Cardiac hypertrophy is an independent risk factor for heart failure. Recent studies on gene regulation of proteins have involved intracellular Ca2+ homeostasis. The Ca2+-sensitive phosphatase, calcineurin, is one potential regulator of the hypertrophic response, so we aimed to investigate the calcineurin-dependent signal pathway at different stages of hypertrophy in human myocardium. We found the calcineurin pathway to be significantly activated in hypertrophic compared with non-hypertrophic myocardium as demonstrated by increased calcineurin activity and expression of calcineurin A-β and B, and GATA-4, and a shift of phosphorylated cytoplasmic NFAT-3 into the nucleus as dephosphorylated nuclear NFAT-3. There was a tendency for these changes to be more pronounced in the decompensated compared with the compensated hypertrophic myocardium. The present study provides evidence for significant activation of the Ca2+-triggered calcineurin pathway in hypertrophic humans. Already present in compensated hypertrophy it showed a tendency to a further increase following transition to decompensated hypertrophy.
LevyDGarrisonRJSavageDD: Prognostic implications of echocardiographically determined left ventricular mass in the Framingham heart study. N Engl J Med1990; 322: 1561–1566.
2.
BalkeCWShorofskySR: Alterations in calcium handling in cardiac hypertrophy and heart failure. Cardiovasc Res1998; 37: 290–299.
3.
BeuckelmannDJNabauerMErdmannE: Intracellular calcium handling in isolated ventricular myocytes from patients with terminal heart failure. Circulation1992; 85: 1046–1055.
4.
SchwingerRHGBöhmMSchmidtU: Unchanged protein levels of SERCA II and phospholamban but reduced Ca2+ uptake and Ca2+-ATPase activity of cardiac sarcoplasmic reticulum from dilated cardiomyopathy patients compared with patients with nonfailing hearts. Circulation1995; 92: 3220–3228.
5.
GwathmeyJKCopelasLMacKinnonR: Abnormal intracellular calcium handling in myocardium from patients with end-stage heart failure. Circ Res1987; 61: 70–76.
6.
FeldmanMDGwathmeyJKPhillipsP: Reversal of the force-frequency relationship in working myocardium from patients with end-stage heart failure. J Appl Cardiol1988; 3: 273–283.
7.
LimasCJOlivariMTGoldenbergIF: Calcium uptake by cardiac sarcoplasmic reticulum in human dilated cardiomyopathy. Cardiovasc Res1987; 21: 601–605.
8.
MolkentinJDLuJRAntosCL: A calcineurin-dependent transcriptional pathway for cardiac hypertrophy. Cell1998; 93: 215–228.
9.
De WindtLJLimHWTaigenT: Calcineurin-mediated hypertrophy protects cardiomyocytes from apoptosis in vitro and in vivo. Circ Res2000; 86: 255–263.
10.
HaqSChoukrounGLimH: Differential activation of signal transduction pathway in human hearts with hypertrophy versus advanced heart failure. Circulation2001; 103: 670–677.
11.
LimHWMolkentinJD: Calcineurin and human heart failure. Nat Med1999; 5: 246–247.
12.
DiedrichsHChiMBoelckB: Increased regulatory activity of the calcineurin/NFAT pathway in human heart failure. Eur J Heart Fail2004; 6: 3–9.
13.
MünchGBoelckBKarczewskiP: Evidence for calcineurin-mediated regulation of SERCA 2a activity in human myocardium. J Mol Cell Cardiol2002; 34: 321–334.
14.
DeryckereFGannonF: A one-hour minipreparation technique for extraction of DNA-binding proteins from animal tissues. Biotechniques1994; 16: 405.
15.
RitterOHackSSchuhK: Calcineurin in human heart hypertophy. Circulation2002; 105: 2265–2269.
16.
SussmanMALimHWGudeN: Prevention of cardiac hypertrophy in mice by calcineurin inhibition. Science1998; 281: 1690–1693.
17.
LimHWDe WindtLJSteinbergL: Calcineurin expression, activation, and function in cardiac pressure-overload hypertrophy. Circulation2000; 101: 2431–2437.
18.
EtoYYonekuraKSondaM: Calcineurin is activated in rat hearts with physiological left ventricular hypertrophy induced by voluntary exercise training. Circulation2000; 101: 2134–2137.
19.
HayashidaWKiharaYYasakaA: Cardiac calcineurin during transition from hypertrophy to heart failure in rats. Biochem Biophys Res Commun2000; 273: 347–351.
20.
ShimoyamaMHayashiDTakimotoE: Calcineurin plays a critical role in pressure overload-induced cardiac hypertrophy. Circulation1999; 100: 2449–2454.
21.
DingBPriceRLBorgTK: Pressure overload induces severe hypertrophy in mice treated with cyclosporin, an inhibitor of calcineurin. Circ Res1999; 84: 729–734.
22.
ZhangWKowalRCRusnakF: Failure of calcineurin inhibitors to prevent pressure-overload left ventricular hypertrophy in rats. Circ Res1999; 84: 722–728.
23.
WilkinsBJMolkentinJD: Calcineurin and cardiac hypertrophy: Where have we been? Where are we going?J Physiol2002; 541: 1–8.
24.
ChowCWRincónMCavanaghJ: Nuclear accumulation of NFAT4 opposed by the JNK signal transduction pathway. Science1997; 278: 1638–1641.
25.
LiangQGDe WindtLJWittSA: The transcription factors GATA4 and GATA6 regulate cardiomyocyte hypertrophy in vitro and in vivo. J Biol Chem2001; 276: 30245–30253.
26.
CrabtreeGR: Generic signals and specific outcomes: Signalling through Ca2+, calcineurin and NFAT. Cell1999; 96: 611–614.
27.
YangTTCXiongQFEnslenH: Phosphorylation of NFATc4 by p38 mitogen-activated protein kinases. Mol Cell Biol2002; 22: 3892–3904.
28.
MorimotoTHasegawaKWadaH: Calcineurin-GATA4 pathway is involved in β-adrenergic agonist-responsive endothelin-1 transcription in cardiac myocytes. J Biol Chem2001; 276: 34983–34989.
29.
HasegawaKLeeSJJobeS: cis-Acting sequences that mediate induction of β-myosin heavy chain gene expression during left ventricular hypertrophy due to aortic constriction. Circulation1997; 96: 3943–3953.
30.
HerzigTCJobeSMAokiH: Angiotensin II type Iα receptor gene expression in the heart: AP-1 and GATA-4 participate in the responses to pressure overload. Proc Natl Acad Sci USA1997; 94: 7543–7548.
31.
KleeCBRenHWangX: Regulation of the calmodulin-stimulated protein phosphatase, calcineurin. J Biol Chem1998; 273: 13367–13370.
BuenoOFBrandtEBRothenbergME: Defective T cell development and function in calcineurin A beta-deficient mice. Proc Natl Acad Sci USA2002; 99: 9398–9403.
