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Restoration of type VII collagen expression and function in dystrophic epidermolysis bullosa

Nature Genetics volume 32, pages 670–675 (2002)Cite this article

Abstract

Dystrophic epidermolysis bullosa (DEB) is a family of inherited mechano-bullous disorders caused by mutations in the human type VII collagen gene (COL7A1). Individuals with DEB lack type VII collagen and anchoring fibrils, structures that attach epidermis and dermis. The current lack of treatment for DEB is an impetus to develop gene therapy strategies that efficiently transfer and stably express genes delivered to skin cells in vivo. In this study, we delivered and expressed full-length type VII collagen using a self-inactivating minimal lentivirus-based vector. Transduction of lentiviral vectors containing the COL7A1 transgene into recessive DEB (RDEB) keratinocytes and fibroblasts (in which type VII collagen was absent) resulted in persistent synthesis and secretion of type VII collagen. Unlike RDEB parent cells, the gene-corrected cells had normal morphology, proliferative potential, matrix attachment and motility. We used these gene-corrected cells to regenerate human skin on immune-deficient mice. Human skin regenerated by gene-corrected RDEB cells had restored expression of type VII collagen and formation of anchoring fibrils at the dermal–epidermal junction in vivo. These studies demonstrate that it is possible to restore type VII collagen gene expression in RDEB skin in vivo.

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Figure 1: Lentiviral vector–mediated gene transfer of type VII collagen to RDEB cells.
Figure 2: Expression of type VII collagen reversed RDEB morphology.
Figure 3: Expression of type VII collagen reverted RDEB hypermotility.
Figure 4: Expression of type VII collagen enhanced RDEB adhesion.
Figure 5: Gene-corrected RDEB cells showed enhanced growth potential.
Figure 6: Restoration of type VII collagen and anchoring fibrils in reconstituted DEB skin in vivo.

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Acknowledgements

This work was supported by grants from the US National Institutes of Health to M.C. and D.T.W. M.C. was supported by a Dermatology Foundation Career Development Award and a Dermatology Foundation Research Grant. C.M. and N.K. are also funded in part by a grant from the US National Institutes of Health through the Molecular Biology Core/Virus Vector Subcore of the University of Southern California Research Center for Liver Diseases.

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Authors and Affiliations

  1. Department of Medicine, Division of Dermatology, University of Southern California, CRL 204, 1303 Mission Road, Los Angeles, 90033, California, USA

    Mei Chen & David T. Woodley

  2. Departments of Pathology and Biochemistry, University of Southern California, CRL 204, 1303 Mission Road, Los Angeles, 90033, California, USA

    Noriyuki Kasahara, Maria Barcova, Paula M. Cannon & Constance Mazurek

  3. Shriners Hospital for Children, Portland, Oregon, USA

    Douglas R. Keene

  4. Department of Dermatology, Northwestern University, Chicago, Illinois, USA

    Lawrence Chan

  5. Xgene Corporation, San Carlos, California, USA

    Warren K. Hoeffler & Deborah Finlay

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  1. Mei Chen
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Correspondence to Mei Chen.

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Chen, M., Kasahara, N., Keene, D. et al. Restoration of type VII collagen expression and function in dystrophic epidermolysis bullosa. Nat Genet 32, 670–675 (2002). https://doi.org/10.1038/ng1041

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