Stimulating cardiac muscle by light: cardiac optogenetics by cell delivery.
Stimulating cardiac muscle by light: cardiac optogenetics by cell delivery.
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DOI：
10.1161/circep.111.964247
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发表时间：
2011-10
期刊：
影响因子：
--
通讯作者：
Entcheva E
中科院分区：
文献类型：
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作者：
Jia Z;Valiunas V;Lu Z;Bien H;Liu H;Wang HZ;Rosati B;Brink PR;Cohen IS;Entcheva E
在最近克隆了光敏离子通道并在哺乳动物细胞中表达之后，神经科学中出现了一个新的领域，即光遗传学，允许光对神经回路进行精确扰动。然而，光遗传学工具的功能尚未在神经科学之外得到充分探索;并且之前尚未显示出用于光遗传学的非病毒、非胚胎发生的策略。我们通过串联细胞单位（TCU）策略证明了光遗传学对心肌的效用，其中非兴奋细胞携带外源性光敏离子通道，并且当电耦合到心肌细胞时，产生光兴奋的心脏组织。开发稳定的通道视紫红质2（ChR 2）表达细胞系，表征并用作细胞递送系统。TCU策略在体外与成年犬肌细胞的细胞对（广泛的耦合强度）和与新生大鼠心肌细胞的心脏合胞体中进行了验证。这是第一次，我们结合了光学激发和光学成像来捕获光触发的肌肉收缩和光触发电波的高分辨率传播图，发现它们与电触发波在数量上无法区分。我们的研究结果证明了使用TCU方法通过光来控制心肌的兴奋和收缩的可行性。在这种情况下，光学起搏使用更少的能量，提供上级时空控制，远程访问，不仅可以作为心律失常研究中的一个优雅的工具，但可能形成新一代的光驱动心脏起搏器和肌肉致动器的基础。TCU策略可扩展到（非病毒）干细胞治疗，并与体内应用直接相关。
After the recent cloning of light-sensitive ion channels and their expression in mammalian cells, a new field, optogenetics, emerged in neuroscience, allowing for precise perturbations of neural circuits by light. However, functionality of optogenetic tools has not been fully explored outside neuroscience; and a non-viral, non-embryogenesis based strategy for optogenetics has not been shown before. We demonstrate the utility of optogenetics to cardiac muscle by a tandem cell unit (TCU) strategy, where non-excitable cells carry exogenous light-sensitive ion channels, and when electrically coupled to cardiomyocytes, produce optically-excitable heart tissue. A stable channelrhodopsin2 (ChR2) expressing cell line was developed, characterized and used as a cell delivery system. The TCU strategy was validated in vitro in cell pairs with adult canine myocytes (for a wide range of coupling strengths) and in cardiac syncytium with neonatal rat cardiomyocytes. For the first time, we combined optical excitation and optical imaging to capture light-triggered muscle contractions and high-resolution propagation maps of light-triggered electrical waves, found to be quantitatively indistinguishable from electrically-triggered waves. Our results demonstrate feasibility to control excitation and contraction in cardiac muscle by light using the TCU approach. Optical pacing in this case uses less energy, offers superior spatiotemporal control, remote access and can serve not only as an elegant tool in arrhythmia research, but may form the basis for a new generation of light-driven cardiac pacemakers and muscle actuators. The TCU strategy is extendable to (non-viral) stem cell therapy and is directly relevant to in vivo applications.