Myocardial fibrosis can contribute to diastolic dysfunction and the development of heart failure. Preliminary results indicate that adenosine (ADO) can inhibit cardiac fibroblast (CF) function. These effects appear mediated via ADO A2b receptors. We will test the hypothesis that: """"""""The enhancement of myocardial adenosine levels reduces the synthesis of cardiac collagens and associated fibrosis through the stimulation of cardiac fibroblast adenosine A2b receptors"""""""". We will examine the following aims:
AIM 1 : Enhanced myocardial ADO levels in the setting of myocardial infarction (MI) inhibits the deposition of cardiac collagens. We will induce the acute deposition of cardiac collagens in rats via MI induced scar formation. To enhance tissue ADO levels we will use the ADO kinase inhibitor compound GP515. We will assess for the functional consequences of the inhibition of scar formation in control untreated and treated MI rats by determining passive and active structure-function and by analysis of the microstructural properties of the scar.
AIM 2 : Enhanced myocardial ADO levels in the setting of MI reduces in vivo cardiac fibroblast proliferation and type I collagen gene transcription. We will use MI to induce cardiac fibroblast proliferation and collagen deposition in transgenic mice that link the expression of the marker green fluorescent protein (GFP) to collagen type I (CI- GFP) gene expression. Myocardial ADO levels will be enhanced as in aim 1. In vivo CF proliferation, type I gene expression and cardiac function, scarring/remodeling will be assessed at predetermined time points.
AIM 3 : The absence of A2b receptors in the MI mouse heart impairs the ability of ADO to diminish cardiac collagen synthesis yielding enhanced scar deposition. We will use mice null (-/-) for A2b receptors. We will induce the acute deposition of cardiac collagens in wild type and in A2b receptor null mice by using MI. Tissue ADO levels will be enhanced as in aim 1. Cardiac passive and active structure- function will be assessed at predetermined time points. The effects of enhanced ADO levels will also be examined in vitro using cultured CF from A2b (-/-) mice and assessing for ADO modulation of cell function.
AIM 4 : ADO inhibits TNF-alpha release and cardiac fibroblast function through the stimulation of A2b receptors and associated increases in cAMP and/or cGMP levels. We will assess for adenosine A2b receptor mediated induced inhibition of CF function and for the role of intracellular cAMP and/or cGMP in mediating ADO induced alterations in CF function.
| Spinale, Francis G; Villarreal, Francisco (2014) Targeting matrix metalloproteinases in heart disease: lessons from endogenous inhibitors. Biochem Pharmacol 90:7-15 |
| Maya, Lisandro; Villarreal, Francisco J (2010) Diagnostic approaches for diabetic cardiomyopathy and myocardial fibrosis. J Mol Cell Cardiol 48:524-9 |
| Villarreal, Francisco; Epperson, Sara A; Ramirez-Sanchez, Israel et al. (2009) Regulation of cardiac fibroblast collagen synthesis by adenosine: roles for Epac and PI3K. Am J Physiol Cell Physiol 296:C1178-84 |
| Epperson, Sara A; Brunton, Laurence L; Ramirez-Sanchez, Israel et al. (2009) Adenosine receptors and second messenger signaling pathways in rat cardiac fibroblasts. Am J Physiol Cell Physiol 296:C1171-7 |
| Romero-Perez, Diego; Agrawal, Arpita; Jacobsen, Jennifer et al. (2009) Effects of novel semiselective matrix metalloproteinase inhibitors on ex vivo cardiac structure-function. J Cardiovasc Pharmacol 53:452-61 |
| Romero-Perez, Diego; Fricovsky, Eduardo; Yamasaki, Katrina Go et al. (2008) Cardiac uptake of minocycline and mechanisms for in vivo cardioprotection. J Am Coll Cardiol 52:1086-94 |
| Yamazaki, Katrina Go; Romero-Perez, Diego; Barraza-Hidalgo, Maraliz et al. (2008) Short- and long-term effects of (-)-epicatechin on myocardial ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol 295:H761-7 |
| Go, Katrina; Horikawa, Yousuke; Garcia, Ricardo et al. (2008) Fluorescent method for detection of cleaved collagens using O-phthaldialdehyde (OPA). J Biochem Biophys Methods 70:878-82 |
| Griffin, Michael O; Jinno, Miki; Miles, Lindsey A et al. (2005) Reduction of myocardial infarct size by doxycycline: a role for plasmin inhibition. Mol Cell Biochem 270:1-11 |
| Asbun, Juan; Manso, Ana Maria; Villarreal, Francisco J (2005) Profibrotic influence of high glucose concentration on cardiac fibroblast functions: effects of losartan and vitamin E. Am J Physiol Heart Circ Physiol 288:H227-34 |
