Optogenetically controlled human functional motor endplate for testing botulinum neurotoxins

Stem Cell Research & Therapy

Camille Nicoleau¹  Juliette Duchesne de Lamotte²  Léa Lesueur³  Florine Roussange³  Cécile Martinat³  Jérôme Polentes³  Pauline Feurgard³  Anselme Perrier⁴ 

[1] IPSEN Innovation, 5 avenue du Canada, 91940, Les Ulis, France;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;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;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;Laboratoire Des Maladies Neurodégénératives: Mécanismes, thérapies, imagerie, Université Paris Saclay/CEA/CNRS UMR9199, MIRCen, Bâtiment 61, CEA-Fontenay-Aux-Roses, 18 route du Panorama, 92265, Fontenay-aux-Roses, France;

关键词: Human-induced pluripotent stem cells;    Motor endplate;    Functional;    Botulinum neurotoxins;    Calcium indicators;    Optogenetics;   

DOI  :  10.1186/s13287-021-02665-3

来源: Springer

【 摘 要 】

BackgroundThe 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.MethodsWe 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.ResultsFunctional 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.ConclusionsAltogether, our findings demonstrate the promise of optogenetically hiPSC-derived controlled muscle-nerve system for pharmaceutical BoNTs testing and development.

【 授权许可】

CC BY