Stat 1 modification for antiviral defense

抗病毒防御的 Stat 1 修改

• 批准号: 8446489
• 负责人: Michael J Holtzman
• 金额: $ 36.24万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8446489
• 关键词: Acute; Address; Antiviral Agents; Antiviral Response; Behavior; Bioinformatics; Biological Assay; Bioterrorism; Candidate Disease Gene; Cell Line; Cell model; Cells; Chronic Disease; Clinical; Collaborations; Complement; Cowpox virus; Cysteine; Data; Dependovirus; Diagnosis; Diagnostic; Disease; Effectiveness; Encephalomyocarditis virus; Engineering; Epidemic; Epithelial Cells; Exhibits; Gene Expression; Gene Transfer; Genes; Genome; Histopathology; Human; Immune response; Immunohistochemistry; In Vitro; Infection; Inflammation; Influenza A Virus, H1N1 Subtype; Influenza A virus; Interferon Activation; Interferon Receptor; Interferons; Laboratories; Mediating; Modification; Molecular; Molecular Profiling; Monitor; Mus; Mutation; Natural Immunity; Nature; Oligonucleotide Microarrays; Outcome; Pattern; Population; Public Health; Respiratory Tract Infections; Route; STAT1 gene; Sendai virus; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Small Interfering RNA; Specimen; Structure of parenchyma of lung; System; Technology; Testing; Time; Tissues; Toxic effect; Transgenes; Transgenic Mice; Transgenic Organisms; Treatment Efficacy; Vaccines; Viral; Virus; Virus Diseases; Virus Replication; Work; base; biodefense; cell type; cellular engineering; cellular transduction; diagnostic accuracy; gene transfer vector; improved; in vivo; mouse model; novel diagnostics; novel therapeutics; overexpression; pathogen; promoter; receptor expression; respiratory; response; vector; vector control

Viruses are among the most frequent causes of acute and chronic illness, and newly discovered viruses continue to cause emergent diseases that resist established vaccines. Despite the scope of this problem, the accuracy of diagnosis and efficacy of treatment for most viral infections, especially new types of infections, is very limited in effectiveness. To address this issue, we have developed an alternative strategy that is aimed at defining and then improving the antiviral host response. We focus particularly on respiratory infection since this is a common route of natural infection and a likely route for delivery of bioterrorist threats. In that regard, we have developed a system for primary-culture and infection of human airway epithelial cells that exhibits high fidelity to behavior found in vivo. Our analysis of this epithelial cell model in concert with a corresponding mouse model indicates that antiviral defense against respiratory viral infection depends critically on interferon (IFN) activation of the Stall signaling molecule in airway epithelial cells. Based on these findings, we proposed that improving Stall function in airway epithelial cells would enhance antiviral defense. Accordingly, we engineered a modified Stall with strategic double-cysteine mutalions (designaled Slal1-CC) that is hyperresponsive to endogenous interferon levels. Expression of Stall-CC should thereby enhance interferon-signaling function and provide for better control of viral replication. Indeed, our Preliminary Studies demonstrate that Stall-CC expression markedly decreases Ihe level of viral replication both in vitro (using transduced cells) and in vivo (using transgenic or gene transfer technology). For example, mice carrying the Stall-CC transgene or treated with a Stall-CC gene-transfer vector are fully protected against otherwise lethal infections due to each of the three viruses studied thus far. Furthermore, we have observed no toxicities of Stall-CC expression, in contrast to the situation for direct administralion or overexpression of interferon itself. In this proposal, we aim to extend our approach to the study of emergent pathogens with the capability for epidemic spread through the human population. In doing so, we aim to establish infection capabilities and diagnostic signatures for these new viruses, a new therapeutic strategy for these pathogens, and a better understanding of innate immunity to these agents.

病毒是急性和慢性病最常见的原因之一，也是新发现的病毒 继续引起抵抗已建立疫苗的新兴疾病。尽管有这个问题的范围， 大多数病毒感染的诊断和治疗功效的准确性，尤其是新型感染，是 有效性非常有限。为了解决这个问题，我们制定了一种针对的替代策略 定义并改善抗病毒宿主反应。我们特别关注呼吸道感染 由于这是自然感染的常见途径，也是生物恐怖威胁的可能途径。在那 考虑到，我们已经开发了一种用于人类气道上皮细胞的初级文化和感染的系统，该系统 表现出对体内发现的行为的高保真度。我们与A的上皮细胞模型的分析 相应的小鼠模型表明抗病毒防御呼吸道病毒感染取决于 对气道上皮细胞中失速信号分子的干扰素（IFN）激活进行严格的激活。基于 这些发现，我们提出，改善气道上皮细胞中的失速功能将增强抗病毒药 防御。因此，我们设计了一个具有战略性双层型突变的改良摊位（设计 slal1-cc）对内源性干扰素水平有效。摊位的表达应因此 增强干扰素信号函数，并更好地控制病毒复制。确实，我们的 初步研究表明，失速-CC表达明显降低了病毒复制水平 体外（使用转导细胞）和体内（使用转基因或基因转移技术）。例如， 携带Stall-CC转基因或用失速CC基因转移载体处理的小鼠受到完全保护 由于迄今为止研究的三种病毒中的每一种，都可以反对致命感染。此外，我们还有 与直接行政或 干扰素本身的过表达。在此提案中，我们旨在扩展我们的新兴方法 具有流行能力的病原体通过人口传播。这样，我们的目标是 建立这些新病毒的感染能力和诊断特征，这是一种新的治疗策略 对于这些病原体，以及对这些药物的先天免疫的更好理解。