Exon-Skipping Oligonucleotides Restore Functional Collagen VI by Correcting a Common COL6A1 Mutation in Ullrich CMD.

Collagen VI Ullrich muscular dystrophy antisense oligonucleotide congenital muscular dystrophy deep intronic mutation exon skipping extra cellular matrix

Journal

Molecular therapy. Nucleic acids
ISSN: 2162-2531
Titre abrégé: Mol Ther Nucleic Acids
Pays: United States
ID NLM: 101581621

Informations de publication

Date de publication:
04 Sep 2020
Historique:
received: 07 04 2020
revised: 12 05 2020
accepted: 26 05 2020
pubmed: 26 6 2020
medline: 26 6 2020
entrez: 26 6 2020
Statut: ppublish

Résumé

Collagen VI-related congenital muscular dystrophies (COL6-CMDs) are the second most common form of congenital muscular dystrophy. Currently, there is no effective treatment available. COL6-CMDs are caused by recessive or dominant mutations in one of the three genes encoding for the α chains of collagen type VI (COL6A1, COL6A2, and COL6A3). One of the most common mutations in COL6-CMD patients is a de novo deep intronic c.930+189C > T mutation in COL6A1 gene. This mutation creates a cryptic donor splice site and induces incorporation of a novel in-frame pseudo-exon in the mature transcripts. In this study, we systematically evaluated the splice switching approach using antisense oligonucleotides (ASOs) to correct this mutation. Fifteen ASOs were designed using the RNA-tiling approach to target the misspliced pseudo-exon and its flanking sequences. The efficiency of ASOs was evaluated at RNA, protein, and structural levels in skin fibroblasts established from four patients carrying the c.930+189C > T mutation. We identified two additional lead ASO candidates that efficiently induce pseudo-exon exclusion from the mature transcripts, thus allowing for the restoration of a functional collagen VI microfibrillar matrix. Our findings provide further evidence for ASO exon skipping as a therapeutic approach for COL6-CMD patients carrying this common intronic mutation.

Identifiants

pubmed: 32585628
pii: S2162-2531(20)30155-4
doi: 10.1016/j.omtn.2020.05.029
pmc: PMC7321786
pii:
doi:

Types de publication

Journal Article

Langues

eng

Pagination

205-216

Subventions

Organisme : Wellcome Trust
Pays : United Kingdom

Informations de copyright

Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.

Références

Neuromuscul Disord. 2012 Jun;22(6):522-7
pubmed: 22480491
Am J Hum Genet. 1988 Mar;42(3):435-45
pubmed: 3348212
Nat Rev Neurol. 2011 Jun 21;7(7):379-90
pubmed: 21691338
Lancet. 2011 Aug 13;378(9791):595-605
pubmed: 21784508
Ann Neurol. 2001 Aug;50(2):261-5
pubmed: 11506412
J Neurol Neurosurg Psychiatry. 2015 Mar;86(3):280-7
pubmed: 24938411
Acta Biomater. 2017 Apr 1;52:21-32
pubmed: 27956360
Hum Mol Genet. 1996 Jul;5(7):1043-6
pubmed: 8817344
Nucleic Acids Res. 2010 Jan;38(1):e3
pubmed: 19854938
Handb Clin Neurol. 2011;101:81-96
pubmed: 21496625
Mol Ther Nucleic Acids. 2014 Feb 11;3:e147
pubmed: 24518369
J Neuropathol Exp Neurol. 2008 Feb;67(2):144-54
pubmed: 18219255
J Cell Biol. 1986 Mar;102(3):703-10
pubmed: 3456350
Hum Gene Ther. 2012 Dec;23(12):1313-8
pubmed: 22992134
N Engl J Med. 2018 Feb 15;378(7):625-635
pubmed: 29443664
J Biol Chem. 1990 Sep 15;265(26):15349-52
pubmed: 2203776
Methods Mol Biol. 2012;867:117-29
pubmed: 22454058
Hum Mol Genet. 2005 Jan 15;14(2):279-93
pubmed: 15563506
Neurology. 2004 Feb 24;62(4):620-3
pubmed: 14981181
Neuromuscul Disord. 2007 Jul;17(7):547-57
pubmed: 17537636
J Cell Sci. 2015 Oct 1;128(19):3525-31
pubmed: 26377767
Neurology. 2009 Jul 7;73(1):25-31
pubmed: 19564581
Neuromuscul Disord. 2006 Oct;16(9-10):571-82
pubmed: 16935502
Hum Mutat. 2015 Jan;36(1):48-56
pubmed: 25204870
Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7516-21
pubmed: 11381124
JCI Insight. 2019 Mar 21;4(6):
pubmed: 30895940
N Engl J Med. 2017 Nov 2;377(18):1723-1732
pubmed: 29091570
Sci Transl Med. 2017 Apr 19;9(386):
pubmed: 28424332
Matrix Biol. 2018 Oct;71-72:348-367
pubmed: 29277723
Mol Ther Nucleic Acids. 2014 Jun 24;3:e171
pubmed: 24959844
Am J Hum Genet. 2002 Jun;70(6):1446-58
pubmed: 11992252
Skelet Muscle. 2011 Sep 23;1:30
pubmed: 21943391
Methods Mol Biol. 2012;815:333-46
pubmed: 22131003
Annu Rev Med. 2019 Jan 27;70:307-321
pubmed: 30691367
Ann Neurol. 2011 Jan;69(1):206-11
pubmed: 21280092
J Mol Biol. 2003 Jul 4;330(2):297-307
pubmed: 12823969
Am J Hum Genet. 2003 Aug;73(2):355-69
pubmed: 12840783
Ann Neurol. 2005 Sep;58(3):400-10
pubmed: 16130093
Adv Drug Deliv Rev. 2015 Jun 29;87:35-45
pubmed: 25881722
Proc R Soc Med. 1908;1(Pathol Sect):157-66
pubmed: 19973308
J Med Genet. 2005 Sep;42(9):673-85
pubmed: 16141002
Mol Ther Nucleic Acids. 2017 Sep 15;8:416-427
pubmed: 28918041

Auteurs

Sara Aguti (S)

The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Research and Teaching Department, Great Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK.

Véronique Bolduc (V)

Neuromuscular and Neurogenetic Disorders of Childhood Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland MD 20892, USA.

Pierpaolo Ala (P)

The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Research and Teaching Department, Great Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK.

Mark Turmaine (M)

Division of Biosciences, University College London, Gower Street, London WC1E 6BT, UK.

Carsten G Bönnemann (CG)

Neuromuscular and Neurogenetic Disorders of Childhood Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland MD 20892, USA.

Francesco Muntoni (F)

The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Research and Teaching Department, Great Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK; NIHR Great Ormond Street Hospital Biomedical Research Centre, London WC1N 1EH, UK. Electronic address: f.muntoni@ucl.ac.uk.

Haiyan Zhou (H)

NIHR Great Ormond Street Hospital Biomedical Research Centre, London WC1N 1EH, UK; Genetics and Genomic Medicine Research and Teaching Department, Great Ormond Street Institute of Child Health, University College London, London WC1N 1EH, UK. Electronic address: haiyan.zhou@ucl.ac.uk.

Classifications MeSH