Restoration of type VII collagen expression and function in dystrophic epidermolysis bullosa.
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.
Chen
Mei
M
Department of Medicine, Division of Dermatology, University of Southern California, CRL 204, 1303 Mission Road, Los Angeles, California 90033, USA.
Kasahara
Noriyuki
N
Keene
Douglas R
DR
Chan
Lawrence
L
Hoeffler
Warren K
WK
Finlay
Deborah
D
Barcova
Maria
M
Cannon
Paula M
PM
Mazurek
Constance
C
Woodley
David T
DT
D002448:Cell Adhesion; D002455:Cell Division; D002460:Cell Line; D002465:Cell Movement; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D024143:Collagen Type VII / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology*; D018076:DNA, Complementary; D000078404:Epidermal Cells; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D018014:Gene Transfer Techniques; D005808:Genes, Recessive; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D007797:Laminin / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D009154:Mutation; D014162:Transfection; D019076:Transgenes
D024143:Collagen Type VII; D018076:DNA, Complementary; D007797:Laminin