Effect of Mechanical Stretch on Sex-Dependent Gene Expression by RNA Sequencing in Mouse Renal Endothelial Cells

Abstract

Increased mechanical endothelial cell stretch contributes to the development of numerous cardiovascular and renal pathologies by orchestration of immune cell activation. While endothelial cells are not recognized as professional antigen presenting cells, they can express both major histocompatibility complexes (MHC) I and II. MHCs are essential to form part of the immunological synapse during antigen presentation to adaptive immune cells. The mechanisms by which increased endothelial cell stretch regulates renal endothelial antigen processing and presentation is unknown. We hypothesized that increased mechanical endothelial stretch promotes upregulation of key antigen processing and presentation genes in mouse renal endothelial cells (MRECs) in a sex-dependent manner. MRECs derived from both sexes underwent 5%, 10%, and 15% (1 Hz) uniaxial cyclical stretch for 48 hours. After stretch, RNA sequencing was performed and bioinformatic and pathway analysis was executed to determine potential inflammatory mechanisms of action. We found that 10% stretch induced downregulation in MHC and costimulatory genes ( H2-Da, H2-K1, H2-T23, H2-T24) and upregulation of antigen processing associated genes ( tap1, psmb2, psmb4, and psmb8) in male MRECs. Females undergoing 10% stretch exhibited inverse gene expression profiles with upregulation of MHC genes and downregulation of antigen processing and presentation genes. At 15% stretch, the sex-specific gene expression tendencies were inverse from 10% stretch. To understand the function of differential gene expression we performed gene ontology enrichment pathway analysis. Male MRECs undergoing 10% stretch exhibited an enrichment for renal and cardiovascular development processes and inflammatory processes including chemokine-mediated signaling and cytokine-mediated signaling. When we compared female MRECs at 10% stretch, we found enrichment for renal and cardiovascular development processes including endothelial cell migration, angiogenesis and glomerular vascular, kidney, and artery development. Interestingly, 10% mechanical stretch enriched pro-inflammatory processes including positive regulation of cell migration and leukocyte migration. These data provide novel insight into how mechanical stretch regulates key antigen processing and presentation pathways in renal endothelial cells in a stretch and sex-dependent manner. This work was funded by MUSC Core Centers for Clinical Research Project Award to JPVB and Biomedical Laboratory Research and Development Service of the VA office of Research and Development Awards IK2BX003922 to KYDP and IK2BX005605 to JPVB. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.