Abstract
An important issue for in vivo gene therapy for cystic fibrosis (CF) is the percentage of cells within the CF airway that will require correction. In this study, we mixed populations of a CF airway cell line expressing either the normal cystic fibrosis transmembrane conductance regulator (CFTR) cDNA (corrected cells) or a reporter gene in defined percentages. As few as 6–10% corrected cells within an epithelial sheet generated Cl− transport properties similar to sheets comprised of 100% corrected cells. Cell–cell coupling may serve as the mechanism for amplification of the functional effects of corrected cells. These data suggest that in vivo correction of all CF airway cells may not be mandatory.
Similar content being viewed by others
References
Boat, T.F., Welsh, M.J. & Beaudet, A.L. Cystic fibrosis. in The Metabolic Basis of Inherited Disease, (eds. Scriver C.R., Beaudet, A.L., Sly, W.S. & D. valle) 2649–2680 (McGraw Hill, New York, 1989).
Quinton, P. Cystic fibrosis: a disease in electrolyte transport. FASEB J. 4, 2709–2717 (1990).
Quinton, P.M. Chloride impermeability in cystic fibrosis. Nature 301, 421–422 (1983).
Quinton, P.M. & Bijman, J. Higher bioelectric potentials due to decreased chloride absorption in the sweat glands of patients with cystic fibrosis. New Engl. J. Med. 308, 1185–1188 (1983).
Knowles, M.R. et al. Abnormal ion permeation through cystic fibrosis respiratory epithelium. Science 221, 1067–1070 (1983).
Knowles, M., Gatzy, J. & Boucher, R. Relative ion permeability of normal & cystic fibrosis nasal epithelium. J. clin. Invest. 71, 1410–1417 (1983).
Drumm, M.L. et al. Correction of the cystic fibrosis defect in vitro by retrovirus-mediated gene transfer. Cell 62, 1227–1233 (1990).
Rich, D.P. et al. Expression of the cystic fibrosis transmembrane conductance regulator corrects defective chloride channel regulation in cystic fibrosis airway epithelial cells. Nature 347, 358–363 (1990).
Olsen, J.C. et al. Correction of the apical membrane chloride permeability defect in polarized cystic fibrosis airway epithelia following retroviral-mediated gene transfer. Hum. Gene Therapy 3, 253–266 (1992).
Rosenfeld, M.A. et al. In vivo transfer of the human cystic fibrosis transmembrane conductance regulator to the airway epithelium. Cell 68, 143–155 (1992).
Willumsen, N.J., Davis, C.W. & Boucher, R.C. Intracellular Cl− activity & cellular pathways in cultured human airway epithelium. Am. J. Physiol. 256, C1033–1044 (1989).
Willumsen, N.J. & Boucher, R.C. Shunt resistance & ion permeabilities in normal & cystic fibrosis airway epithelium. Am. J. Physiol. 256, C1054–1063 (1989).
Willumsen, N.J., Davis, C.W. & Boucher, R.C. Cellular Cl− transport in cultured cystic fibrosis airway epithelium. Am. J. Physiol. 256, C1045–C1053 (1989).
Boucher, R.C., Stutts, M.J., Knowles, M.R., Cantley, L. & Gatzy, J.T. Na+ transport in cystic fibrosis respiratory epithelia: abnormal basal rate & response to adenylate cyclase activation. J. clin. Invest. 78, 1245–1252 (1986).
Yankaskas, J.R., Knowles, M.R., Gatzy, J.T. & Boucher, R.C. Persistence of abnormal chloride ion permeability in cystic fibrosis nasal epithelial cells in heterologous culture. Lancet 1, 954–956 (1985).
Flagg-Newton, J., Simpson, I. & Loewenstein, W.R. Permeability of the cell-to-cell membrane channels in mammalian cell junction. Science 205, 404–407 (1979).
Sarkadi, B. et al. Biochemical characterization of the cystic fibrosis transmembrane conductance regulator in normal & cystic fibrosis epithelial cells. J. biol. Chem. 267, 2087–2095 (1992).
Cotton, C.U., Stutts, M.J., Knowles, M.R., Gatzy, J.T. & Boucher, R.C. Abnormal apical cell membrane in cystic fibrosis respiratory epithelium: an in vitro electrophysiologic analysis. J. clin. Invest. 79, 80–85 (1987).
Cheng, S.H. et al. Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis. Cell 63, 827–834 (1990).
Sanderson, M.J., Chow, I. & Dirksen, E.R. Intercellular communication between ciliated cells in culture. Am. J. Physiol. 254, C63–C74 (1988).
Inoue, S. & Hogg, J.C. Intercellular junctions of the tracheal epithelium in guinea pigs. Lab. Invest. 31, 68–74 (1974).
Carson, J.L., Willumsen, N.J., Gambling, T.M., Hu, S.-C.S. & Collier, A.M. Dynamics of intracellular communications & differentiation in a rapidly developing mammalian airway epithelium. Am. J. Resp. Cell molec. Biol. 1, 385–390 (1989).
Schneeberger, E.E., Walters, D.V. & Olver, R.E. Development of intercellular junctions in the pulmonary epithelium of the foetal lamb. J. Cell Science 32, 307–324 (1978).
Miller, A.D. & Rosman, G.J. Improved retroviral vectors for gene transfer and expression. Biotechniques 7, 980–990 (1990).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Johnson, L., Olsen, J., Sarkadi, B. et al. Efficiency of gene transfer for restoration of normal airway epithelial function in cystic fibrosis. Nat Genet 2, 21–25 (1992). https://doi.org/10.1038/ng0992-21
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/ng0992-21
- Springer Nature America, Inc.
This article is cited by
-
Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
Nature Communications (2019)
-
Reversal of Surfactant Protein B Deficiency in Patient Specific Human Induced Pluripotent Stem Cell Derived Lung Organoids by Gene Therapy
Scientific Reports (2019)
-
Physiological study of pulmonary involvement in adults with cystic fibrosis through simulated modeling of different clinical scenarios
Medical & Biological Engineering & Computing (2019)
-
Impact of gene editing on the study of cystic fibrosis
Human Genetics (2016)
-
Cystic fibrosis genetics: from molecular understanding to clinical application
Nature Reviews Genetics (2015)