Induction of cardiac alternans in human iPS-derived cardiomyocytes through β-adrenergic receptor stimulation

Physiol Rep. 2024 Dec;12(24):e70152. doi: 10.14814/phy2.70152.

Abstract

Cardiac alternans (C-ALT) is a phenomenon of alternating strong and weak contractions in the heart and is considered a risk factor for the development of heart failure and arrhythmias. However, no model has been reported that can induce C-ALT in vitro using human cells, and the developmental mechanism of C-ALT has not been studied using human cells. In this study, we successfully induced C-ALT in vitro using human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). By stimulating β-adrenergic receptor with isoproterenol on hiPSC-CMs cultured in atmospheric condition (with ~0.04% CO2), contractility and calcium transient were observed to alternately increase and decrease with each beat. In contrast, C-ALT was not induced in hiPSC-CMs cultured at 5% CO2 concentration. Since previous studies have linked C-ALT to problems with calcium regulation in the sarcoplasmic reticulum (SR), we exposed hiPSC-CMs to compounds that alter SR Ca2+ loading and analyzed their contractile responses. The results showed that exposure to verapamil, thapsigargin, and ryanodine either suppressed or eliminated C-ALT. In contrast, omecamtiv mecarbil and blebbistatin, which alter contractility without SR Ca2+ loading, did not induce or suppress C-ALT. These results suggest that C-ALT in hiPSC-CMs induced by isoproterenol may be due to abnormal regulation of the ryanodine receptor's opening and closing caused by excessive Ca2+ load in the SR from β-adrenergic receptor stimulation.

Keywords: calcium alternans; cardiac alternans; cardiac contractility; cell sheet technology; human‐induced pluripotent stem cell‐derived cardiomyocytes.

MeSH terms

  • Adrenergic beta-Agonists* / pharmacology
  • Arrhythmias, Cardiac / chemically induced
  • Arrhythmias, Cardiac / metabolism
  • Arrhythmias, Cardiac / physiopathology
  • Calcium / metabolism
  • Calcium Signaling / drug effects
  • Cells, Cultured
  • Humans
  • Induced Pluripotent Stem Cells* / cytology
  • Induced Pluripotent Stem Cells* / drug effects
  • Induced Pluripotent Stem Cells* / metabolism
  • Isoproterenol* / pharmacology
  • Myocardial Contraction / drug effects
  • Myocytes, Cardiac* / drug effects
  • Myocytes, Cardiac* / metabolism
  • Myocytes, Cardiac* / physiology
  • Receptors, Adrenergic, beta* / metabolism
  • Sarcoplasmic Reticulum / drug effects
  • Sarcoplasmic Reticulum / metabolism

Substances

  • Receptors, Adrenergic, beta
  • Isoproterenol
  • Adrenergic beta-Agonists
  • Calcium