Vurro Vito, Federici Beatrice, Ronchi Carlotta, Florindi Chiara, Sesti Valentina, Crasto Silvia, Maniezzi Claudia, Galli Camilla, Antognazza Maria Rosa, Bertarelli Chiara, Di Pasquale Elisa, Lanzani Guglielmo, Lodola Francesco
Center for Nano Science and Technology @PoliMi, Istituto Italiano di Tecnologia, Via Raffaele Rubattino, 81, 20134 Milan, Italy.
Department of Biotechnology and Biosciences, University of Milan-Bicocca, Building U3 - BIOS, Piazza della Scienza, 2, 20126 Milan, Italy.
iScience. 2023 Feb 2;26(3):106121. doi: 10.1016/j.isci.2023.106121. eCollection 2023 Mar 17.
Non-genetic photostimulation is a novel and rapidly growing multidisciplinary field that aims to induce light-sensitivity in living systems by exploiting exogeneous phototransducers. Here, we propose an intramembrane photoswitch, based on an azobenzene derivative (Ziapin2), for optical pacing of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The light-mediated stimulation process has been studied by applying several techniques to detect the effect on the cell properties. In particular, we recorded changes in membrane capacitance, in membrane potential (V), and modulation of intracellular Ca dynamics. Finally, cell contractility was analyzed using a custom MATLAB algorithm. Photostimulation of intramembrane Ziapin2 causes a transient V hyperpolarization followed by a delayed depolarization and action potential firing. The observed initial electrical modulation nicely correlates with changes in Ca dynamics and contraction rate. This work represents the proof of principle that Ziapin2 can modulate electrical activity and contractility in hiPSC-CMs, opening up a future development in cardiac physiology.
非基因光刺激是一个新兴且发展迅速的多学科领域，旨在通过利用外源光传感器诱导生物系统产生光敏感性。在此，我们提出一种基于偶氮苯衍生物（Ziapin2）的膜内光开关，用于对人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）进行光学起搏。通过应用多种技术来检测对细胞特性的影响，研究了光介导的刺激过程。特别地，我们记录了膜电容、膜电位（V）的变化以及细胞内钙动力学的调制。最后，使用定制的MATLAB算法分析细胞收缩性。膜内Ziapin2的光刺激导致V短暂超极化，随后是延迟去极化和动作电位发放。观察到的初始电调制与钙动力学和收缩率的变化密切相关。这项工作证明了Ziapin2可以调节hiPSC-CMs中的电活动和收缩性，为心脏生理学的未来发展开辟了道路。
