Regulation of Motile Cilia Assembly in Lung Disease

项目摘要/摘要 纤毛功能障碍导致慢性肺部疾病，如遗传综合征原发纤毛运动障碍所致 (PCD)，并可能导致阻塞性肺疾病(COPD)的呼吸道清除障碍。不像 囊性纤维化也会导致慢性呼吸道破坏，目前还没有针对纤毛相关的特效治疗方法 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 沿睫状轴丝的纤毛马达的放置。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 确定纤毛运动成分如何从细胞质进入纤毛，然后找到它们的途径 沿纤毛轴丝的特定部位。在这项提案中，我们追溯了多功能、 异源二聚体CCDC39/CCDC40，当突变导致轴丝微管结构紊乱时， 严重的PCD疾病。CCDC39贩运用于识别纤毛部件移动的机制 通过鞭毛内转运(IFT)从细胞质进入纤毛。我们的预赛 数据表明：(1)包括CCDC39在内的睫状物与中心粒卫星蛋白结合，中心粒卫星蛋白的功能是 (2)CCDC39/CCDC40与微管相连 和(3)CCDC39是一种IFT共适配蛋白，与纤毛蛋白MLF1共同作用于 把货物送到cilia。我们假设纤毛货物从细胞质到纤毛的运动 利用卫星蛋白的转运机制，然后是共适配蛋白，包括CCDC39/CCDC40，以 正确组装纤毛。我们将在具体目标上描述这一系列路径：(1)确定如何 活动纤毛的成分从细胞质运输到基本体；(2)表征 通过生物物理和生化手段鉴定多功能蛋白质CCDC39的作用 蛋白质；以及(3)确定CCDC39和MLF1作为IFT共适配蛋白在PCD突变体和 慢性阻塞性肺疾病组织。该项目使用了高度集成的策略，使用了从单细胞生物体到 由华盛顿大学Cilia小组进行的多学科人体组织研究，以确定新的途径 用于纤毛组装，可以转化为治疗。

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.