title: Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector.
abstract: Patients with recessive dystrophic epidermolysis bullosa (RDEB) lack type VII collagen and therefore have severely impaired dermal-epidermal stability causing recurrent skin and mucosal blistering. There is currently no specific approved treatment for RDEB. We present preclinical data showing that intradermal injections of genetically corrected patient-derived RDEB fibroblasts using a Good Manufacturing Practices grade self-inactivating COL7A1 retroviral vector reverse the disease phenotype in a xenograft model in nude mice. We obtained 50% transduction efficiency in primary human RDEB fibroblasts with an average low copy number (range = 1-2) of integrated provirus. Transduced fibroblasts showed strong type VII collagen re-expression, improved adhesion properties, normal proliferative capabilities, and viability in vitro. We show that a single intradermal injection of 3 × 10(6) genetically corrected RDEB fibroblasts beneath RDEB skin equivalents grafted onto mice allows type VII collagen deposition, anchoring fibril formation at the dermal-epidermal junction, and improved dermal-epidermal adherence 2 months after treatment, supporting functional correction in vivo. Gene-corrected fibroblasts previously showed no tumorigenicity. These data show the efficacy and safety of gene-corrected fibroblast therapy using a self-inactivating vector that has now been good manufacturing grade-certified and pave the way for clinical translation to treat nonhealing wounds in RDEB patients.
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France.
authors.forename: Joanna
authors.initials: J
authors.lastname: Jacków
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France.
authors.forename: Matthias
authors.initials: M
authors.lastname: Titeux
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France.
authors.forename: Soizic
authors.initials: S
authors.lastname: Portier
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France.
authors.forename: Soëli
authors.initials: S
authors.lastname: Charbonnier
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France.
authors.forename: Clarisse
authors.initials: C
authors.lastname: Ganier
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France; Department of General Surgery, Plastic Surgery, and Ambulatory Surgery, AP-HP, HUPC, Cochin Hospital, Paris, France.
authors.forename: Sonia
authors.initials: S
authors.lastname: Gaucher
authors.affiliation: Laboratory of Genetic Skin Diseases, Inserm UMR1163 and Imagine Institute of Genetic Diseases, Paris, France; University Paris Descartes-Sorbonne Paris Cite, Paris, France; Department of Genetics, Necker Hospital, Paris, France. Electronic address: alain.hovnanian@inserm.fr.
authors.forename: Alain
authors.initials: A
authors.lastname: Hovnanian
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D049109:Cell Proliferation; D024143:Collagen Type VII / Q000235:genetics* / Q000378:metabolism; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics* / Q000628:therapy*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005808:Genes, Recessive; D015316:Genetic Therapy; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D009368:Neoplasm Transplantation; D011994:Recombinant Proteins / Q000235:genetics; D012190:Retroviridae; D012867:Skin / Q000378:metabolism; D013696:Temperature
chemical_list: C569053:COL7A1 protein, human; D024143:Collagen Type VII; D011994:Recombinant Proteins