IPSEN Innovation, 5 avenue du Canada, 91940, Les Ulis, France.
Université Evry-Paris Saclay/INSERM UMR861, Institut Des Cellules Souches Pour Le Traitement Et L'étude Des Maladies Monogéniques (I-Stem), 2 rue Henri Auguste Desbruères, 91100, Corbeil-Essonne, France.
Stem Cell Res Ther. 2021 Dec 5;12(1):599. doi: 10.1186/s13287-021-02665-3.
The lack of physiologically relevant and predictive cell-based assays is one of the major obstacles for testing and developing botulinum neurotoxins (BoNTs) therapeutics. Human-induced pluripotent stem cells (hiPSCs)-derivatives now offer the opportunity to improve the relevance of cellular models and thus the translational value of preclinical data.
We investigated the potential of hiPSC-derived motor neurons (hMNs) optical stimulation combined with calcium imaging in cocultured muscle cells activity to investigate BoNT-sensitivity of an in vitro model of human muscle-nerve system.
Functional muscle-nerve coculture system was developed using hMNs and human immortalized skeletal muscle cells. Our results demonstrated that hMNs can innervate myotubes and induce contractions and calcium transient in muscle cells, generating an in vitro human motor endplate showing dose-dependent sensitivity to BoNTs intoxication. The implementation of optogenetics combined with live calcium imaging allows to monitor the impact of BoNTs intoxication on synaptic transmission in human motor endplate model.
Altogether, our findings demonstrate the promise of optogenetically hiPSC-derived controlled muscle-nerve system for pharmaceutical BoNTs testing and development.
缺乏与生理相关且具有预测性的基于细胞的检测方法是测试和开发肉毒神经毒素（BoNTs）疗法的主要障碍之一。人诱导多能干细胞（hiPSCs）衍生的细胞现在为提高细胞模型的相关性提供了机会，从而提高了临床前数据的转化价值。
我们研究了 hiPSC 衍生的运动神经元（hMNs）光学刺激与共培养肌肉细胞活性的钙成像相结合，以研究体外人肌肉-神经系统模型中 BoNT 的敏感性。
使用 hMNs 和人永生化骨骼肌细胞开发了功能性肌肉-神经共培养系统。我们的结果表明，hMNs 可以支配肌管并诱导肌肉细胞收缩和钙瞬变，产生体外人运动终板，对 BoNTs 中毒表现出剂量依赖性敏感性。光遗传学与活钙成像的结合实施允许监测 BoNTs 中毒对人运动终板模型中突触传递的影响。
总之，我们的研究结果表明，光遗传学 hiPSC 衍生的受控肌肉-神经系统在药物 BoNTs 测试和开发方面具有广阔的前景。
