title: Antisense Oligonucleotide-mediated Exon Skipping as a Systemic Therapeutic Approach for Recessive Dystrophic Epidermolysis Bullosa.
abstract: The "generalized severe" form of recessive dystrophic epidermolysis bullosa (RDEB-gen sev) is caused by bi-allelic null mutations in COL7A1, encoding type VII collagen. The absence of type VII collagen leads to blistering of the skin and mucous membranes upon the slightest trauma. Because most patients carry exonic point mutations or small insertions/deletions, most exons of COL7A1 are in-frame, and low levels of type VII collagen already drastically improve the disease phenotype, this gene seems a perfect candidate for antisense oligonucleotide (AON)-mediated exon skipping. In this study, we examined the feasibility of AON-mediated exon skipping in vitro in primary cultured keratinocytes and fibroblasts, and systemically in vivo using a human skin-graft mouse model. We show that treatment with AONs designed against exon 105 leads to in-frame exon 105 skipping at the RNA level and restores type VII collagen protein production in vitro. Moreover, we demonstrate that systemic delivery in vivo induces de novo expression of type VII collagen in skin grafts generated from patient cells. Our data demonstrate strong proof-of-concept for AON-mediated exon skipping as a systemic therapeutic strategy for RDEB.
authors.affiliation: Department of Dermatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Bremer
authors.affiliation: Department of Dermatology, Medical Center - University of Freiburg, Freiburg, Germany.
authors.forename: Olivier
authors.initials: O
authors.lastname: Bornert
authors.affiliation: Department of Dermatology, Medical Center - University of Freiburg, Freiburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Nyström
authors.affiliation: Department of Dermatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors.forename: Antoni
authors.initials: A
authors.lastname: Gostynski
authors.affiliation: Department of Dermatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors.forename: Marcel F
authors.initials: MF
authors.lastname: Jonkman
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Annemieke
authors.initials: A
authors.lastname: Aartsma-Rus
authors.affiliation: Department of Dermatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.|Department of Genetics, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors.forename: Peter C
authors.initials: PC
authors.lastname: van den Akker
authors.affiliation: Department of Dermatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors.forename: Anna Mg
authors.initials: AM
authors.lastname: Pasmooij