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Activation of CaMKIIδA promotes Ca2+ leak from the sarcoplasmic reticulum in cardiomyocytes of chronic heart failure rats

Acta Pharmacologica Sinicavolume 39pages16041612 (2018) | Download Citation



Activation of the Ca2+/calmodulin-dependent protein kinase II isoform δA (CaMKIIδA) disturbs intracellular Ca2+ homeostasis in cardiomyocytes during chronic heart failure (CHF). We hypothesized that upregulation of CaMKIIδA in cardiomyocytes might enhance Ca2+ leak from the sarcoplasmic reticulum (SR) via activation of phosphorylated ryanodine receptor type 2 (P-RyR2) and decrease Ca2+ uptake by inhibition of SR calcium ATPase 2a (SERCA2a). In this study, CHF was induced in rats by ligation of the left anterior descending coronary artery. We found that CHF caused an increase in the expression of CaMKIIδA and P-RyR2 in the left ventricle (LV). The role of CaMKIIδA in regulation of P-RyR2 was elucidated in cardiomyocytes isolated from neonatal rats in vitro. Hypoxia induced upregulation of CaMKIIδA and activation of P-RyR2 in the cardiomyocytes, which both were attenuated by knockdown of CaMKIIδA. Furthermore, we showed that knockdown of CaMKIIδA significantly decreased the Ca2+ leak from the SR elicited by hypoxia in the cardiomyocytes. In addition, CHF also induced a downregulation of SERCA2a in the LV of CHF rats. Knockdown of CaMKIIδA normalized hypoxia-induced downregulation of SERCA2a in cardiomyocytes in vitro. The results demonstrate that the inhibition of CaMKIIδA may improve cardiac function by preventing SR Ca2+ leak through downregulation of P-RyR2 and upregulation of SERCA2a expression in cardiomyocytes in CHF.

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  1. 1

    Fabiato A. Calcium-induced release of calcium from the cardiac sarcoplasmic reticulum. Am J Physiol 1983; 245: C1–14.

  2. 2

    Srinivasan M, Edman CF, Schulman H. Alternative splicing introduces a nuclear localization signal that targets multifunctional CaM kinase to the nucleus. J Cell Biol 1994; 126: 839–52.

  3. 3

    Gray CB, Heller Brown J. CaMKIIdelta subtypes: localization and function. Front Pharmacol 2014; 5: 15.

  4. 4

    Yao J, Qin X, Zhu J, Sheng H. Dyrk1A-ASF-CaMKIIdelta signaling Is involved in valsartan inhibition of cardiac hypertrophy in renovascular hypertensive rats. Cardiology 2016; 133: 198–204.

  5. 5

    Schworer CM, Rothblum LI, Thekkumkara TJ, Singer HA. Identification of novel isoforms of the delta subunit of Ca2+/calmodulin-dependent protein kinase II. Differential expression in rat brain and aorta. J Biol Chem 1993; 268: 14443–9.

  6. 6

    Purohit A, Rokita AG, Guan X, Chen B, Koval OM, Voigt N, et al. Oxidized Ca2+/calmodulin-dependent protein kinase II triggers atrial fibrillation. Circulation 2013; 128: 1748–57.

  7. 7

    Zhang T, Guo T, Mishra S, Dalton ND, Kranias EG, Peterson KL, et al. Phospholamban ablation rescues sarcoplasmic reticulum Ca2+ handling but exacerbates cardiac dysfunction in CaMKIIdelta(C) transgenic mice. Circ Res 2010; 106: 354–62.

  8. 8

    Maier LS, Zhang T, Chen L, DeSantiago J, Brown JH, Bers DM. Transgenic CaMKIIdeltaC overexpression uniquely alters cardiac myocyte Ca2+ handling: reduced SR Ca2+ load and activated SR Ca2+ release. Circ Res 2003; 92: 904–11.

  9. 9

    Polakova E, Illaste A, Niggli E, Sobie EA. Maximal acceleration of Ca2+ release refractoriness by beta-adrenergic stimulation requires dual activation of kinases PKA and CaMKII in mouse ventricular myocytes. J Physiol 2015; 583: 1495–507.

  10. 10

    Stokke MK, Tovsrud N, Louch WE, Oyehaug L, Hougen K, Sejersted OM, et al. I CaL inhibition prevents arrhythmogenic Ca2+ waves caused by abnormal Ca2+ sensitivity of RyR or SR Ca2+ accumulation. Cardiovasc Res 2013; 98: 315–25.

  11. 11

    Marks AR. Calcium cycling proteins and heart failure: mechanisms and therapeutics. J Clin Invest 2013; 123: 46–52.

  12. 12

    Netticadan T, Temsah RM, Kawabata K, Dhalla NS. Sarcoplasmic reticulum Ca2+/calmodulin-dependent protein kinase is altered in heart failure. Circ Res 2000; 86: 596–605.

  13. 13

    Bobin P, Varin A, Lefebvre F, Fischmeister R, Vandecasteele G, Leroy J. Calmodulin kinase II inhibition limits the pro-arrhythmic Ca2+ waves induced by cAMP-phosphodiesterase inhibitors. Cardiovasc Res 2016; 110: 151–61.

  14. 14

    Woo AY, Xiao RP. beta-Adrenergic receptor subtype signaling in heart: from bench to bedside. Acta Pharmacol Sin 2012; 33: 335–41.

  15. 15

    Ogrodnik J, Niggli E. Increased Ca2+ leak and spatiotemporal coherence of Ca2+ release in cardiomyocytes during beta-adrenergic stimulation. J Physiol 2010; 588: 225–42.

  16. 16

    Arai M, Alpert NR, MacLennan DH, Barton P, Periasamy M. Alterations in sarcoplasmic reticulum gene expression in human heart failure. A possible mechanism for alterations in systolic and diastolic properties of the failing myocardium. Circ Res 1993; 72: 463–9.

  17. 17

    Yang Y, Rong X, Lv X, Jiang W, Yang Y, Lai D, et al. Inhibition of mevalonate pathway prevents ischemia-induced cardiac dysfunction in rats via RhoA-independent signaling pathway. Cardiovasc Ther 201; 35. doi: 10.1111/1755-5922.12285.

  18. 18

    Lompre AM, Hajjar RJ, Harding SE, Kranias EG, Lohse MJ, Marks AR. Ca2+ cycling and new therapeutic approaches for heart failure. Circulation 2010; 121: 822–30.

  19. 19

    Renaud-Gabardos E, Tatin F, Hantelys F, Lebas B, Calise D, Kunduzova O, et al. Therapeutic benefit and gene network regulation by combined gene transfer of apelin, FGF2, and SERCA2a into ischemic heart. Mol Ther 2017 Nov 16. doi: 10.1016/j.ymthe.2017.11.007

  20. 20

    Vangheluwe P, Raeymaekers L, Dode L, Wuytack F. Modulating sarco(endo)plasmic reticulum Ca2+ ATPase 2 (SERCA2) activity: cell biological implications. Cell Calcium 2005; 38: 291–302.

  21. 21

    Zhang T, Miyamoto S, Brown JH. Cardiomyocyte calcium and calcium/calmodulin-dependent protein kinase II: friends or foes? Recent Prog Horm Res 2004; 59: 141–68.

  22. 22

    Bossuyt J, Helmstadter K, Wu X, Clements-Jewery H, Haworth RS, Avkiran M, et al. Ca2+/calmodulin-dependent protein kinase IIdelta and protein kinase D overexpression reinforce the histone deacetylase 5 redistribution in heart failure. Circ Res 2008; 102: 695–702.

  23. 23

    Gray CB, Suetomi T, Xiang S, Mishra S, Blackwood EA, Glembotski CC, et al. CaMKIIdelta subtypes differentially regulate infarct formation following ex vivo myocardial ischemia/reperfusion through NF-kappaB and TNF-alpha. J Mol Cell Cardiol 2017; 103: 48–55.

  24. 24

    Li C, Cai X, Sun H, Bai T, Zheng X, Zhou XW, et al. The deltaA isoform of calmodulin kinase II mediates pathological cardiac hypertrophy by interfering with the HDAC4-MEF2 signaling pathway. Biochem Biophys Res Commun 2011; 409: 125–30.

  25. 25

    Gui L, Bao Z, Jia Y, Qin X, Cheng ZJ, Zhu J, et al. Ventricular tachyarrhythmias in rats with acute myocardial infarction involves activation of small-conductance Ca2+-activated K+ channels. Am J Physiol Heart Circ Physiol 2013; 304: H118–130.

  26. 26

    Ni YG, Berenji K, Wang N, Oh M, Sachan N, Dey A, et al. Foxo transcription factors blunt cardiac hypertrophy by inhibiting calcineurin signaling. Circulation 2006; 114: 1159–68.

  27. 27

    Bracken CP, Whitelaw ML, Peet DJ. The hypoxia-inducible factors: key transcriptional regulators of hypoxic responses. Cell Mol Life Sci 2003; 60: 1376–93.

  28. 28

    Chen CC, Kuo CY, Chen RF. Role of CAPE on cardiomyocyte protection via connexin 43 regulation under hypoxia. Int J Med Sci 2016; 13: 754–8.

  29. 29

    Pan LJ, Wang X, Ling Y, Gong H. MiR-24 alleviates cardiomyocyte apoptosis after myocardial infarction via targeting BIM. Eur Rev Med Pharmacol Sci 2017; 21: 3088–97.

  30. 30

    Shannon TR, Ginsburg KS, Bers DM. Quantitative assessment of the SR Ca2+ leak-load relationship. Circ Res 2002; 91: 594–600.

  31. 31

    Voigt N, Li N, Wang Q, Wang W, Trafford AW, Abu-Taha I, et al. Enhanced sarcoplasmic reticulum Ca2+ leak and increased Na+-Ca2+ exchanger function underlie delayed afterdepolarizations in patients with chronic atrial fibrillation. Circulation 2012; 125: 2059–70.

  32. 32

    Ji Y, Guo X, Zhang Z, Huang Z, Zhu J, Chen QH, et al. CaMKIIdelta meditates phenylephrine induced cardiomyocyte hypertrophy through store-operated Ca2+ entry. Cardiovasc Pathol 2016; 27: 9–17.

  33. 33

    Litwin SE, Katz SE, Morgan JP, Douglas PS. Serial echocardiographic assessment of left ventricular geometry and function after large myocardial infarction in the rat. Circulation 1994; 89: 345–54.

  34. 34

    Zhang T, Zhang Y, Cui M, Jin L, Wang Y, Lv F, et al. CaMKII is a RIP3 substrate mediating ischemia- and oxidative stress-induced myocardial necroptosis. Nat Med 2016; 22: 175–82.

  35. 35

    Tombes RM, Faison MO, Turbeville JM. Organization and evolution of multifunctional Ca2+/CaM-dependent protein kinase genes. Gene 2003; 322: 17–31.

  36. 36

    Bell JR, Raaijmakers AJ, Janssens JV, Delbridge LM. CaMKIIdelta and cardiomyocyte Ca2+ signalling new perspectives on splice variant targeting. Clin Exp Pharmacol Physiol 2015; 42: 1327–32.

  37. 37

    Liao RJ, Tong LJ, Huang C, Cao WW, Wang YZ, Wang J, et al. Rescue of cardiac failing and remodelling by inhibition of protein phosphatase 1gamma is associated with suppression of the alternative splicing factor-mediated splicing of Ca2+/calmodulin-dependent protein kinase delta. Clin Exp Pharmacol Physiol 2014; 41: 976–85.

  38. 38

    Zhang L, Liu BF, Liang S, Jones RL, Lu YT. Molecular and biochemical characterization of a calcium/calmodulin-binding protein kinase from rice. Biochem J 2002; 368: 145–57.

  39. 39

    Sondergaard MT, Tian X, Liu Y, Wang R, Chazin WJ, Chen SR, et al. Arrhythmogenic calmodulin mutations affect the activation and termination of cardiac ryanodine receptor-mediated Ca2+ release. J Biol Chem 2015; 290: 26151–62.

  40. 40

    Dibb K, Eisner D. A small leak may sink a great ship but what does it do to the heart? J Physiol 2010; 588: 4849.

  41. 41

    Tzimas C, Terrovitis J, Lehnart SE, Kranias EG, Sanoudou D. Calcium/calmodulin-dependent protein kinase II (CaMKII) inhibition ameliorates arrhythmias elicited by junctin ablation under stress conditions. Heart Rhythm 2015; 12: 1599–610.

  42. 42

    Respress JL, Gershovich PM, Wang T, Reynolds JO, Skapura DG, Sutton JP, et al. Long-term simulated microgravity causes cardiac RyR2 phosphorylation and arrhythmias in mice. Int J Cardiol 2014; 176: 994–1000.

  43. 43

    Hayward C, Banner NR, Morley-Smith A, Lyon AR, Harding SE. The current and future landscape of serca gene therapy for heart failure: a clinical perspective. Hum Gene Ther 2015; 26: 293–304.

  44. 44

    Trafford AW, Diaz ME, Sibbring GC, Eisner DA. Modulation of CICR has no maintained effect on systolic Ca2+: simultaneous measurements of sarcoplasmic reticulum and sarcolemmal Ca2+ fluxes in rat ventricular myocytes. J Physiol 2000; 522 Pt 2: 259–70.

  45. 45

    Marx SO, Reiken S, Hisamatsu Y, Jayaraman T, Burkhoff D, Rosemblit N, et al. PKA phosphorylation dissociates FKBP12.6 from the calcium release channel (ryanodine receptor): defective regulation in failing hearts. Cell 2000; 101: 365–76.

  46. 46

    Lyon AR, Bannister ML, Collins T, Pearce E, Sepehripour AH, Dubb SS, et al. SERCA2a gene transfer decreases sarcoplasmic reticulum calcium leak and reduces ventricular arrhythmias in a model of chronic heart failure. Circ Arrhythm Electrophysiol 2011; 4: 362–72.

  47. 47

    Zhang T, Brown JH. Role of Ca2+/calmodulin-dependent protein kinase II in cardiac hypertrophy and heart failure. Cardiovasc Res 2004; 63: 476–86.

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This study was supported by operating grants to Le GUI. from the National Natural Science Foundation of China (No 81570294), to Jiang-hua ZHU. from the National Natural Science Foundation of China (No 81370344) and to Qing-hui CHEN from the National Institute of Health (No HL122952).

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  1. These authors contributed equally to this work.


  1. Institute of Cardiovascular Disease, Department of Cardiology, Nantong University, Nantong, 226000, China

    • Le Gui
    • , Zhe Zhang
    • , Hui Xu
    • , Ya-wei Ji
    •  & Jiang-hua Zhu
  2. Department of Urology, Nantong University, Nantong, 226000, China

    • Xin Guo
  3. Departments of Biotechnology, School of Life Science, Jilin Normal University, Siping, 136000, China

    • Ren-jun Wang
  4. Department of Kinesiology and Integrative Physiology, Michigan Technological University, Houghton, 49931, Michigan, USA

    • Ren-jun Wang
    •  & Qing-hui Chen


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Correspondence to Qing-hui Chen.

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