title: Analysis of the functional consequences of targeted exon deletion in COL7A1 reveals prospects for dystrophic epidermolysis bullosa therapy.
abstract: Genetically evoked deficiency of collagen VII causes dystrophic epidermolysis bullosa (DEB)-a debilitating disease characterized by chronic skin fragility and progressive fibrosis. Removal of exons carrying frame-disrupting mutations can reinstate protein expression in genetic diseases. The therapeutic potential of this approach is critically dependent on gene, protein, and disease intrinsic factors. Naturally occurring exon skipping in COL7A1, translating collagen VII, suggests that skipping of exons containing disease-causing mutations may be feasible for the treatment of DEB. However, despite a primarily in-frame arrangement of exons in the COL7A1 gene, no general conclusion of the aptitude of exon skipping for DEB can be drawn, since regulation of collagen VII functionality is complex involving folding, intra- and intermolecular interactions. To directly address this, we deleted two conceptually important exons located at both ends of COL7A1, exon 13, containing recurrent mutations, and exon 105, predicted to impact folding. The resulting recombinantly expressed proteins showed conserved functionality in biochemical and in vitro assays. Injected into DEB mice, the proteins promoted skin stability. By demonstrating functionality of internally deleted collagen VII variants, our study provides support of targeted exon deletion or skipping as a potential therapy to treat a large number of individuals with DEB.
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: Tobias
authors.initials: T
authors.lastname: Kühl
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, 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
authors.affiliation: Department of Dermatology, Medical Center - University of Freiburg, Freiburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Nyström
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics; D024143:Collagen Type VII / Q000737:chemistry / Q000235:genetics*; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D005091:Exons; D018390:Gene Targeting; D006801:Humans; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000235:genetics; D017510:Protein Folding; D055550:Protein Stability; D016364:Reading Frames; D017384:Sequence Deletion; D012867:Skin / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D024143:Collagen Type VII; D016376:Oligonucleotides, Antisense