REGULATION OF MOTILE CILIA ASSEMBLY IN LUNG DISEASE

 描述（由申请人提供）：气道清除失败是慢性气道疾病的典型特征。纤毛遗传性疾病的研究以及纤毛功能障碍可能导致慢性阻塞性肺疾病（COPD）和哮喘的报道证明，完整的粘膜纤毛装置对于清除至关重要。纤毛基因和蛋白质的全面收集促进了运动纤毛功能障碍综合征（称为原发性纤毛运动障碍（PCD））遗传病因学鉴定的快速进展。然而，对于这种疾病没有特定的治疗方法， 发展为支气管扩张在这里，我们解决一组关键的问题，以获得更完整的了解大会和调节运动纤毛。利用PCD中突变的新基因作为指导，我们发现了对运动纤毛组装至关重要的特定调控步骤，并提供了增强纤毛发生的治疗干预的前景。我们的初步数据定义了运动纤毛发生的途径，该途径（i）在HEATR 2控制的细胞质中马达蛋白预组装的最早点开始，（ii）在功能上与将蛋白质从细胞质运输到由CCDC 11调节的基体和运动纤毛的网络连接，和（iii）需要蛋白质CCDC 39和CCDC 40在运动纤毛内建立马达和微管的适当组织。我们发现，这些分子关键因子的突变或缺失导致不成功的运动纤毛生物发生和功能的预期。我们还发现了在每一个点的途径，我们表明是由新的抑制基因的遗传破坏，以修复纤毛缺陷修改的易处理的机制。我们的初步和拟议的研究利用了原代多纤毛人气道上皮细胞、斑马鱼遗传缺陷株和 衣原体，它们一起提供纤毛组装途径的体外和体内分析和快速遗传操作。利用这些策略，我们将探索动力蛋白马达组装的组装、运输和活性，目的如下：（1）确定动力蛋白马达蛋白复合物递送所需的预组装途径;（2）确定马达蛋白从基体到纤毛运输的调节;（3）确定可以恢复缺乏马达蛋白对接和组装因子的细胞功能的基因。我们认为，操纵这些途径将提供设计分子治疗的缺陷，在运动纤毛的可能性。

 DESCRIPTION (provided by applicant): Failure of airway clearance is a characteristic feature of chronic airway disorders. Evidence that an intact mucociliary apparatus is essential for clearance derives from studies of genetic disease of cilia as well as reports that cilia dysfunctio may contribute to chronic obstructive pulmonary disease (COPD) and asthma. Comprehensive collections of ciliary genes and proteins have contributed to the rapid pace of progress in the identification of genetic etiologies of the syndrome of motile cilia dysfunction, called primary ciliary dyskinesia (PCD). However, there are no specific therapies for this disease that ultimately progresses to bronchiectasis. Here, we address a key set of questions to gain a more complete understanding of the assembly and regulation of motile cilia. Using novel genes mutated in PCD as a guide, we have uncovered specific regulatory steps that are critical for the assembly of motile cilia and offer the prospect of therapeutic intervention to augment ciliogenesis. Our preliminary data define a pathway for motile ciliogenesis that (i) commences at the earliest point of motor protein preassembly in the cytoplasm controlled by HEATR2, (ii) is functionally linked to a network that traffics protein from the cytoplasm to the basal bodies and motile cilia regulated by CCDC11, and (iii) requires the proteins CCDC39 and CCDC40 to establish the proper organization of motors and microtubules within the motile cilium. We show that mutation or depletion of each of these molecular linchpins results in unsuccessful motile cilia biogenesis and function as expected. We also uncover tractable mechanisms at each point of the pathway that we show are modified by the genetic disruption of novel suppressor genes to remediate cilia defects. Our preliminary and proposed studies take advantage of complementary models of primary multiciliated human airway epithelial cells, genetically deficient strains of zebrafish and the alga Chlamydomonas, which together provide in vitro and in vivo analysis and rapid genetic manipulation of cilia assembly pathways. Using these strategies, we will probe the assembly, trafficking and activity of dynein motor assembly in the following aims: (1) Identify the preassembly pathways required for dynein motor protein complex delivery; (2) Identify the regulation of motor protein trafficking from basal bodies to cilia; (3) Identify genes that can restore function to cells lacking docking and assembly factors for the motor proteins. We propose that manipulation of these pathways will offer possibilities of devising molecular therapies for defects in motile cilia.