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Single-Cell RNA-Seq Reveals the Transcriptional Landscape and Heterogeneity of Aortic Macrophages in Murine Atherosclerosis
Abstract
Rationale: It is assumed that atherosclerotic arteries contain several macrophage subsets endowed with specific functions. The precise identity of these subsets is poorly characterized as they have been defined by the expression of a restricted number of markers. Objective: We have applied single-cell RNA sequencing as an unbiased profiling strategy to interrogate and classify aortic macrophage heterogeneity at the single-cell level in atherosclerosis. Method and Results: We performed single-cell RNA sequencing of total aortic CD45 + cells extracted from the nondiseased (chow fed) and atherosclerotic (11 weeks of high-fat diet) aorta of low-density lipoprotein receptor–deficient ( Ldlr −/− ) mice. Unsupervised clustering singled out 13 distinct aortic cell clusters. Among the myeloid cell populations, resident-like macrophages with a gene expression profile similar to aortic resident macrophages were found in healthy and diseased aortas, whereas monocytes, monocyte-derived dendritic cells, and 2 populations of macrophages were almost exclusively detectable in atherosclerotic aortas, comprising inflammatory macrophages showing enrichment in Il1b and previously undescribed TREM2 hi (triggered receptor expressed on myeloid cells 2) macrophages showing enrichment in Trem2 . Differential gene expression and gene ontology enrichment analyses revealed specific gene expression patterns distinguishing these 3 macrophage subsets and monocyte-derived dendritic cells and uncovered putative functions of each cell type. Notably, TREM2 hi macrophages seemed to be endowed with specialized functions in lipid metabolism and catabolism and presented a gene expression signature reminiscent of osteoclasts, suggesting a role in lesion calcification. TREM2 expression was moreover detected in human lesional macrophages. Importantly, these macrophage populations were present also in advanced atherosclerosis and in Apoe −/− aortas, indicating relevance of our findings in different stages of atherosclerosis and mouse models. Conclusions: These data unprecedentedly uncovered the transcriptional landscape and phenotypic heterogeneity of aortic macrophages and monocyte-derived dendritic cells in atherosclerotic and identified previously unrecognized macrophage populations and their gene expression signature, suggesting specialized functions. Our findings will open up novel opportunities to explore distinct myeloid cell populations and their functions in atherosclerosis.
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In the citing paper
“In atherosclerosis, TREM2+ macrophages are enriched in atherosclerotic lesions and specialize in lipid catabolism (Cochain et al., 2018).”
From TREM2 Modulation Remodels the Tumor Myeloid Landscape Enhancing Anti-PD-1 Immunotherapy (2020) · cites this paper as Cochain et al., 2018
In the citing paper
“LD accumulation in macrophages is associated with several inflammatory conditions (e.g., atherosclerosis) (4, 5), infectious diseases (e.g., mycobacterial infections) (6, 7), and cancer (8, 9).”
From Reducing microglial lipid load enhances β amyloid phagocytosis in an Alzheimer’s disease mouse model (2025) · cites this paper as [5]
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“LD accumulation in macrophages has also been documented in lipid-rich microenvironments in the peripheral, such as atherosclerosis plaques (5), white adipose tissue (10), and tumors (9).”
From Reducing microglial lipid load enhances β amyloid phagocytosis in an Alzheimer’s disease mouse model (2025) · cites this paper as [5]
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“Indeed, findings on unique macrophage subsets (such as TREM2+ macrophages) on the basis of single-cell analyses have uncovered unprecedented phenotypic heterogeneity of macrophages and monocyte-derived dendritic cells in atherosclerotic lesions and have defined a novel atlas of the immune cell repertoire in atherosclerosis135,154.”
From Inflammation and its resolution in atherosclerosis: mediators and therapeutic opportunities (2019) · cites this paper as [154]
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Metadata
- Authors
- Cochain, C., Vafadarnejad, E., Arampatzi, P., Pelisek, J., Winkels, H., Ley, K., Wolf, D., Saliba, A.-E., & Zernecke, A.
- Affiliations
- Universitätsklinikum Würzburg (DE)
- Helmholtz Institute for RNA-based Infection Research (DE)
- Core Unit Systems Medicine (P.A.)
- TUM Klinikum (DE)
- La Jolla Institute for Immunology (US)
- From the Institute of Experimental Biomedicine (C.C., A.Z.)
- Corresponding author
- Cochain, Clément
- Vafadarnejad, Ehsan
- ORCID
- Venue
- Circulation Research
- Published
- 2018-06-08
- Volume
- 122
- Issue
- 12
- Pages
- 1661-1674
- ISSN
- 0009-7330 (ISSN-L)
- 1524-4571
- OpenAlex
- W2792855672
- Cited by
- 894
- References
- 54
- Citations by year
- 2026: 99 · 2025: 122 · 2024: 118 · 2023: 126 · 2022: 138 · 2021: 117 · 2020: 90 · 2019: 58 · 2018: 24
- FWCI
- 31.06
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Circulation Research (publishedVersion)
- PubMed (publishedVersion)
- mediaTUM (Technical University of Munich) (submittedVersion)
- Funders
- National Heart, Lung, and Blood Institute (R01HL121697)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Atherosclerosis and Cardiovascular Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Keywords
- atherosclerosis, macrophage heterogeneity, TREM2, monocyte-derived dendritic cells, inflammatory macrophages, tissue-resident macrophages, aortic calcification, lipid metabolism, Ldlr-deficient mice, ApoE-deficient mice, high-fat diet
- MeSH terms
- Animals, Aortic Diseases, Apolipoproteins E, B-Lymphocytes, Dendritic Cells, Humans, Leukocytes, Macrophages, Male, Monocytes, Phenotype, Receptors, LDL, T-Lymphocytes, Biomarkers, Sequence Analysis, RNA, Gene Expression Profiling, Atherosclerosis, Mice, Single-Cell Analysis
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