Light-induced termination of spiral wave arrhythmias by optogenetic engineering of atrial cardiomyocytes
Light-induced termination of spiral wave arrhythmias by optogenetic engineering of atrial cardiomyocytes
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DOI：
10.1093/cvr/cvu179
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发表时间：
2014-10-01
影响因子：
10.8
通讯作者：
de Vries, Antoine A. F.
中科院分区：
文献类型：
--
作者：
Bingen, Brian O.;Engels, Marc C.;de Vries, Antoine A. F.
目的房颤（AF）是最常见的心律失常，常涉及再入性电激活（如螺旋波）。房颤的药物治疗可能有严重的副作用，包括心律失常，而电击治疗与不适和组织损伤有关。假设，心肌细胞中光门控阳离子通道的强制表达和随后的激活可能提供足以除颤的去极化力，从而规避上述缺点。因此，我们通过心脏光遗传学研究了光诱导螺旋波终止的可行性。方法和结果用编码光激活Ca2+易位通道视紫红质（CatCh； LV）的慢病毒载体转染新生大鼠心房心肌细胞单层。CatCh类似于eYFP向上箭头)或eYFP （LV）。eYFP向上箭头)作为控制，并爆发节奏，以诱导螺旋波围绕功能核心旋转。通过光学和多电极阵列（MEA）测绘研究了CatCh激活对再入的影响。Western blot分析和免疫细胞学证实转基因表达。短暂的蓝光脉冲（10 ms/470 nm）仅在左心室触发动作电位。CatCh类似于eYFP向上的箭头转导培养，证实了功能性CatCh介导的电流。延长的光脉冲（500毫秒）导致100%的LV再入终止。捕获类似于eYFP上箭头转导培养（n = 31），而LV为0%。eYFP向上箭头转导培养（n = 11）。在这里，CatCh激活引起均匀去极化，从而降低了整体兴奋性（MEA峰对峰振幅下降251.3 +/- 217.1，而对照组为9.2 +/- 9.5 μ V）。因此，功能核尺寸增加，相位奇点（ps）漂移，导致PS-PS或ps -边界碰撞导致再入终止。结论本研究表明，在光基因工程的基础上，由致心律失常底物本身产生的光诱导去极化电流可以有效地终止心房心肌细胞单层的螺旋波。这些结果为无冲击除颤提供了概念证明。
Aims Atrial fibrillation (AF) is the most common cardiac arrhythmia and often involves reentrant electrical activation (e. g. spiral waves). Drug therapy for AF can have serious side effects including proarrhythmia, while electrical shock therapy is associated with discomfort and tissue damage. Hypothetically, forced expression and subsequent activation of light-gated cation channels in cardiomyocytes might deliver a depolarizing force sufficient for defibrillation, thereby circumventing the aforementioned drawbacks. We therefore investigated the feasibility of light-induced spiral wave termination through cardiac optogenetics.Methods and results Neonatal rat atrial cardiomyocyte monolayers were transduced with lentiviral vectors encoding light-activated Ca2+-translocating channelrhodopsin (CatCh; LV.CatCh similar to eYFP up arrow) or eYFP (LV.eYFP up arrow) as control, and burst-paced to induce spiral waves rotating around functional cores. Effects of CatCh activation on reentry were investigated by optical and multi-electrode array (MEA) mapping. Western blot analyses and immunocytology confirmed transgene expression. Brief blue light pulses (10 ms/470 nm) triggered action potentials only in LV.CatCh similar to eYFP up arrow-transduced cultures, confirming functional CatCh-mediated current. Prolonged light pulses (500 ms) resulted in reentry termination in 100% of LV.CatCh similar to eYFP up arrow-transduced cultures (n = 31) vs. 0% of LV.eYFP up arrow-transduced cultures (n = 11). Here, CatCh activation caused uniform depolarization, thereby decreasing overall excitability (MEA peak-to-peak amplitude decreased 251.3 +/- 217.1 vs. 9.2 +/- 9.5 mu V in controls). Consequently, functional coresize increased and phase singularities (PSs) drifted, leading to reentry termination by PS-PS or PS-boundary collisions.Conclusion This study shows that spiral waves in atrial cardiomyocyte monolayers can be terminated effectively by a light-induced depolarizing current, produced by the arrhythmogenic substrate itself, upon optogenetic engineering. These results provide proof-of-concept for shockless defibrillation.