Arefin Ayesha, Mendoza Melissa, Dame Keri, Garcia M Iveth, Strauss David G, Ribeiro Alexandre J S
Division of Applied Regulatory Science, Office of Clinical Pharmacology, Office of Translational Sciences, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, United States.
Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, United States.
Front Pharmacol. 2023 Jul 4;14:1212092. doi: 10.3389/fphar.2023.1212092. eCollection 2023.
Engineered heart tissues (EHTs) are three-dimensional culture platforms with cardiomyocytes differentiated from human pluripotent stem cells (hPSCs) and were designed for assaying cardiac contractility. For drug development applications, EHTs must have a stable function and provide reproducible results. We investigated these properties with EHTs made with different tissue casting batches and lines of differentiated hPSC-cardiomyocytes and analyzed them at different times after being fabricated. A video-optical assay was used for measuring EHT contractile outputs, and these results were compared with results from motion traction analysis of beating hPSC-cardiomyocytes cultured as monolayers in two-dimensional cultures. The reproducibility of induced contractile variations was tested using compounds with known mechanistic cardiac effects (isoproterenol, EMD-57033, omecamtiv mecarbil, verapamil, ranolazine, and mavacamten), or known to be clinically cardiotoxic (doxorubicin, sunitinib). These drug-induced variations were characterized at different electrical pacing rates and variations in intracellular calcium transients were also assessed in EHTs. To ensure reproducibility in experiments, we established EHT quality control criteria based on excitation-contraction coupling and contractile sensitivity to extracellular calcium concentration. In summary, a baseline contractile force of 0.2 mN and excitation-contraction coupling of EHTs were used as quality control criteria to select suitable EHTs for analysis. Overall, drug-induced contractile responses were similar between monolayers and EHTs, where a close relationship was observed between contractile output and calcium kinetics. Contractile variations at multiple time points after adding cardiotoxic compounds were also detectable in EHTs. Reproducibility of drug-induced effects in EHTs between experiments and relative to published work on these cellular models was generally observed. Future applications for EHTs may require additional mechanistic criteria related to drug effects and cardiac functional outputs to be measured in regard to specific contexts of use.
工程化心脏组织（EHTs）是一种三维培养平台，其中的心肌细胞由人类多能干细胞（hPSCs）分化而来，旨在用于检测心脏收缩性。对于药物研发应用而言，EHTs必须具备稳定的功能并提供可重复的结果。我们使用不同组织铸型批次和分化的hPSC-心肌细胞系制备的EHTs研究了这些特性，并在制备后的不同时间对其进行分析。采用视频光学测定法测量EHTs的收缩输出，并将这些结果与二维培养中单层培养的跳动hPSC-心肌细胞的运动牵引分析结果进行比较。使用具有已知心脏作用机制的化合物（异丙肾上腺素、EMD-57033、奥米卡替麦卡比、维拉帕米、雷诺嗪和马伐卡坦）或已知具有临床心脏毒性的化合物（阿霉素、舒尼替尼）测试诱导收缩变化的可重复性。在不同的电起搏速率下对这些药物诱导的变化进行表征，并在EHTs中评估细胞内钙瞬变的变化。为确保实验的可重复性，我们基于兴奋-收缩偶联和对细胞外钙浓度的收缩敏感性建立了EHT质量控制标准。总之，将EHTs的基线收缩力0.2 mN和兴奋-收缩偶联用作质量控制标准，以选择合适的EHTs进行分析。总体而言，单层培养物和EHTs之间的药物诱导收缩反应相似，其中在收缩输出和钙动力学之间观察到密切关系。在添加心脏毒性化合物后的多个时间点，EHTs中也可检测到收缩变化。一般观察到EHTs中药物诱导效应在实验之间以及相对于这些细胞模型的已发表研究的可重复性。EHTs的未来应用可能需要与药物效应和心脏功能输出相关的额外机制标准，以便在特定使用背景下进行测量。
