Gene transfer to freshly isolated human respiratory epithelial cells in vitro using a replication-deficient adenovirus containing the human cystic fibrosis transmembrane conductance regulator cDNA

Melissa A. Rosenfeld, Chin Shyan Chu, Prem Seth, Claire Danel, Tyrone Banks, Koichi Yoneyama, Kunihiko Yoshimura, Ronald Crystal

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Cystic fibrosis (CF) results from mutations of the CF transmembrane conductance regulator (CFTR) gene and subsequent defective regulation of cAMP-stimulated chloride (Cl-) permeability across the apical membrane of epithelial cells. In vitro transfer of normal CFTR cDNA corrects this defect, and studies in experimental animals have shown successful gene transfer to airway epithelium in vivo using a recombinant adenoviral vector containing the human CFTR cDNA (AdCFTR), supporting the feasibility of in vivo AdCFTR-mediated gene therapy for the respiratory manifestations of CF. One step in applying this therapy to CF patients is to evaluate the safety and efficacy of AdCFTR-mediated gene transfer in the actual target for human gene therapy, human airway epithelium. The present study demonstrates that AdCFTR restores cAMP-stimulated Cl- permeability in human CF bronchial epithelial cells. In addition, the study utilizes freshly isolated human airway epithelial cells from the nose and/or bronchi of normal individuals and/or individuals with CF to demonstrate that after in vitro AdCFTR-mediated gene transfer: (i) AdCFTR DNA does not replicate as a function of dose and time; (ii) CF epithelial cells express AdCFTR-mediated normal human CFTR mRNA; and (iii) CF epithelial cells, including terminally differentiated ciliated cells (the most common airway epithelial cell type), express the normal human CFTR protein. Together, these data support the use of AdCFTR in human gene therapy trials and suggest that biologic efficacy should be achievable in vivo.

Original languageEnglish
Pages (from-to)331-342
Number of pages12
JournalHuman Gene Therapy
Volume5
Issue number3
Publication statusPublished - 1 Mar 1994
Externally publishedYes

Fingerprint

Human Adenoviruses
Cystic Fibrosis Transmembrane Conductance Regulator
Cystic Fibrosis
Complementary DNA
Epithelial Cells
Genes
Genetic Therapy
Permeability
Epithelium
In Vitro Techniques
Bronchi
Regulator Genes
Nose
Chlorides
Safety
Messenger RNA
Mutation
Membranes
DNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Gene transfer to freshly isolated human respiratory epithelial cells in vitro using a replication-deficient adenovirus containing the human cystic fibrosis transmembrane conductance regulator cDNA. / Rosenfeld, Melissa A.; Chu, Chin Shyan; Seth, Prem; Danel, Claire; Banks, Tyrone; Yoneyama, Koichi; Yoshimura, Kunihiko; Crystal, Ronald.

In: Human Gene Therapy, Vol. 5, No. 3, 01.03.1994, p. 331-342.

Research output: Contribution to journalArticle

Rosenfeld, Melissa A. ; Chu, Chin Shyan ; Seth, Prem ; Danel, Claire ; Banks, Tyrone ; Yoneyama, Koichi ; Yoshimura, Kunihiko ; Crystal, Ronald. / Gene transfer to freshly isolated human respiratory epithelial cells in vitro using a replication-deficient adenovirus containing the human cystic fibrosis transmembrane conductance regulator cDNA. In: Human Gene Therapy. 1994 ; Vol. 5, No. 3. pp. 331-342.
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