title: Translation from a DMD exon 5 IRES results in a functional dystrophin isoform that attenuates dystrophinopathy in humans and mice.
abstract: Most mutations that truncate the reading frame of the DMD gene cause loss of dystrophin expression and lead to Duchenne muscular dystrophy. However, amelioration of disease severity has been shown to result from alternative translation initiation beginning in DMD exon 6 that leads to expression of a highly functional N-truncated dystrophin. Here we demonstrate that this isoform results from usage of an internal ribosome entry site (IRES) within exon 5 that is glucocorticoid inducible. We confirmed IRES activity by both peptide sequencing and ribosome profiling in muscle from individuals with minimal symptoms despite the presence of truncating mutations. We generated a truncated reading frame upstream of the IRES by exon skipping, which led to synthesis of a functional N-truncated isoform in both human subject-derived cell lines and in a new DMD mouse model, where expression of the truncated isoform protected muscle from contraction-induced injury and corrected muscle force to the same level as that observed in control mice. These results support a potential therapeutic approach for patients with mutations within the 5' exons of DMD.
authors.affiliation: 1] The Center for Gene Therapy, Nationwide Children's Hospital, The Ohio State University, Columbus, Ohio, USA. [2].
authors.forename: Nicolas
authors.initials: N
authors.lastname: Wein
authors.affiliation: 1] The Center for Gene Therapy, Nationwide Children's Hospital, The Ohio State University, Columbus, Ohio, USA. [2].
authors.forename: Adeline
authors.initials: A
authors.lastname: Vulin
authors.affiliation: Section of Microbiology and Medical Genetics, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Maria S
authors.initials: MS
authors.lastname: Falzarano
authors.affiliation: Department of Proteomics and Nanobiotechnology, School of Biotechnology, KTH Royal Institute of Technology, Stockholm, Sweden.
authors.forename: Christina Al-Khalili
authors.initials: CA
authors.lastname: Szigyarto
authors.affiliation: Department of Neurology, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Baijayanta
authors.initials: B
authors.lastname: Maiti
authors.affiliation: The Center for Gene Therapy, Nationwide Children's Hospital, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Findlay
authors.affiliation: The Center for Gene Therapy, Nationwide Children's Hospital, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Kristin N
authors.initials: KN
authors.lastname: Heller
authors.affiliation: Department of Proteomics and Nanobiotechnology, School of Biotechnology, KTH Royal Institute of Technology, Stockholm, Sweden.
authors.forename: Mathias
authors.initials: M
authors.lastname: Uhlén
authors.affiliation: 1] Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA. [2] Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Baskar
authors.initials: B
authors.lastname: Bakthavachalu
authors.affiliation: Department of Neuroscience, University of Messina and Centro Clinico Nemo Sud, Messina, Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Messina
authors.affiliation: Department of Neuroscience, University of Messina and Centro Clinico Nemo Sud, Messina, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Vita
authors.affiliation: Bambino Gesù Children's Hospital, Rome, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Passarelli
authors.forename: Simona
authors.initials: S
authors.lastname: Brioschi
authors.forename: Matteo
authors.initials: M
authors.lastname: Bovolenta
authors.forename: Marcella
authors.initials: M
authors.lastname: Neri
authors.affiliation: Section of Microbiology and Medical Genetics, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Gualandi
authors.affiliation: Centre for Comparative Genomics, Murdoch University, Perth, Western Australia, Australia.
authors.forename: Steve D
authors.initials: SD
authors.lastname: Wilton
authors.affiliation: 1] The Center for Gene Therapy, Nationwide Children's Hospital, The Ohio State University, Columbus, Ohio, USA. [2] Department of Pediatrics, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Louise R
authors.initials: LR
authors.lastname: Rodino-Klapac
authors.affiliation: Division of Biomedical Informatics, Department of Computer Science, University of Kentucky Lexington, Kentucky, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Human Genetics, The University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Diane M
authors.initials: DM
authors.lastname: Dunn
authors.affiliation: 1] Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA. [2] Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Daniel R
authors.identifier: 0000000345701946
authors.initials: DR
authors.lastname: Schoenberg
authors.affiliation: Department of Human Genetics, The University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Robert B
authors.initials: RB
authors.lastname: Weiss
authors.affiliation: Department of Human Genetics, The University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Howard
authors.affiliation: Section of Microbiology and Medical Genetics, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ferlini
authors.affiliation: 1] The Center for Gene Therapy, Nationwide Children's Hospital, The Ohio State University, Columbus, Ohio, USA. [2] Department of Pediatrics, The Ohio State University, Columbus, Ohio, USA. [3] Department of Neurology, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Flanigan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics*; D005091:Exons; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000473:pathology; D014176:Protein Biosynthesis; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D017386:Sequence Homology, Amino Acid
chemical_list: C484258:DMD protein, human; D016189:Dystrophin; D020033:Protein Isoforms
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