scholarly journals Interfering with VE-PTP stabilizes endothelial junctions in vivo via Tie-2 in the absence of VE-cadherin

2015 ◽  
Vol 212 (13) ◽  
pp. 2267-2287 ◽  
Author(s):  
Maike Frye ◽  
Martina Dierkes ◽  
Verena Küppers ◽  
Matthias Vockel ◽  
Janina Tomm ◽  
...  
Keyword(s):  
Tyrosine Phosphatase ◽  
Endothelial Barrier ◽  
Vascular Endothelial ◽  
Leukocyte Transmigration ◽  
Gene Ablation ◽  
Nonmuscle Myosin Ii ◽  
Endothelial Junctions ◽  
The Stability ◽  
Endothelial Junction

Vascular endothelial (VE)–protein tyrosine phosphatase (PTP) associates with VE-cadherin, thereby supporting its adhesive activity and endothelial junction integrity. VE-PTP also associates with Tie-2, dampening the tyrosine kinase activity of this receptor that can support stabilization of endothelial junctions. Here, we have analyzed how interference with VE-PTP affects the stability of endothelial junctions in vivo. Blocking VE-PTP by antibodies, a specific pharmacological inhibitor (AKB-9778), and gene ablation counteracted vascular leak induction by inflammatory mediators. In addition, leukocyte transmigration through the endothelial barrier was attenuated. Interference with Tie-2 expression in vivo reversed junction-stabilizing effects of AKB-9778 into junction-destabilizing effects. Furthermore, lack of Tie-2 was sufficient to weaken the vessel barrier. Mechanistically, inhibition of VE-PTP stabilized endothelial junctions via Tie-2, which triggered activation of Rap1, which then caused the dissolution of radial stress fibers via Rac1 and suppression of nonmuscle myosin II. Remarkably, VE-cadherin gene ablation did not abolish the junction-stabilizing effect of the VE-PTP inhibitor. Collectively, we conclude that inhibition of VE-PTP stabilizes challenged endothelial junctions in vivo via Tie-2 by a VE-cadherin–independent mechanism. In the absence of Tie-2, however, VE-PTP inhibition destabilizes endothelial barrier integrity in agreement with the VE-cadherin–supportive effect of VE-PTP.

scholarly journals Rasip1 regulates vertebrate vascular endothelial junction stability through Epac1-Rap1 signaling

Blood ◽  
2013 ◽  
Vol 122 (22) ◽  
pp. 3678-3690 ◽  
Author(s):  
Christopher W. Wilson ◽  
Leon H. Parker ◽  
Christopher J. Hall ◽  
Tanya Smyczek ◽  
Judy Mak ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Vascular Endothelial ◽  
Key Points ◽  
Endothelial Junction

Key Points RASIP1 is required for stabilizing nascent patent blood vessels in both mice and zebrafish. RASIP1 is a dynamic effector of EPAC1-RAP1 signaling that controls actin bundling and restricts junction remodeling in vitro and in vivo.

scholarly journals Interference With ESAM (Endothelial Cell-Selective Adhesion Molecule) Plus Vascular Endothelial-Cadherin Causes Immediate Lethality and Lung-Specific Blood Coagulation

2020 ◽  
Vol 40 (2) ◽  
pp. 378-393 ◽  
Author(s):  
Cao Nguyen Duong ◽  
Astrid F. Nottebaum ◽  
Stefan Butz ◽  
Stefan Volkery ◽  
Dagmar Zeuschner ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Vascular Permeability ◽  
Blood Coagulation ◽  
Adhesion Molecule ◽  
Gene Inactivation ◽  
Vascular Endothelial ◽  
Barrier Integrity ◽  
Mouse Tissues ◽  
Endothelial Junctions ◽  
Endothelial Junction

Objective: Vascular endothelial (VE)-cadherin is of dominant importance for the formation and stability of endothelial junctions, yet induced gene inactivation enhances vascular permeability in the lung but does not cause junction rupture. This study aims at identifying the junctional adhesion molecule, which is responsible for preventing endothelial junction rupture in the pulmonary vasculature in the absence of VE-cadherin. Approach and Results: We have compared the relevance of ESAM (endothelial cell-selective adhesion molecule), JAM (junctional adhesion molecule)-A, PECAM (platelet endothelial cell adhesion molecule)-1, and VE-cadherin for vascular barrier integrity in various mouse tissues. Gene inactivation of ESAM enhanced vascular permeability in the lung but not in the heart, skin, and brain. In contrast, deletion of JAM-A or PECAM-1 did not affect barrier integrity in any of these organs. Blocking VE-cadherin with antibodies caused lethality in ESAM −/− mice within 30 minutes but had no such effect in JAM-A −/− , PECAM-1 −/− or wild-type mice. Likewise, induced gene inactivation of VE-cadherin caused rapid lethality only in the absence of ESAM. Ultrastructural analysis revealed that only combined interference with VE-cadherin and ESAM disrupted endothelial junctions and caused massive blood coagulation in the lung. Mechanistically, we could exclude a role of platelet ESAM in coagulation, changes in the expression of other junctional proteins or a contribution of cytoplasmic signaling domains of ESAM. Conclusions: Despite well-documented roles of JAM-A and PECAM-1 for the regulation of endothelial junctions, only for ESAM, we detected an essential role for endothelial barrier integrity in a tissue-specific way. In addition, we found that it is ESAM which prevents endothelial junction rupture in the lung when VE-cadherin is absent.

scholarly journals S-nitrosylation regulates VE-cadherin phosphorylation and internalization in microvascular permeability

2016 ◽  
Vol 310 (8) ◽  
pp. H1039-H1044 ◽  
Author(s):  
Anita Guequén ◽  
Rodrigo Carrasco ◽  
Patricia Zamorano ◽  
Lorena Rebolledo ◽  
Pia Burboa ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Platelet Activating Factor ◽  
Adherens Junction ◽  
Endothelial Barrier ◽  
Main Element ◽  
Nitric Oxide Production ◽  
Vascular Endothelial ◽  
Junctional Proteins

The adherens junction complex, composed mainly of vascular endothelial (VE)-cadherin, β-catenin, p120, and γ-catenin, is the main element of the endothelial barrier in postcapillary venules. S-nitrosylation of β-catenin and p120 is an important step in proinflammatory agents-induced hyperpermeability. We investigated in vitro and in vivo whether or not VE-cadherin is S-nitrosylated using platelet-activating factor (PAF) as agonist. We report that PAF-stimulates S-nitrosylation of VE-cadherin, which disrupts its association with β-catenin. In addition, based on inhibition of nitric oxide production, our results strongly suggest that S-nitrosylation is required for VE-cadherin phosphorylation on tyrosine and for its internalization. Our results unveil an important mechanism to regulate phosphorylation of junctional proteins in association with S-nitrosylation.

scholarly journals Phospholipase D2 restores endothelial barrier function by promoting PTPN14-mediated VE-cadherin dephosphorylation

2020 ◽  
Vol 295 (22) ◽  
pp. 7669-7685 ◽  
Author(s):  
Panfeng Fu ◽  
Ramaswamy Ramchandran ◽  
Mark Shaaya ◽  
Longshuang Huang ◽  
David L. Ebenezer ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Protein Tyrosine Phosphatase ◽  
Barrier Function ◽  
Adherens Junctions ◽  
Tyrosine Phosphatase ◽  
Endothelial Barrier ◽  
Endothelial Barrier Function ◽  
Protein Tyrosine ◽  
Adhesion Junctions

Increased permeability of vascular lung tissues is a hallmark of acute lung injury and is often caused by edemagenic insults resulting in inflammation. Vascular endothelial (VE)-cadherin undergoes internalization in response to inflammatory stimuli and is recycled at cell adhesion junctions during endothelial barrier re-establishment. Here, we hypothesized that phospholipase D (PLD)-generated phosphatidic acid (PA) signaling regulates VE-cadherin recycling and promotes endothelial barrier recovery by dephosphorylating VE-cadherin. Genetic deletion of PLD2 impaired recovery from protease-activated receptor-1–activating peptide (PAR-1–AP)-induced lung vascular permeability and potentiated inflammation in vivo. In human lung microvascular endothelial cells (HLMVECs), inhibition or deletion of PLD2, but not of PLD1, delayed endothelial barrier recovery after thrombin stimulation. Thrombin stimulation of HLMVECs increased co-localization of PLD2-generated PA and VE-cadherin at cell-cell adhesion junctions. Inhibition of PLD2 activity resulted in prolonged phosphorylation of Tyr-658 in VE-cadherin during the recovery phase 3 h post-thrombin challenge. Immunoprecipitation experiments revealed that after HLMVECs are thrombin stimulated, PLD2, VE-cadherin, and protein-tyrosine phosphatase nonreceptor type 14 (PTPN14), a PLD2-dependent protein-tyrosine phosphatase, strongly associate with each other. PTPN14 depletion delayed VE-cadherin dephosphorylation, reannealing of adherens junctions, and barrier function recovery. PLD2 inhibition attenuated PTPN14 activity and reversed PTPN14-dependent VE-cadherin dephosphorylation after thrombin stimulation. Our findings indicate that PLD2 promotes PTPN14-mediated dephosphorylation of VE-cadherin and that redistribution of VE-cadherin at adherens junctions is essential for recovery of endothelial barrier function after an edemagenic insult.

scholarly journals Dissociation of VE-PTP from VE-cadherin is required for leukocyte extravasation and for VEGF-induced vascular permeability in vivo

2011 ◽  
Vol 208 (12) ◽  
pp. 2393-2401 ◽  
Author(s):  
Andre Broermann ◽  
Mark Winderlich ◽  
Helena Block ◽  
Maike Frye ◽  
Jan Rossaint ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Vascular Permeability ◽  
Fusion Proteins ◽  
Tyrosine Phosphatase ◽  
Cell Contact ◽  
Vascular Endothelial ◽  
Cell Contacts ◽  
Leukocyte Extravasation

We have recently shown that vascular endothelial protein tyrosine phosphatase (VE-PTP), an endothelial membrane protein, associates with VE-cadherin and is required for optimal VE-cadherin function and endothelial cell contact integrity. The dissociation of VE-PTP from VE-cadherin is triggered by vascular endothelial growth factor (VEGF) and by the binding of leukocytes to endothelial cells in vitro, suggesting that this dissociation is a prerequisite for the destabilization of endothelial cell contacts. Here, we show that VE-cadherin/VE-PTP dissociation also occurs in vivo in response to LPS stimulation of the lung or systemic VEGF stimulation. To show that this dissociation is indeed necessary in vivo for leukocyte extravasation and VEGF-induced vascular permeability, we generated knock-in mice expressing the fusion proteins VE-cadherin-FK 506 binding protein and VE-PTP-FRB* under the control of the endogenous VE-cadherin promoter, thus replacing endogenous VE-cadherin. The additional domains in both fusion proteins allow the heterodimeric complex to be stabilized by a chemical compound (rapalog). We found that intravenous application of the rapalog strongly inhibited VEGF-induced (skin) and LPS-induced (lung) vascular permeability and inhibited neutrophil extravasation in the IL-1β inflamed cremaster and the LPS-inflamed lung. We conclude that the dissociation of VE-PTP from VE-cadherin is indeed required in vivo for the opening of endothelial cell contacts during induction of vascular permeability and leukocyte extravasation.

scholarly journals p120-Catenin regulates leukocyte transmigration through an effect on VE-cadherin phosphorylation

Blood ◽  
2008 ◽  
Vol 112 (7) ◽  
pp. 2770-2779 ◽  
Author(s):  
Pilar Alcaide ◽  
Gail Newton ◽  
Scott Auerbach ◽  
Seema Sehrawat ◽  
Tanya N. Mayadas ◽  
...  
Keyword(s):  
Surface Expression ◽  
Vascular Endothelial ◽  
Gap Formation ◽  
P120 Catenin ◽  
Leukocyte Transmigration ◽  
Vascular Endothelial Cadherin ◽  
Leukocyte Transendothelial Migration ◽  
Endothelial Junctions ◽  
Functional Consequences ◽  
Time Required

Abstract Vascular endothelial–cadherin (VE-cad) is localized to adherens junctions at endothelial cell borders and forms a complex with α-, β-, γ-, and p120-catenins (p120). We previously showed that the VE-cad complex disassociates to form short-lived “gaps” during leukocyte transendothelial migration (TEM); however, whether these gaps are required for leukocyte TEM is not clear. Recently p120 has been shown to control VE-cad surface expression through endocytosis. We hypothesized that p120 regulates VE-cad surface expression, which would in turn have functional consequences for leukocyte transmigration. Here we show that endothelial cells transduced with an adenovirus expressing p120GFP fusion protein significantly increase VE-cad expression. Moreover, endothelial junctions with high p120GFP expression largely prevent VE-cad gap formation and neutrophil leukocyte TEM; if TEM occurs, the length of time required is prolonged. We find no evidence that VE-cad endocytosis plays a role in VE-cad gap formation and instead show that this process is regulated by changes in VE-cad phosphorylation. In fact, a nonphosphorylatable VE-cad mutant prevented TEM. In summary, our studies provide compelling evidence that VE-cad gap formation is required for leukocyte transmigration and identify p120 as a critical intracellular mediator of this process through its regulation of VE-cad expression at junctions.

scholarly journals Endothelial Barrier Function and Leukocyte Transmigration in Atherosclerosis

Biomedicines ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 328
Author(s):  
Thijs J. Sluiter ◽  
Jaap D. van Buul ◽  
Stephan Huveneers ◽  
Paul H. A. Quax ◽  
Margreet R. de Vries
Keyword(s):  
Endothelial Cells ◽  
Barrier Function ◽  
Endothelial Barrier ◽  
Intraplaque Hemorrhage ◽  
Vascular Endothelial ◽  
Endothelial Barrier Function ◽  
Leukocyte Transmigration ◽  
And Migration ◽  
Plaque Destabilization ◽  
Slow Rolling

The vascular endothelium is a highly specialized barrier that controls passage of fluids and migration of cells from the lumen into the vessel wall. Endothelial cells assist leukocytes to extravasate and despite the variety in the specific mechanisms utilized by different leukocytes to cross different vascular beds, there is a general principle of capture, rolling, slow rolling, arrest, crawling, and ultimately diapedesis via a paracellular or transcellular route. In atherosclerosis, the barrier function of the endothelium is impaired leading to uncontrolled leukocyte extravasation and vascular leakage. This is also observed in the neovessels that grow into the atherosclerotic plaque leading to intraplaque hemorrhage and plaque destabilization. This review focuses on the vascular endothelial barrier function and the interaction between endothelial cells and leukocytes during transmigration. We will discuss the role of endothelial dysfunction, transendothelial migration of leukocytes and plaque angiogenesis in atherosclerosis.

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