P3830Carvedilol treatment diminishes spontaneous calcium release and electrical activity in human atrial myocytes
Authors: Jimenez-Sabado V , Lu T , Casabella S , Tarifa C , Herraiz-Martinez A , Colino H , Llach A , Ginel A , Cinca J , Hove-Madsen L 
Published: October 2019

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