Document Detail

CFTR gene transfer to human cystic fibrosis pancreatic duct cells using a Sendai virus vector.
MedLine Citation:
PMID:  17654517     Owner:  NLM     Status:  MEDLINE    
Cystic fibrosis (CF) is a fatal inherited disease caused by the absence or dysfunction of the CF transmembrane conductance regulator (CFTR) Cl- channel. About 70% of CF patients are exocrine pancreatic insufficient due to failure of the pancreatic ducts to secrete a HCO3- -rich fluid. Our aim in this study was to investigate the potential of a recombinant Sendai virus (SeV) vector to introduce normal CFTR into human CF pancreatic duct (CFPAC-1) cells, and to assess the effect of CFTR gene transfer on the key transporters involved in HCO3- transport. Using polarized cultures of homozygous F508del CFPAC-1 cells as a model for the human CF pancreatic ductal epithelium we showed that SeV was an efficient gene transfer agent when applied to the apical membrane. The presence of functional CFTR was confirmed using iodide efflux assay. CFTR expression had no effect on cell growth, monolayer integrity, and mRNA levels for key transporters in the duct cell (pNBC, AE2, NHE2, NHE3, DRA, and PAT-1), but did upregulate the activity of apical Cl-/HCO3- and Na+/H+ exchangers (NHEs). In CFTR-corrected cells, apical Cl-/HCO3- exchange activity was further enhanced by cAMP, a key feature exhibited by normal pancreatic duct cells. The cAMP stimulated Cl-/HCO3- exchange was inhibited by dihydro-4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (H2-DIDS), but not by a specific CFTR inhibitor, CFTR(inh)-172. Our data show that SeV vector is a potential CFTR gene transfer agent for human pancreatic duct cells and that expression of CFTR in CF cells is associated with a restoration of Cl- and HCO3- transport at the apical membrane.
Zoltán Rakonczay; Péter Hegyi; Mamoru Hasegawa; Makoto Inoue; Jun You; Akihiro Iida; Imre Ignáth; Eric W F W Alton; Uta Griesenbach; Gabriella Ovári; János Vág; Ana C Da Paula; Russell M Crawford; Gábor Varga; Margarida D Amaral; Anil Mehta; János Lonovics; Barry E Argent; Michael A Gray
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of cellular physiology     Volume:  214     ISSN:  1097-4652     ISO Abbreviation:  J. Cell. Physiol.     Publication Date:  2008 Feb 
Date Detail:
Created Date:  2007-11-27     Completed Date:  2007-12-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0050222     Medline TA:  J Cell Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  442-55     Citation Subset:  IM    
Copyright Information:
(c) 2007 Wiley-Liss, Inc.
Institute for Cell and Molecular Biosciences, University of Newcastle, Newcastle upon Tyne, United Kingdom.
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MeSH Terms
Cell Line
Cystic Fibrosis / genetics,  physiopathology
Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
Gene Transfer Techniques*
Genetic Vectors*
Hydrogen-Ion Concentration
Pancreatic Ducts / physiology*
RNA, Messenger / metabolism
Sendai virus / physiology*
beta-Galactosidase / metabolism
Reg. No./Substance:
0/CFTR protein, human; 0/RNA, Messenger; 126880-72-6/Cystic Fibrosis Transmembrane Conductance Regulator; EC

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