Functional Studies of Novel Genes Mutated in Primary Ciliary Dyskinesia II: Genotype to Phenotype

原发性纤毛运动障碍 II 中新突变基因的功能研究：基因型到表型

• 批准号: 9887916
• 负责人: LAWRENCE E OSTROWSKI
• 金额: $ 67.31万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9887916
• 关键词: Affect; Agonist; Animal Model; Asthma; Bronchiectasis; Cell Culture Techniques; Chronic; Chronic Obstructive Airway Disease; Chronic Sinusitis; Cilia; Clinical; Cystic Fibrosis; DNA Sequence Alteration; Defense Mechanisms; Disease; Disease Progression; Epithelial Cells; Frequencies; Gene Proteins; Genes; Genetic; Genetic Heterogeneity; Genotype; Health; Human; Human Cell Line; Impairment; In Vitro; Individual; Infertility; Inhalation; Ion Transport; Liquid substance; Lung; Lung Transplantation; Lung diseases; Lung infections; Measures; Molecular; Morbidity - disease rate; Mucociliary Clearance; Mucous body substance; Mutate; Mutation; Nasal Epithelium; Nitric Oxide; Nose; Otitis Media; Pathogenesis; Patients; Phenotype; Plant Roots; Play; Primary Ciliary Dyskinesias; Process; Production; Proteins; Rare Diseases; Regulation; Reporting; Residual state; Role; Severities; Severity of illness; Situs Inversus; Symptoms; System; Toxin; cilium motility; clinical phenotype; disease heterogeneity; disease phenotype; gene function; improved; in vivo; induced pluripotent stem cell; male; mortality; mouse model; neonatal respiratory distress; novel; novel therapeutics; pathogen; personalized medicine; protein function; stem cell biology

Project Summary/Abstract In primary ciliary dyskinesia (PCD), mutations in proteins that play a role in the proper assembly or function of the cilia result in defective ciliary beating, which leads to greatly reduced or absent mucociliary clearance (MCC). The lack of effective MCC results in chronic lung infections, bronchiectasis, chronic sinusitis, and otitis media. Although all PCD subjects have a similar clinical phenotype, the disease is heterogeneous in severity, with some patients having mild symptoms, while others develop severe bronchiectasis. Because mutations in many of the genes that cause PCD have been shown to result in essentially immotile cilia and are expected to result in no MCC, it is not obvious why there exists such a wide range of clinical phenotypes. Our hypothesis is that much of the heterogeneity of disease severity observed in PCD patients is a result of genetic heterogeneity, and that mutations in different genes result in varying levels of residual MCC and consequently, varying severity of disease. Further, we hypothesize that different mutations in the same gene can also result in different levels of ciliary impairment, MCC, and disease severity. Finally, we propose that understanding the mechanistic basis for the differences in disease phenotype will provide targets and opportunities to develop new, personalized treatments for this rare disease. The specific aims are: 1. To investigate the expression and function of genes and proteins, including CFAP57 and PCDP1, mutations of which have been newly shown to cause PCD. Using patient derived human nasal epithelial (HNE) cells and/or induced pluripotent stem (iPS) cells, we will investigate the effect of the mutations at the level of the protein, ciliary function (waveform and beat frequency), and mucociliary transport (MCT) in vitro. 2. To investigate genotype/phenotype relationships in patient derived iPS cells. We will examine the functional consequences of mutations in different genes (CFAP57, PCDP1, RSPH1, CCDC39, and DNAI1) and of different mutations in the same gene (SPAG1, CCDC114, DNAH5) in iPS cells from PCD patients. 3. To investigate the pathogenesis of disease in a mouse model with a deletion of Ccdc39. We will compare the effects of an inducible deletion of Ccdc39 to an inducible of Dnaic1. 4. To investigate the relationship between genotype, MCC, and clinical phenotype in vivo. MCC will be measured in PCD subjects with RSPH1 mutations and compared to PCD subjects with mutations in other genes (e.g., DNAI1, DNAH5). In addition, MCC will be measured after administration of a beta-agonist to determine if MCC can be stimulated in the RSPH1 subjects.

项目摘要/摘要 在原发性睫状肌运动障碍(PCD)中，在正常组装或功能过程中起作用的蛋白质突变 纤毛导致有缺陷的纤毛跳动，从而导致粘液纤毛清除能力大大降低或缺失。 (MCC)。缺乏有效的MCC会导致慢性肺部感染、支气管扩张、慢性鼻窦炎和中耳炎。 媒体。尽管所有的PCD患者都有相似的临床表型，但该病的严重程度是不同的， 一些患者有轻微的症状，而另一些患者则发展为严重的支气管扩张。因为基因突变 许多导致PCD的基因已被证明导致基本上静止的纤毛，并有望 结果没有MCC，为什么会存在如此广泛的临床表型并不明显。我们的假设是 在PCD患者中观察到的疾病严重程度的异质性在很大程度上是遗传的结果 异质性，且不同基因突变会导致不同水平的残留MCC， 疾病的严重程度各不相同。此外，我们假设同一基因中的不同突变也可能导致 在不同程度的睫毛损害、MCC和疾病严重程度。最后，我们建议理解 疾病表型差异的机制基础将为其发展提供靶点和机遇 针对这种罕见疾病的新的个性化治疗方法。具体目标是： 1.研究CFAP57和PCDP1等基因和蛋白质的表达和功能。 最近发现了导致PCD的基因突变。 使用患者来源的人鼻上皮(HNE)细胞和/或诱导的多能干细胞(IPS)，我们 将在蛋白质、睫毛功能(波形和节拍)的水平上研究突变的影响 频率)和体外粘液纤毛转运(MCT)。 2.研究患者来源iPS细胞的基因/表型关系。 我们将研究不同基因(CFAP57、PCDP1、RSPH1、 CCDC39和DNAI1)和同一基因(SPAG1、CCDC114、DNAH5)的不同突变 来自PCD患者的细胞。 3.建立Ccdc39基因缺失的小鼠模型，探讨疾病的发病机制。 我们将比较Ccdc39的诱导性缺失和DNAIC1的诱导性缺失的效果。 4.探讨基因分型、MCC与体内临床表型的关系。 有RSPH1突变的PCD患者将测量MCC，并与有RSPH1突变的PCD患者进行比较 其他基因(如DNAI1、DNAH5)的突变。此外，在给药后将测量MCC 一种确定RSPH1受试者是否可以刺激MCC的β-激动剂。