VCAM-1 induces signals that stimulate ZO-1 serine phosphorylation and reduces ZO-1 localization at lung endothelial cell junctions

Abstract

Endothelial cell VCAM-1 regulates recruitment of lymphocytes, eosinophils, mast cells, or dendritic cells during allergic inflammation. In this report, we demonstrated that, during allergic lung responses, there was reduced zonula occludens (ZO)-1 localization in lung endothelial cell junctions, whereas there was increased lung endothelial cell expression of VCAM-1, N-cadherin, and angiomotin. In vitro, leukocyte binding to VCAM-1 reduced ZO-1 in endothelial cell junctions. Using primary human endothelial cells and mouse endothelial cell lines, Ab crosslinking of VCAM-1 increased serine phosphorylation of ZO-1 and induced dissociation of ZO-1 from endothelial cell junctions, demonstrating that VCAM-1 regulates ZO-1. Moreover, VCAM-1 induction of ZO-1 phosphorylation and loss of ZO-1 localization at cell junctions was blocked by inhibition of VCAM-1 intracellular signals that regulate leukocyte transendothelial migration, including NOX2, PKCα, and PTP1B. Furthermore, exogenous addition of the VCAM-1 signaling intermediate H O (1 μM) stimulated PKCα-dependent and PTP1B-dependent serine phosphorylation of ZO-1 and loss of ZO-1 from junctions. Overexpression of ZO-1 blocked leukocyte transendothelial migration. In summary, leukocyte binding to VCAM-1 induces signals that stimulated ZO-1 serine phosphorylation and reduced ZO-1 localization at endothelial cell junctions during leukocyte transendothelial migration.

Transcriptional profiling of breast cancer‐associated lymphatic vessels reveals VCAM‐1 as regulator of lymphatic invasion and permeability

“…We next hypothesized that VCAM-1 on LECs might be involved in the organization of cell-to-cell junctions and in the regulation of permeability, as has been reported to be the case in blood vessels. [44][45][46] Indeed, using an in vitro permeability assay of mouse LEC monolayers and a 70kD FITC-dextran tracer, we observed that treatment with 4T1-1F8 CM increased permeability, which could be entirely blocked by preincubation of the LECs with a VCAM-1 blocking antibody (Fig. 5d).…”

“…reported that melanoma cell‐derived SPARC binds to VCAM‐1 on blood vessel endothelial cells, triggering a signaling cascade that leads to actin remodeling and vascular barrier disruption. Furthermore, leukocyte binding to VCAM‐1 via integrin α4/β1 was reported to destabilize endothelial junctions via dissociation of the endothelial phosphatase VE‐PTP from VE‐cadherin complexes and phosphorylation of ZO‐1, leading to loosening of endothelial junctions. Our data suggest that although 4T1 tumor cells express SPARC (data not shown), SPARC is not involved in the regulation of lymphatic permeability, at least in vitro .…”

“…Leukocyte binding to VCAM-1 on blood vessel endothelial cells via integrin α4/β1 has also been reported to stimulate junctional weakening by displacing ZO-1 46 and by reducing VE-cadherin mediated cell adhesion via VE-PTP displacement from VE-cadherin. 44 Cultured mouse LECs expressed α4, α9 and β1 integrins (Fig.…”

The Heat Shock Protein 70 Plays a Protective Role in Sepsis by Maintenance of the Endothelial Permeability

“…Thus, HSP70 can regulate LPS-induced augmentation of cell permeability of vascular endothelial cells in vitro . The cell junction is a key factor of maintenance of normal cell permeability in vascular endothelial cells [ 23 ]. To investigate the mechanism, we detected the expression of several cell junction proteins including E-cadherin, occludin, and ZO-1 using western blot.…”

Tight junctions in the development of asthma, chronic rhinosinusitis, atopic dermatitis, eosinophilic esophagitis, and inflammatory bowel diseases

“…To date, relatively little is known about the regulation of claudins in asthma, but recent studies revealed that the lung expression of claudin‐18 in asthma is decreased in association with Th2 inflammation, and IL‐13 acts to decrease its expression in both human and mice lung tissue 44,45 . During recruitment of leukocytes, VCAM‐1 signaling in endothelial cells induces changes in endothelial cell junction involving ZO‐1 46 . An in vitro culture experiment using SV40‐transformed human bronchial epithelial cell line indicated that the nuclear factor erythroid 2‐related factor 2 (Nrf2)‐aldehyde oxidase 1 (AOX1) pathway is an important regulator of barrier integrity 47 .…”