Protocatechuic acid boosts continual efferocytosis in macrophages by derepressing KLF4 to transcriptionally activate MerTK

Abstract

Macrophages clear apoptotic cells through a process called continual efferocytosis. We found that protocatechuic acid (PCA), a polyphenolic compound abundant in fruits and vegetables, increased the continual efferocytic capacity of macrophages and inhibited the progression of advanced atherosclerosis. PCA reduced the intracellular amounts of microRNA-10b (miR-10b) by promoting its secretion in extracellular vesicles, which led to an increase in the abundance of the miR-10b target Krüppel-like factor 4 (KLF4). In turn, KLF4 transcriptionally induced the gene encoding Mer proto-oncogene tyrosine kinase (MerTK), an efferocytic receptor for the recognition of apoptotic cells, resulting in increased continual efferocytic capacity. However, in naive macrophages, the PCA-induced secretion of miR-10b did not affect KLF4 and MerTK protein abundance or efferocytic capacity. In mice, oral administration of PCA increased continual efferocytosis in macrophages residing in the peritoneal cavities, thymi, and advanced atherosclerotic plaques through the miR-10b–KLF4–MerTK pathway. In addition, pharmacological inhibition of miR-10b with antagomiR-10b also increased the efferocytic capacity of efferocytic but not naive macrophages in vitro and in vivo. Together, these data describe a pathway that promotes continual efferocytosis in macrophages through miR-10b secretion and a KLF4-dependent increase in MerTK abundance, which can be activated by dietary PCA and which has implications for understanding the regulation of continual efferocytosis in macrophages.