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（通过应用程序提供）：气道间隙的故障是慢性气道疾病的特征。证据表明，完整的粘膜膜仪是清除纤毛遗传疾病以及纤毛功能障碍可能有助于慢性阻塞性肺部疾病（COPD）和哮喘的报道所必需的。睫状基因和蛋白质的综合收集有助于鉴定母亲纤毛功能障碍综合征的遗传病因的进步迅速，称为原发性纤毛性运动障碍（PCD）。但是，没有针对这种疾病的具体疗法 发展到支气管腔。在这里，我们解决了一组关键问题，以更完整地了解纤毛的纤毛的组装和调节。使用PCD中突变的新基因作为指导，我们发现了特定的调节步骤，这些步骤对于运动纤毛的组装至关重要，并为增强纤毛生成提供热干预的前景。我们的初步数据定义了运动纤毛发生的途径，该途径（i）在由Heatr2控制的细胞质中最早开始的运动蛋白蛋白质序列开始，（ii）在功能上与一个网络相关，该网络在流量蛋白上与从细胞质量和母亲ccdc11 and ccdc11 and ccd ccd and CCD ccd andcd ccd andcd ccd andcd ccd andcd ccd andcd andcd ccd andcd ccd andcd secd secd链接起来。在母纤毛内建立适当的电动机和微管的组织。我们表明，这些分子Linchpins的突变或部署会导致纤毛生物发生和正常功能不成功。我们还发现了我们所显示的途径的每个点的可拖动机制，这是通过新型抑制基因的遗传破坏来修改的，从而修复了纤毛缺陷。我们的初步和提议的研究利用了原发性多细胞气道上皮细胞的完整模型，斑马鱼和 使用这些策略，我们将在以下目的中探测动力蛋白运动组装的组装，运输和活性：（1）确定Dynein运动蛋白复合物所需的预组装途径； （2）确定从基本物体到纤毛的运动蛋白运输的调节； （3）确定可以恢复缺乏电动机蛋白对接和装配因子的细胞功能的基因。我们建议对这些途径的操纵将为苏里亚母亲的缺陷设计分子疗法提供可能。