4967Effects of atrial fibrillation on the human ventricle

Abstract

Abstract Background The consequence of normofrequent atrial fibrillation (AF) on the ventricle remains largely unknown. Methods and results To elucidate the effects of arrhythmic excitation on human ventricular myocardium we performed contractility experiments using ventricular trabecula from patients with heart failure (HF). Normofrequent AF was simulated using arrhythmic (60 bpm, 40% R-R interval variability) or rhythmic field stimulation (60 bpm). Within 8h of arrhythmic stimulation, human specimen showed an impaired systolic force, while diastolic tension increased pathologically (n=5–7 each/7 HF patients, Fig. 1). The characterization of the ventricular (in-vivo) phenotype in patients with AF was performed by utilizing ventricular myocardium from patients with sinus rhythm (SR) and from patients with AF in the absence of HF (compensated hypertrophy, EF>50%, matched clinical characteristics, LV myocardium obtained from aortic valve replacement surgery). Histological investigation showed increased levels of interstitial fibrosis in myocardium from patients with AF compared to SR (n=10 patients each). Studies of cellular Ca-homeostasis (epifluorescence microscopy, Fura-2) were performed using isolated human ventricular cardiomyocytes. While systolic Ca-transient amplitude (0.5 Hz) was preserved in ventricular cardiomyocytes from patients with AF, we found a significantly prolonged Ca-elimination time (RT80) by 22.0±7.7% and a trend towards increased diastolic Ca-levels (n=17–23 cells/4 patients each). This finding may be explained by a decrease in SERCA2a activity (ksys-kCaff, n=10–12/4 each) and an enhanced phospholamban expression in Western Blot experiments (n=5 patients each). For the standardized investigation of the involved targets/mechanisms mediating the pathological changes upon arrhythmic excitation, we utilized human induced pluripotent stem cell cardiomyocytes (iPSC-CM) from healthy donors for chronic arrhythmic culture stimulation (24h). Arrhythmic paced iPSC-CM showed no changes in systolic Ca-transient amplitude (0.5 Hz), whereas diastolic Ca-levels were increased, which fits nicely to the finding of disturbed trabeculae diastolic function (n=15 cells each). In patch clamp experiments, arrhythmic paced cells showed no alterations of resting membrane potential, upstroke velocity, action-potential amplitude or -duration (n=7–9 cells each). Protein expression levels of key Ca-handling proteins in iPSC-CM as well as regulated genes are already under investigation. Conclusion This study demonstrates that arrhythmic ventricular excitation deteriorates human myocardial contractility early in HF. In biopsies from patients with preserved EF, chronic AF was associated with increased levels of interstitial fibrosis and pathological diastolic Ca-handling, which could be causally confirmed in chronically arrhythmic paced iPSC-CM. Therefore, this study provides first mechanistic characterization of AF mediated effects on the human ventricle.