REGULATION OF MOTILE CILIA ASSEMBLY IN LUNG DISEASE

肺部疾病中活动纤毛组件的调节

• 批准号: 8941243
• 负责人: Steven Brody
• 金额: $ 57.64万
• 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 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8941243
• 关键词: Address; Algae; Apical; Asthma; Bronchiectasis; Carrier Proteins; Cells; Characteristics; Chlamydomonas; Chronic; Chronic Obstructive Airway Disease; Chronic lung disease; Cilia; Collection; Complex; Congenital Heart Defects; Cytoplasm; Cytoplasmic Protein; Data; Defect; Development; Disease; Docking; Dynein ATPase; Epithelial Cells; Event; Failure; Functional disorder; Future; Gene Proteins; Genes; Genetic; Genetic Predisposition to Disease; Goals; Hereditary Disease; Host Defense; Human; In Vitro; Infection; Inherited; Knowledge; Lead; Length; Link; Lung; Lung diseases; Microtubules; Modeling; Molecular; Motor; Mutate; Mutation; Nuclear; Outcome; Particulate; Pathway interactions; Phosphotransferases; Primary Ciliary Dyskinesias; Proteins; Regulation; Regulatory Pathway; Reporting; Role; Staging; Structure; Suppressor Genes; Syndrome; Testing; Therapeutic; Therapeutic Intervention; Zebrafish; arm; cell motility; cilium biogenesis; forward genetics; functional restoration; gain of function; genetic manipulation; improved; in vivo; kinetosome; loss of function mutation; member; mutant; novel; pathogen; programs; protein complex; protein transport; public health relevance; targeted treatment; therapy development; trafficking

 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.

 描述（由申请人提供）：气道清除失败是慢性气道疾病的一个特征。完整的粘液纤毛装置对于清除至关重要的证据来自对纤毛遗传疾病的研究以及纤毛功能障碍可能导致慢性阻塞性呼吸道疾病的报道。纤毛基因和蛋白质的全面收集促进了运动性纤毛功能障碍综合征遗传病因学的快速进展，称为原发性纤毛运动障碍（PCD），但目前尚无针对这种疾病的具体疗法。 在这里，我们解决了一系列关键问题，以更全面地了解运动纤毛的组装和调节，以 PCD 中突变的新基因为指导，我们发现了对组装至关重要的特定调节步骤。我们的初步数据定义了运动纤毛发生的途径，该途径（i）始于细胞质中由运动蛋白预组装的最早点。 HEATR2，(ii) 在功能上连接到一个网络，该网络将蛋白质从细胞质运输到受 CCDC11 调节的基体和运动纤毛，并且 (iii) 需要蛋白质 CCDC39 和 CCDC40 在运动纤毛内建立马达和微管的正确组织。我们表明，这些分子关键蛋白的突变或缺失会导致运动纤毛生物发生和功能不成功，我们还揭示了每个点的易处理机制。我们所展示的途径是通过新的抑制基因的遗传破坏来修复纤毛缺陷的，我们的初步和拟议研究利用了原代多纤毛人气道上皮细胞、遗传缺陷斑马鱼品系的互补模型。 藻类衣藻共同提供纤毛组装途径的体外和体内分析和快速遗传操作，我们将探讨动力蛋白运动组装的组装、运输和活性，目的如下：（1）识别预组装。动力蛋白运动蛋白复合物递送所需的途径；（2）确定运动蛋白从基体到纤毛运输的调节；（3）确定可以恢复缺乏运动蛋白对接和组装因子的细胞的功能的基因。对这些途径的操纵将为设计针对运动纤毛缺陷的分子疗法提供可能性。