New N-aryl-N-alkyl-thiophene-2-carboxamide Compound Enhances Intracellular Ca Dynamics by Increasing SERCA2a Ca Pumping

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2022 Dec 4

PMID 36463408

Authors

Roman Nikolaienko

Elisa Bovo

Samantha L Yuen

Levy M Treinen

Kaja Berg

Courtney C Aldrich

David D Thomas

Razvan L Cornea

Aleksey V Zima

Affiliations

Soon will be listed here.

Abstract

The type 2a sarco/endoplasmic reticulum Ca-ATPase (SERCA2a) plays a central role in the intracellular Ca homeostasis of cardiac myocytes, pumping Ca 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 Ca transport is of great clinical significance. In this study, we characterized the effect of a recently identified N-aryl-N-alkyl-thiophene-2-carboxamide (or compound 1) on SERCA2a Ca-ATPase and Ca transport activities in cardiac SR vesicles, and on Ca regulation in a HEK293 cell expression system and in mouse ventricular myocytes. We found that compound 1 enhances SERCA2a Ca-ATPase and Ca transport in SR vesicles. Fluorescence lifetime measurements of fluorescence resonance energy transfer between SERCA2a and phospholamban indicated that compound 1 interacts with the SERCA-phospholamban complex. Measurement of endoplasmic reticulum Ca dynamics in HEK293 cells expressing human SERCA2a showed that compound 1 increases endoplasmic reticulum Ca load by enhancing SERCA2a-mediated Ca transport. Analysis of cytosolic Ca dynamics in mouse ventricular myocytes revealed that compound 1 increases the action potential-induced Ca transients and SR Ca load, with negligible effects on L-type Ca channels and Na/Ca exchanger. However, during adrenergic receptor activation, compound 1 did not further increase Ca transients and SR Ca load, but it decreased the propensity toward Ca waves. Suggestive of concurrent desirable effects of compound 1 on RyR2, [H]-ryanodine binding to cardiac SR vesicles shows a small decrease in nM Ca and a small increase in μM Ca. Accordingly, compound 1 slightly decreased Ca sparks in permeabilized myocytes. Thus, this novel compound shows promising characteristics to improve intracellular Ca dynamics in cardiomyocytes that exhibit reduced SERCA2a Ca uptake, as found in failing hearts.

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References

Zsebo K, Yaroshinsky A, Rudy J, Wagner K, Greenberg B, Jessup M . Long-term effects of AAV1/SERCA2a gene transfer in patients with severe heart failure: analysis of recurrent cardiovascular events and mortality. Circ Res. 2013; 114(1):101-8. DOI: 10.1161/CIRCRESAHA.113.302421. View

Rebbeck R, Nitu F, Rohde D, Most P, Bers D, Thomas D . S100A1 Protein Does Not Compete with Calmodulin for Ryanodine Receptor Binding but Structurally Alters the Ryanodine Receptor·Calmodulin Complex. J Biol Chem. 2016; 291(30):15896-907. PMC: 4957069. DOI: 10.1074/jbc.M115.713107. View

Gruber S, Cornea R, Li J, Peterson K, Schaaf T, Gillispie G . Discovery of enzyme modulators via high-throughput time-resolved FRET in living cells. J Biomol Screen. 2014; 19(2):215-22. PMC: 4013825. DOI: 10.1177/1087057113510740. View

Jiang M, Lokuta A, Farrell E, Wolff M, Haworth R, Valdivia H . Abnormal Ca2+ release, but normal ryanodine receptors, in canine and human heart failure. Circ Res. 2002; 91(11):1015-22. DOI: 10.1161/01.res.0000043663.08689.05. View

Bovo E, Martin J, Tyryfter J, de Tombe P, Zima A . R-CEPIA1er as a new tool to directly measure sarcoplasmic reticulum [Ca] in ventricular myocytes. Am J Physiol Heart Circ Physiol. 2016; 311(1):H268-75. PMC: 4967208. DOI: 10.1152/ajpheart.00175.2016. View

Bovo E, Lipsius S, Zima A . Reactive oxygen species contribute to the development of arrhythmogenic Ca²⁺ waves during β-adrenergic receptor stimulation in rabbit cardiomyocytes. J Physiol. 2012; 590(14):3291-304. PMC: 3459043. DOI: 10.1113/jphysiol.2012.230748. View

Kranias E, Hajjar R . Modulation of cardiac contractility by the phospholamban/SERCA2a regulatome. Circ Res. 2012; 110(12):1646-60. PMC: 3392125. DOI: 10.1161/CIRCRESAHA.111.259754. View

Qaisar R, Bhaskaran S, Ranjit R, Sataranatarajan K, Premkumar P, Huseman K . Restoration of SERCA ATPase prevents oxidative stress-related muscle atrophy and weakness. Redox Biol. 2018; 20:68-74. PMC: 6174848. DOI: 10.1016/j.redox.2018.09.018. View

Schaaf T, Kleinboehl E, Yuen S, Roelike L, Svensson B, Thompson A . Live-Cell Cardiac-Specific High-Throughput Screening Platform for Drug-Like Molecules that Enhance Ca Transport. Cells. 2020; 9(5). PMC: 7291019. DOI: 10.3390/cells9051170. View

10.

Ferrandi M, Barassi P, Tadini-Buoninsegni F, Bartolommei G, Molinari I, Tripodi M . Istaroxime stimulates SERCA2a and accelerates calcium cycling in heart failure by relieving phospholamban inhibition. Br J Pharmacol. 2013; 169(8):1849-61. PMC: 3753840. DOI: 10.1111/bph.12278. View

11.

Periasamy M, Kalyanasundaram A . SERCA pump isoforms: their role in calcium transport and disease. Muscle Nerve. 2007; 35(4):430-42. DOI: 10.1002/mus.20745. View

12.

Bers D . Cardiac excitation-contraction coupling. Nature. 2002; 415(6868):198-205. DOI: 10.1038/415198a. View

13.

Curran J, Brown K, Santiago D, Pogwizd S, Bers D, R Shannon T . Spontaneous Ca waves in ventricular myocytes from failing hearts depend on Ca(2+)-calmodulin-dependent protein kinase II. J Mol Cell Cardiol. 2010; 49(1):25-32. PMC: 2883657. DOI: 10.1016/j.yjmcc.2010.03.013. View

14.

Dahl R . A new target for Parkinson's disease: Small molecule SERCA activator CDN1163 ameliorates dyskinesia in 6-OHDA-lesioned rats. Bioorg Med Chem. 2016; 25(1):53-57. DOI: 10.1016/j.bmc.2016.10.008. View

15.

Cantilina T, Sagara Y, Inesi G, Jones L . Comparative studies of cardiac and skeletal sarcoplasmic reticulum ATPases. Effect of a phospholamban antibody on enzyme activation by Ca2+. J Biol Chem. 1993; 268(23):17018-25. View

16.

ORourke B, Kass D, Tomaselli G, Kaab S, Tunin R, Marban E . Mechanisms of altered excitation-contraction coupling in canine tachycardia-induced heart failure, I: experimental studies. Circ Res. 1999; 84(5):562-70. DOI: 10.1161/01.res.84.5.562. View

17.

Savignac M, Edir A, Simon M, Hovnanian A . Darier disease : a disease model of impaired calcium homeostasis in the skin. Biochim Biophys Acta. 2010; 1813(5):1111-7. DOI: 10.1016/j.bbamcr.2010.12.006. View

18.

Kuo I, Kwaczala A, Nguyen L, Russell K, Campbell S, Ehrlich B . Decreased polycystin 2 expression alters calcium-contraction coupling and changes β-adrenergic signaling pathways. Proc Natl Acad Sci U S A. 2014; 111(46):16604-9. PMC: 4246301. DOI: 10.1073/pnas.1415933111. View

19.

Currie S, Smith G . Enhanced phosphorylation of phospholamban and downregulation of sarco/endoplasmic reticulum Ca2+ ATPase type 2 (SERCA 2) in cardiac sarcoplasmic reticulum from rabbits with heart failure. Cardiovasc Res. 1999; 41(1):135-46. DOI: 10.1016/s0008-6363(98)00241-7. View

20.

Schmidt U, Hajjar R, Helm P, Kim C, Doye A, Gwathmey J . Contribution of abnormal sarcoplasmic reticulum ATPase activity to systolic and diastolic dysfunction in human heart failure. J Mol Cell Cardiol. 1998; 30(10):1929-37. DOI: 10.1006/jmcc.1998.0748. View