Examining Mechanisms of Synergy between Asthma Exacerbations and RV Infection

项目摘要/摘要 肯尼迪博士是阿肯色大学医学科学系的过敏症专家/免疫学家。 阿肯色州儿童医院研究部儿科和内科的一名年轻研究员 研究所。一个三管齐下的使命，进行尖端研究，提供卓越的临床护理，以及 追求卓越教育总结了他最重要的学术生涯目标。他的训练和 经验使他能够培养提供高质量护理所需的技能和洞察力 对哮喘和过敏性疾病患者的研究，以及为人类研究对象提供基础。 这项有指导的职业发展奖提案的主要目标是促进肯尼迪博士的 具备基础调查和翻译调查方面的知识和能力。这一目标将特别使他能够 完成我的长期研究目标，包括：1)了解免疫应答占 哮喘加重与轮状病毒感染之间的协同作用，2)发展哮喘的生物标志物 RV感染后的疾病严重性和恶化，以及3)将这项研究转化为临床 哮喘患者的相关预防和干预策略。 哮喘在大约12%的美国人口中流行，轮状病毒被认为是最重要的病毒 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可能会出现下呼吸道症状 (如咳嗽、喘息、呼吸急促)。事实上，60%到80%的哮喘加重与RV有关 需要在急诊科接受治疗的儿童9-11。尽管有如此牢固的关系，但一个重要的 关于RV加重哮喘症状的机制，存在知识差距。近期 细胞因子生物学的发展日益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过敏免疫反应进化的环境中的作用。整体而言 这项建议的目的是了解轮状病毒感染和上皮免疫之间的联系。 在哮喘患者中架起感染过敏反应桥梁的反应。我们假设轮状病毒感染 调节上皮细胞因子表达[白介素25和胸腺间质淋巴生成素] 偏向过敏性炎症反应的哮喘患者，这是感染的基础- 中介可增加呼吸道高反应性(AHR)。为了解决这一假设，我们有一个独特的 汇集哮喘患者上皮源性细胞因子体内研究的方法 急性加重、原代上皮细胞培养和一种新的精密切割肺切片(PCLS)移植系统 允许比较哮喘和非哮喘组织中的轮状病毒感染。使用横截面设计 儿科急诊科，我们将比较哮喘患者鼻腔冲洗液中的细胞因子信号与 并将这些水平与哮喘症状进行比较。 在体外方法中，我们将研究Toll样受体驱动IL-25和TSLP产生的机制 (TLR)-3和维甲酸诱导基因-I(RIG-I)样受体(RLR)在RV识别中的重要作用 在上皮细胞内。最后，我们将使用人类呼吸道PCLS来评估IL-25和TSLP的作用 卡巴胆碱对AHR的影响及供体组织对氨基甲胆碱的反应 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信号并描绘涉及 将提供对由RV产生的导致哮喘加重的炎症环境的洞察。 此外，该提案中的研究为研究独立性提供了坚实的基础，允许 通过进一步研究细胞靶点(例如，先天性淋巴样细胞2型细胞(ILC2)、肥大细胞)的进展 这些细胞因子在轮状病毒诱导的哮喘加重中的作用。 为了实现他的研究和学术目标，肯尼迪博士组织了一个多层次的指导 拥有广泛经验的团队。与他的主要科学合作导师的互动(理查德博士 库尔滕、乌莎·波纳潘和雷诺·帕内蒂耶里)和他的科学咨询委员会(SAC)将加强 他对轮状病毒免疫致病的基本机制的理解，发展研究设计技能，以及 扩大他对先进统计技术的了解。与儿科主任Stacie Jones医生的互动 过敏和免疫学和主要职业发展导师，将有助于提高他的翻译能力 敏锐的研究能力和提升职业发展的机会。此外，一名科室临床医生科学家 由他所在机构的成功研究人员组成的指导委员会自 开始他的教职任命。这种多层次的导师制度将为科学和职业提供 发展指导，使肯尼迪博士能够成为一名独立的研究人员和经验丰富的 专门研究轮状病毒引起的哮喘加重的临床科学家。

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.