New N-aryl-N-alkyl-thiophene-2-carboxamide compound enhances intracellular Ca2+ dynamics by increasing SERCA2a Ca2+ pumping.
New N-aryl-N-alkyl-thiophene-2-carboxamide compound enhances intracellular Ca2+ dynamics by increasing SERCA2a Ca2+ pumping.
复制标题
新型 N-芳基-N-烷基-噻吩-2-甲酰胺化合物通过增加 SERCA2a Ca2 泵送来增强细胞内 Ca2 动力学。
DOI：
10.1016/j.bpj.2022.12.002
复制
发表时间：
2023
影响因子：
3.4
通讯作者：
Zima,AlekseyV
中科院分区：
文献类型：
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作者：
Nikolaienko,Roman;Bovo,Elisa;Yuen,SamanthaL;Treinen,LevyM;Berg,Kaja;Aldrich,CourtneyC;Thomas,DavidD;Cornea,RazvanL;Zima,AlekseyV
2a型肌浆网/内质网Ca 2 +-ATP酶（SERCA 2a）在心肌细胞的细胞内Ca 2+稳态中起核心作用，将Ca 2+从细胞质泵入肌浆网（SR）腔以维持舒张（舒张）并为收缩（收缩）做准备。在包括心力衰竭在内的几种病理状况中已经报道了SERCA 2a功能减弱。因此，开发改善SERCA 2a钙转运的新药具有重要的临床意义。在这项研究中，我们表征了最近鉴定的N-芳基-N-烷基-噻吩-2-甲酰胺（或化合物1）对心脏SR囊泡中SERCA 2a Ca 2 +-ATP酶和Ca 2+转运活性的影响，以及对HEK 293细胞表达系统和小鼠心室肌细胞中Ca 2+调节的影响。结果表明，化合物1能增强SR囊泡的SERCA 2a Ca ~（2+）-ATP酶和Ca ~（2+）转运。荧光共振能量转移的荧光寿命的测量表明，化合物1与SERCA-phospholamban复合物相互作用。在表达人SERCA 2a的HEK 293细胞中内质网Ca 2+动力学的测量显示，化合物1通过增强SERCA 2a介导的Ca 2+转运来增加内质网Ca 2+负荷。对小鼠心室肌细胞胞浆Ca 2+动力学的分析表明，化合物1增加动作电位诱导的Ca 2+瞬变和SR Ca 2+负荷，对L型Ca 2+通道和Na+/Ca 2+交换器的影响可忽略不计。然而，在肾上腺素能受体激活过程中，化合物1没有进一步增加Ca 2+瞬变和SR Ca 2+负荷，但它降低了Ca 2+波的倾向。[3 H]-ryanodine与心脏SR囊泡的结合表明化合物1对RyR 2的同时理想作用，显示nM Ca 2+的小幅降低和μM Ca 2+的小幅增加。因此，化合物1轻微降低透化的肌细胞中的Ca 2+火花。因此，这种新型化合物显示出改善心肌细胞中细胞内Ca 2+动力学的有希望的特性，所述心肌细胞表现出降低的SERCA 2a Ca 2+摄取，如在衰竭心脏中发现的。
The type 2a sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2a) plays a central role in the intracellular Ca2+homeostasis of cardiac myocytes, pumping Ca2+from the cytoplasm into the sarcoplasmic reticulum (SR) lumen to maintain relaxation (diastole) and prepare for contraction (systole). Diminished SERCA2a function has been reported in several pathological conditions, including heart failure. Therefore, development of new drugs that improve SERCA2a Ca2+transport is of great clinical significance. In this study, we characterized the effect of a recently identifiedN-aryl-N-alkyl-thiophene-2-carboxamide(or compound1) on SERCA2a Ca2+-ATPase and Ca2+transport activities in cardiac SR vesicles, and on Ca2+regulation in a HEK293 cell expression system and in mouse ventricular myocytes. We found that compound1enhances SERCA2a Ca2+-ATPase and Ca2+transport in SR vesicles. Fluorescence lifetime measurements of fluorescence resonance energy transfer between SERCA2a and phospholamban indicated that compound1interacts with the SERCA-phospholamban complex. Measurement of endoplasmic reticulum Ca2+dynamics in HEK293 cells expressing human SERCA2a showed that compound1increases endoplasmic reticulum Ca2+load by enhancing SERCA2a-mediated Ca2+transport. Analysis of cytosolic Ca2+dynamics in mouse ventricular myocytes revealed that compound1increases the action potential-induced Ca2+transients and SR Ca2+load, with negligible effects on L-type Ca2+channels and Na+/Ca2+exchanger. However, during adrenergic receptor activation, compound1did not further increase Ca2+transients and SR Ca2+load, but it decreased the propensity toward Ca2+waves. Suggestive of concurrent desirable effects of compound1on RyR2, [3H]-ryanodine binding to cardiac SR vesicles shows a small decrease in nM Ca2+and a small increase in μM Ca2+. Accordingly, compound1slightly decreased Ca2+sparks in permeabilized myocytes. Thus, this novel compound shows promising characteristics to improve intracellular Ca2+dynamics in cardiomyocytes that exhibit reduced SERCA2a Ca2+uptake, as found in failing hearts.