Modulation of Genes Responsible for Cilia Length by Exposure to Cigarette Smoke

通过暴露于香烟烟雾来调节负责纤毛长度的基因

基本信息

批准号：
8505023
负责人：
Ann E Tilley
金额：
$ 13.65万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-12 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8505023
关键词：
Address Air Alternative Splicing Basic Science Bioinformatics Biometry Bronchoscopy Cell Culture Techniques Cells Centrioles Chronic Obstructive Airway Disease Cilia Clinical Trials Critical Care Cultured Cells Data Development Dynein ATPase Environment Epithelial Epithelial Cells Evaluation Exposure to Fellowship Fiber Funding Gene Expression Generations Genes Genetic Medicine Goals Growth Human In Vitro Individual Infection Instruction Lead Length Liquid substance Lung Lung diseases Maintenance Medicine Mentored Patient-Oriented Research Career Development Award Mentors Mentorship Modeling Molecular Biology Mucociliary Clearance Mucous body substance Mus Play Process Pulmonary function tests RNA Splicing Regulation Research Research Personnel Resources Respiratory physiology Risk Factors Role Sampling Smoke Smoker Smoking Sperm Tail Stress Structure Surface Symptoms Therapeutic Intervention Training Variant Work adverse outcome airway epithelium airway obstruction career cigarette smoking cigarette smoking cilium biogenesis experience ezrin gene discovery in vitro Model kinetosome knock-down light microscopy medical schools meetings non-smoker novel patient oriented research professor small hairpin RNA statistics

项目摘要

DESCRIPTION (provided by applicant): Candidate. I am an Assistant Professor of Genetic Medicine and of Medicine, Division of Pulmonary and Critical Care Medicine, at Weill Cornell Medical College. I have been working in Dr. Ronald G. Crystal's lab since my fellowship, focused on evaluation of gene expression in the airway epithelium in healthy nonsmokers, smokers, and smokers with COPD. For the last two years I have taken courses leading to a Master's in Clinical Investigation degree to gain formal instruction relevant to a career in patient-oriented research. I am seeking the K23 award to gain the mentored, practical experience that I need to attain my long-term goal of being an independent investigator in the field of smoking-related lung disease. Environment. My primary mentor for the proposal is Dr. Ronald G. Crystal. Dr. Crystal has an outstanding record of mentoring young investigators to independent careers, and has sufficient funding to provide resources for my proposed project. Co-mentors are Dr. Andrew Clark, providing expertise in biostatistics and bioinformatics, and Dr. Neil Hackett, providing expertise in molecular biology and the basic science approaches used in the project. Consultants are Dr. Jason Mezey for additional statistics expertise and Dr. Charleen Hollman who will provide guidance related to the conduct of patient-oriented research. Through a combination of structured tutorials, mentorship committee meetings, informal interactions and relevant didactic coursework, I will gain the additional training I need to succeed as an independent investigator. Research. Cigarette smoking is the major risk factor for the development of chronic obstructive pulmonary disease (COPD). One manifestation of COPD is diminished mucociliary clearance, the process by which motile cilia on the surface of the airway epithelium function to sweep mucus and debris in a cephalad direction to clear the airways. Even before the development of COPD, individuals who smoke may demonstrate a reduction in mucociliary clearance. Diminishment in mucociliary clearance leads to mucus plugging, forming a nidus for infection and eventually worsening airway obstruction. We have observed that clinically healthy smokers, who have no signs or symptoms of lung disease and who have normal pulmonary function tests, have cilia that are an average of 10% shorter than the cilia of normal nonsmokers. This decrease in length is likely to have adverse consequences for mucociliary clearance. We hypothesize that the stress of smoking modulates the airway epithelial expression of genes critical to the formation and maintenance of cilia of normal length. We will use bronchoscopy to obtain airway epithelial cells and will study gene expression in those cells both ex vivo and after culture at air-liquid interface, with and without exposure to cigarette smoke. Preliminary data suggests smoking- induced alterations in expression of 6 genes likely to play a role in the growth and maintenance of normal-length cilia. In this context, the current proposal has 3 specific aims: (1) to evaluate the hypothesis that exposure to cigarette smoke will result in shorter cilia in human airway epithelial cells grown in culture at air-liquid interface; (2) to evaluate the hypothesis that in vitro suppression of expression of genes involved in the regulation of ciliogenesis and cilia length will result in short cilia in a culture model; and (3) to evaluate the hypothesis that cigarette smoke exposure induces changes in alternative splicing of ODF2, resulting in dysfunctional cilia formation. An understanding of one of the mechanisms underlying dysfunctional mucociliary clearance in smokers may lead to targets for therapeutic intervention.

描述（由申请人提供）：候选人。我是威尔康奈尔医学院肺科和重症监护医学部遗传医学和医学助理教授。自获得奖学金以来，我一直在 Ronald G. Crystal 博士的实验室工作，专注于评估健康不吸烟者、吸烟者和患有慢性阻塞性肺病的吸烟者气道上皮的基因表达。在过去的两年里，我修读了临床研究硕士学位课程，以获得与以患者为导向的研究职业相关的正式指导。我正在寻求 K23 奖，以获得指导和实践经验，以实现成为吸烟相关肺部疾病领域独立研究者的长期目标。环境。我的提案的主要导师是 Ronald G. Crystal 博士。克里斯托博士在指导年轻研究人员走向独立职业方面有着出色的记录，并且有足够的资金为我提议的项目提供资源。共同导师是安德鲁·克拉克博士（提供生物统计学和生物信息学方面的专业知识）和尼尔·哈克特博士（提供分子生物学和项目中使用的基础科学方法方面的专业知识）。顾问包括 Jason Mezey 博士（提供额外的统计专业知识）和 Charleen Hollman 博士（提供有关以患者为导向的研究的指导）。通过结构化教程、指导委员会会议、非正式互动和相关教学课程的结合，我将获得作为独立调查员取得成功所需的额外培训。研究。吸烟是导致慢性阻塞性肺病（COPD）的主要危险因素。 COPD 的表现之一是粘液纤毛清除功能减弱，这是气道上皮表面的活动纤毛发挥作用，沿头侧方向清除粘液和碎片以清除气道的过程。甚至在慢性阻塞性肺病发生之前，吸烟的人就可能表现出粘液纤毛清除能力的降低。粘液纤毛清除功能的减少导致粘液堵塞，形成感染病灶，最终加剧气道阻塞。我们观察到，临床上健康的吸烟者，没有肺部疾病的体征或症状，肺功能测试正常，其纤毛比正常不吸烟者的纤毛平均短 10%。长度的减少可能会对粘液纤毛的清除产生不利影响。我们假设吸烟的压力调节气道上皮基因的表达，这些基因对于纤毛正常长度的形成和维持至关重要。我们将使用支气管镜检查来获取气道上皮细胞，并研究这些细胞的基因表达，无论是在体外还是在气液界面培养后，无论是否暴露于香烟烟雾。初步数据表明，吸烟引起的 6 种基因表达变化可能在正常长度纤毛的生长和维持中发挥作用。在这种背景下，当前的提案有3个具体目标：（1）评估暴露于香烟烟雾会导致在气液界面培养的人气道上皮细胞中纤毛缩短的假设； (2) 评估体外抑制参与纤毛发生和纤毛长度调节的基因表达将导致培养模型中纤毛短的假设； (3) 评估以下假设：香烟烟雾暴露会引起 ODF2 选择性剪接的变化，导致纤毛形成功能失调。了解吸烟者粘液纤毛清除功能障碍的机制之一可能会导致治疗干预的目标。