Age-related changes in tissue macrophages precede cardiac functional impairment

Cardiac tissue macrophages (cTMs) are abundant in the murine heart but the extent to which the cTM phenotype changes with age is unknown. This study characterizes aging-dependent phenotypic changes in cTM subsets. Using the Cx₃cr1^GFP/+ mouse reporter line where GFP marks cTMs, and the tissue macrophage marker Mrc1, we show that two major cardiac tissue macrophage subsets, Mrc1⁻GFP^hi and Mrc1⁺GFP^hi cTMs, are present in the young (<10 week old) mouse heart, and a third subset, Mrc1⁺GFP^lo, comprises ~50% of total Mrc1⁺ cTMs from 30 weeks of age. Immunostaining and functional assays show that Mrc1⁺ cTMs are the principal myeloid sentinels in the mouse heart and that they retain proliferative capacity throughout life. Gene expression profiles of the two Mrc1⁺ subsets also reveal that Mrc1⁺GFP^lo cTMs have a decreased number of immune response genes (Cx₃cr1, Lpar6, CD9, Cxcr4, Itga6 and Tgfβr1), and an increased number of fibrogenic genes (Ltc4s, Retnla, Fgfr1, Mmp9 and Ccl24), consistent with a potential role for cTMs in cardiac fibrosis. These findings identify early age-dependent gene expression changes in cTMs, with significant implications for cardiac tissue injury responses and aging-associated cardiac fibrosis.