Cftr gene transfer corrects defective glycoconjugate secretion in
human cystic fibrosis epithelial tracheal cells.
Mergey, Martine, Mustapha Lemnaouar, Danielle Veissiere, #michel
Perricaudet, Dieter C Gruenert, Jacques Picard, Jacqueline Capeau, M
-Christiane Brahimi-Horn, and Annick Paul.
Laboratoire de Biologie Cellulaire, I.N.S.E.R.M. U 402,
Facult[acute]e de M[acute]edecine Saint-Antoine 27, rue Chaligny,
75571 Paris Cedex 12, France, #CNRS UA1301 Institut Gustave Roussy,
Villejuif Cedex, France, Gene Therapy Core Center, Cardiovascular
Research Institute and Department of Laboratory Medicine, University
of California, San Francisco CA 94143, USA
APStracts 2:0152L, 1995.
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 pfu/cell, high levels of CFTR mRNA were detected by RT-PCR and de
novo synthesis of CFTR protein was demonstrated by immunoblotting.
Gene transfer to CF cells restored defective 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.
Received 30 March 1995; accepted in final form 12 July 1995.
APS Manuscript Number L100-5.
Article publication pending Am. J. Physiol. (Lung Cell. Mol.
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 15 September 1995.