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Single cell RNA sequencing (scRNAseq) of severely atherosclerotic coronary artery from Ossabaw miniature swine with metabolic syndrome
Author(s) -
Eggenberger Caleb,
Badin Jill K.,
Alloosh Mouhamad,
Sturek Michael
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.05177
Subject(s) - medicine , artery , neointima , coronary arteries , fatty streak , cardiology , endothelium , fibrous cap , population , pathology , restenosis , aorta , stent , environmental health
Coronary atherogenesis involves multiple signaling pathways and underlying gene expression. The atherogenic plasma milieu (excess lipoproteins, glucose, etc.) injures the endothelium, which is the initiating event of coronary plaque development. Subsequently, smooth muscle cells proliferate and migrate from the wall into the neointima, and blood cells and macrophages infiltrate the vessel wall and contribute to the growing plaque. Our goal was to characterize the gene expression in the heterogeneous cell population in this dynamic coronary artery pathology. Methods We performed single cell RNA sequencing (scRNAseq) on cells dispersed from the proximal 45 mm of the left anterior descending coronary artery of healthy pigs compared to separate plaque and wall cell dispersions from arteries of Ossabaw swine that had metabolic syndrome and were atherosclerotic for 23 months. Primary analysis scRNAseq data used t‐distributed stochastic neighbor embedding (T‐SNE) plots for an integrated analysis of the 3 cell preparations. Results Metabolic syndrome was confirmed by obesity, dyslipidemia, and hypertension. In vivo intravascular ultrasound (IVUS) of the entire artery revealed every stage of atherosclerosis from fatty streak to complex lesions having up to 70% occlusion with fibrofatty and calcified lesions. Cells (3,510) from healthy coronary artery expressed predominantly markers for endothelial (PECAM1), endocardial (CDH5, POSTN), and smooth muscle cells (ACTA2, TAGLN, CNN1, MYL9, CALD1, RGS5, MHY11). Overall, the healthy coronary artery cells were much more homogeneous than atherosclerotic plaque and wall cells. Wall cells (8,224) in atherosclerotic arteries expressed ~no (0.1%) endothelial, many smooth muscle, and fibroblast genes. Plaque cells (8,049) expressed ~no (0.1%) endothelial genes, robust smooth muscle marker genes, and a high abundance of early marker genes for chondrocytes, osteoblasts, monocytes, neutrophils, and macrophages. Conclusion Profound gene expression patterns were shown with scRNAseq in a clinically relevant model of coronary atherosclerosis. The striking heterogeneity of gene expression and marker genes for chondrocytes and osteoblasts in plaque cells indicate significant transdifferentiation and suggest further progression to severe coronary artery calcification.