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.

Dysferlinopathy 是一种由dysferlin ( DYSF ) 基因突变引起的进行性肌病。Dysferlin 蛋白在质膜重新密封中起主要作用。一些带有DYSF缺失突变的患者表现出轻微的症状，这表明可以去除DYSF 的某些区域而不会显着影响蛋白质功能。反义介导的外显子跳跃疗法使用称为反义寡核苷酸的合成分子来调节剪接，从而去除携带或接近基因突变的外显子并纠正开放阅读框。以前的研究集中在DYSF外显子 32 跳跃作为一种潜在的治疗方法，基于轻度表型与外显子 32 框内缺失的关联。迄今为止，尚未确定其他DYSF外显子跳跃靶点，以及DYSF外显子缺失模式和蛋白质功能在很大程度上仍未得到表征。在这项研究中，我们利用膜损伤分析来评估携带突变体DYSF以及反义寡核苷酸的质粒构建体在患者细胞中挽救膜重新密封的能力。我们报告了DYSF 的多外显子跳跃外显子 26-27 和 28-29 挽救了质膜重新密封。将这些发现成功转化为临床反义药物的开发将建立新的治疗方法，适用于全球约 5%–7%（外显子 26–27 跳跃）和约 8%（外显子 28–29 跳跃）的铁蛋白病患者.