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Lentiviral transduction of the murine lung provides efficient pseudotype and developmental stage-dependent cell-specific transgene expression

Gene Therapy volume 15, pages 1167–1175 (2008)Cite this article

Abstract

Gene transfer for cystic fibrosis (CF) airway disease has been hampered by the lung's innate refractivity to pathogen infection. We hypothesized that early intervention with an integrating gene transfer vector capable of transducing the lung via the lumen may be a successful therapeutic approach. An HIV-based lentiviral vector pseudotyped with the baculovirus gp64 envelope was applied to the fetal, neonatal or adult airways. Fetal intra-amniotic administration resulted in transduction of approximately 14% of airway epithelial cells, including both ciliated and non-ciliated epithelia of the upper, mid and lower airways; there was negligible alveolar or nasal transduction. Following neonatal intra-nasal administration we observed significant transduction of the airway epithelium (approximately 11%), although mainly in the distal lung, and substantial alveolar transduction. This expression was still detectable at 1 year after application. In the adult, the majority of transduction was restricted to the alveoli. In contrast, vesicular stomatitis virus glycoprotein pseudotyped virus transduced only alveoli after adult and neonatal application and no transduction was observed after fetal administration. Repeat administration did not increase transduction levels of the conducting airway epithelia. These data demonstrate that application at early developmental stages in conjunction with an appropriately pseudotyped virus provides efficient, high-level transgene expression in the murine lung. This may provide a modality for treatment for lung disease in CF.

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Acknowledgements

We thank Dr David Parsons and Associate Professor Don Anson, Women and Children's Children's Hospital, Adelaide, Australia for the kind gift of the gp64 plasmid. Plasmid was generated by Plasmid Factory (Bielefeld, Germany). TRM was funded by combined MRC/BBSRC/EPSRC as part of the UKCTE, University of Manchester. SNW is recipient of the Philip Gray Memorial Fellowship, Katharine Dormandy Trust. SMKB is a senior research fellow of the Clive Knight Laboratories and is part-funded by an FP6 grant from the European Union (LSHB-CT-2004-00213). SJH is supported by a European Commission Contract (CONSERT 005242) and AJT is a Wellcome Trust Senior Clinical Fellow.

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Author notes

  1. S M K Buckley and S J Howe: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Haematology, Haemophilia Centre and Haemostasis Unit, Royal Free and University College Medical School, London, UK

    S M K Buckley & S N Waddington

  2. Molecular Immunology Unit, Institute of Child Health, University College London, London, UK

    S J Howe, N J Ward & A J Thrasher

  3. Department of Life Sciences, North West Embryonic Stem Cell Centre, Core Technology Facility, University of Manchester, Manchester, UK

    V Sheard & T R McKay

  4. Gene Therapy Research Group, Molecular Medicine, National Heart and Lung Institute, Imperial College, London, UK

    C Coutelle

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Correspondence to T R McKay.

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Buckley, S., Howe, S., Sheard, V. et al. Lentiviral transduction of the murine lung provides efficient pseudotype and developmental stage-dependent cell-specific transgene expression. Gene Ther 15, 1167–1175 (2008). https://doi.org/10.1038/gt.2008.74

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