Stat 1 modification for antiviral defense

抗病毒防御的 Stat 1 修改

• 批准号: 8037557
• 负责人: Michael J Holtzman
• 金额: $ 40.05万
• 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
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8037557
• 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; 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; Therapeutic; Time; Tissues; Toxic effect; Transgenes; Transgenic Mice; Transgenic Organisms; Treatment Efficacy; Vaccines; Viral; Virus; Virus Diseases; Virus Replication; Work; base; 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.

病毒是急性和慢性疾病最常见的原因之一，新发现的病毒 继续引发对现有疫苗具有抵抗力的突发疾病。尽管这个问题的范围很广， 大多数病毒感染，特别是新型感染的诊断和治疗效果的准确性 效果非常有限。为了解决这个问题，我们制定了一个替代策略，旨在 定义并改善抗病毒宿主反应。我们特别关注呼吸道感染 因为这是一种常见的自然感染途径，也是传播生物恐怖威胁的可能途径。在那 在这方面，我们开发了一种用于人气道上皮细胞原代培养和感染的系统， 对体内发现的行为表现出高保真度。我们对该上皮细胞模型的分析与 相应的小鼠模型表明，针对呼吸道病毒感染的抗病毒防御取决于 关键是干扰素 (IFN) 激活气道上皮细胞中的 Stall 信号分子。基于 根据这些发现，我们提出改善气道上皮细胞的失速功能将增强抗病毒作用 防御。因此，我们设计了一种具有战略性双半胱氨酸突变体（指定为 Slal1-CC）对内源性干扰素水平高度敏感。因此，Stall-CC 的表达应 增强干扰素信号传导功能并更好地控制病毒复制。确实，我们的 初步研究表明 Stall-CC 表达显着降低病毒复制水平 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 Stall-CC 基因转移载体治疗的小鼠受到充分保护 对抗迄今为止研究的三种病毒中的每一种造成的致命感染。此外，我们还有 与直接施用或 干扰素本身的过度表达。在本提案中，我们的目标是将我们的方法扩展到紧急情况的研究 具有在人群中流行传播能力的病原体。在此过程中，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，以及更好地了解这些病原体的先天免疫。