Human Atrial Cardiac Microtissues for Chamber-Specific Arrhythmic Risk Assessment
Human Atrial Cardiac Microtissues for Chamber-Specific Arrhythmic Risk Assessment
复制标题
用于心室特异性心律失常风险评估的人心房心脏微组织
DOI：
10.1007/s12195-021-00703-x
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发表时间：
2021
影响因子：
2.8
通讯作者：
Coulombe, Kareen L.
中科院分区：
文献类型：
--
作者：
Soepriatna, Arvin H.;Kim, Tae Yun;Daley, Mark C.;Song, Elena;Choi, Bum-Rak;Coulombe, Kareen L.
前言虽然房颤是最常见的电传导障碍，但房性心律失常背后的机制仍不清楚。为了应对这一挑战，我们从人诱导的多能干细胞(HiPSC)来源的心肌细胞中建立了三维心房微组织的robustin体外模型，并通过实验和计算评估了房间特异性的化学反应。方法我们从表达GCaMP6f的hiPSC中区分出心房和心室心肌细胞(ACM/VCM)，并使用荧光成像技术评估自发AP的活性。用乳酸纯化的CMS和5%的人心脏成纤维细胞形成自组装的三维微组织，电刺激一周后进行高分辨动作电位(AP)光学标测。AP对心房特异性钾复极电流IKur受体阻滞剂4-氨基吡啶(4-AP)和有趣的If受体阻滞剂伊夫拉定的反应在其治疗窗内进行了表征。结果高纯度CMS(>；75%cTnT+)通过MLC2v的表达证明了亚型的特异性。3D微组织形成后，自发性搏动频率显著降低，心房微组织自发性搏动频率较快，AP上升时间较慢，AP时程较短。我们测量了心房特异性反应，包括4-AP治疗后剂量依赖性的时程延长和伊夫拉定治疗后自发活动的剂量依赖性减少。结论本实验建立的筛选心房特异性反应的体外平台既可靠又灵敏，具有高通量，使研究集中于阐明房性心律失常的机制。
IntroductionAlthough atrial fibrillation is the most prevalent disorder of electrical conduction, the mechanisms behind atrial arrhythmias remain elusive. To address this challenge, we developed a robustin vitromodel of 3D atrial microtissue from human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and evaluated chamber-specific chemical responses experimentally and computationally.MethodsWe differentiated atrial and ventricular cardiomyocytes (aCMs/vCMs) from GCaMP6f-expressing hiPSCs and assessed spontaneous AP activity using fluorescence imaging. Self-assembling 3D microtissues were formed with lactate purified CMs and 5% human cardiac fibroblasts and electrically stimulated for one week before high resolution action potential (AP) optical mapping. AP responses to the atrial-specific potassium repolarizing currentIKur-blocker 4-Aminopyridine (4-AP) and funny currentIf-blocker Ivabradine were characterized within their therapeutic window. Finally, we expanded upon a published hiPSC-CM computational model by incorporating the atrial-specificIKurcurrent, modifying ion channel conductances to match the AP waveforms of our microtissues, and employing the updated model to reinforce our experimental findings.ResultsHigh purity CMs (> 75% cTnT+) demonstrated subtype specification by MLC2v expression. Spontaneous beating rates significantly decreased following 3D microtissue formation, with atrial microtissues characterized by their faster spontaneous beating rate, slower AP rise time, and shorter AP duration (APD) compared to ventricular microtissues. We measured atrial-specific responses, including dose-dependent APD prolongation with 4-AP treatment and dose-dependent reduction in spontaneous activity post-Ivabradine treatment.ConclusionThe presentedin vitroplatform for screening atrial-specific responses is both robust and sensitive, with high throughput, enabling studies focused at elucidating the mechanisms underlying atrial arrhythmias.
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影响因子：
5
作者：
Kim, Jong J.;Yang, Lei;Lin, Bo;Zhu, Xiaodong;Sun, Bin;Kaplan, Aaron D.;Bett, Glenna C. L.;Rasmusson, Randall L.;London, Barry;Salama, Guy
通讯作者：
Salama, Guy
影响因子：
5
作者：
Walden, A. P.;Dibb, K. M.;Trafford, A. W.
通讯作者：
Trafford, A. W.
影响因子：
3.4
作者：
Xie, Yuanfang;Sato, Daisuke;Weiss, James N.
通讯作者：
Weiss, James N.
伊伐布雷定和房颤：巧合、相关还是新的治疗方法？
影响因子：
3
作者：
M. Abdelnabi;Ashraf Ahmed;A. Almaghraby;Y. Saleh;H. Badran
通讯作者：
H. Badran
心脏成纤维细胞
DOI：
--
发表时间：
2018
期刊：
Herzschrittmachertherapie & Elektrophysiologie
影响因子：
--
作者：
A. Klesen;D. Jakob;R. Emig;P. Kohl;U. Ravens;R. Peyronnet
通讯作者：
R. Peyronnet