TY - JOUR
T1 - Protein Kinase C epsilon-Calcineurin Cosignaling Downstream of Toll-Like Receptor 4 Downregulates Fibrosis and Induces Wound Healing Gene Expression in Cardiac Myofibroblasts
AU - Mesquita, Rui F. D. S.
AU - Paul, Margaret A.
AU - Valmaseda, Aida
AU - Francois, Asvi
AU - Jabr, Rita
AU - Anjum, Shahzia
AU - Marber, Michael S.
AU - Budhram-Mahadeo, Vishwanie
AU - Heads, Richard J.
PY - 2014/2
Y1 - 2014/2
N2 - The pathways which regulate resolution of inflammation and contribute to positive remodeling of the myocardium following injury are poorly understood. Here we show that protein kinase C epsilon (PKC epsilon) cooperates with the phosphatase calcineurin (CN) to potentiate induction of cardioprotective gene expression while suppressing expression of fibrosis markers. This was achieved by detailed analysis of the regulation of cyclooxygenase 2 (COX-2) expression as a marker gene and by using gene expression profiling to identify genes regulated by coexpression of CN-A alpha/PKC epsilon in adult rat cardiac myofibroblasts (ARVFs) on a larger scale. GeneChip analysis of CN-A alpha/PKC epsilon-coexpressing ARVFs showed that COX-2 provides a signature for wound healing and is associated with downregulation of fibrosis markers, including connective tissue growth factor (CTGF), fibronectin, and collagens Col1a1, Col3a1, Col6a3, Col11a1, Col12a1, and Col14a1, with concomitant upregulation of cardioprotection markers, including COX-2 itself, lipocalin 2 (LCN2), tissue inhibitor of metalloproteinase 1 (TIMP-1), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS). In primary rat cardiomyocyte cultures Toll-like receptor 4 (TLR4) agonist-or PKC epsilon/CN-dependent COX-2 induction occurred in coresident fibroblasts and was blocked by selective inhibition of CN or PKC alpha/epsilon or elimination of fibroblasts. Furthermore, ectopic expression of PKC epsilon and CN in ARVFs showed that the effects on COX-2 expression are mediated by specific NFAT sites within the COX-2 promoter as confirmed by site-directed mutagenesis and chromatin immunoprecipitation (ChIP). Therefore, PKC epsilon may negatively regulate adverse myocardial remodeling by cooperating with CN to downregulate fibrosis and induce transcription of cardioprotective wound healing genes, including COX-2.
AB - The pathways which regulate resolution of inflammation and contribute to positive remodeling of the myocardium following injury are poorly understood. Here we show that protein kinase C epsilon (PKC epsilon) cooperates with the phosphatase calcineurin (CN) to potentiate induction of cardioprotective gene expression while suppressing expression of fibrosis markers. This was achieved by detailed analysis of the regulation of cyclooxygenase 2 (COX-2) expression as a marker gene and by using gene expression profiling to identify genes regulated by coexpression of CN-A alpha/PKC epsilon in adult rat cardiac myofibroblasts (ARVFs) on a larger scale. GeneChip analysis of CN-A alpha/PKC epsilon-coexpressing ARVFs showed that COX-2 provides a signature for wound healing and is associated with downregulation of fibrosis markers, including connective tissue growth factor (CTGF), fibronectin, and collagens Col1a1, Col3a1, Col6a3, Col11a1, Col12a1, and Col14a1, with concomitant upregulation of cardioprotection markers, including COX-2 itself, lipocalin 2 (LCN2), tissue inhibitor of metalloproteinase 1 (TIMP-1), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS). In primary rat cardiomyocyte cultures Toll-like receptor 4 (TLR4) agonist-or PKC epsilon/CN-dependent COX-2 induction occurred in coresident fibroblasts and was blocked by selective inhibition of CN or PKC alpha/epsilon or elimination of fibroblasts. Furthermore, ectopic expression of PKC epsilon and CN in ARVFs showed that the effects on COX-2 expression are mediated by specific NFAT sites within the COX-2 promoter as confirmed by site-directed mutagenesis and chromatin immunoprecipitation (ChIP). Therefore, PKC epsilon may negatively regulate adverse myocardial remodeling by cooperating with CN to downregulate fibrosis and induce transcription of cardioprotective wound healing genes, including COX-2.
KW - GELATINASE-ASSOCIATED LIPOCALIN
KW - TISSUE GROWTH-FACTOR
KW - MYOCARDIAL-INFARCTION
KW - ANGIOTENSIN-II
KW - PKC-EPSILON
KW - SIMULATED ISCHEMIA
KW - HEART-FAILURE
KW - T-LYMPHOCYTES
KW - UP-REGULATION
KW - NITRIC-OXIDE
U2 - 10.1128/MCB.01098-13
DO - 10.1128/MCB.01098-13
M3 - Article
SN - 0270-7306
VL - 34
SP - 574
EP - 594
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
IS - 4
ER -