Approval of antisense oligonucleotide eteplirsen highlights the promise of exon-skipping therapeutics for Duchenne muscular dystrophy patients. However, the limited efficacy of eteplirsen underscores the importance to improve systemic delivery and efficacy. Recently, we demonstrated that a glucose and fructose (GF) delivery formulation effectively potentiates phosphorodiamidate morpholino oligomer (PMO). Considering the clinical potential of GF, it is important to determine the long-term compatibility and efficacy with PMO in mdx mice prior to clinical translation. Here, we report that yearlong administration of a clinically applicable PMO dose (50 mg/kg/week for 3 weeks followed by 50 mg/kg/month for 11 months) with GF elicited sustainably high levels of dystrophin expression in mdx mice, with up to 45% of the normal level of dystrophin restored in most peripheral muscles without any detectable toxicity. Importantly, PMO-GF resulted in phenotypical rescue and mitochondrial biogenesis with functional improvement. Carbohydrate metabolites measurements revealed improved metabolic and energetic conditions after PMO-GF treatment in mdx mice without metabolic anomaly. Collectively, our study shows PMO-GF’s ability to elicit long-lasting therapeutic effects with tolerable toxicity and represents a new treatment modality for Duchenne muscular dystrophy, and provides guidelines for antisense oligonucleotides with GF in clinical use.

反义寡核苷酸eteplirsen的批准突显了杜兴氏肌营养不良患者跳过外显子疗法的前景。然而，依替普林有限的疗效强调了改善全身递送和疗效的重要性。最近，我们证明了葡萄糖和果糖（GF）输送配方可以有效增强二氨基磷酸二氢吗啡啉低聚物（PMO）的强度​​。考虑到GF的临床潜力，重要的是在临床翻译之前确定mdx小鼠中PMO的长期相容性和疗效。在这里，我们报告，一年服用临床适用的PMO剂量（3周50 mg / kg /周，然后连续11周服用50 mg / kg /月，共11个月）会引起mdx中肌营养不良蛋白的持续高水平表达在大多数外周肌肉中，肌萎缩蛋白的正常水平高达正常水平的45％的小鼠恢复了，没有任何可检测到的毒性。重要的是，PMO-GF导致表型抢救和线粒体生物发生，功能得到改善。碳水化合物代谢物测量显示，在无代谢异常的mdx小鼠中，经PMO-GF处理后，代谢和能量状况得到改善。总体而言，我们的研究表明PMO-GF具有可耐受的毒性引发长期治疗效果的能力，代表了杜氏肌营养不良症的一种新治疗方式，并为临床上使用GF的反义寡核苷酸提供了指导。