CL-316243 facilitates stable atherosclerotic plaque phenotypes in association with suppression of perivascular adipose tissue ferroptosis via upregulating C/EBPβ

Atherosclerotic plaque rupture is the leading cause of most acute cardiovascular events and death. However, effective treatment strategies targeting plaque stability remain elusive. Perivascular adipose tissue (PVAT) dysfunction is increasingly recognized as a contributor to vulnerable plaque, implying that elucidating and targeting its underlying mechanism could represent a novel therapeutic strategy for atherosclerosis. This study identifies Ferroptosis as a critical pathological feature in PVAT of atherosclerotic artery, establishing a previously unrecognized link between PVAT dysfunction and Ferroptosis. The β3-adrenoceptor agonist CL-316243 (CL) significantly suppressed ferroptotic markers in both cultured adipocytes and atherosclerotic PVAT. Furthermore, CL treatment reduced PVAT inflammation and dysfunction, as shown by histological, gene expression, and functional analyses, thereby facilitating stable plaque phenotypes, reflected in reduced necrotic core size, lower CD68 and MMP expression, and increased Collagen content. Notably, the decline in Ferroptosis markers correlated significantly with these phenotypic improvements. Mechanistically, C/EBPβ was identified as a direct transcriptional regulator of GPX4 through CUT&Tag and dual-luciferase reporter assays, by which the critical promoter binding motif was also identified. Subsequently, CL was shown to upregulate GPX4 via C/EBPβ, and this effect was abolished by Cebpb knockdown. Further analysis indicated that CL-induced upregulation of C/EBPβ depends on the cAMP-PKA-CREB pathway. Finally, Cebpb knockdown abolished the anti-ferroptosis effect of CL, confirming C/EBPβ as a critical component of this mechanism. Our study reveals that CL stabilizes atherosclerotic plaques by suppressing PVAT Ferroptosis through the C/EBPβ-GPX4 axis, uncovering novel therapeutic targets and providing a potential treatment strategy for atherosclerotic diseases.