Department of Human Genetics, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Hum Gene Ther. 2012 Mar;23(3):262-73. doi: 10.1089/hum.2011.127. Epub 2012 Jan 26.
In Duchenne muscular dystrophy (DMD), dystrophin deficiency leading to progressive muscular degeneration is caused by frame-shifting mutations in the DMD gene. Antisense oligonucleotides (AONs) aim to restore the reading frame by skipping of a specific exon(s), thereby allowing the production of a shorter, but semifunctional protein, as is found in the mostly more mildly affected patients with Becker muscular dystrophy. AONs are currently being investigated in phase 3 placebo-controlled clinical trials. Most of the participating patients are treated symptomatically with corticosteroids (mainly predniso[lo]ne) to stabilize the muscle fibers, which might affect the uptake and/or efficiency of AONs. Therefore the effect of prednisolone on 2'-O-methyl phosphorothioate AON efficacy in patient-derived cultured muscle cells and the mdx mouse model (after local and systemic AON treatment) was assessed in this study. Both in vitro and in vivo skip efficiency and biomarker expression were comparable between saline- and prednisolone-cotreated cells and mice. After systemic exon 23-specific AON (23AON) treatment for 8 weeks, dystrophin was detectable in all treated mice. Western blot analyses indicated slightly higher dystrophin levels in prednisolone-treated mice, which might be explained by better muscle condition and consequently more target dystrophin pre-mRNA. In addition, fibrotic and regeneration biomarkers were normalized to some extent in prednisolone- and/or 23AON-treated mice. Overall these results show that the use of prednisone forms no barrier to participation in clinical trials with AONs.
在杜氏肌营养不良症(DMD)中,肌营养不良蛋白的缺乏导致进行性肌肉退化,这是由于 DMD 基因的移码突变引起的。反义寡核苷酸(AONs)旨在通过跳过特定的外显子(多个外显子)来恢复阅读框,从而产生较短但具有部分功能的蛋白质,正如在大多数受影响较轻的贝克肌营养不良症患者中发现的那样。AONs 目前正在进行 3 期安慰剂对照临床试验。大多数参与的患者接受皮质类固醇(主要是泼尼松龙)的对症治疗,以稳定肌肉纤维,这可能会影响 AON 的摄取和/或效率。因此,本研究评估了泼尼松龙对患者来源的培养肌肉细胞和 mdx 小鼠模型(局部和全身 AON 治疗后)中 2'-O-甲基磷硫代酸酯 AON 疗效的影响。在体外和体内,盐皮质激素和泼尼松龙共同处理的细胞和小鼠之间的跳跃效率和生物标志物表达均无差异。在接受 8 周系统性外显子 23 特异性 AON(23AON)治疗后,所有治疗小鼠均可检测到肌营养不良蛋白。Western blot 分析表明,泼尼松龙治疗小鼠中的肌营养不良蛋白水平略高,这可能是由于肌肉状况更好,从而导致更多的靶标肌营养不良蛋白前体 mRNA。此外,在接受泼尼松龙和/或 23AON 治疗的小鼠中,纤维性和再生生物标志物在一定程度上得到了正常化。总体而言,这些结果表明,使用泼尼松龙不会成为 AON 临床试验参与的障碍。