Correction of the apical membrane chloride permeability defect in polarized cystic fibrosis airway epithelia following retroviral-mediated gene transfer.
We are studying the introduction and expression of the normal cystic fibrosis transmembrane conductance regulator (CFTR) cDNA into cultured human airway epithelial cells as a model for gene therapy of cystic fibrosis. In this paper, we show that the chloride transport defect at the apical membrane is corrected in vitro in differentiated ion-transporting CF airway epithelial cells that exhibit polarized properties similar to those found in vivo. Using a retroviral vector containing a copy of the normal CFTR cDNA, we infected cultures of proliferating, cystic fibrosis CFT1 cells and found that correction was maintained following differentiation into a polarized epithelial sheet. At least partial correction of the Cl- transport defect was preserved in CFT1 cells for periods of up to 6 months without selection for maintenance of the retroviral provirus. These results suggest that it may be feasible to target proliferating cells in the lung using retroviral vectors for treatment of CF lung disease.
Olsen
J C
JC
UNC Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill 27599.
Johnson
L G
LG
Stutts
M J
MJ
Sarkadi
B
B
Yankaskas
J R
JR
Swanstrom
R
R
Boucher
R C
RC
D016475:3T3 Cells; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D002463:Cell Membrane Permeability; D018118:Chloride Channels; D002712:Chlorides / Q000378:metabolism*; D003001:Cloning, Molecular; D003550:Cystic Fibrosis / Q000235:genetics* / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004277:DNA, Single-Stranded; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism; D008565:Membrane Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010641:Phenotype; D012190:Retroviridae / Q000235:genetics; D014132:Trachea / Q000166:cytology / Q000378:metabolism; D014161:Transduction, Genetic; D014162:Transfection
C505032:CFTR protein, human; D018118:Chloride Channels; D002712:Chlorides; D004277:DNA, Single-Stranded; D007473:Ion Channels; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000584:Amiloride