Molecular understanding of the GSDMB-regulated innate immune response

项目摘要 呼吸道病毒感染本身就是一种日益加重的健康和社会负担，也增加了发展风险 以及其他呼吸道疾病的恶化，包括影响1700万美国人的哮喘。病毒 感染引发先天免疫反应，这是一种重要的防御机制，通过激活 干扰素信号传导：抗病毒蛋白的稳健诱导，包括干扰素（IFN）和干扰素刺激的 通过识别病毒核酸和激活随后的信号级联反应，将病毒基因（ISG）与病毒结合。 具体而言，细胞内病毒RNA被RNA传感器感测和识别，随后激活 一系列的信号级联以诱导IFN的产生。分泌的干扰素反过来激活转录 数以百计的ISG来放大先天免疫反应，这是一把双刃剑，既限制了病毒 复制和没有适当的控制，导致过度的炎症反应，损害气道 病毒感染引起的哮喘恶化期间的上皮细胞。此外，这种病毒诱导的哮喘 恶化具有很强的遗传基础。染色体17 q21 Gasdermin B（GSDMB）区域已被 与儿童哮喘的易感性和严重程度显著相关，主要是在有既往哮喘史的儿童中， 呼吸道病毒感染然而，病毒感染如何决定哮喘的机制 具有不同遗传背景的个体之间的易感性还不完全清楚，这是 提案的重点。值得注意的是，17 q21哮喘风险等位基因的基因型与哮喘风险增加相关。 GSDMB在人气道上皮细胞中的表达。我们未发表的工作表明，GSDMB 不仅是足够的，而且是促进IFN信号传导和诱导人ISGs表达所必需的 用RNA病毒或其类似物：poly（I：C）处理的气道上皮细胞。我们现在建议描述 GSDMB决定RNA病毒感染后哮喘风险的生物学和分子机制 感染我们已经提出了一系列整合和互补的体外，离体和体内的方法 1）剖析GSDMB促进IFN信号传导的分子机制（目的1）; 2）建立 GSDMB诱导人鼻病毒感染细胞模型的长期炎症反应 支气管上皮细胞与GSDMB相反的基因型（目的2）; 3）在体内的结果， 在小鼠中用人GSDMB的条件性和诱导性表达和重复性呼吸道病毒感染， 气道上皮细胞（Aim 3）。我们对这种基因与环境相互作用的深刻机制理解 将阐明新的治疗和干扰策略，以防止严重哮喘急性发作的易感 科目

Project Summary Respiratory virus infection, an increasing health and social burden alone, also increases risk for development and exacerbation of other respiratory diseases, including asthma that affects 17 million U.S. people. Viral infection triggers innate immune response, an important defensive mechanism through activation of the interferon signaling: robust induction of antiviral proteins including interferons (IFNs) and interferon-stimulated genes (ISGs) through the recognition of viral nucleic acid and activation of subsequent signal cascades. Specifically, intracellular viral RNA is sensed and recognized by RNA sensors, which subsequently activates series of signaling cascade to induce the production of IFNs. Secreted IFNs, in turn, activates the transcription of hundreds of ISGs to amplify innate immune response, a double-edged sword, both constraining viral replication and without proper control, leading to exuberant inflammatory response that damages airway epithelium during asthma exacerbation caused by viral infection. Furthermore, such virus-induced asthmatic exacerbation has a strong genetic basis. Chromosome 17q21 Gasdermin B (GSDMB) region has been significantly associated with susceptibility and severity of childhood asthma, primarily in children who had prior respiratory virus infection. However, the mechanism by which how viral infection determines the asthmatic susceptibility among individuals with various genetic background is incompletely understood, which is the major focus of the proposal. Notably, the genotype of 17q21 asthma risk allele is associated with increased expression of GSDMB in human airway epithelial cells. Our unpublished work has demonstrated that GSDMB is not only sufficient but also required to promote IFNs signaling and induce expression of ISGs in human airway epithelial cells treated with RNA viruses or their analogue: poly (I:C). We now propose to characterize the biological and molecular mechanism by which GSDMB determines the risk for asthma following RNA virus infection. We have proposed a series of integrative and complementary in vitro, ex vivo and in vivo approaches to 1) dissect the molecular mechanisms by which GSDMB promotes IFNs signaling (Aim 1); 2) establish the Rhinovirus infection cellular model with prolonged inflammatory response induced by GSDMB in human bronchial epithelial cells with opposing genotypes of GSDMB (Aim 2); 3) in vivo consequence of prolonged and repetitive respiratory virus infection in mice with conditional and inducible expression of human GSDMB in airway epithelial cells (Aim 3). Our deep mechanistic understanding of such gene by environment interaction will illuminate novel treatment and perturbation strategy to prevent severe asthma exacerbation in susceptible subjects.