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性疾病的研究以及纤毛功能障碍可能导致慢性阻塞性肺病 (COPD) 和哮喘的报道。纤毛基因和蛋白质的全面收集有助于快速识别运动纤毛功能障碍综合征（称为原发性纤毛运动障碍（PCD））的遗传病因。然而，目前还没有针对这种疾病的具体疗法，最终 进展为支气管扩张。在这里，我们解决了一系列关键问题，以便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，我们发现了对运动纤毛组装至关重要的特定调控步骤，并为增强纤毛发生的治疗干预提供了前景。我们的初步数据定义了运动纤毛发生的途径，该途径（i）开始于由 HEATR2 控制的细胞质中运动蛋白预组装的最早点，（ii）在功能上与将蛋白质从细胞质运输到受 CCDC11 调节的基体和运动纤毛的网络相关，以及（iii）需要蛋白质 CCDC39 和 CCDC40 在运动纤毛内建立适当的马达和微管组织。我们表明，这些分子关键蛋白中的每一个的突变或缺失都会导致运动纤毛生物发生和功能不成功。我们还发现了该途径每个点的易处理机制，这些机制通过新抑制基因的遗传破坏进行修改，以修复纤毛缺陷。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和 藻类衣藻，它们共同提供纤毛组装途径的体外和体内分析以及快速遗传操作。利用这些策略，我们将探讨动力蛋白运动组装的组装、运输和活性，以实现以下目标：（1）确定动力蛋白运动蛋白复合物递送所需的预组装途径； (2) 确定运动蛋白从基体到纤毛运输的调控； (3) 鉴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，操纵这些途径将为设计针对运动纤毛缺陷的分子疗法提供可能性。