Skarsfeldt Mark A, Jepps Thomas A, Bomholtz Sofia H, Abildgaard Lea, Sørensen Ulrik S, Gregers Emilie, Svendsen Jesper H, Diness Jonas G, Grunnet Morten, Schmitt Nicole, Olesen Søren-Peter, Bentzen Bo H
Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark.
Acesion Pharma, Copenhagen, Denmark.
Pflugers Arch. 2016 Apr;468(4):643-54. doi: 10.1007/s00424-015-1779-0. Epub 2016 Jan 5.
In isolated human atrial cardiomyocytes, inhibition of K2P3.1 K(+) channels results in action potential (action potential duration (APD)) prolongation. It has therefore been postulated that K2P3.1 (KCNK3), together with K2P9.1 (KCNK9), could represent novel drug targets for the treatment of atrial fibrillation (AF). However, it is unknown whether these findings in isolated cells translate to the whole heart. The purposes of this study were to investigate the expression levels of KCNK3 and KCNK9 in human hearts and two relevant rodent models and determine the antiarrhythmic potential of K2P3.1 inhibition in isolated whole-heart preparations. By quantitative PCR, we found that KCNK3 is predominantly expressed in human atria whereas KCNK9 was not detectable in heart human tissue. No differences were found between patients in AF or sinus rhythm. The expression in guinea pig heart resembled humans whereas rats displayed a more uniform expression of KCNK3 between atria and ventricle. In voltage-clamp experiments, ML365 and A293 were found to be potent and selective inhibitors of K2P3.1, but at pH 7.4, they failed to prolong atrial APD and refractory period (effective refractory period (ERP)) in isolated perfused rat and guinea pig hearts. At pH 7.8, which augments K2P3.1 currents, pharmacological channel inhibition produced a significant prolongation of atrial ERP (11.6 %, p = 0.004) without prolonging ventricular APD but did not display a significant antiarrhythmic effect in our guinea pig AF model (3/8 hearts converted on A293 vs 0/7 hearts in time-matched controls). These results suggest that when K2P3.1 current is augmented, K2P3.1 inhibition leads to atrial-specific prolongation of ERP; however, this ERP prolongation did not translate into significant antiarrhythmic effects in our AF model.
在分离的人心房心肌细胞中,抑制K2P3.1钾通道会导致动作电位(动作电位时程(APD))延长。因此,有人推测K2P3.1(KCNK3)与K2P9.1(KCNK9)一起可能是治疗心房颤动(AF)的新型药物靶点。然而,这些在分离细胞中的发现是否适用于整个心脏尚不清楚。本研究的目的是调查KCNK3和KCNK9在人心脏及两种相关啮齿动物模型中的表达水平,并确定在分离的全心标本中抑制K2P3.1的抗心律失常潜力。通过定量PCR,我们发现KCNK3主要在人心房中表达,而在人心脏组织中未检测到KCNK9。房颤患者和窦性心律患者之间未发现差异。豚鼠心脏中的表达与人类相似,而大鼠心房和心室之间KCNK3的表达更为均匀。在电压钳实验中,发现ML365和A293是K2P3.1的有效且选择性抑制剂,但在pH 7.4时,它们未能延长分离的灌注大鼠和豚鼠心脏的心房APD和不应期(有效不应期(ERP))。在pH 7.8时,K2P3.1电流增强,药理学通道抑制使心房ERP显著延长(11.6%,p = 0.004),而未延长心室APD,但在我们的豚鼠房颤模型中未显示出显著的抗心律失常作用(A293处理后8只心脏中有3只转复,而时间匹配对照组7只心脏中0只转复)。这些结果表明,当K2P3.1电流增强时,抑制K2P3.1会导致心房特异性ERP延长;然而,这种ERP延长在我们的房颤模型中并未转化为显著的抗心律失常作用。