ORMDL3 Stimulated ICAM1 and RVA-induced Childhood Asthma Exacerbations

项目摘要/摘要 哮喘发生在大约12%的美国人口中，鼻病毒(RV)被认为是主要病毒 在世界范围内产生了普通感冒综合症。与没有哮喘的患者不同，后者通常会发展为上 感冒期间的呼吸道症状，患有轮状病毒感染的哮喘患者表现出严重的下呼吸道症状 (如咳嗽、喘息、呼吸急促)。事实上，轮状病毒，特别是A和C种，与60%到 80%的哮喘恶化是需要在急诊科治疗的儿童。近现代文学 研究表明，RVC的受体钙粘素相关家族成员3(CDHR3)的突变增加 哮喘发作期对RVC感染的易感性。RVA病毒同样会导致肺炎的恶化 哮喘；然而，与疾病的基因联系仍不清楚。中国汉族人的单核苷酸多态性(SNPs) 17q21/ORM1-like 3(ORMDL3)与哮喘的加重和发展有关。 值得注意的是，约60%的儿童哮喘患者将在该基因座上有危险等位基因。最近， ORMDL3可调节A549细胞细胞间黏附分子1(ICAM1)的表达。ICAM1是 RVA用来感染上皮细胞的受体。综上所述，这项提案推测 17q21/ORMDL3基因座(风险等位基因)上的功能(GOF)SNPs调节ICAM1的表达 哮喘患者对RVA加重的遗传易感性。对这一假设的评估将在两年内进行 使肯尼迪实验室的研究优势最大化的具体目标。对于目标1，肯尼迪实验室 将评估风险等位基因对ICAM1表达的影响和下游影响，包括炎症 哮喘恶化儿童的细胞浸润物和介质。《目标2》将聚焦于一位新人 精密切割肺切片(PCLS)平台，来自哮喘和风险等位基因的捐赠者。对ICAM1的评价 表达和炎症介质，以及衡量呼吸道高反应性(AHR) 在有和没有危险等位基因的PCLS中，RV16感染后的卡巴胆碱是这一目标的基础。据了解，房车 在小鼠系统中复制不好，人类细胞培养缺乏研究生理学的能力 病毒感染时组织和宿主免疫的反应。肯尼迪实验室有能力准备 并维护来自人类捐赠者的PCL，以保存肺结构和生理反应，我们的 实验室可以在正确的宿主和靶组织内检查驱动AHR的免疫机制， 哮喘患者供者肺组织中哮喘加重的替代物和RVA中的风险等位基因 体外感染。所描述的项目将在#年产生关于RVA疾病免疫学的重要数据 并将建立一个框架，以便在此基础上进行进一步的 17q21/ORMDL3风险等位基因、ICAM1与RVA免疫应答的相关性研究 这会引发RVA诱导的哮喘加重。

PROJECT SUMMARY/ABSTRACT Asthma occurs in ~12% of the US population, and rhinovirus (RV) is recognized as the principal virus producing the common cold syndrome worldwide. Unlike patients without asthma who generally develop upper respiratory symptoms during colds, asthmatics with an RV infection exhibit severe lower respiratory symptoms (e.g., cough, wheeze, shortness of breath). In fact, RV, especially species A and C, are associated with 60% to 80% of asthma exacerbations in children requiring treatment in the emergency department. Recent literature has shown that mutations in cadherin-related family member 3 (CDHR3), the receptor for RVC, increases susceptibility to RVC infection during exacerbations of asthma. RVA viruses similarly lead to exacerbations of asthma; however, a genetic link to disease remains unclear. Single nucleotide polymorphisms (SNPs) in 17q21/ORM1-like 3 (ORMDL3) have been associated with both exacerbation and development of asthma. Strikingly, ~60% of individuals with childhood onset asthma will have risk alleles at this locus. Recently, ORMDL3 was shown to regulate intercellular adhesion molecule 1 (ICAM1) expression in A549 cells. ICAM1 is the receptor utilized by RVA to infect epithelial cells. Taken together, this proposal speculates that gain of function (GOF) SNPs in the 17q21/ORMDL3 locus (risk alleles) modulating ICAM1 expression underlie the genetic susceptibility to RVA exacerbations in those with asthma. Evaluation of this hypothesis will occur in two specific aims that maximize research strengths of the Kennedy Laboratory. For Aim 1, the Kennedy Laboratory will evaluate the effects of risk alleles on ICAM1 expression and downstream effects, including inflammatory cell infiltrates and mediators in children with asthma exacerbations. Aim 2 will focus on a novel human precision-cut lung slice (PCLS) platform from donors with asthma and risk alleles. Evaluations of ICAM1 expression and inflammatory mediators, as well as measures of airway hyper-responsiveness (AHR) to carbachol after RV16 infection in PCLS with and without risk alleles underlie this aim. It is understood that RV does not replicate well in murine systems and that human cell cultures lack the ability to investigate physiologic responses of tissue and host immunity during viral infections. With the Kennedy Laboratory’s ability to prepare and maintain PCLS from human donors that preserve lung architecture and physiologic responses, our laboratory can examine, within the correct host and target tissue, immunologic mechanisms driving AHR, a surrogate for asthma exacerbations, in lung tissue from donors with asthma and risk alleles during RVA infections ex vivo. The project described will generate important data about the immunology of RVA disease in high-risk asthma populations and will establish a framework on which to conduct further translational investigations into the relevance of 17q21/ORMDL3 risk alleles, ICAM1, and RVA-induced immune responses that trigger RVA-induced asthma exacerbations.