Single-Nuclear RNA Sequencing Reveals Regional Specialization and Cellular Interactions in Epicardial and Perivascular Adipose Tissue

Figures

• 

  Figure 1.. Single cell profiling of epicardial adipose tissue. a, Diagram showing locations of adipose tissue samples. b, UMAP plot of integrated snRNA-seq analysis of adipose tissue, split by depot and subsampled to the same number of cells (4781) per depot. c, Proportion of cell types in adipose samples split by depot.

• 

  Figure 2.. Adipocyte subclusters and gene expression vary across adipose depot. a, UMAP plot of integrated adipocyte subclusters (n = 7513 cells). b, Subcluster distribution across depot. c, Marker genes for adipocyte subclusters. d, Selected GO pathways from pathway analysis of adipocyte subcluster marker genes. e, Selected marker genes highlighted in pathway analysis.

• 

  Figure 3:. ASPC subclusters and gene expression vary across adipose depot. a, UMAP plot of integrated ASPC subclusters (n = 3593 cells). b, Subcluster distribution across depot. c, Marker genes for ASPC subclusters. d, Selected GO pathways from pathway analysis of ASPC subcluster marker genes. e, Selected marker genes highlighted in pathway analysis.

• 

  Figure 4. Pseudobulk analysis of adipocytes and ASPCs by depot. a, PCA analysis of adipocyte pseudobulk data. b, Volcano plots comparing gene expression in adipocytes from each epicardial depot compared to subcutaneous adipocytes. c, UpSet plot showing the intersection between upregulated (left) or downregulated (right) genes in epicardial adipocytes compared to subcutaneous adipocytes. d, Pathway analysis of genes upregulated (green) or downregulated (yellow) in both left atrium and right coronary artery adipocytes compared to subcutaneous adipocytes. e, PCA analysis of ASPC pseudobulk data. f, Volcano plots comparing gene expression in each ASPCs from epicardial depot compared to subcutaneous ASPCs. g, UpSet plot showing the intersection between upregulated (left) or downregulated (right) genes in epicardial ASPCs compared to subcutaneous ASPCs. h, Pathway analysis of genes upregulated (green) or downregulated (yellow) in both left atrium and right coronary artery ASPCs compared to subcutaneous ASPCs.

• 

  Figure 5:. Cellchat analysis predicts epicardal fat-selective interactions with cardiomyocytes. a, Diagram describing the cellchat analysis experiments. Data from each epicardial depot was combined with cardiomyocyte or vascular smooth muscle cell data from a corresponding cardiac or vascular dataset and cellchat was run on this combined dataset. Subcutaneous data was similarly combined with each cardiac and vascular dataset to serve as a control. b, Heatmaps showing the predicted interaction strength in each epicardial dataset compared to subcutaneous. c, Comparison of predicted interactions that were significantly upregulated between epicardial and cardiovascular cell types compared to subcutaneous and cardiovascular cell types. d, Chord plot showing the predicted EPHA signaling pathway network in subcutaneous versus Left Atrium fat. e, Violin plot showing expression of the receptor gene EPHA3 in all cell types, split by depot.

• 

  Figure 6.. Cellchat analysis predicts changing interactions between fat and cardiovascular cells in the disease state. a, Diagram of performed cellchat analysis. Single nuclear data from left atrium fat was combined with cardiomyocytes from either healthy or AFib left atrium hearts, then predicted interactions were compared across disease state. b, Heatmap showing the strength of predicted interactions with AFib versus healthy cardiomyocytes. c, Select signaling networks predicted between left atrium epicardial fat and healthy versus AFib cardiomyocytes. d, Diagram of performed cellchat analysis. Single nuclear data from ascending aorta fat was combined with VSMCs from healthy aorta versus aortic aneurysm. e, Heatmap of strength of predicted interactions in aortic aneurysm compared to healthy aorta. f, Selected signaling networks predicted between ascending aorta epicardial fat and VSMCs from healthy or aneurysm aortas.