Effects of Adrenaline on Beat-to-Beat Variability of hiPSC-CMs with Hypertrophic Cardiomyopathy

Venla Koivunen, Ville Ali-Hokka, Chandra Prajapati, Teemu Pukkila, Katriina Aalto-Setälä, Esa Rasanen
Tampere University


Abstract

Hypertrophic cardiomyopathy (HCM) is one of the major causes of sudden cardiac death in young individuals, often triggered by adrenergic stress. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offer an in vitro platform for investigating patient-specific electrophysiology. Here, we studied how adrenaline affects beat-to-beat variability (B2BV) in spontaneously beating hiPSC-CMs with heart rate variability (HRV)–based metrics in cells carrying HCM-related mutations.

The dataset consists of different hiPSC-CM lines: two control lines, two lines with a MYBPC3-Gln1061X mutation (HCMM), and three lines with a TPM1-Asp175Asn mutation (HCMT). Previous studies have shown that hiPSC-CMs carrying HCMM or HCMT mutations exhibit, respectively, a higher incidence of arrhythmias. Interbeat intervals were extracted from patch-clamp recordings, preprocessed, and analyzed across baseline (BL), adrenaline (ADR), and washout (WO) phases. Rhythm dynamics were characterized using a range of HRV methods, with a focus on time-domain and nonlinear metrics.

Adrenaline induced mutation‑dependent changes in rhythm variability. As shown in Fig. 1A-B, both mean RR and RMSSD were altered, with high adrenaline concentrations producing effects that persisted into the washout phase, suggesting incomplete recovery of spontaneous rhythm regulation. Δ-values showed a similar pattern (Fig. 1C-D), whereas measures such as DFA α1 showed no genotype-dependent differences. Overall, the findings demonstrate genotype-specific modulation of beat-to-beat variability under adrenergic stimulation, suggesting that HRV-based analysis of hiPSC-CMs may provide useful insights into arrhythmia-related cellular phenotypes in HCM.