Single-Cell Analysis of the Normal Mouse Aorta Reveals Functionally Distinct Endothelial Cell Populations

• Published in: Circulation (New York, N.Y.) N.Y.), 2019-07, Vol.140 (2), p.147-163
• Main Authors: Kalluri, Aditya S, Vellarikkal, Shamsudheen K, Edelman, Elazer R, Nguyen, Lan, Subramanian, Ayshwarya, Ellinor, Patrick T, Regev, Aviv, Kathiresan, Sekar, Gupta, Rajat M
• Format: Article
• Language: English
• Subjects: Animals; Aorta - cytology; Aorta - physiology; Endothelial Cells - physiology; Female; Gene Expression Profiling - methods; Mice; Mice, Inbred C57BL; Single-Cell Analysis - methods
• ISSN: 0009-7322; 1524-4539
• DOI: 10.1161/CIRCULATIONAHA.118.038362

BACKGROUND:The cells that form the arterial wall contribute to multiple vascular diseases. The extent of cellular heterogeneity within these populations has not been fully characterized. Recent advances in single-cell RNA-sequencing make it possible to identify and characterize cellular subpopulations. METHODS:We validate a method for generating a droplet-based single-cell atlas of gene expression in a normal blood vessel. Enzymatic dissociation of 4 whole mouse aortas was followed by single-cell sequencing of >10 000 cells. RESULTS:Clustering analysis of gene expression from aortic cells identified 10 populations of cells representing each of the main arterial cell typesfibroblasts, vascular smooth muscle cells, endothelial cells (ECs), and immune cells, including monocytes, macrophages, and lymphocytes. The most significant cellular heterogeneity was seen in the 3 distinct EC populations. Gene set enrichment analysis of these EC subpopulations identified a lymphatic EC cluster and 2 other populations more specialized in lipoprotein handling, angiogenesis, and extracellular matrix production. These subpopulations persist and exhibit similar changes in gene expression in response to a Western diet. Immunofluorescence for Vcam1 and Cd36 demonstrates regional heterogeneity in EC populations throughout the aorta. CONCLUSIONS:We present a comprehensive single-cell atlas of all cells in the aorta. By integrating expression from >1900 genes per cell, we are better able to characterize cellular heterogeneity compared with conventional approaches. Gene expression signatures identify cell subpopulations with vascular disease–relevant functions.