Gene Transfer to Freshly Isolated Human Respiratory Epithelial CellsIn VitroUsing a Replication-Deficient Adenovirus Containing the Human Cystic Fibrosis Transmembrane Conductance Regulator cDNA
| Autor: | Chin-Shyan Chu, Claire Danel, Tyrone C. Banks, Ronald G. Crystal, Prem Seth, Kunihiko Yoshimura, Melissa A. Rosenfeld, Koichi Yoneyama |
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| Rok vydání: | 1994 |
| Předmět: |
DNA Replication
DNA Complementary Cystic Fibrosis Recombinant Fusion Proteins Genetic enhancement Genetic Vectors Cystic Fibrosis Transmembrane Conductance Regulator Bronchi Biology Cystic fibrosis Adenoviridae Viral vector Genetics medicine Humans Molecular Biology Cells Cultured Cell Line Transformed Gene Transfer Techniques Membrane Proteins Epithelial Cells respiratory system Apical membrane medicine.disease Molecular biology Epithelium Cystic fibrosis transmembrane conductance regulator respiratory tract diseases Nasal Mucosa medicine.anatomical_structure Cell culture DNA Viral biology.protein Molecular Medicine Respiratory epithelium |
| Zdroj: | Human Gene Therapy. 5:331-342 |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/hum.1994.5.3-331 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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