Role of A2AR-D2R Dimerization and D2R-Biased Signaling in PDE10A-Mediated Cardiac Hypertrophy.

BACKGROUND Pathological cardiomyocyte (CM) hypertrophy is a hallmark of dilated cardiomyopathy and heart failure, driven by mechanical or neurohumoral stress. Although we previously demonstrated PDE10A (phosphodiesterase 10A) as a critical contributor and potential therapeutic target in pathological cardiac remodeling and dysfunction, the underlying mechanisms remain unclear. Here, we investigated the specific signaling pathways and sources of cyclic nucleotides modulated by PDE10A in CM hypertrophy. METHODS CM hypertrophy was induced by angiotensin II in isolated adult mouse CMs, with hypertrophy assessed by cell surface area and protein synthesis. PDE10A inactivation was achieved through PDE10A knockout or inhibition with TP-10. Various pharmacological agonists/antagonists, genetic mutants, and knockout models of D2R (dopamine receptor D2) and A2AR (adenosine receptor 2A) were used to study the role of A2AR-D2R heterodimer and downstream signaling in CM hypertrophy in vitro and in vivo. Viral vectors were used to manipulate protein or shRNA expression targeting key signaling molecules. RESULTS We demonstrated that the antihypertrophic effect of PDE10A inactivation is specifically dependent on cAMP/PKA (protein kinase A) signaling. Importantly, we discovered an A2AR-D2R heterodimer and its association with PDE10A in CMs. The A2AR-D2R heterodimer mediates the βarr2 (beta-arrestin 2)-biased D2R signaling, and activating D2R-biased signaling antagonizes cardiac hypertrophy and dysfunction. PDE10A deficiency or inhibition enhanced A2AR-D2R heterodimerization, promoting βarr2-biased D2R signaling and downstream PP2A (protein phosphatase 2A)-B56δ activation through increased A2AR/cAMP/PKA-mediated D2R phosphorylation. PDE10A inactivation inhibits stress signal-induced HDAC5 (histone deacetylase 5) nuclear export and phosphorylation, which are dependent on PP2A. Genetic and pharmacological approaches in animal models confirmed the critical roles of A2AR-D2R heterodimerization, D2R phosphorylation, and biased D2R signaling in vivo. Furthermore, combining PDE10A inhibition with A2AR or D2R-biased agonism produced synergistic antihypertrophic effects, highlighting their novel therapeutic potential. CONCLUSIONS Our findings reveal A2AR-D2R dimerization and βarr2-biased D2R signaling as novel and critical mechanisms for counteracting pathological CM hypertrophy and cardiac dysfunction. PDE10A acts as a pivotal negative regulator of this signaling axis. Targeting PDE10A, A2AR-D2R/βarr2 signaling, or both offers potentially novel therapeutic strategies for combating pathological cardiac remodeling and cardiac dysfunction.