Examining Mechanisms of Synergy between Asthma Exacerbations and RV Infection

项目总结/摘要 Kennedy博士是阿肯色州大学医学科学系的过敏症专家/免疫学家 一位儿科和内科的教授和一位阿肯色州儿童医院研究的年轻研究员 院一个三管齐下的使命，以执行尖端研究，提供卓越的临床护理， 追求卓越的教育概括了他的总体学术生涯目标。他的训练和 经验使他能够发展提供高质量护理所需的技能和见解， 哮喘和过敏性疾病患者，以及为人类受试者研究提供基础。 这个指导职业发展奖提案的主要目标是进一步肯尼迪博士的 基础研究和翻译研究的知识和能力。这一目标将具体使他能够 实现我的长期研究目标，包括：1）了解免疫反应占 哮喘急性发作和RV感染之间的协同作用，2）开发哮喘的生物标志物 RV感染后的疾病严重程度和恶化，以及3）将本研究转化为临床 哮喘患者的相关预防和干预策略。 哮喘在约12%的美国人口中流行，RV被认为是最重要的病毒 在世界范围内引起普通感冒综合症。不像没有哮喘的患者， 呼吸道症状在感冒期间，RV感染的哮喘患者可能会出现下呼吸道症状 (e.g.,咳嗽、喘鸣、呼吸急促）。事实上，RV与60%至80%的哮喘急性发作相关 需要在急诊室治疗的儿童9 -11。尽管有如此强大的关系，一个重要的 关于RV加重哮喘症状的机制存在知识缺口。最近 细胞因子生物学的发展越来越强调呼吸道上皮细胞的重要性， 衍生的细胞因子在创造促进变应性免疫应答演变的环境中的作用。整体 本提案的目的是了解RV感染与上皮免疫之间的关系， 连接哮喘患者对感染的过敏反应的反应。我们假设RV感染 调节上皮细胞因子表达[白细胞介素（IL）-25和胸腺基质淋巴细胞生成素（TSLP）] 哮喘患者倾向于过敏性炎症反应，这是感染的基础- 介导的增加气道高反应性（AHR）。为了解决这个假设，我们有一个独特的 该方法将哮喘受试者的上皮源性细胞因子的体内研究结合在一起 急性加重、原代上皮细胞培养和新型精密切割肺切片（PCLS）外植体系统 允许比较哮喘和非哮喘组织中的RV感染。使用横截面设计， 在儿科急诊科，我们将比较哮喘患者鼻洗液中的细胞因子特征， RV诱导的急性发作和对照组的感冒症状，并将这些水平与哮喘症状进行比较。 在离体方法中，我们将研究Toll样受体驱动IL-25和TSLP产生的机制 （TLR）-3和视黄酸诱导基因-I（RIG-I）样受体（RLR），两者在RV识别中都很重要 在上皮细胞内。最后，我们将使用人气道PCLS来评估IL-25和TSLP的作用 对AHR对卡巴胆碱的反应，并比较来自供体的组织之间的这些反应， 哮喘通过评估上皮源性细胞因子与RV之间的协同作用机制， 感染和哮喘恶化，我们将阐明细胞因子的签名和描绘的途径， 将提供由RV产生的导致哮喘恶化的炎症环境的洞察。 此外，本提案中的研究为研究独立性提供了坚实的基础， 通过进一步研究细胞靶（例如，先天性淋巴样2型细胞（ILC 2），肥大 细胞）的这些细胞因子在RV诱导的哮喘恶化。 为了实现他的研究和学术目标，肯尼迪博士组建了一个多层次的指导团队， 拥有广泛经验的团队。与他的主要科学合作导师（理查德博士）的互动 库尔滕，乌莎Ponnappan，和Escholold Panettieri）和他的科学顾问委员会（SAC）将加强 他对RV免疫发病机制的基本机制的理解，发展研究设计技能， 扩展他对高级统计技术的知识。与儿科主任Stacie Jones博士的互动 过敏和免疫学和主要的职业发展导师，将有助于提高他的翻译 研究敏锐度和增加职业发展机会。此外，部门临床医生科学家 自2000年以来，由他所在机构的成功研究人员组成的指导委员会已经到位。 开始他的教师任命。这种多层次的指导系统将提供科学和职业 发展指导，这将使肯尼迪博士成为一个独立的研究人员和经验丰富的 专门研究RV引起的哮喘恶化的临床科学家。

PROJECT SUMMARY/ABSTRACT Dr. Kennedy is an Allergist/Immunologist at the University of Arkansas for Medical Sciences Departments of Pediatrics and Internal Medicine and a young investigator at Arkansas Children's Hospital Research Institute. A three-pronged mission to perform cutting-edge research, provide outstanding clinical care, and pursue educational excellence summarizes his overarching academic career objectives. His training and experience have enabled him to develop the skills and insight necessary to provide high-quality care to patients with asthma and allergic disorders, as well as providing a foundation for human subjects research. The primary objective for this mentored career development award proposal is to further Dr. Kennedy's knowledge and abilities in basic and translational investigation. This objective will specifically enable him to achieve my long-term research goals, including: 1) understanding the immune responses accounting for synergy between asthma exacerbations and infection with RV, 2) developing biomarkers of asthma disease severity and exacerbation following RV infection, and 3) translating this research into clinically relevant prevention and intervention strategies for patients with asthma. Asthma is prevalent in ~12% of the US population, and RV is recognized as the most important virus producing the common cold syndrome worldwide. Unlike patients without asthma who generally develop upper respiratory symptoms during colds, asthmatics with an RV infection may exhibit lower respiratory symptoms (e.g., cough, wheeze, shortness of breath). In fact, RV is associated with 60% to 80% of asthma exacerbations in children requiring treatment in the emergency department9-11. Despite such strong relationships, a significant knowledge gap exists with regards to the mechanisms whereby RV exacerbates asthma symptoms. Recent developments in cytokine biology have increasingly emphasized the importance of respiratory epithelial- derived cytokines in creating the milieu that promotes the evolution of allergic immune responses. The overall goal of this proposal is to understand the association between RV infection and the epithelial immune responses that bridge the allergic response to infection in asthmatics. We hypothesize that RV infection modulates epithelial cytokine expression [Interleukin (IL)-25 and thymic stromal lymphopoietin (TSLP)] in asthmatics with bias towards an allergic inflammatory response, and this underlies infection- mediated increases airway hyper-responsiveness (AHR). To address this hypothesis, we have a unique approach that brings together an in vivo study of epithelial-derived cytokines from subjects with asthma exacerbations, primary epithelial cell cultures, and a novel precision cut lung slice (PCLS) explant system allowing comparison of RV infections in asthmatic and non-asthmatic tissues. Using a cross-sectional design in a pediatric emergency department, we will compare cytokine signatures within nasal washes of asthmatics with RV-induced exacerbations and controls with cold symptoms and compare these levels with asthma symptoms. In ex vivo approaches, we will study mechanisms driving the production of IL-25 and TSLP by Toll-like receptor (TLR)-3 and Retinoic Acid Inducible Gene-I (RIG-I)-like receptors (RLR), both important in recognition of RV within epithelial cells. Finally, we will use the human airways PCLS to evaluate the effects of IL-25 and TSLP on AHR to carbachol and compare these responses between tissue derived from donors with and without asthma. By evaluating mechanisms of synergy that bridge the role of epithelial-derived cytokines to RV infection and asthma exacerbations, we will elucidate cytokine signatures and delineate pathways involved that will provide insight into the inflammatory environment produced by RV leading to exacerbations of asthma. Further, the studies within this proposal provide a firm foundation for research independence, allowing progression through further investigation of cellular targets (e.g., innate lymphoid type 2 cells (ILC2), mast cells) for these cytokines in RV-induced asthma exacerbations. To accomplish his research and academic goals, Dr. Kennedy has assembled a multi-tiered mentoring group with a wide breadth of experience. Interactions with his primary scientific co-mentors (Drs. Richard Kurten, Usha Ponnappan, and Reynold Panettieri) and his Scientific Advisory Committee (SAC) will enhance his understanding of the basic mechanisms of RV immunopathogenesis, develop research design skills, and expand his knowledge of advanced statistical techniques. Interactions with Dr. Stacie Jones, Chief of Pediatric Allergy and Immunology and primary career development mentor, will serve to improve his translational research acumen and enhance career development opportunities. Further, a Departmental Clinician Scientist Mentoring Committee composed of successful researchers at his institution has been in place since the beginning of his faculty appointment. This multi-tiered mentoring system will provide scientific and career development guidance that will enable Dr. Kennedy to become an independent researcher and an experienced clinician scientist specializing in RV-induced exacerbations of asthma.