title: Sustained phenotypic reversion of junctional epidermolysis bullosa dog keratinocytes: Establishment of an immunocompetent animal model for cutaneous gene therapy.
abstract: Gene transfer represents the unique therapeutic issue for a number of inherited skin disorders including junctional epidermolysis bullosa (JEB), an untreatable genodermatose caused by mutations in the adhesion ligand laminin 5 (alpha3beta3gamma2) that is secreted in the extracellular matrix by the epidermal basal keratinocytes. Because gene therapy protocols require validation in animal models, we have phenotypically reverted by oncoretroviral transfer of the curative gene the keratinocytes isolated from dogs with a spontaneous form of JEB associated with a genetic mutation in the alpha3 chain of laminin 5. We show that the transduced dog JEB keratinocytes: (1) display a sustained secretion of laminin 5 in the extracellular matrix; (2) recover the adhesion, proliferation, and clonogenic capacity of wild-type keratinocytes; (3) generate fully differentiated stratified epithelia that after grafting on immunocompromised mice produce phenotypically normal skin and sustain permanent expression of the transgene. We validate an animal model that appears particularly suitable to demonstrate feasibility, efficacy, and safety of genetic therapeutic strategies for cutaneous disorders before undertaking human clinical trials.
authors.affiliation: INSERM U634, Faculty of Medicine, Nice, France.
authors.forename: Flavia
authors.initials: F
authors.lastname: Spirito
authors.forename: Annabelle
authors.initials: A
authors.lastname: Capt
authors.forename: Marcela
authors.initials: M
authors.lastname: Del Rio
authors.forename: Fernando
authors.initials: F
authors.lastname: Larcher
authors.forename: Eric
authors.initials: E
authors.lastname: Guaguere
authors.forename: Olivier
authors.initials: O
authors.lastname: Danos
authors.forename: Guerrino
authors.initials: G
authors.lastname: Meneguzzi
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004285:Dogs; D004820:Epidermolysis Bullosa / Q000235:genetics* / Q000628:therapy*; D018390:Gene Targeting / Q000379:methods; D015316:Genetic Therapy / Q000379:methods*; D007121:Immunocompetence / Q000235:genetics; D015603:Keratinocytes / Q000378:metabolism*; D007797:Laminin / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use*; D051379:Mice; D011994:Recombinant Proteins / Q000378:metabolism; D012867:Skin / Q000378:metabolism; D016896:Treatment Outcome
chemical_list: D007797:Laminin; D011994:Recombinant Proteins; C095567:laminin alpha 3