Dubowitz Neuromuscular Centre, UCL Great Ormond Street Institute of Child Health, 30 Guilford St, London, WC1N 1EH, UK.
NIHR Great Ormond Street Hospital Biomedical Research Centre, UCL Great Ormond Street Institute of Child Health and Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK.
Acta Neuropathol Commun. 2021 Jan 6;9(1):7. doi: 10.1186/s40478-020-01106-1.
During the last decade, multiple clinical trials for Duchenne muscular dystrophy (DMD) have focused on the induction of dystrophin expression using different strategies. Many of these trials have reported a clear increase in dystrophin protein following treatment. However, the low levels of the induced dystrophin protein have raised questions on its functionality. In our present study, using an unbiased, high-throughput digital image analysis platform, we assessed markers of regeneration and levels of dystrophin associated protein via immunofluorescent analysis of whole muscle sections in 25 DMD boys who received 48-weeks treatment with exon 53 skipping morpholino antisense oligonucleotide (PMO) golodirsen. We demonstrate that the de novo dystrophin induced by exon skipping with PMO golodirsen is capable of conferring a histological benefit in treated patients with an increase in dystrophin associated proteins at the dystrophin positive regions of the sarcolemma in post-treatment biopsies. Although 48 weeks treatment with golodirsen did not result in a significant change in the levels of fetal/developmental myosins for the entire cohort, there was a significant negative correlation between the amount of dystrophin and levels of regeneration observed in different biopsy samples. Our results provide, for the first time, evidence of functionality of induced dystrophin following successful therapeutic intervention in the human.
在过去的十年中，多项针对杜氏肌营养不良症（DMD）的临床试验都集中在使用不同策略诱导肌营养不良蛋白的表达上。这些试验中的许多都报告了治疗后肌营养不良蛋白的明显增加。然而，诱导的肌营养不良蛋白水平较低，引起了其功能的质疑。在我们目前的研究中，我们使用一种无偏的、高通量的数字图像分析平台，通过对 25 名接受外显子 53 跳跃修饰型反义寡核苷酸（PMO）golodirsen 治疗 48 周的 DMD 男孩的整个肌肉切片进行免疫荧光分析，评估了再生标志物和肌营养不良蛋白相关蛋白的水平。我们证明，PMO golodirsen 外显子跳跃诱导的新肌营养不良蛋白能够在治疗后的患者中赋予组织学益处，即在治疗后的活检中，在肌膜的肌营养不良蛋白阳性区域增加肌营养不良蛋白相关蛋白。尽管 golodirsen 治疗 48 周并未导致整个队列的胎儿/发育性肌球蛋白水平发生显著变化，但在不同活检样本中观察到的肌营养不良蛋白和再生水平之间存在显著的负相关。我们的结果首次提供了在人类成功治疗干预后诱导的肌营养不良蛋白具有功能的证据。
