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

• 批准号: 8111560
• 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
• 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/8111560
• 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

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. PUBLIC HEALTH RELEVANCE: Project Narrative One of the ways in which smoking leads to lung disease is by causing is dysfunction of cilia, the hair-like projections on cells lining the airways of the lungs, that function to sweep pathogens and debris out of the lung. We have found that the respiratory cilia of smokers are approximately 10% shorter than those of nonsmokers, which likely has consequences for ciliary function. We propose to study the genetic mechanisms involved in control of cilia length, which could potentially lead to discovering novel drug targets for smoking-related lung disease.

描述（由申请人提供）：候选人。我是威尔·康奈尔医学院（Weill Cornell Medical College）的遗传医学和医学，肺和重症监护医学司的助理教授。自从我的团契以来，我就一直在罗纳德·G·克里斯特尔（Ronald G. Crystal）的实验室工作，重点是评估健康的非吸烟者，吸烟者和患有COPD的吸烟者中气道上皮中的基因表达。在过去的两年中，我参加了导致临床研究硕士学位的课程，以获得与以患者为导向的研究职业有关的正式教学。我正在寻求K23奖，以获得指导的实践经验，我需要实现我的长期目标，即成为与吸烟有关的肺部疾病领域的独立研究者。环境。该提案的主要导师是Ronald G. Crystal博士。 Crystal博士在指导年轻的研究人员到独立职业方面拥有出色的记录，并有足够的资金为我的拟议项目提供资源。联合官员是安德鲁·克拉克（Andrew Clark）博士，他提供了生物统计学和生物信息学方面的专业知识，尼尔·哈克特（Neil Hackett）博士提供了分子生物学方面的专业知识以及项目中使用的基础科学方法。顾问是Jason Mezey博士的其他统计专业知识，Charleen Hollman博士将提供与以患者为导向的研究有关的指导。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，我将获得作为独立研究人员成功的额外培训。研究。吸烟是慢性阻塞性肺疾病（COPD）发展的主要危险因素。 COPD的一种表现降低了粘膜纤毛清除率，这是气道上皮功能表面上的纤毛纤毛在沿头球方向扫除粘液和碎屑以清除气道的过程。甚至在开发COPD之前，吸烟的个体可能会显示出粘膜扫除度清除率的降低。粘膜缩减清除的减少会导致粘液塞，形成一种用于感染的Nidus，并最终导致气道阻塞。我们已经观察到，临床健康的吸烟者没有肺部疾病的迹象或症状并且患有正常肺功能检查的吸烟者的纤毛平均比正常非吸烟者的纤毛短10％。这种长度的减小可能会对粘膜毛的清除产生不利的后果。我们假设吸烟的压力调节了对正常长度纤毛的形成和维持至关重要的基因的气道上皮表达。我们将使用支气管镜检查获得气道上皮细胞，并将研究这些细胞中的基因表达，并在空气界面处培养，并在有或没有暴露于香烟烟雾的情况下进行培养。初步数据表明，吸烟引起的6个基因表达改变可能在正常长度纤毛的生长和维持中起作用。在这种情况下，当前的建议具有3个具体目的：（1）评估以下假设：暴露于香烟烟雾将导致人类气道上皮细胞在空气液体界面培养中生长的纤毛较短； （2）评估以下假设：在体外抑制参与纤毛生成和纤毛长度调节的基因表达将导致培养模型中的简短纤毛； （3）评估以下假设：烟烟暴露会引起ODF2替代剪接的变化，从而导致纤毛功能障碍。对吸烟者中功能失调的粘膜纤维化清除率的一种理解可能导致治疗干预的靶标。 公共卫生相关性：项目叙事一种导致肺部疾病的方法之一是导致纤毛的功能障碍，纤毛的功能障碍，纤毛（肺气道的细胞上的头发样突起），可以将病原体和碎屑从肺中扫除。我们发现，吸烟者的呼吸纤毛比非吸烟者短约10％，这可能会对睫状功能产生影响。我们建议研究控制纤毛长度的遗传机制，这有可能导致发现与吸烟相关的肺部疾病的新型药物靶标。