Matricellular Protein CCN5 Reverses Established Cardiac Fibrosis

Abstract

BACKGROUND Cardiac fibrosis (CF) is associated with increased ventricular stiffness and diastolic dysfunction and is an independent predictor of long-term clinical outcomes of patients with heart failure (HF). We previously showed that the matricellular CCN5 protein is cardioprotective via its ability to inhibit CF and preserve cardiac contractility. OBJECTIVES This study examined the role of CCN5 in human heart failure and tested whether CCN5 can reverse established CF in an experimental model of HF induced by pressure overload. METHODS Human hearts were obtained from patients with end-stage heart failure. Extensive CF was induced by applying transverse aortic constriction for 8 weeks, which was followed by adeno-associated virus-mediated transfer of CCN5 to the heart. Eight weeks following gene transfer, cellular and molecular effects were examined. RESULTS Expression of CCN5 was significantly decreased in failing hearts from patients with end-stage heart failure compared to nonfailing hearts. Trichrome staining and myofibroblast content measurements revealed that the established CF had been reversed by CCN5 gene transfer. Anti-CF effects of CCN5 were associated with inhibition of the transforming growth factor beta signaling pathway. CCN5 significantly inhibited endothelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation, which are 2 critical processes for CF progression, both in vivo and in vitro. In addition, CCN5 induced apoptosis in myofibroblasts, but not in cardiomyocytes or fibroblasts, both in vivo and in vitro. CCN5 provoked the intrinsic apoptotic pathway specifically in myofibroblasts, which may have been due the ability of CCN5 to inhibit the activity of NFκB, an antiapoptotic molecule. CONCLUSIONS CCN5 can reverse established CF by inhibiting the generation of and enhancing apoptosis of myofibroblasts in the myocardium. CCN5 may provide a novel platform for the development of targeted anti-CF therapies.

Mitochondrial quality-control dysregulation in conditional HO-1–/– mice

“…We found increased fibrosis in HO-1(CM) -/-mice, accompanied by increases in TGF-β1 expression, which stimulates the accumulation of extracellular matrix proteins and fibrosis (84,85). In WT/Cre mice, TGF-β1 was not significantly induced after hyperoxia, and there was a significant increase in CCN5, a negative regulator of α-SMA and collagen I expression and a potent repressor of TGF-β1 (86,87). CCN5 expression, however, was reduced in HO-1(CM) -/-mice.…”

Cytokine-Like 1 Regulates Cardiac Fibrosis via Modulation of TGF-β Signaling

“…The recombinant AAV was produced by transfecting 293T cells as previously described [19]. The AAV particles in the cell culture media were precipitated with ammonium sulfate and purified by ultracentrifugation on an iodixanol gradient.…”

Enhancing atrial‐specific gene expression using a calsequestrin cis‐regulatory module 4 with a sarcolipin promoter

“…Cardiac gene therapy has emerged as a promising method for combating cardiac diseases. Currently, adeno‐associated virus serotype 9 (AAV9) is the most promising delivery vehicle for transgene expression and the most widely used in cardiac preclinical trials because of its tropism for the heart . Although the AAV9 vector in conjunction with the ubiquitous cytomegalovirus (CMV) promoter has shown cardiac specificity, a major limitation is the expression of transgenes in other organs, such as liver, lung, thymus, brain and kidney .…”