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Background Optogenetics, using light-sensitive proteins, emerged as a unique experimental paradigm to modulate cardiac excitability. We aimed to develop high-resolution optogenetic approaches to modulate electrical activity in 2- and 3-dimensional cardiac tissue models derived from human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes. Methods and Results To establish light-controllable cardiac tissue models, opsin-carrying HEK293 cells, expressing the light-sensitive cationic-channel CoChR, were mixed with hiPSC-cardiomyocytes to generate 2-dimensional hiPSC-derived cardiac cell-sheets or 3-dimensional engineered heart tissues. Complex illumination patterns were designed with a high-resolution digital micro-mirror device. Optical mapping and force measurements were used to evaluate the tissues' electromechanical properties. The ability to optogenetically pace and shape the tissue's conduction properties was demonstrated by using single or multiple illumination stimulation sites, complex illumination patterns, or diffuse illumination. This allowed to establish in vitro models for optogenetic-based cardiac resynchronization therapy, where the electrical activation could be synchronized (hiPSC-derived cardiac cell-sheets and engineered heart tissue models) and contractile properties improved (engineered heart tissues). Next, reentrant activity (rotors) was induced in the hiPSC-derived cardiac cell-sheets and engineered heart tissue models through optogenetics programmed- or cross-field stimulations. Diffuse illumination protocols were then used to terminate arrhythmias, demonstrating the potential to study optogenetics cardioversion mechanisms and to identify optimal illumination parameters for arrhythmia termination. Conclusions By combining optogenetics and hiPSC technologies, light-controllable human cardiac tissue models could be established, in which tissue excitability can be modulated in a functional, reversible, and localized manner. This approach may bring a unique value for physiological/pathophysiological studies, for disease modeling, and for developing optogenetic-based cardiac pacing, resynchronization, and defibrillation approaches.

背景

光遗传学利用光敏感蛋白，成为一种独特的实验范例，可调节心脏兴奋性。我们旨在开发高分辨率的光遗传学方法，以调节源自人诱导多能干细胞（hiPSC）衍生的心肌细胞的 2 维和 3 维心脏组织模型中的电活动。

方法和结果

为了建立光可控的心脏组织模型，携带 opsin 的 HEK293 细胞表达光敏感阳离子通道 CoChR，与 hiPSC 心肌细胞混合，生成 2 维 hiPSC 衍生的心脏细胞片或 3 维工程心脏组织。使用高分辨率数字微镜设备设计复杂的照明模式。光学映射和力测量用于评估组织的机电特性。通过使用单个或多个照明刺激点、复杂的照明模式或漫射照明，证明了光遗传学起搏和塑造组织传导特性的能力。这允许建立基于光遗传学的心脏再同步治疗的体外模型，其中电激活可以被同步（hiPSC 衍生的心脏细胞片和工程心脏组织模型），并且收缩性能得到改善（工程心脏组织）。接下来，通过光遗传学程控或交叉场刺激在 hiPSC 衍生的心脏细胞片和工程心脏组织模型中诱导折返活动（转子）。然后使用漫射照明方案终止心律失常，证明了研究光遗传学转复机制和确定心律失常终止的最佳照明参数的潜力。