Regulation of Motile Cilia Assembly in Lung Disease

项目总结/摘要 纤毛功能障碍导致慢性肺部疾病，如发生在遗传综合征原发性纤毛运动障碍 (PCD)并且可能导致阻塞性肺病（COPD）中气道清除受损。不像 囊性纤维化，这也会导致慢性气道破坏，目前还没有特异性治疗纤毛相关的 疾病我们现在知道，大多数PCD突变会导致生产、运输或适当的缺陷。 纤毛运动沿着纤毛轴丝的放置。然而，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。因此，我们的目标是 确定纤毛运动成分是如何从细胞质进入纤毛的，然后找到它们的方式， 沿着纤毛轴丝的特定位点。在这个建议中，我们跟踪多功能的通道， 异源二聚体CCDC 39/CCDC 40，当突变时导致轴丝微管结构紊乱， 严重的PCD疾病。CCDC 39运输用于识别纤毛成分移动的机制 从细胞质，途中到基体，并进入纤毛内鞭毛运输（IFT）。我们的初步 数据表明：（1）纤毛货物，包括CCDC 39，结合中心粒卫星蛋白，其功能是 “汽车”，以促进贩运到基体;（2）CCDC 39/CCDC 40连接到微管协会 CCDC 39是IFT共衔接蛋白，与纤毛蛋白MLF 1， 运送货物到纤毛。我们假设纤毛货物从细胞质到纤毛的运动 转运机制使用卫星蛋白，然后共衔接蛋白，包括CCDC 39/CCDC 40， 正确组装纤毛。我们将在具体目标中描述这一系列途径：（1）确定如何 运动纤毛的成分从细胞质运输到基体;（2）表征 多功能蛋白质CCDC 39的生物物理和生化手段，将确定新的主食的作用， （3）鉴定CCDC 39和MLF 1作为IFT共衔接蛋白在PCD突变体中的作用， COPD组织。该项目采用高度集成的策略，采用单细胞生物模型， 由多学科的华盛顿大学纤毛组进行的人体组织，以确定新的途径， 用于纤毛组装，可以转化为治疗方法。

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.