Cystic Fibrosis (CF) has been at the forefront of gene transfer research for the last decade. This disease, resulting from a single gene defect, affects the epithelia of multiple organs of the body including the lung and gastrointestinal tract. The lack of effective long-term treatment for the pulmonary manifestations of CF, the accessibility of the lung via the airway lumen, and the fact that viruses known to infect the lung were being developed into non-replicating gene transfer vectors led investigators to believe that administration of gene transfer vectors to the lung could potentially result in an effective treatment of this disease. The assessment of the efficacy of gene transfer for CF requires a routine and reliable assay to verify expression of functional CFTR. This Correction Core will provide multiple techniques to detect the efficacy of CFTR gene transfer at correcting the ion, fluid and mucus transport defects in both the airway and gut. Cultured airway epithelia from the CF human and mouse will be used extensively in Ussing chamber studies to demonstrate correction of the CFTR mediated Cl- secretion and Na+ hyperabsorption. These preparations will also be used to determine if the altered rate of fluid transport has been corrected by CFTR gene transfer using confocal microscopy and measurement of blue dextran absorption. The pH and mucus biophysical properties will be measured on CF cultured airway epithelia to ascertain CFTR correction. Both human and murine nasal PD assays will be provided to assess CFTR gene transfer in vivo. Ion transport across CF human rectal biopsy studied in Ussing chambers will also be available as will freshly excised epithelial tissue from neonatal murine airways and gut to measure the effectiveness of in vivo vector dosing in neonatal CF mice. Finally, in anticipation of a new animal model for CF (a CF pig) we have begun to characterize the bioelectric properties of the porcine epithelium so that we will be poised for those possible studies.

Public Health Relevance

CF is the most common fatal genetic disorder of the Caucasian population. Gene transfer approaches provide one of the best possibilities to cure this disease. The novel techniques described in this proposal will provide gene transfer investigators routine access to high-quality, high-sensitivity, well-controlled, robust measures of CFTR correction.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Center Core Grants (P30)
Project #
3P30DK065988-10S1
Application #
8851231
Study Section
Special Emphasis Panel (ZDK1-GRB-1)
Project Start
Project End
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
10
Fiscal Year
2014
Total Cost
$157,320
Indirect Cost
$53,820
Name
University of North Carolina Chapel Hill
Department
Type
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Ostrowski, L E; Yin, W; Patel, M et al. (2014) Restoring ciliary function to differentiated primary ciliary dyskinesia cells with a lentiviral vector. Gene Ther 21:253-61
Morales Johansson, Helena; Newman, Donna R; Sannes, Philip L (2014) Whole-genome analysis of temporal gene expression during early transdifferentiation of human lung alveolar epithelial type 2 cells in vitro. PLoS One 9:e93413
Henderson, Ashley G; Ehre, Camille; Button, Brian et al. (2014) Cystic fibrosis airway secretions exhibit mucin hyperconcentration and increased osmotic pressure. J Clin Invest 124:3047-60
Saini, Yogesh; Dang, Hong; Livraghi-Butrico, Alessandra et al. (2014) Gene expression in whole lung and pulmonary macrophages reflects the dynamic pathology associated with airway surface dehydration. BMC Genomics 15:726
Esther Jr, Charles R; Boucher, Richard C; Johnson, M Ross et al. (2014) Airway drug pharmacokinetics via analysis of exhaled breath condensate. Pulm Pharmacol Ther 27:76-82
Cholon, Deborah M; Quinney, Nancy L; Fulcher, M Leslie et al. (2014) Potentiator ivacaftor abrogates pharmacological correction of ?F508 CFTR in cystic fibrosis. Sci Transl Med 6:246ra96
Tadokoro, Tomomi; Wang, Yang; Barak, Larry S et al. (2014) IL-6/STAT3 promotes regeneration of airway ciliated cells from basal stem cells. Proc Natl Acad Sci U S A 111:E3641-9
Bove, Peter F; Dang, Hong; Cheluvaraju, Chaitra et al. (2014) Breaking the in vitro alveolar type II cell proliferation barrier while retaining ion transport properties. Am J Respir Cell Mol Biol 50:767-76
Guo, Xueliang; Zheng, Shuo; Dang, Hong et al. (2014) Genome reference and sequence variation in the large repetitive central exon of human MUC5AC. Am J Respir Cell Mol Biol 50:223-32
Schwab, Ute; Abdullah, Lubna H; Perlmutt, Olivia S et al. (2014) Localization of Burkholderia cepacia complex bacteria in cystic fibrosis lungs and interactions with Pseudomonas aeruginosa in hypoxic mucus. Infect Immun 82:4729-45

Showing the most recent 10 out of 57 publications