Elucidating the Roles of the Membrane-Binding Proteins ciBAR1 and ciBAR2 in Ciliogenesis

阐明膜结合蛋白 ciBAR1 和 ciBAR2 在纤毛发生中的作用

基本信息

批准号：
10677252
负责人：
Eunice Na Young Kim
金额：
$ 4.01万
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：
2023
资助国家：
美国
起止时间：
2023-08-23 至 2026-08-22
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677252
关键词：
Address Affect Age Antibodies Apical Binding Binding Proteins Biochemical Biogenesis Biological Biological Models Biological Process Biology Biosensor Cell Differentiation process Cell membrane Cell surface Cells Centrioles Chronic Chronic lung disease Cilia Complex DNA Sequence Alteration Data Defect Defense Mechanisms Development Dimerization Disease Distal Docking Endocytosis Epithelial Cells Exhibits Functional disorder Goals Hair Histology Homeostasis Human Immunofluorescence Immunologic Impairment In Vitro Inhalation Knockout Mice Knowledge Label Laboratories Link Lipids Liposomes Literature Lung Lung diseases Maintenance Mammalian Cell Mediating Membrane Membrane Lipids Methods Microtubules Molecular Morphology Mucociliary Clearance Mucous body substance Mus Natural Immunity Organ Organelles Patients Pharmaceutical Preparations Phenotype Phospholipids Physiological Play Preventive treatment Primary Cell Cultures Process Property Proteins Reporting Research Respiration Disorders Respiratory System Role SARS-CoV-2 infection Scanning Electron Microscopy Sedimentation process Shapes Stains Structure System Tertiary Protein Structure Testing Trachea Tracheal Epithelium Transmission Electron Microscopy Vesicle amphiphysin apical membrane base body system ciliopathy cilium biogenesis cilium motility clinical phenotype dimer in vitro Assay in vivo insight kinetosome loss of function membrane assembly mouse model novel pathogen recruit symptom treatment wasting

项目摘要

PROJECT SUMMARY Airway motile cilia play an important role in mucociliary clearance, the primary innate defense mechanism of the lung, by propelling inhaled pathogens trapped in mucus up and out of the respiratory tract. Impaired ciliary function is commonly found in patients with chronic pulmonary diseases, and genetic mutations that result in aberrant cilia formation and loss of function often lead to the development of genetically heterogenous, multisystem disorders called ciliopathies. The current body of literature suggests that the unique composition and morphology of the ciliary membrane that envelopes the cilium are involved in maintaining cilia function and homeostasis. However, the molecular players and mechanisms underlying the formation and maintenance of the ciliary membrane remain poorly understood. The overarching objective of this proposal is to define the physiological roles of the membrane-binding proteins ciBAR1 and 2 (formerly known as FAM92A and B) in ciliogenesis and airway ciliated cell differentiation by using novel knockout mouse models. Both ciBAR proteins possess a single Bin/Amphiphysin/Rvs (BAR) domain, which allows them to dimerize and form crescent- shaped structures to preferentially sense and sculpt curved membranes. Previous studies have shown that ciBAR1 and 2 bind to the ciliary protein Chibby1 (Cby1), which acts in preciliary vesicle recruitment and basal body docking during early stages of ciliated cell differentiation. ciBAR 1 and 2 colocalize with Cby1 at the ciliary base and play crucial roles in ciliogenesis, but their in vivo functions are currently unknown. The central hypothesis of this proposal is that ciBAR1 and 2 play key roles in ciliogenesis and differentiation of airway ciliated cells via their membrane-binding and shaping activity. To address this hypothesis, I will use novel ciBAR1-/- and ciBAR2-/- knockout mouse models developed by our laboratory and propose the following Specific Aims. In Aim 1, I will investigate if the loss of ciBAR1 or ciBAR2 in mice results in ciliary defects in the trachea. In Aim 2, I will examine the roles of ciBAR proteins in formation and maintenance of ciliary membranes during ciliogenesis using primary cell cultures, biochemical methods, and cell biological approaches. Collectively, my proposed project will shed light on the in vivo functions of ciBAR1 and 2 in ciliogenesis and airway ciliated cell differentiation and advance our existing knowledge of how the ciliary membrane is generated and maintained throughout the process of ciliogenesis.

项目摘要气道纤毛纤毛在粘膜校利清除率中起着重要作用，这是主要的先天防御机制肺部通过驱动吸入的病原体被困在呼吸道上的粘液中。睫状体受损功能通常在患有慢性肺部疾病的患者和导致的基因突变中发现异常的纤毛形成和功能丧失通常会导致遗传异质的发展，多系统疾病称为纤毛病。目前的文献表明独特的构图包裹纤毛的睫状膜的形态参与维持纤毛功能和稳态。但是，构成和维护的基础的分子参与者和机制睫状膜的理解仍然很差。该提议的总体目标是定义膜结合蛋白CIBAR1和2（以前称为FAM92A和B）的生理作用纤毛生成和气道通过使用新型敲除小鼠模型的细胞分化。两种Cibar蛋白具有单个bin/Amphiphysin/rvs（bar）域，这使它们能够二聚并形成新月形 - 形状的结构优先感应和雕刻弯曲的膜。先前的研究表明 CIBAR1和2与睫状蛋白奇迹1（CBY1）结合，该纤毛蛋白质作用于预科囊泡募集和基础上纤毛细胞分化的早期阶段的身体对接。 CILAIRY在Ciliary处的CIBAR 1和2与CBY1共定位基础和在纤毛发生中起关键作用，但它们的体内功能目前尚不清楚。中央该提议的假设是CIBAR1和2在纤毛发生和分化中起关键作用气道纤毛细胞通过其膜结合和塑造活性。为了解决这一假设，我将使用新颖的Cibar1 - / - 和Cibar2 - / - 由我们的实验室开发的小鼠模型，并提出遵循特定目标。在AIM 1中，我将调查小鼠中CIBAR1或CIBAR2的损失是否导致睫状气管中的缺陷。在AIM 2中，我将研究Cibar蛋白在形成和维护中的作用纤毛膜在纤毛生成过程中使用原代细胞培养物，生化方法和细胞生物学方法。总的来说，我提议的项目将阐明Cibar1和2的体内功能纤毛生成和气道纤毛细胞分化，并促进了我们现有的睫状体知识在整个纤毛发生过程中，产生和维持膜。