4966Targeting INaL by a neuronal sodium channel isoform improves survival in a CaMKII-transgenic heart failure mouse model

Abstract Background Cardiac pathologies like hypertrophy and heart failure are known to be associated with proarrhythmogenic triggers like early- (EADs) and delayed afterdepolarizations (DADs) that can be partly attributed to an augmentation of late sodium current (INaL). Enhanced INaL is closely connected with increased activity of Ca2+/calmodulin dependent-kinase II (CaMKII) in pathology as it is enhanced by CaMKII on the one hand but can also indirectly increase CaMKII-activity on the other. We recently found neuronal sodium channel NaV1.8 to be involved in INaL-augmentation in heart failure and cardiac hypertrophy. Here, we studied possible antiarrhythmic effects of NaV1.8-inhibition in a transgenic mouse model with enhanced CaMKII-expression by selectively knocking out NaV1.8. Methods/Results To investigate antiarrhythmic effects of NaV1.8-depletion in-vivo and in-vitro we crossbred CaMKII-transgenic mice (CaMKII+/T) with NaV1.8-knock-out mice (SCN10A−/−). Surprisingly, CaMKII+/T-mice lacking NaV1.8 (CaMKII+/T & SCN10A−/−) showed a significantly improved survival compared to CaMKII+/T alone (97.5 vs 72.0 days, p<0.05). Heart weight to tibia length ratio was significantly increased in CaMKII+/T-mice compared to wild-type, without any differences between CaMKII+/T and CaMKII+/T & SCN10A−/−. To investigate the underlying mechanisms out of this observation we isolated single cardiomyocytes and performed patch-clamp experiments as well as confocal microscopy to measure Ca2+-transients and diastolic Ca2+-waves. INaL-integral was significantly smaller in cardiomyocytes from CaMKII+/T & SCN10A−/−-mice compared to CaMKII+/T alone. During action potential recordings, significantly less afterdepolarizations occurred in CaMKII+/T & SCN10A−/− compared to cardiomyocytes from CaMKII+/T -mice (16.7/min vs 34.9/min, p<0.05). There was a trend of less cells exhibiting diastolic Ca2+-waves in Ca2+-measurements from CaMKII+/T & SCN10A−/− compared to CaMKII+/T (15% vs 25%, p=0.09). As some cells showed more than one event, we calculated the frequency of Ca2+-waves and found a significant reduction of Ca2+-waves in CaMKII+/T & SCN10A−/− vs. CaMKII+/T (22.8/min vs 43.0/min, p<0.05). Moreover, the time to the first event was significantly longer in CaMKII+/T & SCN10A−/−. Ca2+-transient amplitude (F/F0) was significantly lower in CaMKII+/T compared to CaMKII+/T & SCN10A−/− (4.6 vs. 5.3, p=0.05). Further, Ca2+-extrusion from the cytosol was significantly faster in CaMKII+/T & SCN10A−/−. Conclusion Our data demonstrates, that inhibition of INaL by targeting NaV1.8 has a potent antiarrhythmic potential as we found a reduction of EADs, DADs and diastolic Ca2+-waves in CaMKII+/T & SCN10A−/−-cardiomyocytes. This antiarrhythmic potential appears to be potent enough to improve survival and to rescue the proarrhythmogenic phenotype of CaMKII-overexpression. However, further in-vivo experiments are necessary to investigate NaV1.8-inhibition for a possible therapeutic approach.