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: 8509548
• 负责人: Erin D. Gordon
• 金额: $ 13.31万
• 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/8509548
• 关键词: Acute; Affect; Agonist; American; Animal Model; Apoptosis; Asthma; Award; Basic Science; Biological Assay; Biological Models; Biology; Biometry; Biopsy; Blood; California; Cell Culture Techniques; Cell Death; Cell Line; Cell Nucleus; Cells; Cellular biology; Chronic; Chronic lung disease; Clinical; Clinical Research; Code; Collaborations; Commit; 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; Staging; Techniques; Testing; Time; Tissue Banking; Tissue Banks; Training; Training Programs; Training Support; Transfection; Universities; Virus; Virus Diseases; Work; airway epithelium; airway remodeling; analog; career; career development; cohort; cytokine; effective therapy; experience; extracellular; genome wide association study; inhibitor/antagonist; interest; medical schools; member; molecular phenotype; mortality; mouse model; novel; periostin; professor; promoter; receptor; research and development; response; skills; success; 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）：在这项修订的申请中，题为“哮喘中的气道上皮细胞中IL-33的功能和调节”，为指导的临床科学家研究职业发展奖，我提出了加利福尼亚大学旧金山分校的研究和职业发展的全面研究计划。该建议不仅将使我能够发展建立独立的学术研究职业所需的技能，技术和协作，而且还将推进哮喘生物学领域。我是这个指导职业发展奖的理想候选人，因为我强烈致力于建立独立的研究职业。我对加利福尼亚大学伯克利分校的本科生的研究科学感兴趣，在那里我从事病毒学实验室工作了三年。我以荣誉毕业，获得了全额学费奖学金，可以在南加州大学学习医学。我首先在我的医学院课程中毕业，我选择在圣地亚哥加州大学加州大学圣地亚哥分校完成内科医学培训，在那里我在帕特里夏·芬恩（Patricia Finn）博士的实验室进行了基础科学研究。在那里，我对哮喘生物学产生了浓厚的兴趣，并参加了美国内科研究培训委员会。这使我能够尽早开始在UCSF的肺重症监护奖，并花更多的时间进行研究。在UCSF期间，我继续在约翰·法希（John Fahy）实验室学习哮喘。在法希（Fahy），埃尔（Erle），伍德拉夫（Woodruff）和谢泼德（Sheppard）博士的研究和职业指导下，我完成了一个针对骨膜在哮喘中作用的项目。我于2012年1月发表了这些结果。此后，我关注IL-33在人类哮喘中的作用，这是我当前建议的主题。我对研究生涯以及坚持不懈和成功的往绩使我能够从这个奖项中受益。我概述了该应用程序中的研究计划，该计划的重点是了解人类哮喘中IL-33的功能和调节。哮喘是一种慢性肺部疾病，在全球范围内影响近3000万美国人，估计有3亿人。尽管存在有效的治疗方法，但许多哮喘患者对可用方案没有最佳反应。因此，需要新颖的治疗方法。长期以来，被认为是免疫系统的疾病，人们对气道上皮在策划哮喘的免疫反应的重要性越来越多。新描述的上皮细胞细胞因子IL-33及其受体ST2在多个大型基因组广泛的关联研究中与哮喘发病机理有很大的影响。小鼠模型表明，通过ST2受体作用的IL-33促进了TH2炎症。但是，人类哮喘中IL-33的研究受到限制。在这笔赠款中，我将重点介绍IL-33生物学中的三个重要问题：1）如何从气道上皮细胞中释放IL-33，2）2）I-33与人类哮喘中I-33与Th2炎症的关系是什么，3）3）SNP在ST2基因座对IL-33在IL-33 Bi Biy The Alluuble bi Bi Bi Bi Bi Biusitior Inbleble Inbleble Inphibble Inbible ssst2中的功能是什么？ IL-33在气道上皮细胞的核中组成型表达，但缺乏信号序列，其释放的机制 是未知的。我通过在气道上皮细胞系中表达GFP标记的IL-33来开发一种新型的模型系统，这将使我能够解剖导致IL-33释放的途径。 一旦释放，IL-33假定会在气道中诱导Th2炎症。使用UCSF气道临床研究中心的庞大人类生物循环库，我将确定IL-33与急性和慢性哮喘中Th2炎症和粘液重塑的关系的关系。最后，细胞外IL-33可以由其可溶抑制剂SST2调节。我会 序列来自大量哮喘患者的ST2基因座，并识别与改变的ST2上皮细胞和痰液颗粒基因表达相关的SNP。我将使用启动子记者测定法和功能性生物测定法测试念珠菌SNP在人类原发性上皮细胞培养中的功能。该研究计划不仅将推进哮喘生物学领域，而且还可以使我发展新的研究技能以及合作，这对于我作为独立研究者的成功至关重要。最后，我概述了一项职业发展计划，该计划利用了UCSF的许多机会。我的主要导师约翰·法希（John Fahy）非常适合担任我的顾问。他在哮喘中带来了气道上皮细胞生物学和分子表型的专业知识，并且是大型UCSF人类哮喘组织库的主任。我组建了一支由Sheppard，Erle，Woodruff和Seibold博士在内的其他顾问和合作者团队，他们具有互补的技能。他们将为我提供有关细胞生物学，原发气道上皮细胞转染，分子克隆，生物统计学和晚期人类遗传学的理想。我将通过UCSF在遗传学，细胞生物学，免疫学和生物统计学以及免疫学和通用肺医学的每周研讨会上提供的正式课程来为该团队的指导提供指导。医学系和肺重症监护局的支持和培训的良好记录是早期医师科学家，并拥有资源丰富的实验室和核心设施。这些资源对于执行我的研究计划将是无价的，并确保我的成功。医学系强烈支持我的职业道路和这一应用。 2012年7月，我被任命为肺重症监护室内的助理教授，并受到80％的保护时间。有了这些资源和支持，我将有能力建立成功的独立研究职业。