Vascular Bed–specific Expression of an Endothelial Cell Gene Is Programmed by the Tissue Microenvironment

TLDR

The endothelium adapts morphologically and functionally to the demands of underlying tissues, yet the molecular basis of endothelial cell diversity remains largely unknown. The study aims to investigate the mechanisms governing differential expression of the endothelial cell–restricted von Willebrand factor (vWF) gene. Transgenic mice carrying a 2,182‑bp vWF promoter fragment linked to a LacZ reporter were generated to assess tissue‑specific expression. β‑galactosidase driven by the vWF promoter was detected in endothelial cells of brain, heart, and skeletal muscle, and in transplantation and coculture assays cardiac microvascular endothelial cells upregulated vWF in response to cardiac myocytes, whereas thrombomodulin and vWF expression were unchanged by fibroblasts or hepatocytes, indicating that vWF is regulated by vascular bed–specific pathways responding to local microenvironment signals.

Abstract

The endothelium is morphologically and functionally adapted to meet the unique demands of the underlying tissue. At the present time, little is known about the molecular basis of endothelial cell diversity. As one approach to this problem, we have chosen to study the mechanisms that govern differential expression of the endothelial cell–restricted von Willebrand factor (vWF) gene. Transgenic mice were generated with a fragment of the vWF gene containing 2,182 bp of 5′ flanking sequence, the first exon and first intron coupled to the LacZ reporter gene. In multiple independent lines of mice, β-galactosidase expression was detected within endothelial cells in the brain, heart, and skeletal muscle. In isogeneic transplantation models, LacZ expression in host-derived auricular blood vessels was specifically induced by the microenvironment of the heart. In in vitro coculture assays, expression of both the transgene and the endogenous vWF gene in cardiac microvascular endothelial cells (CMEC) was upregulated in the presence of cardiac myocytes. In contrast, endothelial cell levels of thrombomodulin protein and mRNA were unchanged by the addition of ventricular myocytes. Moreover, CMEC expression of vWF was not influenced by the addition of 3T3 fibroblasts or mouse hepatocytes. Taken together, the results suggest that the vWF gene is regulated by vascular bed–specific pathways in response to signals derived from the local microenvironment.