P3830Carvedilol treatment diminishes spontaneous calcium release and electrical activity in human atrial myocytes

Abstract

Abstract Background Atrial fibrillation (AF) has been associated with an increase in spontaneous calcium release induced electrical activity, which could potentially be reversed by carvedilol, a nonselective beta-blocker that also inhibits the cardiac ryanodine receptor (RyR2). Interestingly the enantiomer R-carvedilol inhibits the RyR2 but not beta-adrenergic receptors, allowing it to effectively prevent calcium release-induced spontaneous electrical activity without inducing bradycardia and hypotension. Purpose The purpose of this study was to determine how carvedilol treatment affects calcium release-induced transient inward currents (ITI) in human atrial myocytes from patients with AF; and to test the effects of R-carvedilol on spontaneous calcium release in order to assess its therapeutical utility. Methods Human atrial myocytes were isolated from patients undergoing cardiac surgery and subjected to patch-clamp technique (n=60) or confocal calcium imaging (n=6). Beta-2 adrenergic receptors were activated with the selective agonist fenoterol (3μM) and 1μM R-carvedilol was used to inhibit spontaneous calcium release events. Results Recordings of calcium release-induced transient inward currents (ITI) revealed that carvedilol treatment reduced the ITI frequency in patients with AF from 2.2±0.4 events/min in untreated patients to 0.59±0.35 events/min (p<0.01), which was even lower than the incidence in patients without AF (1.0±0.1 events/min; p<0.01). To assess the effects of R-carvedilol, myocytes were first simulated with fenoterol. This increased the calcium spark frequency from 23±15 to 960±336 events/s/1000μm2 in 16 cells from 6 patients (p<0.05). This was due to an increase in the spark site density (from 0.50±0.24 to 12.1±2.4 sites/1000μm2, p<0.001) rather than in the firing rate (0.068±0.14 vs. 0.035±0.012 sparks/s in control, p=0.14). Fenoterol also increased the spark duration from 50.9±5.4 to 77.3±4.1ms (p<0.001) without affecting the amplitude. Importantly, fenoterol also induced global calcium release events such as calcium waves and transients (2.8±1.1 vs. 0 events/min in control, p<0.05). When R-carvedilol was added, the effects of fenoterol were abolished, reducing the incidence of calcium sparks to 69±51 events/s/1000μm2 (p<0.05), the spark site density to 1.68±1.04 sites/1000μm2 (p<0.01), the spark duration to 63.4±4.3ms (p<0.05), and calcium waves and transients were reduced to 0.21±0.14 events/min (p<0.05). Conclusions Carvedilol treatment reduces the ITI frequency in patients with AF to levels below that observed at baseline in patients without AF. Furthermore, the non-beta-blocking R-carvedilol enantiomer abolishes spontaneous calcium release events induced by beta-2 adrenergic stimulation in human atrial myocytes, proposing a therapeutical utility for this compound in patients with AF linked to excessive spontaneous calcium release. Acknowledgement/Funding SAF2017-88019; Marato2015-20-30; SGR2017-1769; CIBERCV