Elucidating the Role of the Distal Appendage Protein CEP164 in Multiciliated Cell Differentiation

阐明远端附属蛋白 CEP164 在多纤毛细胞分化中的作用

基本信息

批准号：
9356318
负责人：
Saul Sylvan Siller
金额：
$ 4.9万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-15 至 2018-09-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9356318
关键词：
Address Affect Apical Asthma Bardet-Biedl Syndrome Binding Binding Proteins Biochemical Biological Assay Biological Models Brain Categories Cell Count Cell Differentiation process Cell Line Cell surface Cells Centrioles Chronic Chronic Obstructive Airway Disease Chronic Rhinitis Chronic Sinusitis Cilia Congenital Disorders Cultured Cells Cystic Fibrosis Data Defect Development Disease Disease model Distal Docking Edema Epithelial Cells Fiber Genes Genetic Goals Growth Hydrocephalus Imagery Individual Infertility Injectable Joubert syndrome Knock-out Knockout Mice Laboratories Location Lung Mammalian Oviducts Mature Centriole Methodology Microscopy Modeling Molecular Movement Mucociliary Clearance Mucous body substance Mus Mutation Nasal Epithelium Nature Nephronophthisis Outcome Pathogenesis Pathologic Pathology Patients Phenotype Physiological Population Primary Ciliary Dyskinesias Process Production Proteins Publishing Recruitment Activity Reporting Research Resolution Role Signal Transduction Structure Structure of respiratory epithelium Symptoms Syndrome System Techniques Time Tissues Ventricular Vesicle Work Zebrafish airway epithelium appendage cell motility ciliopathy cilium biogenesis cilium motility falls human disease human tissue in vivo insight interest kinetosome mouse model novel novel therapeutics public health relevance respiratory therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Multiciliated cells are found in abundance in the respiratory epithelium. The hundreds of motile cilia on each multiciliated cell aids in both signaling processes and mucociliary clearance. Defects in the differentiation of these cells or in cilia formation result in congenital disorders, such as primary ciliary dyskinesia (PCD), and chronic respiratory conditions, such as cystic fibrosis (CF), asthma, and chronic obstructive pulmonary disease. Despite the significant portion of the population affected by these chronic pulmonary conditions as well as the devastating nature of PCD, CF, and similar congenital disorders, little is known about the molecular mechanisms dictating multiciliated cell differentiation. In 2007, an absolute requirement in primary ciliogenesis was discovered for the ciliary protein centrosomal protein 164 (CEP164). Since this time, great interest has followed as CEP164 was revealed to be part of the distal appendage, a structure that extends from the distal portion of the centriole/basal body and is critical for vesicle recruitment and basal body docking. While prior work has made progress in uncovering the role of CEP164 in the formation of primary cilia in cultured cells and morpholino studies in zebrafish, no study has yet to examine its functionality in airway multiciliated cells in the physiological setting of a mammalian model system. The overall goal of this proposal is to define the function of CEP164 during multiciliated cell differentiation in normal and diseased airway epithelium. To accomplish this goal, this proposal employs a novel multiciliated cell-specific CEP164 knockout mouse model and a primary culture system of mouse tracheal epithelial cells (MTECs). By harnessing both of these tools, the roles of CEP164, its interacting partners, and the distal appendage will be elucidated through genetic means, biochemical assays, and direct visualization (SEM, TEM, confocal, and super-resolution microscopy). I expect that these methodologies will provide novel insights into the processes of motile ciliogenesis and multiciliated cell development. I also anticipate that, by knocking out CEP164 in multiciliated cells, a powerful mouse model of PCD will be obtained. This work will identify novel molecular networks regulating ciliogenesis and potential therapeutic targets for patients afflicted with diseases of dysfunctional motile cilia.

描述（由适用提供）：在呼吸上皮中发现了多精细的细胞。每个多毛细胞在信号传导过程和粘膜清除率中的数百个纤毛纤毛有助于。这些细胞分化或纤毛形成的缺陷导致先天性疾病，例如原发性纤毛运动障碍（PCD）和慢性呼吸疾病，例如囊性纤维化（CF），哮喘和慢性阻塞性阻塞性肺疾病。尽管受这些慢性肺部状况影响的人群中有很大的部分以及PCD，CF和类似先天性疾病的毁灭性性质，但对决定多纤维化细胞分化的分子机制知之甚少。在2007年，发现针对纤毛蛋白中心蛋白164（CEP164）的原发性纤毛生成的绝对需求。从那时以来，人们一直是CEP164的极大兴趣，而先前的工作在揭示了CEP164在培养的细胞中形成原发性纤毛中的作用和斑马鱼中的morpholino研究的作用，但没有任何研究尚未研究其在哺乳动物物理环境中的空气道中的功能该提案的总体目标是在正常气道上皮中定义CEP164在多重细胞分化过程中的功能。为了实现这一目标，该提案采用了一种新型的多重细胞特异性CEP164基因敲除小鼠模型和小鼠气管上皮细胞（MTEC）的主要培养系统。通过利用这两种工具，CEP164的角色，其相互作用的伙伴和远端附属物将被阐明。通过遗传手段，生化测定和直接可视化（SEM，TEM，共聚焦和超分辨率显微镜）。我预计这些方法将为动向纤毛发生和多重细胞发育的过程提供新的见解。我还预计，通过将CEP164拆除在多重细胞中，将获得强大的PCD鼠标模型。这项工作将确定针对患有功能障碍纤毛疾病的患者的纤毛生成和潜在治疗靶标的新型分子网络。