AUTHOR=Bögeholz Nils , Schulte Jan S. , Kaese Sven , Bauer B. Klemens , Pauls Paul , Dechering Dirk G. , Frommeyer Gerrit , Goldhaber Joshua I. , Kirchhefer Uwe , Eckardt Lars , Pott Christian , Müller Frank U. TITLE=The Effects of SEA0400 on Ca2+ Transient Amplitude and Proarrhythmia Depend on the Na+/Ca2+ Exchanger Expression Level in Murine Models JOURNAL=Frontiers in Pharmacology VOLUME=8 YEAR=2017 URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2017.00649 DOI=10.3389/fphar.2017.00649 ISSN=1663-9812 ABSTRACT=

Background/Objective: The cardiac Na⁺/Ca²⁺ exchanger (NCX) has been identified as a promising target to counter arrhythmia in previous studies investigating the benefit of NCX inhibition. However, the consequences of NCX inhibition have not been investigated in the setting of altered NCX expression and function, which is essential, since major cardiac diseases (heart failure/atrial fibrillation) exhibit NCX upregulation. Thus, we here investigated the effects of the NCX inhibitor SEA0400 on the Ca²⁺ transient amplitude and on proarrhythmia in homozygous NCX overexpressor (OE) and heterozygous NCX knockout (hetKO) mice compared to corresponding wild-types (WT_OE/WT_hetKO).

Methods/Results: Ca²⁺ transients of field-stimulated isolated ventricular cardiomyocytes were recorded with fluo-4-AM or indo-1-AM. SEA0400 (1 μM) significantly reduced NCX forward mode function in all mouse lines. SEA0400 (1 μM) significantly increased the amplitude of field-stimulated Ca²⁺ transients in WT_OE, WT_hetKO, and hetKO, but not in OE (% of basal; OE = 98.7 ± 5.0; WT_OE = 137.8 ± 5.2^*; WT_hetKO = 126.3 ± 6.0^*; hetKO = 140.6 ± 12.8^*; ^*p < 0.05 vs. basal). SEA0400 (1 μM) significantly reduced the number of proarrhythmic spontaneous Ca²⁺ transients (sCR) in OE, but increased it in WT_OE, WT_hetKO and hetKO (sCR per cell; basal/+SEA0400; OE = 12.5/3.7; WT_OE = 0.2/2.4; WT_hetKO = 1.3/8.8; hetKO = 0.2/5.5) and induced Ca²⁺ overload with subsequent cell death in hetKO.

Conclusion: The effects of SEA0400 on Ca²⁺ transient amplitude and the occurrence of spontaneous Ca²⁺ transients as a proxy measure for inotropy and cellular proarrhythmia depend on the NCX expression level. The antiarrhythmic effect of SEA0400 in conditions of increased NCX expression promotes the therapeutic concept of NCX inhibition in heart failure/atrial fibrillation. Conversely, in conditions of reduced NCX expression, SEA0400 suppressed the NCX function below a critical level leading to adverse Ca²⁺ accumulation as reflected by an increase in Ca²⁺ transient amplitude, proarrhythmia and cell death. Thus, the remaining NCX function under inhibition may be a critical factor determining the inotropic and antiarrhythmic efficacy of SEA0400.