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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

基本信息

批准号：
10570977
负责人：
LAWRENCE E OSTROWSKI
金额：
$ 63.39万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-15 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10570977
关键词：
Affect Agonist Animal Model Asthma Bronchiectasis Cell Culture Techniques Chronic Chronic Obstructive Pulmonary Disease Chronic Sinusitis Cilia Clinical Cystic Fibrosis DNA Sequence Alteration Defense Mechanisms Disease Disease Progression Epithelial Cells Frequencies 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 Neonatal Respiratory Distress Nitric Oxide Nose Otitis Media Pathogenesis Patients Persons Phenotype Primary Ciliary Dyskinesias Process Production Proteins Rare Diseases Regulation Reporting Reproducibility Residual state Respiratory Disease 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 novel novel therapeutics pathogen personalized medicine protein function stem cell biology

项目摘要

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.

项目概要/摘要在原发性纤毛运动障碍 (PCD) 中，在正常组装或功能中发挥作用的蛋白质突变纤毛导致纤毛跳动缺陷，从而导致粘液纤毛清除能力大大减少或消失（中冶集团）。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 PCD 受试者都具有相似的临床表型，但该疾病的严重程度存在异质性，一些患者症状较轻，而另一些患者则出现严重的支气管扩张。因为突变许多导致 PCD 的基因已被证明会导致纤毛基本上不动，预计会虽然没有导致 MCC，但为什么存在如此广泛的临床表型尚不清楚。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很大程度上是遗传因素的结果异质性，不同基因的突变会导致不同水平的残留 MCC，因此，疾病的严重程度不同。此外，我们假设同一基因的不同突变也可能导致不同程度的纤毛损伤、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开发提供目标和机会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等基因和蛋白的表达和功能，新发现其突变可导致 PCD。使用患者来源的人鼻上皮 (HNE) 细胞和/或诱导多能干 (iPS) 细胞，我们将研究突变在蛋白质水平、纤毛功能（波形和节拍）的影响频率）和体外粘膜纤毛转运（MCT）。 2. 研究患者来源的 iPS 细胞的基因型/表型关系。我们将检查不同基因（CFAP57、PCDP1、RSPH1、 CCDC39 和 DNAI1）以及 iPS 中同一基因（SPAG1、CCDC114、DNAH5）的不同突变来自 PCD 患者的细胞。 3. 研究Ccdc39缺失小鼠模型的疾病发病机制。我们将比较 Ccdc39 诱导型缺失与 Dnaic1 诱导型缺失的效果。 4. 体内研究基因型、MCC及临床表型之间的关系。将在具有 RSPH1 突变的 PCD 受试者中测量 MCC，并与具有 RSPH1 突变的 PCD 受试者进行比较其他基因突变（例如 DNAI1、DNAH5）。此外，MCC 将在给药后进行测量。 β-激动剂以确定是否可以在 RSPH1 受试者中刺激 MCC。