title: Pathogenic exon-trapping by SVA retrotransposon and rescue in Fukuyama muscular dystrophy.
abstract: Fukuyama muscular dystrophy (FCMD; MIM253800), one of the most common autosomal recessive disorders in Japan, was the first human disease found to result from ancestral insertion of a SINE-VNTR-Alu (SVA) retrotransposon into a causative gene. In FCMD, the SVA insertion occurs in the 3' untranslated region (UTR) of the fukutin gene. The pathogenic mechanism for FCMD is unknown, and no effective clinical treatments exist. Here we show that aberrant messenger RNA (mRNA) splicing, induced by SVA exon-trapping, underlies the molecular pathogenesis of FCMD. Quantitative mRNA analysis pinpointed a region that was missing from transcripts in patients with FCMD. This region spans part of the 3' end of the fukutin coding region, a proximal part of the 3' UTR and the SVA insertion. Correspondingly, fukutin mRNA transcripts in patients with FCMD and SVA knock-in model mice were shorter than the expected length. Sequence analysis revealed an abnormal splicing event, provoked by a strong acceptor site in SVA and a rare alternative donor site in fukutin exon 10. The resulting product truncates the fukutin carboxy (C) terminus and adds 129 amino acids encoded by the SVA. Introduction of antisense oligonucleotides (AONs) targeting the splice acceptor, the predicted exonic splicing enhancer and the intronic splicing enhancer prevented pathogenic exon-trapping by SVA in cells of patients with FCMD and model mice, rescuing normal fukutin mRNA expression and protein production. AON treatment also restored fukutin functions, including O-glycosylation of α-dystroglycan (α-DG) and laminin binding by α-DG. Moreover, we observe exon-trapping in other SVA insertions associated with disease (hypercholesterolemia, neutral lipid storage disease) and human-specific SVA insertion in a novel gene. Thus, although splicing into SVA is known, we have discovered in human disease a role for SVA-mediated exon-trapping and demonstrated the promise of splicing modulation therapy as the first radical clinical treatment for FCMD and other SVA-mediated diseases.
authors: Division of Neurology/Molecular Brain Science, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.
authors: Mariko
authors: M
authors: Taniguchi-Ikeda
authors: Kazuhiro
authors: K
authors: Kobayashi
authors: Motoi
authors: M
authors: Kanagawa
authors: Chih-chieh
authors: CC
authors: Yu
authors: Kouhei
authors: K
authors: Mori
authors: Tetsuya
authors: T
authors: Oda
authors: Atsushi
authors: A
authors: Kuga
authors: Hiroki
authors: H
authors: Kurahashi
authors: Hasan O
authors: HO
authors: Akman
authors: Salvatore
authors: S
authors: DiMauro
authors: Ryuji
authors: R
authors: Kaji
authors: Toshifumi
authors: T
authors: Yokota
authors: Shin'ichi
authors: S
authors: Takeda
authors: Tatsushi
authors: T
authors: Toda
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D017398:Alternative Splicing / Q000187:drug effects / Q000235:genetics*; D000818:Animals; D004195:Disease Models, Animal; D049030:Dystroglycans / Q000378:metabolism; D005091:Exons / Q000235:genetics*; D055879:Gene Knock-In Techniques; D006031:Glycosylation; D006801:Humans; D007438:Introns / Q000235:genetics; D007564:Japan; D007797:Laminin / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000187:drug effects / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use; D059368:RNA Isoforms / Q000235:genetics; D022821:RNA Splice Sites / Q000235:genetics; D018626:Retroelements / Q000235:genetics*; D058494:Walker-Warburg Syndrome / Q000235:genetics* / Q000473:pathology* / Q000628:therapy
chemical_list: D020413:3' Untranslated Regions; C113587:FKTN protein, human; D007797:Laminin; D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D059368:RNA Isoforms; D022821:RNA Splice Sites; D018626:Retroelements; D049030:Dystroglycans