Correction of Recessive Dystrophic Epidermolysis Bullosa by Transposon-Mediated Integration of COL7A1 in Transplantable Patient-Derived Primary Keratinocytes

• Published in: Journal of investigative dermatology Vol. 137; no. 4; pp. 836 - 844
• Main Authors: Latella, Maria Carmela, Cocchiarella, Fabienne, De Rosa, Laura, Turchiano, Giandomenico, Gonçalves, Manuel A.F.V., Larcher, Fernando, De Luca, Michele, Recchia, Alessandra
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.04.2017
• Subjects: Animals; Cells, Cultured; Collagen Type VII - genetics; Disease Models, Animal; Epidermolysis Bullosa Dystrophica - genetics; Epidermolysis Bullosa Dystrophica - therapy; Fluorescent Antibody Technique; Genetic Predisposition to Disease; Genetic Therapy - methods; Humans; Keratinocytes - transplantation; Mice; Mice, Mutant Strains; Mutation; Random Allocation; Reproducibility of Results; Risk Assessment; Transplantation, Heterologous - methods; Treatment Outcome; AdV; RT; C7; iRDEB; FRT; RDEB; FLPe; HD; SB
• ISSN: 0022-202X; 1523-1747; 1523-1747
• DOI: 10.1016/j.jid.2016.11.038

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.