Deletion of Osteopontin Enhances β2-Adrenergic Receptor-Dependent Anti-Fibrotic Signaling in Cardiomyocytes

Cardiac (2)-adrenergic receptors (ARs) are known to inhibit collagen production and fibrosis in cardiac fibroblasts and myocytes. The (2)AR is a Gs protein-coupled receptor (GPCR) and, upon its activation, stimulates the generation of cyclic 3,5-adenosine monophosphate (cAMP). cAMP has two effectors: protein kinase A (PKA) and the exchange protein directly activated by cAMP (Epac). Epac1 has been shown to inhibit cardiac fibroblast activation and fibrosis. Osteopontin (OPN) is a ubiquitous pro-inflammatory cytokine, which also mediates fibrosis in several tissues, including the heart. OPN underlies several cardiovascular pathologies, including atherosclerosis and cardiac adverse remodeling. We found that the cardiotoxic hormone aldosterone transcriptionally upregulates OPN in H9c2 rat cardiac myoblastsan effect prevented by endogenous (2)AR activation. Additionally, CRISPR-mediated OPN deletion enhanced cAMP generation in response to both (1)AR and (2)AR activation in H9c2 cardiomyocytes, leading to the upregulation of Epac1 protein levels. These effects rendered (2)AR stimulation capable of completely abrogating transforming growth factor (TGF)--dependent fibrosis in OPN-lacking H9c2 cardiomyocytes. Finally, OPN interacted constitutively with Gs subunits in H9c2 cardiac cells. Thus, we uncovered a direct inhibitory role of OPN in cardiac (2)AR anti-fibrotic signaling via cAMP/Epac1. OPN blockade could be of value in the treatment and/or prevention of cardiac fibrosis.