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.

项目概要/摘要 纤毛功能障碍会导致慢性肺部疾病，如遗传综合征原发性纤毛运动障碍中所发生的那样 (PCD)，并可能导致阻塞性肺疾病 (COPD) 气道清除受损。 囊性纤维化，也会导致慢性气道破坏，目前没有针对纤毛相关的具体治疗方法 我们现在知道，大多数 PCD 突变都会导致生产、运输或正常运转方面的缺陷。 沿着纤毛轴丝放置纤毛马达然而，发生这种情况的确切机制是。 不被理解，阻碍了纤毛特异性治疗策略的发展因此，我们的目标是 确定纤毛运动成分如何从细胞质引导到纤毛，然后找到它们的途径 沿着纤毛轴丝的特定位点在这个提议中，我们追踪多功能的通道， 异二聚体 CCDC39/CCDC40，突变时会导致轴丝微管结构紊乱， 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，到基体，并通过鞭毛内运输（IFT）进入纤毛​​。 数据表明：(1) 纤毛货物，包括 CCDC39，结合中心粒卫星蛋白，其功能为 促进向基底体运输的“汽车”；（2）CCDC39/CCDC40与微管相关联； 具有两种新的“主食”蛋白；(3) CCDC39 是一种 IFT 辅接头蛋白，具有纤毛蛋白 MLF1，用于 我们与纤毛货物从细胞质到纤毛的运动进行了斗争。 使用卫星蛋白的转运机制，然后使用辅助接头蛋白（包括 CCDC39/CCDC40） 我们将在具体目标中描述这一系列途径的特征：（1）确定如何组装纤毛。 运动纤毛的成分从细胞质运输到基体；（2）表征 通过生物物理和生化手段鉴定多功能蛋白CCDC39将鉴定新型主食的作用 (3) 确定 CCDC39 和 MLF1 作为 IFT 辅助接头蛋白在 PCD 突变体中的作用， 该项目采用高度集成的策略，采用从单细胞生物到 由华盛顿大学多学科纤毛小组开展的人体组织研究，以确定新的途径 用于可转化为治疗的纤毛组装。