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Mononuclear phagocytes, including monocytes and macrophages within the vessel wall, are major players in atherosclerosis. Single-cell RNA sequencing studies revealed that atherosclerotic artery walls encompass several monocytes and vascular macrophages, defining at least nine distinct subsets potentially serving diverse functions in disease progression. This study investigates the functional phenotype and ontogeny of Olfr2-expressing vascular macrophages in atherosclerosis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods and results<\/jats:title>\n                    <jats:p>Olfr2+ macrophages rapidly increase in Apoe\u2212\/\u2212 mice\u2019s aorta when fed a Western diet (WD). Mass cytometry showed that Olfr2+ cells are clustered within the CD64 high population and enriched for CD11c and Ccr2 markers. Olfr2+ macrophages express many pro-inflammatory cytokines, including Il1b, Il6, Il12, and Il23, and chemokines, including Ccl5, Cx3cl1, Cxcl9, and Ccl22. By extracting differentially expressed genes from bulk RNA sequencing (RNA-seq) of Olfr2+ vs. Olfr2\u2212 macrophages, we defined a signature that significantly mapped to single-cell data of plaque myeloid cells, including monocytes, subendothelial MacAir, and Trem2Gpnmb foamy macrophages. By adoptive transfer experiments, we identified that Olfr2 competent monocytes from CD45.1Apoe\u2212\/\u2212Olfr2+\/+ mice transferred into CD45.2Apoe\u2212\/\u2212Olfr2\u2212\/\u2212 recipient mice fed WD for 12 weeks, accumulate in the atherosclerotic aorta wall already at 72 h, and differentiate in macrophages. Olfr2+ macrophages showed significantly increased BrdU incorporation compared to Olfr2\u2212 macrophages. Flow cytometry confirmed that at least 50% of aortic Olfr2+ macrophages are positive for BODIPY staining and have increased expression of both tumour necrosis factor and interleukin 6 compared to Olfr2\u2212 macrophages. Gene set enrichment analysis of the Olfr2+ macrophage signature revealed a similar enrichment pattern in human atherosclerotic plaques, particularly within foamy\/TREM2hi-M\u03c6 and monocytes.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>In summary, we conclude that Olfr2+ macrophages in the aorta originate from monocytes and can accumulate at the early stages of disease progression. These cells can undergo differentiation into MacAir and Trem2Gpnmb foamy macrophages, exhibiting proliferative and pro-inflammatory potentials. This dynamic behaviour positions them as key influencers in shaping the myeloid landscape within the atherosclerotic plaque.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/cvr\/cvae153","type":"journal-article","created":{"date-parts":[[2024,7,18]],"date-time":"2024-07-18T17:16:50Z","timestamp":1721323010000},"page":"1577-1589","source":"Crossref","is-referenced-by-count":20,"title":["Olfr2-positive macrophages originate from monocytes proliferate\n                    <i>in situ<\/i>\n                    and present a pro-inflammatory foamy-like phenotype"],"prefix":"10.1093","volume":"120","author":[{"given":"Sujit Silas","family":"Armstrong Suthahar","sequence":"first","affiliation":[{"name":"Division of Inflammation Biology, La Jolla Institute for Immunology , 9420 Athena Cir, La Jolla, CA 92037 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felix 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