Long QT syndrome caveolin‐3 mutations differentially modulate K_v4 and Ca_v1.2 channels to contribute to action potential prolongation

Abstract

Mutations in the CAV3 gene encoding caveolin-3 (Cav3), a scaffolding protein integral to caveolae in cardiomyocytes, have been associated with the congenital long-QT syndrome (LQT9). Initial studies demonstrated that LQT9-associated Cav3 mutations, F97C and S141R, increase late sodium current as a potential mechanism to prolong action potential duration (APD) and cause LQT9. Whether these Cav3 LQT9 mutations impact other caveolae related ion channels remains unknown. We used the whole-cell, patch clamp technique to characterize the effect of Cav3-F97C and Cav3-S141R mutations on heterologously expressed Ca_v 1.2+Ca_v β_2cN4 channels, as well as K_v 4.2 and K_v 4.3 channels, in HEK 293 cells. Expression of Cav3-S141R increased I_Ca,L density without changes in gating properties, whereas expression of Cav3-F97C reduced Ca²⁺ -dependent inactivation of I_Ca,L without changing current density. The Cav3-F97C mutation reduced current density and altered the kinetics of I_Kv4.2 and I_Kv4.3 and also slowed recovery from inactivation. Cav3-S141R decreased current density and also slowed activation kinetics and recovery from inactivation of I_Kv4.2 but had no effect on I_Kv4.3 . Using the O'Hara-Rudy computational model of the human ventricular myocyte action potential, the Cav3 mutation-induced changes in I_to are predicted to have negligible effect on APD, whereas blunted Ca²⁺ -dependent inactivation of I_Ca,L by Cav3-F97C is predicted to be primarily responsible for APD prolongation, although increased I_Ca,L and late I_Na by Cav3-S141R contribute equally to APD prolongation. Thus, LQT9 Cav3-associated mutations, F97C and S141R, produce mutation-specific changes in multiple ionic currents leading to different primary causes of APD prolongation, which suggests the use of mutation-specific therapeutic approaches in the future.