Development of In Vitro Drug-Induced Cardiotoxicity Assay by Using Three-Dimensional Cardiac Tissues Derived from Human Induced Pluripotent Stem Cells.
Development of In Vitro Drug-Induced Cardiotoxicity Assay by Using Three-Dimensional Cardiac Tissues Derived from Human Induced Pluripotent Stem Cells.
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DOI：
10.1089/ten.tec.2017.0247
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发表时间：
2018-01
期刊：
影响因子：
--
通讯作者：
Sawa Y
中科院分区：
文献类型：
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作者：
Takeda M;Miyagawa S;Fukushima S;Saito A;Ito E;Harada A;Matsuura R;Iseoka H;Sougawa N;Mochizuki-Oda N;Matsusaki M;Akashi M;Sawa Y
体外药物诱导心脏毒性试验是药物发现和临床应用的关键步骤。人类诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）在这方面很有前景。然而，单个hiPSC-CMs在结构和功能上模仿天然心脏组织的能力有限，使用hiPSC-CMs生成人工心脏组织是一个持续的挑战。因此，我们开发了一种通过在细胞表面涂覆细胞外基质（ECM）成分在短时间内构建三维（3D）人工组织的新方法。我们假设hiPSC-CMs （3D- hipsc - ct）衍生的3D心脏组织可用于体外药物诱导的心脏毒性试验。3D-hiPSC-CT是由纤维连接蛋白和明胶纳米膜包被单个hiPSC-CMs生成的。组织学上，3D-hiPSC-CT显示肌细胞和ECM蛋白（如纤维连接蛋白、I/III型胶原蛋白和层粘连蛋白）中有肌瘤结构。5.0 μM剂量的阿霉素可诱导乳酸脱氢酶的释放，2.0 μM剂量的阿霉素可降低细胞活力。E-4031、hERG型钾通道阻滞剂和异丙肾上腺素均诱导Ca瞬态参数和收缩参数发生显著变化，且呈剂量依赖性。3D-hiPSC-CT在体外表现出对阿霉素敏感的细胞毒性和对hERG通道阻断剂/异丙肾上腺素敏感的电活性，表明其在药物诱导的心脏毒性试验或药物筛选系统中用于药物发现。
An in vitro drug-induced cardiotoxicity assay is a critical step in drug discovery for clinical use. The use of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is promising for this purpose. However, single hiPSC-CMs are limited in their ability to mimic native cardiac tissue structurally and functionally, and the generation of artificial cardiac tissue using hiPSC-CMs is an ongoing challenging. We therefore developed a new method of constructing three-dimensional (3D) artificial tissues in a short time by coating extracellular matrix (ECM) components on cell surfaces. We hypothesized that 3D cardiac tissues derived from hiPSC-CMs (3D-hiPSC-CT) could be used for an in vitro drug-induced cardiotoxicity assay. 3D-hiPSC-CT were generated by fibronectin and gelatin nanofilm coated single hiPSC-CMs. Histologically, 3D-hiPSC-CT exhibited a sarcomere structure in the myocytes and ECM proteins, such as fibronectin, collagen type I/III, and laminin. The administration of cytotoxic doxorubicin at 5.0 μM induced the release of lactate dehydrogenase, while that at 2.0 μM reduced the cell viability. E-4031, human ether-a-go-go related gene (hERG)-type potassium channel blocker, and isoproterenol induced significant changes both in the Ca transient parameters and contractile parameters in a dose-dependent manner. The 3D-hiPSC-CT exhibited doxorubicin-sensitive cytotoxicity and hERG channel blocker/isoproterenol-sensitive electrical activity in vitro, indicating its usefulness for drug-induced cardiotoxicity assays or drug screening systems for drug discovery.