Correction of pseudoexon splicing caused by a novel intronic dysferlin mutation
Authors
Dominov, Janice A.Uyan, Ozgun
McKenna-Yasek, Diane
Nallamilli, Babi Ramesh Reddy
Kergourlay, Virginie
Bartoli, Marc
Levy, Nicolas
Hudson, Judith
Evangelista, Teresinha
Lochmuller, Hanns
Krahn, Martin
Rufibach, Laura
Hegde, Madhuri
Brown, Robert H. Jr.
UMass Chan Affiliations
Department of NeurologyDocument Type
Journal ArticlePublication Date
2019-03-03Keywords
Congenital, Hereditary, and Neonatal Diseases and AbnormalitiesGenetic Phenomena
Musculoskeletal Diseases
Nervous System Diseases
Neurology
Neuroscience and Neurobiology
Nucleic Acids, Nucleotides, and Nucleosides
Translational Medical Research
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Show full item recordAbstract
Objective: Dysferlin is a large transmembrane protein that functions in critical processes of membrane repair and vesicle fusion. Dysferlin-deficiency due to mutations in the dysferlin gene leads to muscular dystrophy (Miyoshi myopathy (MM), limb girdle muscular dystrophy type 2B (LGMD2B), distal myopathy with anterior tibial onset (DMAT)), typically with early adult onset. At least 416 pathogenic dysferlin mutations are known, but for approximately 17% of patients, one or both of their pathogenic variants remain undefined following standard exon sequencing methods that interrogate exons and nearby flanking intronic regions but not the majority of intronic regions. Methods: We sequenced RNA from myogenic cells to identify a novel dysferlin pathogenic variant in two affected siblings that previously had only one disease-causing variant identified. We designed antisense oligonucleotides (AONs) to bypass the effects of this mutation on RNA splicing. Results: We identified a new pathogenic point mutation deep within dysferlin intron 50i. This intronic variant causes aberrant mRNA splicing and inclusion of an additional pseudoexon (PE, we term PE50.1) within the mature dysferlin mRNA. PE50.1 inclusion alters the protein sequence, causing premature translation termination. We identified this mutation in 23 dysferlinopathy patients (seventeen families), revealing it to be one of the more prevalent dysferlin mutations. We used AON-mediated exon skipping to correct the aberrant PE50.1 splicing events in vitro, which increased normal mRNA production and significantly restored dysferlin protein expression. Interpretation: Deep intronic mutations can be a common underlying cause of dysferlinopathy, and importantly, could be treatable with AON-based exon-skipping strategies.Source
Ann Clin Transl Neurol. 2019 Mar 3;6(4):642-654. doi: 10.1002/acn3.738. eCollection 2019 Apr. Link to article on publisher's site
DOI
10.1002/acn3.738Permanent Link to this Item
http://hdl.handle.net/20.500.14038/40987PubMed ID
31019989Related Resources
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Copyright 2019 The Authors. Annals of Clinical and Translational Neurology published by Wiley Periodicals, Inc on behalf of American Neurological Association. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.Distribution License
http://creativecommons.org/licenses/by-nc-nd/4.0/ae974a485f413a2113503eed53cd6c53
10.1002/acn3.738
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Except where otherwise noted, this item's license is described as Copyright 2019 The Authors. Annals of Clinical and Translational Neurology published by Wiley Periodicals, Inc on behalf of American Neurological Association. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.