title: Targeted Exon Skipping Restores Type VII Collagen Expression and Anchoring Fibril Formation in an In Vivo RDEB Model.
abstract: Dystrophic epidermolysis bullosa is a group of orphan genetic skin diseases dominantly or recessively inherited, caused by mutations in COL7A1 encoding type VII collagen, which forms anchoring fibrils. Individuals with recessive dystrophic epidermolysis bullosa develop severe skin and mucosal blistering after mild trauma. The exon skipping strategy consists of modulating splicing of a pre-mRNA to induce skipping of a mutated exon. We have targeted COL7A1 exons 73 and 80, which carry recurrent mutations and whose excision preserves the open reading frame. We first showed the dispensability of these exons for type VII collagen function in vivo. We then showed that transfection of primary recessive dystrophic epidermolysis bullosa keratinocytes and fibroblasts carrying null mutations in exon 73 and/or 80, with 2'-O-methyl antisense oligoribonucleotides, led to efficient ex vivo skipping of these exons (50-95%) and resulted in a significant level (up to 36%) of type VII collagen re-expression. Finally, one or two subcutaneous injections of antisense oligoribonucleotides at doses ranging from 400 μg up to 1 mg restored type VII collagen expression and anchoring fibril formation in vivo in a xenograft model of recessive dystrophic epidermolysis bullosa skin equivalent. This work provides a proof of principle for the treatment of patients with recessive dystrophic epidermolysis bullosa by exon skipping using subcutaneous administration of antisense oligoribonucleotides.
authors: Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche 1163, Paris, France; Imagine Institute, Paris, France; Paris Descartes University, Sorbonne Cité, Paris, France.
authors: Sandrina
authors: S
authors: Turczynski
authors: Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche 1163, Paris, France; Imagine Institute, Paris, France; Paris Descartes University, Sorbonne Cité, Paris, France.
authors: Matthias
authors: M
authors: Titeux
authors: Paul Sabatier University, Toulouse, France; Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5288, Toulouse, France.
authors: Laure
authors: L
authors: Tonasso
authors: Paul Sabatier University, Toulouse, France.
authors: Audrey
authors: A
authors: Décha
authors: Department of Dermatology, Asahikawa Medical University, Asahikawa, Japan.
authors: Akemi
authors: A
authors: Ishida-Yamamoto
authors: Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche 1163, Paris, France; Imagine Institute, Paris, France; Paris Descartes University, Sorbonne Cité, Paris, France; Department of Genetics, Necker Hospital for Sick Children, Paris, France. Electronic address: alain.hovnanian@inserm.fr.
authors: Alain
authors: A
authors: Hovnanian
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics*; D024143:Collagen Type VII / Q000235:genetics*; D003184:Complement C7; D004195:Disease Models, Animal; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics* / Q000503:physiopathology; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D012326:RNA Splicing / Q000235:genetics; D011897:Random Allocation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
chemical_list: C583886:Col7a1 protein, mouse; D024143:Collagen Type VII; D003184:Complement C7; D003094:Collagen