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Early intra-amniotic gene transfer using lentiviral vector improves skin blistering phenotype in a murine model of Herlitz junctional epidermolysis bullosa

Gene Therapy volume 19, pages 561–569 (2012)Cite this article

A Corrigendum to this article was published on 27 October 2011

Abstract

Mutations of the LAMB3 gene cause a lethal form of junctional epidermolysis bullosa (JEB). We hypothesized that early intra-amniotic gene transfer in a severe murine model of JEB would improve or correct the skin phenotype. Time-dated fetuses from heterozygous LAMB3IAP breeding pairs underwent ultrasound guided intra-amniotic injection of lentiviral vector encoding the murine LAMB3 gene at embryonic day 8 (E8). Gene expression was monitored by immunohistochemistry. The transgenic laminin-β3 chain was shown to assemble with its endogenous partner chains, resulting in detectable amounts of laminin-332 in the basement membrane zone of skin and mucosa. Ultrastructually, the restoration of ∼60% of hemidesmosomal structures was also noted. Although we could correct the skin phenotype in 11.9% of homozygous LAMB3IAP mice, none survived beyond 48 h. However, skin transplants from treated E18 homozygous LAMB3IAP fetuses maintained normal appearance for 6 months with persistence of normal assembly of laminin-332. These results demonstrate for the first time long-term phenotypic correction of the skin pathology in a severe model of JEB by in vivo prenatal gene transfer. Although survival remained limited due to the limitations of this mouse model, this study supports the potential for treatment of JEB by prenatal gene transfer.

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Acknowledgements

We thank Keith Alcorn (The Children's Hospital of Philadelphia) for animal support, Ray Meade (The Biomedical Imaging Core/Electron Microscopy Resource Laboratory of the University of Pennsylvania) for electron microscopy support and Xiaohua Chen (The Children's Hospital of Philadelphia) for qPCR support. These studies were supported by NIH/NIAMS Grant R01 AR54876 (JU) and by funds from the Department of Surgery and the Ruth and Tristram C Colket Chair in Pediatric Surgery, Children's Hospital of Philadelphia (AWF).

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Authors and Affiliations

  1. Department of Surgery, The Children's Center for Fetal Research, The Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Abramson Research Center, Philadelphia, PA, USA

    M Endo, P W Zoltick, A Radu & A W Flake

  2. Department of Dermatology and Cutaneous Biology, Thomas Jefferson University, Philadelphia, PA, USA

    J Qiujie & J Uitto

  3. Department of Dermatology, University of Toyama, Toyama, Japan

    C Matsui

  4. Department of Medical Dermatology, Stanford University, Redwood City, CA, USA

    P M Marinkovich

  5. Division of Genetics and Molecular Medicine, Genetic Skin Disease Group, St John's Institute of Dermatology, The Guy's, King's College, and St Thomas’ School of Medicine, London, UK

    J McGrath

  6. Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Osaka, Japan

    K Tamai

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Correspondence to A W Flake.

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Endo, M., Zoltick, P., Radu, A. et al. Early intra-amniotic gene transfer using lentiviral vector improves skin blistering phenotype in a murine model of Herlitz junctional epidermolysis bullosa. Gene Ther 19, 561–569 (2012). https://doi.org/10.1038/gt.2011.135

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