Blinded Contractility Analysis in hiPSC-Cardiomyocytes in Engineered Heart Tissue Format: Comparison With Human Atrial Trabeculae
Blinded Contractility Analysis in hiPSC-Cardiomyocytes in Engineered Heart Tissue Format: Comparison With Human Atrial Trabeculae
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DOI：
10.1093/toxsci/kfx081
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发表时间：
2017-07-01
影响因子：
3.8
通讯作者：
Hansen, Arne
中科院分区：
文献类型：
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作者：
Mannhardt, Ingra;Eder, Alexandra;Hansen, Arne
人诱导的多能干细胞衍生的心肌细胞（hiPSC-CM）可以作为一种新的检测方法，在人类的背景下，药物测试，但其有效性，特别是对正性肌力药物的作用的评价仍然不清楚。在该盲法分析中，我们比较了10种指示剂化合物在电刺激（1.5 Hz）hiPSC-CM衍生的三维工程心脏组织（EHT）和人心房小梁（hAT）中的作用与已知的变力作用。从在常规心脏手术中获得的iCell hiPSC-CM、hAT制备人EHT。基线力测量的平均批内变异系数为17%（EHT）和49%（hAT）。PDE抑制剂米力农不影响EHT收缩力，但增加了hAT的收缩力。西酞普兰（选择性5-羟色胺再摄取抑制剂）、硝苯地平（LTCC阻断剂）和利多卡因（Na+通道阻断剂）对EHT和hAT具有负性肌力作用。福莫特罗（β-2激动剂）在EHT中具有正性肌力作用，但无正性肌力作用，在hAT中具有正性斜向、正性肌力和正性肌力作用。他克莫司（钙调神经磷酸酶抑制剂）对EHT具有负性肌力作用，但对hAT没有影响。地高辛（Na+-K+-ATP酶抑制剂）仅在EHT中显示正性肌力作用，但在hAT中无作用，可能是由于孵育时间短。Ryanodine（Ryanodine受体抑制剂）降低了两种模型的收缩力。咯利普兰和乙酰水杨酸在hAT中显示不可解释的结果。hiPSC-EHT的收缩幅度和动力学随时间推移更稳定，变化小于hAT。HiPSC-EHT忠实地检测到cAMP依赖性和非依赖性正性和负性肌力作用，但有限的β-2肾上腺素能或PDE 3作用，与未成熟CM表型相容。
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) may serve as a new assay for drug testing in a human context, but their validity particularly for the evaluation of inotropic drug effects remains unclear. In this blinded analysis, we compared the effects of 10 indicator compounds with known inotropic effects in electrically stimulated (1.5 Hz) hiPSC-CM-derived 3-dimensional engineered heart tissue (EHT) and human atrial trabeculae (hAT). Human EHTs were prepared from iCell hiPSC-CM, hAT obtained at routine heart surgery. Mean intra-batch variation coefficient in baseline force measurement was 17% for EHT and 49% for hAT. The PDE-inhibitor milrinone did not affect EHT contraction force, but increased force in hAT. Citalopram (selective serotonin reuptake inhibitor), nifedipine (LTCC-blocker) and lidocaine (Na+ channel-blocker) had negative inotropic effects on EHT and hAT. Formoterol (beta-2 agonist) had positive lusitropic but no inotropic effect in EHT, and positive clinotropic, lusitropic, and inotropic effects in hAT. Tacrolimus (calcineurin-inhibitor) had a negative inotropic effect in EHTs, but no effect in hAT. Digoxin (Na+-K+-ATPase-inhibitor) showed a positive inotropic effect only in EHTs, but no effect in hAT probably due to short incubation time. Ryanodine (ryanodine receptor-inhibitor) reduced contraction force in both models. Rolipram and acetylsalicylic acid showed noninterpretable results in hAT. Contraction amplitude and kinetics were more stable over time and less variable in hiPSC-EHTs than hAT. HiPSC-EHT faithfully detected cAMP-dependent and -independent positive and negative inotropic effects, but limited beta-2 adrenergic or PDE3 effects, compatible with an immature CM phenotype.