分布的可能变化。年龄和免疫抑制是增加 SARS 严重程度的重要宿主因素，我们将通过比较不同年龄的猕猴或接受不同水平皮质类固醇治疗的 SARS 的发展来检查其作用。我们将在猕猴模型的基础上，使用传染性分子克隆的野生型 SARS-CoV 和含有可能导致毒力的特定辅助基因突变的 SARS-CoV。具体目标 2 将侧重于建立适当的细胞培养系统，以评估细胞对野生型和工程化 SARS-CoV 的反应。这将包括对人类肺细胞和小气道上皮细胞的研究。我们还将尝试感染肺 A549 细胞和猕猴原代巨噬细胞和树突状细胞。最后，具体目标 3 将利用全局基因表达分析和功能基因组学确定 SARS 的分子机制和致病特征。我们将使用寡核苷酸微阵列和先进的信息技术来检查针对 SARS-CoV 感染的基因表达变化。实验将使用从体外感染以及实验感染的猕猴的细胞和组织中分离出的 RNA 进行。总的来说，从猕猴模型、细胞培养系统和基因表达分析中获得的数据应该为 SARS 的发病机制提供新的见解，有助于诊断和预后分析的开发，并确定预防或治疗干预的新靶点。