CFTR gene transfer corrects defective glycoconjugate secretion in human CF epithelial tracheal cells.
We demonstrate that in immortalized normal human tracheal epithelial cells (NT-1 and 56FHTE8o-) 14C-labeled glycoconjugate secretion may be regulated independently by agonists of the protein kinase A (PKA) and protein kinase C (PKC) signaling pathways. In contrast, in immortalized cystic fibrosis (CF) human tracheal epithelial cells (CFT-1 and CFT-2), regulation is defective for agonists specific for the PKA but not for the PKC pathway. To characterize the involvement of the cystic fibrosis transmembrane conductance regulator (CFTR) in regulated glycoconjugate secretion, we examined the effect of adenovirus-mediated gene transfer of CFTR to CF and control cells. Forty-eight hours after infection, at a multiplicity of infection of 50 plaque-forming units per cell, high levels of CFTR mRNA were detected by reverse transcription-polymerase chain reaction, and de novo synthesis of CFTR protein was demonstrated by immunoblotting. Gene transfer to CF cells restored defective adenosine 3',5'-cyclic monophosphate (cAMP)-dependent secretion not only of chloride but also of glycoconjugates. Taken together, these results argue for a role for CFTR in cAMP-mediated glycoconjugate secretion.
Mergey
M
M
Institut National de la Santé et de la Recherche Médicale U 402, Faculté de Médecine Saint-Antoine, Paris, France.
Lemnaouar
M
M
Veissiere
D
D
Perricaudet
M
M
Gruenert
D C
DC
Picard
J
J
Capeau
J
J
Brahimi-Horn
M C
MC
Paul
A
A
D000256:Adenoviridae / Q000235:genetics; D001483:Base Sequence; D002712:Chlorides / Q000502:physiology; D000242:Cyclic AMP / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004553:Electric Conductivity; D004848:Epithelium / Q000378:metabolism / Q000473:pathology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006001:Glycoconjugates / Q000378:metabolism*; D006801:Humans; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D011493:Protein Kinase C / Q000378:metabolism; D014132:Trachea / Q000378:metabolism* / Q000473:pathology
C505032:CFTR protein, human; D002712:Chlorides; D018076:DNA, Complementary; D006001:Glycoconjugates; D015335:Molecular Probes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C