The function and regulation of IL-33 in the airway epithelium in asthma

IL-33在哮喘气道上皮中的功能及调控

• 批准号: 9266460
• 负责人: Erin D. Gordon
• 金额: $ 17.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9266460
• 关键词: Acute; Affect; Agonist; American; Animal Model; Apoptosis; Asthma; Award; Basic Science; Biological Assay; Biological Models; Biology; Biometry; Biopsy; Blood; California; Career Choice; Cell Culture Techniques; Cell Death; Cell Line; Cell Nucleus; Cells; Cellular biology; Chronic; Chronic lung disease; Clinical; Clinical Research; Code; Collaborations; Core Facility; Critical Care; Data; Dedications; Development Plans; Disease; Disease susceptibility; Enrollment; Ensure; Eosinophilia; Epithelial; Epithelial Cells; Epithelium; Event; Family; Fellowship; Gene Expression; Genes; Genetic; Grant; Health; Hour; Human; Human Genetics; Hyperplasia; Immune; Immune System Diseases; Immune response; Immunofluorescence Immunologic; Immunology; In Vitro; Infection; Inflammation; Inflammatory; Interleukin-1; Interleukin-13; Interleukin-5; Interleukins; Internal Medicine; Irrigation; K-Series Research Career Programs; Laboratories; Link; Luc Gene; Lung; Measures; Mediator of activation protein; Medicine; Mentors; Modeling; Molecular; Molecular Cloning; Morbidity - disease rate; Mucins; Mucous body substance; Necrosis; Pathogenesis; Pathway interactions; Peptide Signal Sequences; Physicians; Plasmids; Poly I-C; Positioning Attribute; Predisposition; Production; Proteins; Proteolysis; Public Health; Publishing; Pulmonology; Regimen; Regulation; Reporter; Research; Research Personnel; Research Training; Residencies; Resources; Rhinovirus; Role; San Francisco; Scholarship; Science; Scientist; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Source; Sputum; Techniques; Testing; Time; Tissue Banks; Training; Training Programs; Transfection; Universities; Virus; Virus Diseases; Vocational Guidance; Work; airway epithelium; airway remodeling; analog; asthmatic; career; career development; cohort; cytokine; effective therapy; experience; extracellular; genetic signature; genome wide association study; genome-wide; inhibitor/antagonist; interest; laboratory facility; medical schools; member; molecular phenotype; mortality; mouse model; novel; periostin; professor; promoter; receptor; research and development; response; skills; success; undergraduate student; viral RNA; virology

DESCRIPTION (provided by applicant): In this revised application entitled "Function and Regulation of IL-33 in Airway Epithelial Cells in Asthma" for the Mentored Clinical Scientist Research Career Development Award, I propose a comprehensive plan of research and career development at the University of California, San Francisco in the Division of Pulmonary Medicine. This proposal will not only enable me to develop the skills, techniques and collaborations necessary to establish an independent academic research career, but it will also advance the field of asthma biology. I am an ideal candidate for this mentored career development award, as I am strongly committed to establishing an independent research career. I became interested in research science as an undergraduate at the University of California, Berkeley where I worked in a virology laboratory for three years. I graduated with honors earning a full tuition scholarship to study medicine at the University of Southern California. Graduating first in my medical school class, I chose to complete my Internal Medicine residency training at UC San Diego where I performed basic science research in the laboratory of Dr. Patricia Finn. There I developed a strong interest in asthma biology and enrolled in the American Board of Internal Medicine Research Training Program. This allowed me to begin fellowship in Pulmonary Critical Care at UCSF early and spend additional years performing research. While at UCSF, I continued studying asthma in the laboratory of John Fahy. Under the research and career guidance of Dr. Fahy, Erle, Woodruff, and Sheppard, I completed a project focused on the role of periostin in asthma. I published these results in January of 2012. I have since focused on the role of IL- 33 in human asthma, and this is the topic of my current proposal. My dedication to a research career along with persistence and a track record of success will enable me to maximally benefit from this award. I have outlined a research plan in this application, which focuses on understanding the function and regulation of IL-33 in human asthma. Asthma is a chronic lung disease, which affects nearly 30 million Americans and an estimated 300 million people worldwide. Although effective treatments exist, many asthmatics do not respond optimally to available regimens; thus, novel treatments are needed. Long considered a disease of the immune system, there is increasing recognition of the importance of the airway epithelium in orchestrating the immune responses in asthma. The newly described epithelial cell cytokine IL-33 and its receptor ST2 have been strongly implicated in asthma pathogenesis in multiple large-scale genome wide association studies. Mouse models suggest that IL-33, acting through the ST2 receptor, promotes Th2 inflammation. Studies of IL-33 in human asthma are limited however. In this grant, I will focus on three important questions in IL-33 biology: 1) how is IL-33 released from airway epithelial cells, 2) what is the relationship of I-33 to Th2 inflammation in human asthma, and 3) what is the function of SNPs in the ST2 locus on the regulation of IL-33 by the soluble inhibitor sST2? IL-33 is constitutively expressed in the nuclei of airway epithelial cells, but it lacks a signal sequence, and the mechanism of its release is unknown. I have developed a novel model system by expressing GFP-tagged IL-33 in an airway epithelial cell line, which will enable me to dissect the pathways leading to IL-33 release. Once released, IL-33 is postulated to induce Th2 inflammation in the airway. Using the vast human biospecimen bank at the UCSF Airway Clinical Research Center, I will determine the relationship of IL-33 to markers of Th2 inflammation and mucous remodeling in both acute and chronic asthma. Finally, extracellular IL-33 can be regulated by its soluble inhibitor sST2. I will sequence the ST2 locus from a large number of asthmatics and identify SNPs associated with altered ST2 epithelial cell and sputum pellet gene expression. I will test the function of candidat SNPs in human primary epithelial cell culture using promoter reporter assays and a functional bioassay. This research plan will not only advance the field of asthma biology, but will enable me to develop new research skills as well as collaborations that will be essential for my success as an independent investigator. Finally, I have outlined a career development plan that takes advantage of the many opportunities here at UCSF. My primary mentor, John Fahy, is ideally suited to serve as my advisor. He brings expertise in airway epithelial cell biology and molecular phenotyping in asthma and is the director of the large UCSF human asthma tissue bank. I have assembled a team of additional advisors and collaborators, including Dr. Sheppard, Erle, Woodruff and Seibold, who have complementary skills. They will be ideal in advising me on cell biology, primary airway epithelial cell transfection, molecular cloning, biostatistics, and advanced human genetics. I will supplement the guidance of this team with formal coursework offered at UCSF in Genetics, Cell Biology, Immunology, and Biostatistics as well as weekly seminars in Immunology and general pulmonary medicine. The Department of Medicine and the Division of Pulmonary Critical Care has a strong track record of support and training for early-stage physician scientists and boasts well-resourced laboratories and core facilities. These resources will be invaluable in carrying out my research plan and will ensure my success. The Department of Medicine strongly supports my career path and this application. In July 2012, I was appointed to Assistant Professor within the Division of Pulmonary Critical Care with 80% protected research time. With these resources and support, I will be well positioned to establish a successful independent research career.

描述（由申请人提供）：在这份题为“哮喘中气道上皮细胞中IL-33的功能和调节”的修订申请中，我提出了一项在加州大学弗朗西斯科分校肺部医学部进行研究和职业发展的综合计划。这项提议不仅使我能够发展建立独立学术研究生涯所需的技能，技术和合作，而且还将推动哮喘生物学领域的发展。我是这个指导职业发展奖的理想候选人，因为我坚定地致力于建立一个独立的研究生涯。我在加州大学伯克利分校读本科时就对研究科学产生了兴趣，我在那里的一个病毒学实验室工作了三年。我以优异的成绩毕业，获得了全额奖学金，在南加州大学学习医学。我以医学院第一名的成绩毕业，选择在加州大学圣地亚哥分校完成内科住院医师培训，在Patricia Finn博士的实验室进行基础科学研究。在那里，我对哮喘生物学产生了浓厚的兴趣，并参加了美国内科医学研究培训计划委员会。这使我能够开始奖学金在肺重症监护早期在加州大学旧金山分校和花费额外的几年进行研究。在加州大学旧金山分校期间，我继续在约翰·法希的实验室研究哮喘。在Fahy、Erle、Woodruff和Sheppard博士的研究和职业指导下，我完成了一个专注于骨膜蛋白在哮喘中的作用的项目。我在2012年1月发表了这些结果。从那以后，我一直专注于IL- 33在人类哮喘中的作用，这也是我目前提案的主题。我对研究事业的奉献沿着坚持不懈和成功的记录将使我能够最大限度地从这个奖项中受益。我在本申请中概述了一个研究计划，重点是了解IL-33在人类哮喘中的功能和调节。哮喘是一种慢性肺部疾病，影响着近3000万美国人和全球约3亿人。虽然存在有效的治疗方法，但许多哮喘患者对现有的治疗方案没有最佳反应;因此，需要新的治疗方法。长期以来被认为是一种免疫系统疾病，越来越多的人认识到气道上皮细胞在哮喘免疫反应中的重要性。新近描述的上皮细胞细胞因子IL-33及其受体ST 2在多个大规模全基因组关联研究中被强烈地暗示在哮喘发病机制中。小鼠模型表明，IL-33通过ST 2受体起作用，促进Th 2炎症。然而，IL-33在人类哮喘中的研究有限。在这个基金中，我将专注于IL-33生物学中的三个重要问题：1）IL-33是如何从气道上皮细胞释放的，2）在人类哮喘中，I-33与Th 2炎症的关系是什么，以及3）ST 2位点的SNPs对可溶性抑制剂sST 2调节IL-33的功能是什么？IL-33在气道上皮细胞核中组成性表达，但缺乏信号序列，其释放机制 不明我通过在气道上皮细胞系中表达GFP标记的IL-33开发了一种新的模型系统，这将使我能够剖析导致IL-33释放的途径。 一旦释放，IL-33被假定诱导气道中的Th 2炎症。我将利用加州大学旧金山分校气道临床研究中心庞大的人类生物标本库，确定IL-33与急性和慢性哮喘中Th 2炎症和粘液重塑标志物的关系。最后，细胞外IL-33可以由其可溶性抑制剂sST 2调节。我会 对大量哮喘患者的ST 2基因座进行测序，并鉴定与改变的ST 2上皮细胞和痰颗粒基因表达相关的SNPs。我将使用启动子报告基因分析和功能性生物测定来测试candidat SNP在人类原代上皮细胞培养中的功能。这项研究计划不仅将推进哮喘生物学领域，而且将使我能够发展新的研究技能以及合作，这对我作为独立研究者的成功至关重要。最后，我概述了一个职业发展计划，利用加州大学旧金山分校的许多机会。我的主要导师，约翰·法伊，是担任我的顾问的理想人选。他带来了气道上皮细胞生物学和哮喘分子表型方面的专业知识，是加州大学旧金山分校大型人类哮喘组织库的主任。我已经召集了一个由其他顾问和合作者组成的团队，包括谢泼德博士、厄尔、伍德拉夫和塞博尔德，他们都有互补的技能。他们将在细胞生物学，原代气道上皮细胞转染，分子克隆，生物统计学和先进的人类遗传学方面为我提供理想的建议。我将补充在遗传学，细胞生物学，免疫学和生物统计学，以及在免疫学和一般肺部医学每周研讨会UCSF提供的正式课程的指导这个团队。医学系和肺部重症监护科在为早期医生科学家提供支持和培训方面有着良好的记录，并拥有资源充足的实验室和核心设施。这些资源将是无价的，在执行我的研究计划，并将确保我的成功。医学部强烈支持我的职业道路和这一应用程序。2012年7月，我被任命为肺重症监护科的助理教授，有80%的研究时间受到保护。有了这些资源和支持，我将有能力建立一个成功的独立研究生涯。