Efficient gene transfer to airway epithelium using recombinant Sendai virus

Abstract

Clinical studies of gene therapy for cystic fibrosis (CF) suggest that the key problem is the efficiency of gene transfer to the airway epithelium. The availability of relevant vector receptors, the transient contact time between vector and epithelium, and the barrier function of airway mucus contribute significantly to this problem. We have recently developed recombinant Sendai virus (SeV) as a new gene transfer agent. Here we show that SeV produces efficient transfection throughout the respiratory tract of both mice and ferrets in vivo, as well as in freshly obtained human nasal epithelial cells in vitro. Gene transfer efficiency was several log orders greater than with cationic liposomes or adenovirus. Even very brief contact time was sufficient to produce this effect, and levels of expression were not significantly reduced by airway mucus. Our investigations suggest that SeV may provide a useful new vector for airway gene transfer.

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Figure 1: Comparison of SeV and liposome-mediated gene transfer to the mouse lung in vivo.
Figure 2: SeV-mediated gene transfer to the ferret airways in vivo.
Figure 3: Gene transfer to freshly obtained human nasal epithelium ex vivo.
Figure 4: Effect of contact time on gene expression in the mouse nose in vivo.
Figure 5: Gene transfer to mucus-covered and mucus-depleted sheep tracheae ex vivo.

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Acknowledgements

We thank Graciela Sala-Newby and Andrew Baker (Bristol Heart Institute, University of Bristol, UK) for providing the Adeno-luc, Seng Cheng (Genzyme Corp., Framingham, MA) for providing GL-67/DOPE/DMPE-PEG5000, and George Lai for help with propagating SeV. This study was supported by the Cystic Fibrosis Research Trust, the Uehara Memorial Bio-Medical Research Foundation (Y.Y.), Daiwa Anglo-Japanese Foundation (Y.Y.), a Wellcome Trust Travelling Research Fellowship (Y.Y.). and by a Wellcome Trust Senior Clinical Fellowship (E.A.).

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Correspondence to Eric W.F.W. Alton.

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Yonemitsu, Y., Kitson, C., Ferrari, S. et al. Efficient gene transfer to airway epithelium using recombinant Sendai virus. Nat Biotechnol 18, 970–973 (2000). https://doi.org/10.1038/79463

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