739SERCA2 Cys674 modification lead to ventricular arrhythmia due to impaired sarcoplasmic reticulum Ca2+ handling

Abstract Background Sarcoplasmic reticulum Ca2+-ATPase2 (SERCA2) plays an important role in intracellular Ca2+ handling. Under pathological conditions, oxidative stress leads to irreversible oxidation of Cys674 on SERCA2 which causes intracellular Ca2+ overload. Intracellular Ca2+ overload is known as the cause of ventricular arrhythmia, but the relation between SERCA2 function and ventricular arrhythmia remains unclear. Purpose To investigate the role of Cys674 on SERCA2 in the intracellular Ca2+ handling and the induction of ventricular arrhythmia. Methods We employed SERCA2 Cys674Ser heterozygote knock-in mice (SKI) which mimics oxidative modification of Cys674 on SERCA2. Continuous infusion of angiotensin (ANG) (3mg/kg/day) or distilled water were performed both in wild type mice (WT) and SKI for a week. After 1 week, electrophysiological study and intracellular Ca2+ transient measurement were performed. Results ANG elevated blood pressure and represented cardiac hypertrophy with fibrosis similarly both in WT and SKI. The mRNA expression of calcium/calmodulin-dependent protein kinase-II (CaMKII), ryanodine receptor (RyR) and sodium-calcium exchanger (NCX) was increased in SKI heart compared with WT. QTc interval was prolonged in SKI compared with WT, which was markedly prolonged with ANG infusion. Under programmed electrical stimulation, only SKI with ANG showed high incidence of pacing induced ventricular arrhythmia (0/11 in WT/SKI control, 0/14 in WT with ANG vs. 8/14 in SKI with ANG, P < 0.01). In Ca2+ transient measurement, the peak Ca2+ transient amplitude (F/F0) was decreased (WT: 1.81 vs. SKI: 1.46, P < 0.01) and the time to 50% of cytosolic Ca2+ extrusion (T50) was prolonged (WT: 152.6ms vs. SKI: 202.3ms, P < 0.05) in SKI with ANG, suggesting decreased sarcoplasmic reticulum Ca2+ content and impaired SERCA2 activity in SKI with ANG. Intraperitoneal administration of dantrolene sodium (DAN) which inhibit Ca2+ leakage from RyR receptor normalized decreased F/F0 and prolonged T50 in SKI with ANG (Fig. 1 A, B). DAN also prevented QTc prolongation and decreased the incidence of ventricular arrhythmia in SKI with ANG (8/14 in SKI with ANG vs. 2/13 in SKI with ANG + DAN, P < 0.05) (Fig. 2). Conclusions The loss of thiol on Cys674 under pathological condition resulted in impaired Ca2+ handling and high incidence of ventricular arrhythmia which were ameliorated by inhibition of Ca2+ leakage through RyR. Oxidative modification of Cys674 on SERCA2 might contribute to Ca2+ mishandling and arrhythmogenesis. Abstract Figure.