Pharmacological enhancement of repolarization reserve in human induced pluripotent stem cells derived cardiomyocytes

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are used for many applications including safety pharmacology. However, a deficiency or complete absence of several K⁺ currents suggests repolarization reserve is low in hiPSC-CMs. We determined whether a dual I_to and I_Kr activator can improve repolarization reserve in hiPSC-CMs resulting in a more electrophysiologically mature phenotype.

Human iPSC were maintained on growth factor and differentiated into the cardiac phenotype by addition of selective Wnt molecules. Current and voltage clamp recordings in single cells were made using patch electrodes. Extracellular field potentials were made using a microelectrode array on hiPSC monolayers. Action potential recordings from hiPSC-CMs following application of an I_Kr inhibitor resulted in depolarization of the membrane potential and prolongation of the APD. A flattening of the T-wave was noted on the pseudo-ECG. In contrast, application of the I_Kr and I_to agonist, NS3623, resulted in hyperpolarization of the membrane, slowing of the spontaneous rate and shortening of the APD. Voltage clamp recording showed a significant increase in I_Kr; no enhancement of I_to in hiPSC-CMs was noted. AP clamp experiments revealed that I_Kr plays a role in both phase 3 repolarization and phase 4 depolarization. mRNA analysis revealed that KCNH2 is abundantly expressed in hiPSC-CM, consistent with electrophysiological recordings.

Although NS3623 is a dual I_to and I_Kr activator in ventricular myocytes, application of this compound to hiPSC-CMs enhanced only I_Kr and no effect on I_to was noted. Our results suggest I_Kr enhancement can improve repolarization reserve in this cell type. The disconnect between a dramatic increase in I_to in adult myocytes versus the lack of effect in hiPSC-CMs suggest that the translation of pharmacological effects in hiPSC-CM to adult myocytes should be viewed with caution.