Human-induced pluripotent stem cell-derived cardiomyocytes have limited IKs for repolarization reserve as revealed by specific KCNQ1/KCNE1 blocker.

Abstract

Objective: We investigated if there is I_Ks, and if there is repolarization reserve by I_Ks in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).

Design: We used a specific KCNQ1/KCNE1 channel blocker, L-000768673, with an IC₅₀ of 9 nM, and four hERG-specific blockers, astemizole, cisapride, dofetilide, and E-4031 to investigate the issue.

Results: L-000768673 concentration-dependently prolonged feature point duration (FPD)-a surrogate signal of action potential duration-from 1 to 30 nM without pacing or paced at 1.2 Hz, resulting from I_Ks blockade in hiPSC-CMs. At higher concentrations, the effect of L-000768673 on I_Ks was mitigated by its effect on I_Ca-L, resulting in shortened FPD, reduced impedance amplitude, and increased beating rate at 1 µM and above, recapitulating the self-limiting properties of L-000768673 on action potentials. All four hERG-specific blockers prolonged FPD as expected. Co-application of L-000768673 at sub-threshold (0.1 and 0.3 nM) and threshold (1 nM) concentrations failed to synergistically enhance the effects of hERG blockers on FPD prolongation, rather it showed additive effects, inconsistent with the repolarization reserve role of I_Ks in mature human myocytes that enhanced I_Kr response, implying a difference between hiPSC-CMs used in this study and mature human cardiomyocytes.

Conclusion: There was I_Ks current in hiPSC-CMs, and blockade of I_Ks current caused prolongation of action potential of hiPSC-CMs. However, we could not demonstrate any synergistic effects on action potential duration prolongation of hiPSC-CMs by blocking hERG current and I_Ks current simultaneously, implying little or no repolarization reserve by I_Ks current in hiPSC-CMs used in this study.