Abstract：
Duchenne muscular dystrophy (DMD) is the most prevalent neuromuscular disease diagnosed in childhood. It is a progressive and wasting disease, characterized by a degeneration of skeletal and cardiac muscles caused by the lack of dystrophin protein. The absence of this crucial structural protein leads to sarcolemmal fragility, resulting in muscle fiber damage during contraction. Despite ongoing efforts, there is no cure available for DMD patients. One of the primary challenges is the limited efficacy of current preclinical tools, which fail in modeling the biological complexity of the disease. Human-based three-dimensional (3D) cell culture methods appear as a novel approach to accelerate preclinical research by enhancing the reproduction of pathophysiological processes in skeletal muscle. In this work, we developed a patient-derived functional 3D skeletal muscle model of DMD that reproduces the sarcolemmal damage found in the native DMD muscle. These bioengineered skeletal muscle tissues exhibit contractile functionality, as they responded to electrical pulse stimulation. Sustained contractile regimes induced the loss of myotube integrity, mirroring the pathological myotube breakdown inherent in DMD due to sarcolemmal instability. Moreover, damaged DMD tissues showed disease functional phenotypes, such as tetanic fatigue. We also evaluated the therapeutic effect of utrophin upregulator drug candidates on the functionality of the skeletal muscle tissues, thus providing deeper insight into the real impact of these treatments. Overall, our findings underscore the potential of bioengineered 3D skeletal muscle technology to advance DMD research and facilitate the development of novel therapies for DMD and related neuromuscular disorders.
摘要：
杜氏肌营养不良症(DMD)是儿童时期诊断的最普遍的神经肌肉疾病。这是一种进行性和消耗性疾病，其特征在于缺乏肌营养不良蛋白引起的骨骼肌和心肌退化。这种关键结构蛋白的缺失会导致肌膜脆性，导致收缩时肌纤维受损。尽管不断努力,DMD患者没有治愈方法.主要挑战之一是当前临床前工具的功效有限，未能对疾病的生物复杂性进行建模。基于人的3D细胞培养方法似乎是通过增强骨骼肌病理生理过程的繁殖来加速临床前研究的新方法。在这项工作中,我们开发了一种DMD的患者衍生的功能性3D骨骼肌模型,该模型再现了在天然DMD肌肉中发现的肌膜损伤。这些生物工程骨骼肌组织表现出收缩功能，当他们对电脉冲刺激（EPS）做出反应时。持续的收缩机制导致肌管完整性的丧失，反映了由于肌膜不稳定而导致的DMD固有的病理性肌管破坏。此外，受损的DMD组织显示疾病功能表型，如强直性疲劳。我们还评估了营养素上调剂候选药物对骨骼肌组织功能的治疗作用，从而更深入地了解这些治疗的真正影响。总的来说,我们的发现强调了生物工程3D骨骼肌技术在推进DMD研究和促进DMD和相关神经肌肉疾病新疗法的开发方面的潜力.
