Increasing Expression of the Normal Human CFTR cDNA in Cystic Fibrosis Epithelial Cells Results in a Progressive Increase in the Level of CFTR Protein Expression, but a Limit on the Level of cAMP-Stimulated Chloride Secretion

Melissa A. Rosenfeld, Stephen J. Rosenfeld, Claire Danel, Tyrone C. Banks, Ronald Crystal

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Cystic fibrosis (CF) results from mutations of the CF transmembrane conductance regulator (CFTR) gene and the consequent defective regulation of cAMP-stimulated Cl permeability across epithelial cell apical membranes. Given that in vitro transfer of normal CFTR cDNA corrects this defect and that recombinant adenovirus (Ad) vectors can transfer the normal human CFTR cDNA in vivo, Ad vectors have significant potential in the development of effective strategies for CF gene therapy. One concern is whether CFTR overexpression achievable with Ad vectors may have untoward effects on cAMP-stimulated Cl efflux. To address this, the CF pancreatic epithelial cell line CFPAC-1 was infected with increasing doses of AdCFTR, a recombinant Ad containing the normal CFTR cDNA, and analyzed for CFTR mRNA and protein levels and CFTR function. As the AdCFTR dose increased [multiplicity of infection (moi) 0–1,000], CFTR mRNA and protein levels increased. However, while CFTR function measured by cAMP-stimulated 36Cl efflux was observed with low doses of the vector (moi 20), there was no further increase in CFTR function with increasing doses of AdCFTR (moi from 20 to 1,000). These data suggest that after AdCFTR-mediated gene transfer, epithelial cells limit the level of cAMP-stimulated Cl secretion despite increasing levels of CFTR protein.

Original languageEnglish
Pages (from-to)1121-1129
Number of pages9
JournalHuman Gene Therapy
Volume5
Issue number9
DOIs
Publication statusPublished - 1 Sep 1994
Externally publishedYes

Fingerprint

Adenoviridae
Cystic Fibrosis
Chlorides
Complementary DNA
Epithelial Cells
Proteins
Infection
Cystic Fibrosis Transmembrane Conductance Regulator
Messenger RNA
Regulator Genes
Genetic Therapy
Permeability
Cell Membrane
Cell Line
Mutation
Genes

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Increasing Expression of the Normal Human CFTR cDNA in Cystic Fibrosis Epithelial Cells Results in a Progressive Increase in the Level of CFTR Protein Expression, but a Limit on the Level of cAMP-Stimulated Chloride Secretion. / Rosenfeld, Melissa A.; Rosenfeld, Stephen J.; Danel, Claire; Banks, Tyrone C.; Crystal, Ronald.

In: Human Gene Therapy, Vol. 5, No. 9, 01.09.1994, p. 1121-1129.

Research output: Contribution to journalArticle

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