Molecular characterization of outer dynein arm defects in Primary Ciliary Dyskinesia (PCD)

点击翻译按钮获取中文摘要

Primary ciliary dyskinesia (PCD) is a genetically heterogeneous disorder characterized bychronic airway disease, randomization of left-right body asymmetry and male infertility due todefects of motile cilia/flagella function. The disease often results in permanent lung damageand can progress to respiratory failure. Currently diagnosis mainly relies on demonstration ofultrastructural defects by electron microscopy, which often reveals outer dynein arm (ODA)deficiency. We demonstrated that mutations in DNAH5, encoding an ODA heavy chain, areresponsible for half of all ODA defect cases. With the use of high-resolution immunofluorescenceimaging (IF) we identified in patients with ODA defects two distinct types of DNAH5mislocalization in nasal airway cells, introducing a novel diagnostic tool in PCD. Here, wewant to genetically characterize a large international cohort of PCD patients for presence ofmutations in both of the known PCD genes, DNAH5 and DNAI1. To analyze composition andgeneration of human ODAs we will generate antibodies directed against other yet uncharacterizedODA heavy chains and apply protein techniques. To improve understanding ofthe disease causing mecanisms and diagnosis in PCD we will correlate genetic results withclinical, ultrastructural and IF findings. To evaluate novel therapeutic options we will performin vitro ciliogenesis and apply pharmaco-gene therapy in mutant respiratory cells.