Abstract P110: Differential Gene Expression Associated With Ccn1-positive Renal Cells In Murine Hypertensive Models

Abstract

Hypertension affects roughly half of the U.S. population and is characterized by detrimental pro-inflammatory changes that alter renal function and the cardiovascular system over time. The kidney is a major regulator of blood pressure, however, the specific contributions of renal cell types, including lymphatic endothelial cells (LECs), to hypertension have not been thoroughly explored. Previous work from our lab has shown an increase in renal lymphangiogenesis in response to hypertension and that augmentation of renal lymphangiogenesis can reduce blood pressure. CCN1/Cyr61 is an extracellular matrix protein secreted by multiple cell types, particularly LECs and fibroblasts, which contributes to integrin signaling pathways. CCN1 is also known to be involved in lymphangiogenesis, and has previously been implicated in GWAS studies of hypertension. Given these connections, we performed single cell RNA sequencing using angiotensin II and salt sensitive mouse models of hypertension, which were analyzed separately as well as aggregated into one hypertensive group. Kidneys were turned to a single cell suspension and separated for CD31+/Podoplanin+ cells meant to better represent LECs, which are a small fraction of renal cells. Sequencing was performed and the gene reads were run through the Cell Ranger, Seurat, FGSEA, and Slingshot pipelines, then CCN1+ cells were isolated digitally and analyzed separately. In the AngII model, a total of 60 genes (28 up, 32 down) were differentially expressed (p<0.01) between the renal CCN1+ cells in the hypertensive mice compared to controls, while the salt sensitive model had a total of 331 differentially expressed genes (305 up, 26 down). When the hypertension groups were merged, there were a total of 600 differentially expressed genes (503 up, 97 down) with GSEA showing 48 pathways with enrichment scores >3 or <-3. Additionally, the cellular profile of CCN1+ cells shifted, branching out from a fibroblast-like population to other cell types such as endothelium and tubular epithelium. Overall, the expression of CCN1+ renal cells demonstrates notable changes in response to hypertension in murine models and encourages further study of the mechanisms behind CCN1’s role in the development and maintenance of hypertension.