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Lentiviral-mediated phenotypic correction of cystic fibrosis pigs
Ashley L. Cooney, … , Patrick L. Sinn, Paul B. McCray Jr.
Ashley L. Cooney, … , Patrick L. Sinn, Paul B. McCray Jr.
Published September 8, 2016
Citation Information: JCI Insight. 2016;1(14):e88730. https://doi.org/10.1172/jci.insight.88730.
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Research Article Pulmonology

Lentiviral-mediated phenotypic correction of cystic fibrosis pigs

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Abstract

Cystic Fibrosis (CF) is an autosomal recessive disease caused by mutations in CF transmembrane conductance regulator (CFTR), resulting in defective anion transport. Regardless of the disease-causing mutation, gene therapy is a strategy to restore anion transport to airway epithelia. Indeed, viral vector–delivered CFTR can complement the anion channel defect. In this proof-of-principle study, functional in vivo CFTR channel activity was restored in the airways of CF pigs using a feline immunodeficiency virus–based (FIV-based) lentiviral vector pseudotyped with the GP64 envelope. Three newborn CF pigs received aerosolized FIV-CFTR to the nose and lung. Two weeks after viral vector delivery, epithelial tissues were analyzed for functional correction. In freshly excised tracheal and bronchus tissues and cultured ethmoid sinus cells, we observed a significant increase in transepithelial cAMP-stimulated current, evidence of functional CFTR. In addition, we observed increases in tracheal airway surface liquid pH and bacterial killing in CFTR vector–treated animals. Together, these data provide the first evidence to our knowledge that lentiviral delivery of CFTR can partially correct the anion channel defect in a large-animal CF model and validate a translational strategy to treat or prevent CF lung disease.

Authors

Ashley L. Cooney, Mahmoud H. Abou Alaiwa, Viral S. Shah, Drake C. Bouzek, Mallory R. Stroik, Linda S. Powers, Nick D. Gansemer, David K. Meyerholz, Michael J. Welsh, David A. Stoltz, Patrick L. Sinn, Paul B. McCray Jr.

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Figure 1

Correction of chloride (Cl–) transport in cystic fibrosis (CF) pig primary epithelial with a lentiviral vector.

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Correction of chloride (Cl–) transport in cystic fibrosis (CF) pig prima...
Feline immunodeficiency virus–based viral vector expressing cystic fibrosis transmembrane conductance regulator (FIV-CFTR; MOI = 5) was delivered to the apical surface of well-differentiated primary cultures of airway epithelial cells from CF pigs. Transepithelial Cl– currents were measured in Ussing chambers. (A) An example of CFTR-dependent Cl– current is shown. The average change in Cl– current upon addition of forskolin/3-isobutyl-1-methylxanthine (F&I) (B) and GlyH-101 (C) in treated and naive cultures are shown. n = 8 epithelial sheets/treatment (collected from 3 donor pigs). *P < 0.01, Mann-Whitney nonparametric t test. (D) IHC using a CFTR antibody reveals apical localization of CFTR protein in ciliated cells (arrows). c, ciliated cells; nc, nonciliated cells. (E) IHC using a CFTR antibody on untreated CF primary airway epithelia. Scale bar: 250 μM (D and E).

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