On-chip 3D neuromuscular model for drug screening and precision medicine in neuromuscular disease
On-chip 3D neuromuscular model for drug screening and precision medicine in neuromuscular disease
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DOI：
10.1038/s41596-019-0248-1
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发表时间：
2020-01-13
期刊：
影响因子：
14.8
通讯作者：
Kamm, Roger D.
中科院分区：
文献类型：
--
作者：
Osaki, Tatsuya;Uzel, Sebastien G. M.;Kamm, Roger D.
该协议描述了一个3D生理和病理生理运动单元模型的设计，制造和使用，该模型由运动神经元耦合到骨骼肌，通过微流体装置内的神经肌肉接头（NMJ）相互作用。该模型有助于成像和定量功能评估。“NMJ芯片”能够对轴突生长、NMJ形成和肌肉成熟进行实时、实时成像，以及在光遗传学控制下同步运动神经元活动和肌肉收缩，以研究正常生理事件。拟议的协议需要大约2-3个月的时间才能实施。与各种神经肌肉疾病相关的病理行为，例如运动神经元轴突的退化、运动神经元死亡以及肌肉退化和萎缩也可以在该系统中重现。疾病模型可以通过使用患者来源的诱导多能干细胞来产生所使用的运动神经元和骨骼肌细胞来创建。这通过使用来自患有散发性肌萎缩性侧索硬化症的患者的细胞来证明，但是可以更普遍地应用于神经肌肉疾病的模型，例如脊髓性肌萎缩症、NMJ病症和肌营养不良症。这样的模型在精准医学、药物筛选和疾病风险评估方面具有相当大的应用潜力。
This protocol describes the design, fabrication and use of a 3D physiological and pathophysiological motor unit model consisting of motor neurons coupled to skeletal muscles interacting via the neuromuscular junction (NMJ) within a microfluidic device. This model facilitates imaging and quantitative functional assessment. The 'NMJ chip' enables real-time, live imaging of axonal outgrowth, NMJ formation and muscle maturation, as well as synchronization of motor neuron activity and muscle contraction under optogenetic control for the study of normal physiological events. The proposed protocol takes similar to 2-3 months to be implemented. Pathological behaviors associated with various neuromuscular diseases, such as regression of motor neuron axons, motor neuron death, and muscle degradation and atrophy can also be recapitulated in this system. Disease models can be created by the use of patient-derived induced pluripotent stem cells to generate both the motor neurons and skeletal muscle cells used. This is demonstrated by the use of cells from a patient with sporadic amyotrophic lateral sclerosis but can be applied more generally to models of neuromuscular disease, such as spinal muscular atrophy, NMJ disorder and muscular dystrophy. Models such as this hold considerable potential for applications in precision medicine, drug screening and disease risk assessment.