Abstract
The major cause of mortality in patients with cystic fibrosis (CF) is lung disease. Expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene product in the airways is a potential treatment. Clinical studies in which the CFTR cDNA was delivered to the respiratory epithelia of CF patients have resulted in modest, transient gene expression. It seems likely that repeated administration of the gene transfer vector will be required for long-term gene expression. We have undertaken a double-blinded study in which multiple doses of a DNA/liposome formulation were delivered to the nasal epithelium of CF patients. Ten subjects received plasmid DNA expressing the CFTR cDNA complexed with DC-Chol/DOPE cationic liposomes, whilst two subjects received placebo. Each subject received three doses, administered 4 weeks apart. There was no evidence of inflammation, toxicity or an immune response towards the DNA/liposomes or the expressed CFTR. Nasal epithelial cells were collected 4 days after each dose for a series of efficacy assays including quantitation of vector-specific DNA and mRNA, immunohistochemistry of CFTR protein, bacterial adherence, and detection of halide efflux ex vivo. Airway ion transport was also assessed in vivo by repeated nasal potential difference (PD) measurements. On average, six of the treated subjects were positive for CFTR gene transfer after each dose. All subjects positive for CFTR function were also positive for plasmid DNA, plasmid-derived mRNA and CFTR protein. The efficacy measures suggest that unlike high doses of recombinant adenoviral vectors, DNA/liposomes can be successfully re-administered without apparent loss of efficacy.
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Acknowledgements
We thank all the volunteers who participated in the study and the staff at the Wythenshawe Hospital, Manchester, particularly Mary Dodd who helped with patient recruitment. We are grateful to Jim Littlewood for assisting with clinical scores and to Jane Davies, David Ferguson and Helen Turley for help with protocols. We also thank Andrew Wyre of Improvision for technical assistance with fluorescence microscopy, John Marshall for the gift of anti-CFTR antibodies and the Imperial Cancer Research Fund's Biotherapeutic and Hybridoma Development Unit for manufacture of plasmid DNA. This work was supported by grants from the Medical Research Council, the Cystic Fibrosis Research Trust, the Imperial Cancer Research Fund and the Universities of Leeds and Oxford.
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Hyde, S., Southern, K., Gileadi, U. et al. Repeat administration of DNA/liposomes to the nasal epithelium of patients with cystic fibrosis. Gene Ther 7, 1156–1165 (2000). https://doi.org/10.1038/sj.gt.3301212
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DOI: https://doi.org/10.1038/sj.gt.3301212
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