Regulation of Motile Cilia Assembly in Lung Disease

肺部疾病中活动纤毛组装的调节

• 批准号: 9887501
• 负责人: Steven Brody
• 金额: $ 78.69万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-17 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9887501
• 关键词: Actins; Adaptor Signaling Protein; Address; Binding; Biochemical; Biophysics; Carrier Proteins; Cells; Chlamydomonas; Chronic; Chronic lung disease; Cilia; Cities; Complex; Cryoelectron Microscopy; Cystic Fibrosis; Cytoplasm; Data; Development; Disease; Dynein ATPase; Elements; Functional disorder; Funding; Genes; Genetic; Genetic Diseases; Genotype; Goals; Host Defense; Human; Link; Lung; Lung diseases; Lung infections; Mechanics; Microtubules; Modeling; Motor; Movement; Mucous body substance; Mus; Mutation; Organism; Particulate; Pathway interactions; Patients; Phenotype; Positioning Attribute; Production; Proteins; Proteomics; Regulation; Reporter; Reporting; Role; Route; Secure; Series; Set protein; Site; Specificity; Structure; Syndrome; Therapeutic; Tissues; Translating; Transportation; Tubulin; Universities; Washington; arm; base; biophysical properties; cell motility; cilium biogenesis; cilium motility; comparative; genetic regulatory protein; human tissue; injured airway; kinetosome; multidisciplinary; mutant; novel; particle; pathogen; protein complex; protein function; scaffold; targeted treatment; trafficking

Project Summary/Abstract Cilia dysfunction leads to chronic lung disease, as occurs in the genetic syndrome primary ciliary dyskinesis (PCD) and may contribute to impaired airway clearance in obstructive pulmonary disease (COPD). Unlike cystic fibrosis, which also causes chronic airway destruction, there is no specific therapy for cilia-related diseases. We now know that most PCD mutations result in defective production, transport, or proper placement of ciliary motors along the ciliary axoneme. However, the exact mechanism by which this occurs is not understood, impeding the development of cilia-specific therapeutic strategies. Thus, our goal is to determine how ciliary motor components are directed from the cytoplasm, into the cilia, then find their way to specific sites along the ciliary axoneme. In this proposal, we trace the passage of the multifunctional, heterodimer CCDC39/CCDC40, which when mutant result in disorganized axonemal microtubule structure and severe PCD disease. CCDC39 trafficking serves to identify the mechanisms by which ciliary components move from the cytoplasm, en route to the basal body, and into cilia by intraflagellar transport (IFT). Our preliminary data indicate that: (1) Ciliary cargoes, including CCDC39, bind centriolar satellite proteins that function as “cars” to facilitate trafficking to the basal body; (2) CCDC39/CCDC40 are linked to microtubules in association with two novel “staple” proteins; and (3) CCDC39 is an IFT co-adaptor protein, with ciliary protein MLF1, for cargo delivery to cilia. We hypothesize that movement of ciliary cargo from the cytoplasm to cilia engages transport mechanisms using satellite proteins, then co-adapter proteins, including CCDC39/CCDC40, to properly assemble cilia. We will characterize this series of pathways in the Specific Aims: (1) Determine how components of motile cilia are trafficked from the cytoplasm to basal bodies; (2) Characterize the multifunctional protein CCDC39 by biophysical and biochemical means that will identify the role of novel staple proteins; and (3) Identify the role for CCDC39 and MLF1 as IFT co-adapter proteins in PCD mutants and COPD tissues. The project uses highly integrated strategies that employ models from single cell organisms to human tissues carried out by the multidisciplinary Washington University Cilia Group, to define new pathways for cilia assembly that can be translated to therapies.

项目摘要/摘要 纤毛功能障碍会导致慢性肺病 （PCD），可能导致阻塞性肺部疾病（COPD）中的气道清除受损。与众不同 囊性纤维化也会导致慢性气道破坏，没有针对纤毛有关的特定疗法 疾病。我们现在知道，大多数PCD突变会导致产生，运输或正常 沿睫状轴突的睫状电机放置。但是，发生这种情况的确切机制是 不了解，阻碍了纤毛特异性治疗策略的发展。那是我们的目标 确定如何从细胞质引导，进入纤毛，然后找到通往纤毛的方式 沿睫状轴突的特定位点。在此提案中，我们追踪多功能的通过 异二聚体CCDC39/CCDC40，当突变体导致轴突微管结构混乱时， 严重的PCD疾病。 CCDC39贩运有助于确定睫状成分移动的机制 从细胞质到基本体的途径，并通过flagellar内转运（IFT）进入纤毛​​。我们的初步 数据表明：（1）睫状货物，包括CCDC39，结合Centriolar卫星蛋白，起作用 “汽车”促进贩运基本机构； （2）CCDC39/CCDC40与关联的微管链接 带有两个新颖的“主食”蛋白； （3）CCDC39是一种IFT共同适应器蛋白，具有纤毛蛋白MLF1，用于 货物运送到纤毛。我们假设睫状货物从细胞质到纤毛的运动 使用卫星蛋白，然后是包括CCDC39/CCDC40在内的共同适应蛋白的传输机制， 正确组装纤毛。我们将在特定目的中表征这一系列途径：（1）确定如何 纤毛母亲的成分从细胞质贩运到基本体。 （2）表征 生物物理和生化手段的多功能蛋白CCDC39将确定新型主食的作用 蛋白质； （3）将CCDC39和MLF1的作用确定为PCD突变体和 COPD组织。该项目使用高度集成的策略，使员工从单细胞生物模型到 由多学科华盛顿大学西里亚集团（Cilia Group）进行的人体组织，以定义新途径 对于可以翻译成疗法的纤毛组装。