Atlas of the Immune Cell Repertoire in Mouse Atherosclerosis Defined by Single-Cell RNA-Sequencing and Mass Cytometry

• DOI: 10.1161/circresaha.117.312513
• Publication Date: 2018-03-15T09:15:23Z
• Authors: Holger Winkels; Erik Ehinger; Melanie Vassallo; Konrad Buscher; Huy Q. Dinh; Kouji Kobiyama; Anouk A.J. Hamers; Clément Cochain; Ehsan Vafadarnejad; Antoine-Emmanuel ...; Alma Zernecke; Akula Bala Pramod; Amlan K. Ghosh; Nathaly Anto Michel; Natalie Hoppe; Ingo Hilgendorf; Andreas Zirlik; Catherine C. Hedrick; Klaus Ley; Dennis Wolf
• Keywords: Mass cytometry

Atherosclerosis is a chronic inflammatory disease that driven by the interplay of pro- and anti-inflammatory leukocytes in aorta. Yet, phenotypic transcriptional diversity aortic poorly understood.We characterized from healthy atherosclerotic mouse aortas in-depth single-cell RNA-sequencing mass cytometry (cytometry time flight) to define an atlas immune cell landscape atherosclerosis.Using chow diet- Western diet-fed Apoe-/- Ldlr-/- mice, we detected 11 principal leukocyte clusters with distinct spatial characteristics while cellular repertoire was less diverse. Gene set enrichment analysis on level established multiple pathways, such as for lipid metabolism, proliferation, cytokine secretion, were confined particular clusters. Leukocyte populations differentially regulated mice. We confirmed these novel 35-marker panel metal-labeled antibodies conventional flow cytometry. Cell retrieved protein-based approaches highly correlated transcriptionally defined In integrated screening strategy RNA-sequencing, cytometry, fluorescence-activated sorting, 3 B-cell subsets alterations surface markers, functional vitro secretion. cluster gene signatures revealed frequencies 126 human plaques genetic deconvolution strategy. This approach carotid microdissected mostly populated macrophages, T-cells, monocytes. addition, frequency genetically predicted cardiovascular events patients.The definition high-dimensional analyses enables fine-grained subsets, reveals new immunologic mechanisms cell-type-specific establishes relevance lesional atherosclerosis.