Identification of novel antisense-mediated exon skipping targets in DYSF for therapeutic treatment of dysferlinopathy

Joshua Lee, Rika Maruyama, William Duddy, Hidetoshi Sakurai, Toshifumi Yokota

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Dysferlinopathy is a progressive myopathy caused by mutations in the dysferlin (DYSF) gene.
Dysferlin protein plays a major role in plasma membrane resealing. Some patients with DYSF
deletion mutations exhibit mild symptoms, suggesting some regions of DYSF can be removed
without significantly impacting protein function. Antisense-mediated exon skipping therapy uses
synthetic molecules called antisense oligonucleotides to modulate splicing, allowing exons
harboring or near genetic mutations to be removed and the open reading frame corrected.
Previous studies have focused on DYSF exon 32 skipping as a potential therapeutic approach,
based on the association of a mild phenotype with the in-frame deletion of exon 32. To date, no
other DYSF exon skipping targets have been identified and the relationship between DYSF exon
deletion pattern and protein function remains largely uncharacterized. In this study, we utilized a
membrane wounding assay to evaluate the ability of plasmid constructs carrying mutant DYSF,
as well as antisense oligonucleotides, to rescue membrane resealing in patient cells. We report
that multi-exon skipping of DYSF exons 26-27 and 28-29 rescues plasma membrane resealing.
Successful translation of these findings into the development of clinical antisense drugs would
establish new therapeutic approaches that would be applicable to ~5-7% (exons 26-27 skipping)
and ~8% (exons 28-29 skipping) of dysferlinopathy patients worldwide.
LanguageEnglish
JournalMolecular Therapy - Nucleic Acids
DOIs
Publication statusPublished - 11 Oct 2018

Fingerprint

Exons
Antisense Oligonucleotides
Therapeutics
Mutation
Cell Membrane
Proteins
Dysferlinopathy
Muscular Diseases
Open Reading Frames
Plasmids
Phenotype
Membranes
Pharmaceutical Preparations
Genes

Keywords

  • Exon skipping
  • antisense
  • morpholino
  • dysferlin
  • dysferlinopathy
  • limb girdle muscular dystrophy type 2B
  • LGMD-2B

Cite this

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title = "Identification of novel antisense-mediated exon skipping targets in DYSF for therapeutic treatment of dysferlinopathy",
abstract = "Dysferlinopathy is a progressive myopathy caused by mutations in the dysferlin (DYSF) gene.Dysferlin protein plays a major role in plasma membrane resealing. Some patients with DYSFdeletion mutations exhibit mild symptoms, suggesting some regions of DYSF can be removedwithout significantly impacting protein function. Antisense-mediated exon skipping therapy usessynthetic molecules called antisense oligonucleotides to modulate splicing, allowing exonsharboring or near genetic mutations to be removed and the open reading frame corrected.Previous studies have focused on DYSF exon 32 skipping as a potential therapeutic approach,based on the association of a mild phenotype with the in-frame deletion of exon 32. To date, noother DYSF exon skipping targets have been identified and the relationship between DYSF exondeletion pattern and protein function remains largely uncharacterized. In this study, we utilized amembrane wounding assay to evaluate the ability of plasmid constructs carrying mutant DYSF,as well as antisense oligonucleotides, to rescue membrane resealing in patient cells. We reportthat multi-exon skipping of DYSF exons 26-27 and 28-29 rescues plasma membrane resealing.Successful translation of these findings into the development of clinical antisense drugs wouldestablish new therapeutic approaches that would be applicable to ~5-7{\%} (exons 26-27 skipping)and ~8{\%} (exons 28-29 skipping) of dysferlinopathy patients worldwide.",
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author = "Joshua Lee and Rika Maruyama and William Duddy and Hidetoshi Sakurai and Toshifumi Yokota",
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}

Identification of novel antisense-mediated exon skipping targets in DYSF for therapeutic treatment of dysferlinopathy. / Lee, Joshua; Maruyama, Rika; Duddy, William; Sakurai, Hidetoshi; Yokota, Toshifumi.

In: Molecular Therapy - Nucleic Acids, 11.10.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of novel antisense-mediated exon skipping targets in DYSF for therapeutic treatment of dysferlinopathy

AU - Lee, Joshua

AU - Maruyama, Rika

AU - Duddy, William

AU - Sakurai, Hidetoshi

AU - Yokota, Toshifumi

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N2 - Dysferlinopathy is a progressive myopathy caused by mutations in the dysferlin (DYSF) gene.Dysferlin protein plays a major role in plasma membrane resealing. Some patients with DYSFdeletion mutations exhibit mild symptoms, suggesting some regions of DYSF can be removedwithout significantly impacting protein function. Antisense-mediated exon skipping therapy usessynthetic molecules called antisense oligonucleotides to modulate splicing, allowing exonsharboring or near genetic mutations to be removed and the open reading frame corrected.Previous studies have focused on DYSF exon 32 skipping as a potential therapeutic approach,based on the association of a mild phenotype with the in-frame deletion of exon 32. To date, noother DYSF exon skipping targets have been identified and the relationship between DYSF exondeletion pattern and protein function remains largely uncharacterized. In this study, we utilized amembrane wounding assay to evaluate the ability of plasmid constructs carrying mutant DYSF,as well as antisense oligonucleotides, to rescue membrane resealing in patient cells. We reportthat multi-exon skipping of DYSF exons 26-27 and 28-29 rescues plasma membrane resealing.Successful translation of these findings into the development of clinical antisense drugs wouldestablish new therapeutic approaches that would be applicable to ~5-7% (exons 26-27 skipping)and ~8% (exons 28-29 skipping) of dysferlinopathy patients worldwide.

AB - Dysferlinopathy is a progressive myopathy caused by mutations in the dysferlin (DYSF) gene.Dysferlin protein plays a major role in plasma membrane resealing. Some patients with DYSFdeletion mutations exhibit mild symptoms, suggesting some regions of DYSF can be removedwithout significantly impacting protein function. Antisense-mediated exon skipping therapy usessynthetic molecules called antisense oligonucleotides to modulate splicing, allowing exonsharboring or near genetic mutations to be removed and the open reading frame corrected.Previous studies have focused on DYSF exon 32 skipping as a potential therapeutic approach,based on the association of a mild phenotype with the in-frame deletion of exon 32. To date, noother DYSF exon skipping targets have been identified and the relationship between DYSF exondeletion pattern and protein function remains largely uncharacterized. In this study, we utilized amembrane wounding assay to evaluate the ability of plasmid constructs carrying mutant DYSF,as well as antisense oligonucleotides, to rescue membrane resealing in patient cells. We reportthat multi-exon skipping of DYSF exons 26-27 and 28-29 rescues plasma membrane resealing.Successful translation of these findings into the development of clinical antisense drugs wouldestablish new therapeutic approaches that would be applicable to ~5-7% (exons 26-27 skipping)and ~8% (exons 28-29 skipping) of dysferlinopathy patients worldwide.

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