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

基本信息

批准号：
9062491
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
项目类别：
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/9062491
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 Staging Techniques Testing Time Tissue Banking Tissue Banks Training Training Programs Training Support 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 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

项目摘要

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，我提出了一份大学研究和职业发展的全面计划。加利福尼亚州旧金山的肺科。该提案不仅使我能够培养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。我是这个指导性职业发展奖的理想候选人，因为我坚定地致力于建立独立的研究职业。当我在加州大学伯克利分校读本科时，我对研究科学研究产生了兴趣，并在病毒学实验室工作了三年。我以优异的成绩毕业，获得全额学费奖学金，在南加州大学学习医学。我以医学院第一名的成绩毕业后，选择在加州大学圣地亚哥分校完成内科住院医师培训，并在帕特里夏·芬恩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委员会研究培训计划。这使我能够尽早开始在加州大学旧金山分校进行肺部重症监护研究，并花费额外的时间进行研究。在加州大学旧金山分校期间，我继续在约翰·法伊的实验室研究哮喘。在 Fahy 博士、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素在哮喘中的作用的项目。我于 2012 年 1 月发表了这些结果。此后我一直关注 IL-33 在人类哮喘中的作用，这也是我当前提案的主题。我对研究事业的奉献精神以及坚持不懈和成功的记录将使我能够从这个奖项中获得最大的受益。我在此申请中概述了一项研究计划，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，全球估计有 3 亿人。尽管存在有效的治疗方法，但许多哮喘患者对现有治疗方案的反应并不理想。因此，需要新的治疗方法。长期以来，哮喘被认为是一种免疫系统疾病，人们越来越认识到气道上皮在协调哮喘免疫反应中的重要性。在多项大规模全基因组关联研究中，新描述的上皮细胞因子 IL-33 及其受体 ST2 与哮喘发病机制密切相关。小鼠模型表明，IL-33 通过 ST2 受体发挥作用，促进 Th2 炎症。然而，IL-33 在人类哮喘中的研究有限。在这笔资助中，我将重点关注 IL-33 生物学中的三个重要问题：1) IL-33 如何从气道上皮细胞中释放，2) I-33 与人类哮喘中 Th2 炎症的关系是什么，以及 3) ST2 位点中的 SNP 对可溶性抑制剂 sST2 对 IL-33 的调节有何作用？ IL-33在气道上皮细胞核中组成型表达，但缺乏信号序列，其释放机制未知。我通过在气道上皮细胞系中表达 GFP 标记的 IL-33 开发了一种新型模型系统，这将使我能够剖析导致 IL-33 释放的途径。一旦释放，IL-33 就会诱发气道中的 Th2 炎症。利用 UCSF 气道临床研究中心庞大的人类生物样本库，我将确定 IL-33 与急性和慢性哮喘中 Th2 炎症和粘膜重塑标志物的关系。最后，细胞外 IL-33 可以通过其可溶性抑制剂 sST2 进行调节。我会对大量哮喘患者的 ST2 基因座进行测序，并鉴定与 ST2 上皮细胞和痰颗粒基因表达改变相关的 SNP。我将使用启动子报告基因检测和功能生物检测来测试候选 SNP 在人原代上皮细胞培养物中的功能。这项研究计划不仅将推进哮喘生物学领域的发展，而且将使我能够开发新的研究技能以及合作，这对于我作为一名独立研究者的成功至关重要。最后，我概述了一个利用加州大学旧金山分校众多机会的职业发展计划。我的主要导师 John Fahy 非常适合担任我的顾问。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，并且是大型 UCSF 人类哮喘组织库的主任。我组建了一个由其他顾问和合作者组成的团队，包括 Sheppard 博士、Erle、Woodruff 和 Seibold，他们的技能互补。他们非常适合为我提供细胞生物学、原代气道上皮细胞转染、分子克隆、生物统计学和先进人类遗传学方面的建议。我将通过加州大学旧金山分校提供的遗传学、细胞生物学、免疫学和生物统计学的正式课程以及每周一次的免疫学和普通肺医学研讨会来补充该团队的指导。医学系和肺重症监护科在对早期医师科学家的支持和培训方面拥有良好的记录，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非常宝贵，并将确保我的成功。医学系大力支持我的职业道路和这次申请。 2012 年 7 月，我被任命为肺重症监护科助理教授，80% 的研究时间都受到保护。有了这些资源和支持，我将能够建立成功的独立研究生涯。