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.

项目概要气道运动纤毛在粘液纤毛清除中发挥重要作用，这是粘液纤毛的主要先天防御机制肺部，通过将粘液中的吸入病原体向上推动并排出呼吸道。睫状体受损功能常见于慢性肺部疾病患者，基因突变导致异常的纤毛形成和功能丧失常常导致遗传异质性的发展，称为纤毛病的多系统疾病。目前的文献表明，独特的成分包裹纤毛的纤毛膜的形态和形态参与维持纤毛功能体内平衡。然而，形成和维持的分子参与者和机制对睫状膜的了解仍知之甚少。该提案的总体目标是定义膜结合蛋白 ciBAR1 和 2（以前称为 FAM92A 和 B）在使用新型基因敲除小鼠模型进行纤毛发生和气道纤毛细胞分化。两种 ciBAR 蛋白拥有单个 Bin/Amphiphyn/Rvs (BAR) 结构域，这使得它们能够二聚化并形成新月体形状结构优先感知和雕刻弯曲的膜。先前的研究表明 ciBAR1 和 2 与纤毛蛋白 Chibby1 (Cby1) 结合，该蛋白在前囊泡募集和基底细胞中发挥作用纤毛细胞分化早期阶段的身体对接。 ciBAR 1 和 2 与 Cby1 在睫状体处共定位基础并在纤毛发生中发挥关键作用，但其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发挥关键作用气道纤毛细胞通过其膜结合和塑造活性。为了解决这个假设，我将使用我们实验室开发的新型 ciBAR1-/- 和 ciBAR2-/- 敲除小鼠模型，并提出遵循具体目标。在目标 1 中，我将研究小鼠中 ciBAR1 或 ciBAR2 的缺失是否会导致睫状体损伤气管缺陷。在目标 2 中，我将研究 ciBAR 蛋白在形成和维持使用原代细胞培养物、生化方法和细胞生物学方法研究纤毛发生过程中的纤毛膜接近。总的来说，我提出的项目将阐明 ciBAR1 和 2 在体内的功能纤毛发生和气道纤毛细胞分化，并增进我们对纤毛如何形成的现有知识膜在纤毛发生的整个过程中产生并维持。