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Evaluation of prenatal intra-amniotic LAMB3 gene delivery in a mouse model of Herlitz disease

Gene Therapy volume 13, pages 1665–1676 (2006)Cite this article

Abstract

Prenatal gene therapy has been considered for Herlitz junctional epidermolysis bullosa (H-JEB), a lethal genodermatosis caused by the absence of any of the three subunits of laminin-5, resulting from birth in widespread blistering and erosions of skin and mucosae. To investigate this strategy in an animal model, adenovirus type 5- and adeno-associated virus (AAV) type 2-derived vectors carrying a β-galactosidase reporter gene or LAMB3 cDNA encoding the β3 chain of laminin-5 were generated, tested for stability in amniotic fluid and evaluated in vitro on murine H-JEB keratinocytes, and in vivo by prenatal injection into the amniotic cavities of laminin-5 β3-deficient mice. The different vectors were administered individually or combined at maximum doses on day 14 post coitum. Adenoviral vectors infected preferentially the foetal epidermis, whereas AAV delivered the transgene mainly to mucous membranes of the airways and the upper digestive tract. The LAMB3 transgene was expressed in target epithelia of newborn laminin-5 β3-deficient mice, and the transgenic β3 chain was shown to assemble with its endogenous partner chains, resulting in detectable amounts of laminin-5 in the basement membranes of skin and mucosae and in a lower extent of tissue separation in the skin. However, only combined delivery of the two vector types led to a minor increase of the life span of H-JEB mice. Failure to rescue diseased animals was, at least in part, due to abandonment of any conspicuous pup by the heterozygous mother. This is the first study of a prenatal gene therapy approach to a heritable blistering disorder. Although our findings indicate that prenatal combined administration of adenoviral and adeno-associated LAMB3 vectors provides therapeutic benefit to H-JEB mice, this animal model appears unsuitable for long-term investigations of the therapeutic concept.

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Acknowledgements

We thank Erika Ayllón Ortiz for excellent technical assistance, Elisabeth Koppmann for animal care, Dr Philippe Moullier (Genethon, Nantes, France) for providing the plasmid pRepCap, Dr Richard Samulski (University of North Carolina, USA) for the plasmid pXX6-80, Dr Ernst Beinder (Women's Hospital, University of Erlangen-Nuernberg) for providing human amniotic fluid samples, Dr Guerrino Meneguzzi (INSERM U385, Nice, France) for the cDNA of the human laminin-5 β3 chain and the antibody K140, Dr Susan Fischer (MD Anderson Cancer Center, TX, USA) for the JWF2 cells and Dr Anne-Marie Douar (Genethon, Evry, France) for help with the generation of AAVLAMB3. This study was supported by the ELAN programme of the University of Erlangen-Nuernberg and by a grant from the German Research Foundation (SCHN 569/3-1).

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  1. C Mühle and A Neuner: These authors contributed equally to this work.

Authors and Affiliations

  1. University of Erlangen-Nuernberg, Children's Hospital, Erlangen, Germany

    C Mühle, A Neuner, J Park, Q Jiang & H Schneider

  2. Department of Experimental Medicine I, Nikolaus Fiebiger Centre of Molecular Medicine, Erlangen, Germany

    C Mühle, J Park, F Pacho & H Schneider

  3. Division of Biomedical Sciences, Cystic Fibrosis Gene Therapy Research Group, Imperial College School of Medicine, University of London, London, UK

    S N Waddington

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  1. C Mühle
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Correspondence to H Schneider.

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Mühle, C., Neuner, A., Park, J. et al. Evaluation of prenatal intra-amniotic LAMB3 gene delivery in a mouse model of Herlitz disease. Gene Ther 13, 1665–1676 (2006). https://doi.org/10.1038/sj.gt.3302832

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