Project 3

University Of Iowa, Iowa City IA

Investigators

Abstract

Project Summary/Abstract: Primary ciliary dyskinesia (PCD) comprises a group of rare genetic diseases that cause reduced or absent function of motile cilia within the airways, reproductive tracts, choroid plexus, and ependymal cells. Respiratory manifestations of PCD can include: neonatal respiratory distress, chronic rhinosinusitis and otitis media, impaired mucociliary clearance, persistent productive cough, airway obstruction, bacterial colonization, inflammation, and bronchiectasis. Most PCD cases follow a genetically heterogeneous autosomal recessive inheritance pattern with biallelic pathologic mutations in the same gene. Over 50 different PCD causing genes are now identified. Mutations in DNAI1, which encodes the intermediate chain of the axonemal dynein, are among the most common, accounting for ~5-10% of PCD cases. Because of its recessive inheritance and knowledge that carriers of DNAI1 mutations have no known disease manifestations, it is a candidate for correction by gene therapy. Current clinical PCD management consists of symptomatic treatments. There is a pressing need for new preventive and corrective treatments. Our studies focus on the use of a novel porcine DNAI1 null pig model generated by CRISPR genome editing termed PCD pig. The overall goal of these proposed studies is to use gene addition technology to complement DNAI1 mutations in somatic cells of the respiratory tract and correct the clinical phenotype. Here we propose to use lentiviral and adeno-associated viral vectors to: 1) show that gene addition will correct motile cilia defects in DNAI in PCD pig airway epithelial cells in vitro, 2) show that gene addition will correct mucociliary transport (MCT) in DNAI in PCD pig airway epithelial cells in vitro, and 3) correct the motile cilia and MCT defects associated with sinopulmonary disease in the PCD pig model in vivo. Projects 1, 2, and 3 (along with the valuable cores) work closely together to directly address the Program's goal of understanding the natural history, disease pathogenesis, and developing new therapeutic strategies. The Program effectively focuses a team of talented laboratories to address a shared ambition in a highly collaborative environment. Our track record supports our commitment to improving the lives of people with inherited lung diseases and increasing our understanding of lung biology. Our goal is to provide a life-long gene addition strategy that could be adapted to for many PCD causing mutations. This proposed research is highly innovative. The reagents, methods, and data generated by these experiments will significantly advance the gene therapy field.