Abstract 15483: Building a New Drug Screening System for Evaluating Drug Response and Toxicity by Three Dimensional Cardiac Constructs Derived From Human Induced Pluripotent Stem Cells
Background: The establishment of innovative drug screening system may be crucial for the development of new therapeutic agents in terms of costs and acceleration in the pace of drug discovery. Although recent studies indicate human induced pluripotent stem cells (hiPSc) may have some potentials for drug screening in vitro, it may be unclear whether cardiomyocytes cultured in three-dimensional (3D) environment show physiological or electrical behaviors, and a response to cardiovascular agents like native cardiac tissue, which superior to mono-layered cardiac construct. In this study, we examined 3D cardiac tissue derived from hiPSc (3D-hiPSc-CT) could show drug induced cardiotoxity and response to cardiovascular agents and compared with those of the two-dimensional cardiac tissues (2D-hiPSc-CT).
Methods and Results: For construction of 3D-hiPSc-CT, fibronectin and gelatin nanofilms were coated over single hiPSc-CMs and cells were seeded in 96well plate. We examined the toxicity of doxorubicin by LDH and CKK8 assay and the proarrhythmic effect of E-4031, hERG type potassium channel blocker, by cytoplasmic Ca2+ transient. Addition of cytotoxic doxorubicin at 1uM in 2D-hiPSc-CT induced release of LDH (1.33-fold increase) and reduction of cell viability (0.60-fold) (P<0.01 vs vehicle control), while in 3D-hiPSc-CT did not show significant change. Instead, that at 5uM in 3D-hiPSc-CT induced release of LDH (2.25-fold) and reduction of cell viability (0.77-fold) (P<0.01 vs vehicle control). Addition of E-4031 at 10nM in both of 2D and 3D-hiPSc-CT induced significant proarrhythmic responses, with a decrease in falling slope (2D:0.92-fold, 3D:0.79-fold) (P<0.01 vs vehicle control) and an increase in peak width duration of Ca2+ transient (2D:1.06-fold, 3D:1.25-fold) (P<0.01 vs vehicle control). However, 2D-hiPSc-CT showed those proarrhythmic effects till 60 minutes after adding E4031, while 3D-hiPSc-CT maintained them continuously.
Conclusion: The 3D-hiPSc-CT have great potential for in vitro drug screening with possible superiority to 2D-hiPSc-CT, suggesting that 3D-hiPSc-CT may play crucial roles for drug repositioning or development in new cardiovascular agents.
Author Disclosures: M. Takeda: None. S. Miyagawa: None. S. Fukushima: None. A. Saito: None. E. Ito: None. A. Harada: None. R. Matsuura: None. Y. Matsunaga: None. N. Mochizuki-Oda: None. M. Matsusaki: None. M. Akashi: None. Y. Sawa: None.
- © 2016 by American Heart Association, Inc.