CFTR gene transfer to human cystic fibrosis pancreatic duct cells using a Sendai virus vector

Z. Rakonczay, P. Hegyi, Mamoru Hasegawa, Makoto Inoue, Jun You, Akihiro Iida, Imre Ignáth, Eric W F W Alton, Uta Griesenbach, Gabriella Óvári, János Vag, Ana C. Da Paula, Russell M. Crawford, G. Varga, Margarida D. Amaral, Anil Mehta, J. Lonovics, Barry E. Argent, Michael A. Gray

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

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-I) 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-I), but did upregulate the activity of apical Cl -/HCO3- and Na+/H+ exchangers (NHEs). In CFTR-corrected cells, apical Cl-/HCO 3- 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′-disulfonicacid (H 2-DIDS), but not by a specific CFTR inhibitor, CFTR inh-172. Our data show that SeV vector s 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 HCO 3- transport at the apical membrane.

Original languageEnglish
Pages (from-to)442-455
Number of pages14
JournalJournal of Cellular Physiology
Volume214
Issue number2
DOIs
Publication statusPublished - Feb 2008

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Gene transfer
Sendai virus
Pancreatic Ducts
Regulator Genes
Viruses
Cystic Fibrosis
Ducts
Cystic Fibrosis Transmembrane Conductance Regulator
Membranes
Sodium-Hydrogen Antiporter
Cell growth
Iodides
Restoration
Monolayers
Assays
Messenger RNA
Fluids
Membrane Potentials
Up-Regulation
Epithelium

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

CFTR gene transfer to human cystic fibrosis pancreatic duct cells using a Sendai virus vector. / Rakonczay, Z.; Hegyi, P.; Hasegawa, Mamoru; Inoue, Makoto; You, Jun; Iida, Akihiro; Ignáth, Imre; Alton, Eric W F W; Griesenbach, Uta; Óvári, Gabriella; Vag, János; Da Paula, Ana C.; Crawford, Russell M.; Varga, G.; Amaral, Margarida D.; Mehta, Anil; Lonovics, J.; Argent, Barry E.; Gray, Michael A.

In: Journal of Cellular Physiology, Vol. 214, No. 2, 02.2008, p. 442-455.

Research output: Contribution to journalArticle

Rakonczay, Z, Hegyi, P, Hasegawa, M, Inoue, M, You, J, Iida, A, Ignáth, I, Alton, EWFW, Griesenbach, U, Óvári, G, Vag, J, Da Paula, AC, Crawford, RM, Varga, G, Amaral, MD, Mehta, A, Lonovics, J, Argent, BE & Gray, MA 2008, 'CFTR gene transfer to human cystic fibrosis pancreatic duct cells using a Sendai virus vector', Journal of Cellular Physiology, vol. 214, no. 2, pp. 442-455. https://doi.org/10.1002/jcp.21220
Rakonczay, Z. ; Hegyi, P. ; Hasegawa, Mamoru ; Inoue, Makoto ; You, Jun ; Iida, Akihiro ; Ignáth, Imre ; Alton, Eric W F W ; Griesenbach, Uta ; Óvári, Gabriella ; Vag, János ; Da Paula, Ana C. ; Crawford, Russell M. ; Varga, G. ; Amaral, Margarida D. ; Mehta, Anil ; Lonovics, J. ; Argent, Barry E. ; Gray, Michael A. / CFTR gene transfer to human cystic fibrosis pancreatic duct cells using a Sendai virus vector. In: Journal of Cellular Physiology. 2008 ; Vol. 214, No. 2. pp. 442-455.
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