Disrupted Ca_V1.2 selectivity causes overlapping long QT and Brugada syndrome phenotypes in the CACNA1C-E1115K iPS cell model

Background: A missense mutation in the α1c subunit of voltage-gated L-type Ca²⁺ channel-coding CACNA1C-E1115K, located in the Ca²⁺ selectivity site, causes a variety of arrhythmogenic phenotypes.

Objective: We aimed to investigate the electrophysiological features and pathophysiological mechanisms of CACNA1C-E1115K in patient-specific induced pluripotent stem cell (iPSC)-derived cardiomyocytes (CMs).

Methods: We generated iPSCs from a patient carrying heterozygous CACNA1C-E1115K with overlapping phenotypes of long QT syndrome, Brugada syndrome, and mild cardiac dysfunction. Electrophysiological properties were investigated using iPSC-CMs. We used iPSCs from a healthy individual and an isogenic iPSC line corrected using CRISPR-Cas9-mediated gene editing as controls. A mathematical E1115K-CM model was developed using a human ventricular cell model.

Results: Patch-clamp analysis revealed that E1115K-iPSC-CMs exhibited reduced peak Ca²⁺ current density and impaired Ca²⁺ selectivity with an increased permeability to monovalent cations. Consequently, E1115K-iPSC-CMs showed decreased action potential plateau amplitude, longer action potential duration (APD), and a higher frequency of early afterdepolarization compared with controls. In optical recordings examining the antiarrhythmic drug effect, late Na⁺ channel current (I_NaL) inhibitors (mexiletine and GS-458967) shortened APDs specifically in E1115K-iPSC-CMs. The AP-clamp using a voltage command obtained from E1115K-iPSC-CMs with lower action potential plateau amplitude and longer APD confirmed the upregulation of I_NaL. An in silico study recapitulated the in vitro electrophysiological properties.

Conclusion: Our iPSC-based analysis in CACNA1C-E1115K with disrupted Ca_V1.2 selectivity demonstrated that the aberrant currents through the mutant channels carried by monovalent cations resulted in specific action potential changes, which increased endogenous I_NaL, thereby synergistically contributing to the arrhythmogenic phenotype.