Abstract
Cystic fibrosis is a common lethal genetic disease caused by functional absence of the cystic fibrosis transmembrane conductance regulator (CFTR). Although a candidate disease for in utero gene therapy, demonstration of potentially therapeutic levels of transgene expression in the fetal airways after minimally invasive gene delivery is a mandatory prerequisite before application of this approach in humans can be considered. We report here on the delivery of a β-galactosidase expressing adenovirus directly to the airways of fetal sheep in utero using ultrasound-guided percutaneous injection of the trachea in the fetal chest. Injection of adenoviral particles to the fetal airways was not associated with mortality and resulted in low-level expression in the peripheral airways. However, complexation of the virus with DEAE dextran, which confers a positive charge to the virus, and pretreatment of the airways with Na-caprate, which opens tight junctions, increased transgene expression, and a combination of these two enhancers resulted in widespread and efficient gene transfer of the fetal trachea and bronchial tree. Using a percutaneous ultrasound-guided injection technique, we have clearly demonstrated proof of principle for substantial transgene delivery to the fetal airways providing levels of gene expression that could be relevant for a therapeutic application of CFTR expressing vectors.
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Acknowledgements
This work is supported by the British Medical Research Council (G9901081). LG is supported by an F.D. Roosevelt grant from the March of Dimes Birth Defects Foundation and SW by a fellowship from the Katherine Dormandy Haemophilia Trust. We wish to express our gratitude to Ms Irene Hopton-Scott, Ms Melanie Hill, Dr Suzie Miller, Dr Richard Smith, Dr Niki Thiruchelvam, Dr Joy Archer, Mr David Manners, Mr Tony Jones, Ms Lynn Dorsett, Mr Alistair Wallis, Mr Paul Levy and Ms Suzy Buckley for their much appreciated organizational or experimental support during these experiments.
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Peebles, D., Gregory, L., David, A. et al. Widespread and efficient marker gene expression in the airway epithelia of fetal sheep after minimally invasive tracheal application of recombinant adenovirus in utero. Gene Ther 11, 70–78 (2004). https://doi.org/10.1038/sj.gt.3302130
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DOI: https://doi.org/10.1038/sj.gt.3302130
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