title: Correction of Recessive Dystrophic Epidermolysis Bullosa by Transposon-Mediated Integration of COL7A1 in Transplantable Patient-Derived Primary Keratinocytes.
abstract: Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects in type-VII collagen (C7), a protein encoded by the COL7A1 gene and essential for anchoring fibril formation at the dermal-epidermal junction. Gene therapy of RDEB is based on transplantation of autologous epidermal grafts generated from gene-corrected keratinocytes sustaining C7 deposition at the dermal-epidermal junction. Transfer of the COL7A1 gene is complicated by its very large size and repetitive sequence. This article reports a gene delivery approach based on the Sleeping beauty transposon, which allows integration of a full-length COL7A1 cDNA and secretion of C7 at physiological levels in RDEB keratinocytes without rearrangements or detrimental effects on their clonogenic potential. Skin equivalents derived from gene-corrected RDEB keratinocytes were tested in a validated preclinical model of xenotransplantation on immunodeficient mice, where they showed normal deposition of C7 at the dermal-epidermal junction and restoration of skin adhesion properties. These results indicate the feasibility and efficacy of a transposon-based gene therapy approach to RDEB.
authors.affiliation: Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Maria Carmela
authors.initials: MC
authors.lastname: Latella
authors.affiliation: Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Cocchiarella
authors.affiliation: Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: De Rosa
authors.affiliation: Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Giandomenico
authors.initials: G
authors.lastname: Turchiano
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Manuel A F V
authors.initials: MAFV
authors.lastname: Gonçalves
authors.affiliation: Epithelial Biomedicine Division, CIEMAT-CIBERER (Centre for Biomedical Research on Rare Diseases), Madrid, Spain; Department of Bioengineering, Universidad Carlos III de Madrid, Madrid, Spain; Instituto de Investigación Sanitaria de la Fundación Jiménez Díaz, Madrid, Spain.
authors.forename: Fernando
authors.initials: F
authors.lastname: Larcher
authors.affiliation: Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: De Luca
authors.affiliation: Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy. Electronic address: alessandra.recchia@unimore.it.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Recchia
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D024143:Collagen Type VII / Q000235:genetics*; D004195:Disease Models, Animal; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics* / Q000628:therapy*; D005455:Fluorescent Antibody Technique; D020022:Genetic Predisposition to Disease; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D015603:Keratinocytes / Q000637:transplantation*; D051379:Mice; D008817:Mice, Mutant Strains; D009154:Mutation; D011897:Random Allocation; D015203:Reproducibility of Results; D018570:Risk Assessment; D014183:Transplantation, Heterologous / Q000379:methods; D016896:Treatment Outcome
chemical_list: C583886:Col7a1 protein, mouse; D024143:Collagen Type VII