The Experts below are selected from a list of 3354 Experts worldwide ranked by ideXlab platform
Elisabetta Dejana - One of the best experts on this subject based on the ideXlab platform.
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new insights in the control of Vascular permeability Vascular Endothelial Cadherin and other players
Current Opinion in Hematology, 2015Co-Authors: Marianna Trani, Elisabetta DejanaAbstract:PURPOSE OF REVIEW: The control of the Endothelial barrier function is essential for Vascular homeostasis and is mainly mediated by cell-to-cell junctions that tightly regulate permeability to plasma solutes and circulating cells such as leukocytes and tumor cells. While in some circumstances the transient dismantling of Endothelial cell junctions might be beneficial, in pathological conditions, such as cancer, severe alterations of Endothelial junction composition and function are detrimental, causing massive edema and increased interstitial pressure. Here, we aim to discuss the newly and most recently identified molecular mechanisms that cooperate in the control of Vascular permeability. RECENT FINDINGS: Although the involvement of Vascular Endothelial-Cadherin in the regulation of Vascular leakage is well known, recent findings shed light on additional molecules involved in the control of Vascular Endothelial-Cadherin phosphorylation in physiological and pathological conditions, and identified new unknown regulators of the Endothelial barrier function. SUMMARY: In the past years, several studies explored the contribution of various signaling pathways in the regulation of Vascular leakage. Despite encouraging results, a more comprehensive understanding of the molecular mechanisms involved in this process will define druggable targets for new therapeutic interventions to limit Endothelial barrier dysfunctions.
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Vascular Endothelial Cadherin and Vascular stability
Current Opinion in Hematology, 2012Co-Authors: Elisabetta Dejana, Costanza GiampietroAbstract:Purpose of reviewVascular integrity is characterized by a tight control of permeability to cells and solutes and by resistance to blood flow. In several pathologies including tumor angiogenesis, Vascular malformations, hemorrhagic stroke and others, there is the need to stabilize the vessels and pre
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Vascular Endothelial Cadherin controls vegfr 2 internalization and signaling from intracellular compartments
Journal of Cell Biology, 2006Co-Authors: Maria Grazia Lampugnani, Elisabetta Dejana, Fabrizio Orsenigo, Maria Cristina Gagliani, Carlo TacchettiAbstract:Receptor endocytosis is a fundamental step in controlling the magnitude, duration, and nature of cell signaling events. Confluent Endothelial cells are contact inhibited in their growth and respond poorly to the proliferative signals of Vascular Endothelial growth factor (VEGF). In a previous study, we found that the association of Vascular Endothelial Cadherin (VEC) with VEGF receptor (VEGFR) type 2 contributes to density-dependent growth inhibition (Lampugnani, G.M., A. Zanetti, M. Corada, T. Takahashi, G. Balconi, F. Breviario, F. Orsenigo, A. Cattelino, R. Kemler, T.O. Daniel, and E. Dejana. 2003. J. Cell Biol. 161:793–804). In the present study, we describe the mechanism through which VEC reduces VEGFR-2 signaling. We found that VEGF induces the clathrin-dependent internalization of VEGFR-2. When VEC is absent or not engaged at junctions, VEGFR-2 is internalized more rapidly and remains in endosomal compartments for a longer time. Internalization does not terminate its signaling; instead, the internalized receptor is phosphorylated, codistributes with active phospholipase C–γ, and activates p44/42 mitogen-activated protein kinase phosphorylation and cell proliferation. Inhibition of VEGFR-2 internalization reestablishes the contact inhibition of cell growth, whereas silencing the junction-associated density-enhanced phosphatase-1/CD148 phosphatase restores VEGFR-2 internalization and signaling. Thus, VEC limits cell proliferation by retaining VEGFR-2 at the membrane and preventing its internalization into signaling compartments.
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a monoclonal antibody to Vascular Endothelial Cadherin inhibits tumor angiogenesis without side effects on Endothelial permeability
Blood, 2002Co-Authors: Monica Corada, Ferruccio Breviario, Francescafang Liao, Lucia Zanetta, Maria Grazia Lampugnani, Fabrizio Orsenigo, Sergio Bernasconi, Daniel J Hicklin, Peter Bohlen, Elisabetta DejanaAbstract:Vascular Endothelial Cadherin (VE-Cadherin) is an Endothelial-specific, trans-membrane protein that promotes homophilic cell adhesion. Inhibition of VE-Cadherin by the blocking monoclonal antibody (mAb) BV13 inhibited angiogenesis and tumor growth in vivo. However, this effect was accompanied by a marked increase in lung and heart permeability. In the present paper, we characterize a different VE-Cadherin mAb (BV14) that is able to inhibit angiogenesis without affecting Vascular permeability. In vitro studies show that BV14, in contrast to BV13, did not increase paracellular permeability of Endothelial monolayers and did not disrupt VE-Cadherin clusters at junctions. However, both antibodies could inhibit formation of Vascularlike structures in collagen gels and increase migration of Endothelial cells into wounded areas. In vivo, BV14 and BV13 were equally active in inhibiting angiogenesis in the mouse cornea and in reducing the growth of hemangioma and C6 glioma. In contrast to BV13, BV14 did not change Vascular permeability in all the organs tested and at any dose used. BV14 and BV13 bind to VE-Cadherin extracellular repeats EC4 and EC1, respectively. We propose that, in resting vessels, where junctions are stable and well-structured, antibody binding to EC1 but not EC4 disrupts their organization and increases permeability. In contrast, in growing vessels, where Endothelial cells are migrating and junctions are weaker, antibody binding to EC4 may be sufficient to disrupt cell-to-cell adhesion and inhibit assembly of new Vascular structures.
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selective targeting of angiogenic tumor vasculature by Vascular Endothelial Cadherin antibody inhibits tumor growth without affecting Vascular permeability
Cancer Research, 2002Co-Authors: Francescafang Liao, Elisabetta Dejana, Rajiv Bassi, Jay Overholser, Peter Bohlen, Jacqueline F Doody, Bridget Finnerty, Yan Wu, Paul Kussie, Daniel J HicklinAbstract:Vascular Endothelial-Cadherin (VE-Cadherin) is an Endothelial cell-specific adhesion molecule that is localized exclusively at cell-cell contacts referred to as adherens junctions. VE-Cadherin-mediated adhesion is crucial for proper assembly of Vascular structures during angiogenesis as well as for maintenance of a normal Vascular integrity. We have shown previously that a monoclonal antibody (BV13) to VE-Cadherin not only inhibits the formation of Vascular tubes during tumor angiogenesis but also disrupts adherens junctions of normal vasculature with a concomitant increase in Vascular permeability. The goal of the current studies was to block VE-Cadherin function during angiogenesis without disrupting existing junctions on normal endothelium. Using in vitro screening assays to test for functional blocking of adherens junction formation and in vivo assays to detect antibody effects on Vascular permeability in normal tissues, we have identified a novel blocking antibody (E4G10) that inhibits VE-Cadherin function during angiogenesis but does not disrupt existing adherens junctions on normal vasculature. E4G10 inhibited formation of Vascular tubes in vivo in the Matrigel plug and corneal micropocket assays. E4G10 also inhibited tumor growth in three models of mouse and human tumors via an antiangiogenic mechanism. Examination of normal mouse and tumor tissues showed that E4G10 bound to Endothelial cells in a subset of tumor vasculature but not to normal vasculature. Bromodeoxyuridine labeling experiments showed that E4G10 specifically targeted a subset of tumor endothelium that is undergoing active cell proliferation, which likely reflects the activated, angiogenic endothelium. These findings indicate that VE-Cadherin can be selectively targeted during states of pathological angiogenesis, despite its ubiquitous distribution throughout the entire vasculature. Our data also suggest that antibody E4G10 recognizes VE-Cadherin epitopes that are only accessible on Endothelial cells forming new adherens junctions, such as in angiogenic tumor vasculature.
Philippe Huber - One of the best experts on this subject based on the ideXlab platform.
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Vascular Endothelial Cadherin tyrosine phosphorylation in angiogenic and quiescent adult tissues
Circulation Research, 2005Co-Authors: Nathalie Lambeng, Isabelle Vilgrain, Yann Wallez, Christine Rampon, Francine Cand, Georges Christe, Danielle Gulinodebrac, Philippe HuberAbstract:Vascular Endothelial–Cadherin (VE-Cadherin) plays a key role in angiogenesis and in Vascular permeability. The regulation of its biological activity may be a central mechanism in normal or pathological angiogenesis. VE-Cadherin has been shown to be phosphorylated on tyrosine in vitro under various conditions, including stimulation by VEGF. In the present study, we addressed the question of the existence of a tyrosine phosphorylated form of VE-Cadherin in vivo, in correlation with the quiescent versus angiogenic state of adult tissues. Phosphorylated VE-Cadherin was detected in mouse lung, uterus, and ovary but not in other tissues unless mice were injected with peroxovanadate to block protein phosphatases. Remarkably, VE-Cadherin tyrosine phosphorylation was dramatically increased in uterus and ovary, and not in other organs, during PMSG/hCG-induced angiogenesis. In parallel, we observed an increased association of VE-Cadherin with Flk1 (VEGF receptor 2) during hormonal angiogenesis. Additionally, Src kinase was constitutively associated with VE-Cadherin in both quiescent and angiogenic tissues and increased phosphorylation of VE-Cadherin–associated Src was detected in uterus and ovary after hormonal treatment. Src-VE-Cadherin association was detected in cultured Endothelial cells, independent of VE-Cadherin phosphorylation state and Src activation level. In this model, Src inhibition impaired VEGF-induced VE-Cadherin phosphorylation, indicating that VE-Cadherin phosphorylation was dependent on Src activation. We conclude that VE-Cadherin is a substrate for tyrosine kinases in vivo and that its phosphorylation, together with that of associated Src, is increased by angiogenic stimulation. Physical association between Flk1, Src, and VE-Cadherin may thus provide an efficient mechanism for amplification and perpetuation of VEGF-stimulated angiogenic processes.
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ACTH depletion represses Vascular Endothelial-Cadherin transcription in mouse adrenal endothelium in vivo
Journal of Molecular Endocrinology, 2005Co-Authors: Philippe Huber, Christine Rampon, Christine Mallet, Elodie Faure, Marie-hélène Prandini, Olivier Feraud, Stéphanie Bouillot, Isabelle VilgrainAbstract:Vascular Endothelial-Cadherin (VE-Cadherin) is an Endothelial cell-specific adhesion protein that is localised at cell–cell contacts. This molecule is an important determinant of Vascular architecture and Endothelial cell survival. In the adrenal cortex, steroidogenic and Endothelial cells form a complex architecture. The adrenocorticotrophin hormone (ACTH) regulates gland homeostasis whose secretion is subjected to a negative feedback by adrenocorticosteroids. The aim of the present study was to determine whether VE-Cadherin expression in the adrenal gland was regulated by hormonal challenge. We demonstrated that VE-Cadherin protein levels were dramatically decreased (23·5±3·7%) by dexamethasone injections in the mouse and were restored by ACTH within 7 days (94·9±18·6%). Flow cytometry analysis of adrenal cells showed that the ratios of Endothelial versus total adrenal cells were identical (35%) in dexamethasone- or ACTH-treated or untreated mice, suggesting that VE-Cadherin expression could be regulated by ACTH. We demonstrate the existence of a transcriptional regulation of the VE-Cadherin gene using transgenic mice carrying the chloramphenicol acetyl transferase gene under the control of the VE-Cadherin promoter. Indeed, the promoter activity in the adrenals, but not in the lung or liver, was decreased in response to dexamethasone treatment (40±1·3%) and was partially restored after gland regeneration by ACTH injection (82±3%). In conclusion, our results show that transcription of a specific Endothelial gene is controlled by the hypothalamo–pituitary axis and the data expand the knowledge regarding the role of ACTH in the regulation of the adrenal Vascular network.
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Vascular Endothelial–Cadherin Tyrosine Phosphorylation in Angiogenic and Quiescent Adult Tissues
Circulation Research, 2005Co-Authors: Nathalie Lambeng, Isabelle Vilgrain, Yann Wallez, Christine Rampon, Francine Cand, Georges Christe, Danielle Gulino-debrac, Philippe HuberAbstract:Vascular Endothelial–Cadherin (VE-Cadherin) plays a key role in angiogenesis and in Vascular permeability. The regulation of its biological activity may be a central mechanism in normal or pathological angiogenesis. VE-Cadherin has been shown to be phosphorylated on tyrosine in vitro under various conditions, including stimulation by VEGF. In the present study, we addressed the question of the existence of a tyrosine phosphorylated form of VE-Cadherin in vivo, in correlation with the quiescent versus angiogenic state of adult tissues. Phosphorylated VE-Cadherin was detected in mouse lung, uterus, and ovary but not in other tissues unless mice were injected with peroxovanadate to block protein phosphatases. Remarkably, VE-Cadherin tyrosine phosphorylation was dramatically increased in uterus and ovary, and not in other organs, during PMSG/hCG-induced angiogenesis. In parallel, we observed an increased association of VE-Cadherin with Flk1 (VEGF receptor 2) during hormonal angiogenesis. Additionally, Src kinase was constitutively associated with VE-Cadherin in both quiescent and angiogenic tissues and increased phosphorylation of VE-Cadherin–associated Src was detected in uterus and ovary after hormonal treatment. Src-VE-Cadherin association was detected in cultured Endothelial cells, independent of VE-Cadherin phosphorylation state and Src activation level. In this model, Src inhibition impaired VEGF-induced VE-Cadherin phosphorylation, indicating that VE-Cadherin phosphorylation was dependent on Src activation. We conclude that VE-Cadherin is a substrate for tyrosine kinases in vivo and that its phosphorylation, together with that of associated Src, is increased by angiogenic stimulation. Physical association between Flk1, Src, and VE-Cadherin may thus provide an efficient mechanism for amplification and perpetuation of VEGF-stimulated angiogenic processes.
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the Vascular Endothelial Cadherin promoter directs Endothelial specific expression in transgenic mice
Blood, 1999Co-Authors: S Gory, Elisabetta Dejana, M Vernet, M Laurent, J Dalmon, Philippe HuberAbstract:Vascular Endothelial-Cadherin (VE-Cadherin) is a calcium-dependent adhesive molecule, exclusively and constitutively expressed in Endothelial cells. Analysis of the VE-Cadherin promoter fused to a reporter gene in bovine aortic Endothelial cells showed three major functional regions. The proximal region alone (−139, +24) promoted nonspecific transcription; the addition of the (−289, −140) and (−2226, −1190) domains abolished transcription in fibroblasts while expression in Endothelial cells remained unchanged, suggesting that fragments (−2226, +24) and longer contain the full endogenous promoter activity. To study the transcriptional specificity of the promoter region in vivo, we generated transgenic mice carrying the chimeric construct containing the (−2486, +24) region. The promoter directed reporter expression in all examined organs of adult transgenic mice. During embryonic development, transgene expression was detected at the early steps of vasculogenesis. Later, the expression persisted during development of the Vascular system and was restricted to the Endothelial layer of the vessels. Together, these data provide evidence for specific regulatory regions within the VE-Cadherin promoter. Furthermore, the identification of DNA sequences restricting gene expression to the endothelium has many potential applications for the development of animal models of cardioVascular or angiogenic diseases or for the delivery of therapeutic molecules.
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identification of a novel Cadherin Vascular Endothelial Cadherin 2 located at intercellular junctions in Endothelial cells
Journal of Biological Chemistry, 1998Co-Authors: Paola Telo, Philippe Huber, Ferruccio Breviario, Carla Panzeri, Elisabetta DejanaAbstract:Abstract Endothelial cells express two major Cadherins, VE- and N-Cadherins, but only the former consistently participates in adherens junction organization. In heart microVascular Endothelial cells, we identified a new member of the Cadherin superfamily using polymerase chain reaction. The entire putative coding sequence was determined. Similarly to protoCadherins, while the extracellular domain presented homology with other members of the Cadherin superfamily, the intracellular region was unrelated either to Cadherins or to any other known protein. We propose for this new protein the name of Vascular Endothelial Cadherin-2. By Northern blot analysis, the mRNA was present only in cultured Endothelial cell lines but not in other cell types such as NIH 3T3, Chinese hamster ovary, or L cells. In addition, mRNA was particularly abundant in highly Vascularized organs such as lung or kidney. In Endothelial cells and transfectants, this Cadherin was unable to bind catenins and presented a weak association with the cytoskeleton. This new molecule shares some functional properties with VE-Cadherin and other members of the Cadherin family. In Chinese hamster ovary transfectants it promoted homotypic Ca2+ dependent aggregation and adhesion and clustered at intercellular junctions. However, in contrast to VE-Cadherin, it did not modify paracellular permeability, cell migration, and density-dependent cell growth. These observations suggest that different Cadherins may promote homophilic cell-to-cell adhesion but that the functional consequences of this interaction depend on their binding to specific intracellular signaling/cytoskeletal proteins.
Mehran Haidari - One of the best experts on this subject based on the ideXlab platform.
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Correction to: Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β-dependent Vascular Endothelial Cadherin tyrosine phosphorylation.
Cardiovascular diabetology, 2017Co-Authors: Mehran Haidari, Wei Zhang, James T Willerson, Richard A F DixonAbstract:This article was unintentionally published twice in this journal, by the same authors. The following should be considered the version of record and used for citation purposes: "Mehran Haidari, Wei Zhang, James T Willerson and Richard AF Dixon, Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β-dependent Vascular Endothelial Cadherin tyrosine phosphorylation, CardioVascular Diabetology, Volume 13, Issue 1, doi: 10.1186/1475-2840-13-112 ". The duplicate "Mehran Haidari, Wei Zhang, James T Willerson and Richard AF Dixon, Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β-dependent Vascular Endothelial Cadherin tyrosine phosphorylation, CardioVascular Diabetology, Volume 13, Issue 1, doi: 10.1186/1475-2840-13-105 " is to be ignored. The Publisher apologizes to the readers of the journal for not detecting the duplication during the publication process.
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disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase c β dependent Vascular Endothelial Cadherin tyrosine phosphorylation
Cardiovascular Diabetology, 2014Co-Authors: Mehran Haidari, Wei Zhang, James T Willerson, Richard A F DixonAbstract:This article was unintentionally published twice in this journal, by the same authors. The following should be considered the version of record and used for citation purposes: “Mehran Haidari, Wei Zhang, James T Willerson and Richard AF Dixon, Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β–dependent Vascular Endothelial Cadherin tyrosine phosphorylation, CardioVascular Diabetology, Volume 13, Issue 1, doi: 10.1186/1475-2840-13-112 ". The duplicate “Mehran Haidari, Wei Zhang, James T Willerson and Richard AF Dixon, Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β–dependent Vascular Endothelial Cadherin tyrosine phosphorylation, CardioVascular Diabetology, Volume 13, Issue 1, doi: 10.1186/1475-2840-13-105 " is to be ignored. The Publisher apologizes to the readers of the journal for not detecting the duplication during the publication process.
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Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β–dependent Vascular Endothelial Cadherin tyrosine phosphorylation
Cardiovascular Diabetology, 2014Co-Authors: Mehran Haidari, Wei Zhang, James T Willerson, Richard A F DixonAbstract:This article was unintentionally published twice in this journal, by the same authors. The following should be considered the version of record and used for citation purposes: “Mehran Haidari, Wei Zhang, James T Willerson and Richard AF Dixon, Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β–dependent Vascular Endothelial Cadherin tyrosine phosphorylation, CardioVascular Diabetology, Volume 13, Issue 1, doi: 10.1186/1475-2840-13-112 ". The duplicate “Mehran Haidari, Wei Zhang, James T Willerson and Richard AF Dixon, Disruption of Endothelial adherens junctions by high glucose is mediated by protein kinase C-β–dependent Vascular Endothelial Cadherin tyrosine phosphorylation, CardioVascular Diabetology, Volume 13, Issue 1, doi: 10.1186/1475-2840-13-105 " is to be ignored. The Publisher apologizes to the readers of the journal for not detecting the duplication during the publication process.
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integrin α2β1 mediates tyrosine phosphorylation of Vascular Endothelial Cadherin induced by invasive breast cancer cells
Journal of Biological Chemistry, 2012Co-Authors: Mehran Haidari, Wei Zhang, Zhenping Chen, Leila Ganjehei, Ahmadreza Mortazavi, Amy R Caivano, Christopher Stroud, Darren G Woodside, James T WillersonAbstract:The molecular mechanisms that regulate the Endothelial response during transEndothelial migration (TEM) of invasive cancer cells remain elusive. Tyrosine phosphorylation of Vascular Endothelial Cadherin (VE-cad) has been implicated in the disruption of Endothelial cell adherens junctions and in the diapedesis of metastatic cancer cells. We sought to determine the signaling mechanisms underlying the disruption of Endothelial adherens junctions after the attachment of invasive breast cancer cells. Attachment of invasive breast cancer cells (MDA-MB-231) to human umbilical vein Endothelial cells induced tyrosine phosphorylation of VE-cad, dissociation of β-catenin from VE-cad, and retraction of Endothelial cells. Breast cancer cell-induced tyrosine phosphorylation of VE-cad was mediated by activation of the H-Ras/Raf/MEK/ERK signaling cascade and depended on the phosphorylation of Endothelial myosin light chain (MLC). The inhibition of H-Ras or MLC in Endothelial cells inhibited TEM of MDA-MB-231 cells. VE-cad tyrosine phosphorylation in Endothelial cells induced by the attachment of MDA-MB-231 cells was mediated by MDA-MB-231 α2β1 integrin. Compared with highly invasive MDA-MB-231 breast cancer cells, weakly invasive MCF-7 breast cancer cells expressed lower levels of α2β1 integrin. TEM of MCF-7 as well as induction of VE-cad tyrosine phosphorylation and dissociation of β-catenin from the VE-cad complex by MCF-7 cells were lower than in MDA-MB-231 cells. These processes were restored when MCF-7 cells were treated with β1-activating antibody. Moreover, the response of Endothelial cells to the attachment of prostatic (PC-3) and ovarian (SKOV3) invasive cancer cells resembled the response to MDA-MB-231 cells. Our study showed that the MDA-MB-231 cell-induced disruption of Endothelial adherens junction integrity is triggered by MDA-MB-231 cell α2β1 integrin and is mediated by H-Ras/MLC-induced tyrosine phosphorylation of VE-cad.
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atorvastatin preserves the integrity of Endothelial adherens junctions by inhibiting Vascular Endothelial Cadherin tyrosine phosphorylation
Experimental Cell Research, 2012Co-Authors: Mehran Haidari, Wei Zhang, Zhenping Chen, Leila Ganjehei, Ahmadreza Mortazavi, Navin Warier, Peter Vanderslice, Richard A F DixonAbstract:Abstract Vascular Endothelial Cadherin (VE-cad) tyrosine (Tyr) phosphorylation has been implicated in the disruption of adherens junctions (AJs) induced by inflammatory reactions. The impacts of statins on integrity of AJs and VE-cad Tyr phosphorylation have not been explored. The effects of atorvastatin on IL-1β and monocyte-induced VE-cad Tyr phosphorylation in human umbilical vein Endothelial cells (ECs) were studied. In ECs treated with interleukin (IL)-1β for 30 min, VE-cad Tyr phosphorylation, dissociation of the VE-cad/β-catenin complex and transEndothelial migration (TEM) of monocytes were increased. These processes were mediated by activation of HRas and RhoA that leads to phosphorylation of myosin light chain (MLC). Atorvastatin inhibited IL-1β-induced Tyr phosphorylation of VE-cad by inhibiting RhoA and by dephosphorylating MLC. The attenuating effect of atorvastatin on VE-cad Tyr phosphorylation was reversed when RhoA was activated or MLC phosphatase was inhibited. Furthermore, inhibiting farnesyl transferase or geranylgeranyl transferase reproduced the inhibitory effects of atorvastatin on VE-cad Tyr phosphorylation. In addition, atorvastatin inhibited monocyte-induced VE-cad Tyr phosphorylation in ECs and attenuated IL-1β-induced TEM of monocytes. Our study introduces a novel pleiotropic effect of atorvastatin and suggests that statins protect the integrity of AJs in ECs by inhibiting RhoA-mediated Tyr phosphorylation of VE-cad.
Monica Corada - One of the best experts on this subject based on the ideXlab platform.
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Phosphorylation of VE-Cadherin is modulated by haemodynamic forces and contributes to the regulation of Vascular permeability in vivo
Nature Communications, 2012Co-Authors: Fabrizio Orsenigo, Monica Corada, Costanza Giampietro, Aldo Ferrari, Ariane Galaup, Sara Sigismund, Giuseppe Ristagno, Luigi Maddaluno, Davide FrancoAbstract:Endothelial adherens junctions maintain Vascular integrity. Arteries and veins differ in their permeability but whether organization and strength of their adherens junctions vary has not been demonstrated in vivo. Here we report that Vascular Endothelial Cadherin, an Endothelial specific adhesion protein located at adherens junctions, is phosphorylated in Y658 and Y685 in vivo in veins but not in arteries under resting conditions. This difference is due to shear stress-induced junctional Src activation in veins. Phosphorylated Vascular Endothelial-Cadherin is internalized and ubiquitinated in response to permeability-increasing agents such as bradykinin and histamine. Inhibition of Src blocks Vascular Endothelial Cadherin phosphorylation and bradykinin-induced permeability. Point mutation of Y658F and Y685F prevents Vascular Endothelial Cadherin internalization, ubiquitination and an increase in permeability by bradykinin in vitro. Thus, phosphorylation of Vascular Endothelial Cadherin contributes to a dynamic state of adherens junctions, but is not sufficient to increase Vascular permeability in the absence of inflammatory agents.
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contact inhibition of vegf induced proliferation requires Vascular Endothelial Cadherin β catenin and the phosphatase dep 1 cd148
Journal of Cell Biology, 2003Co-Authors: Maria Grazia Lampugnani, Ferruccio Breviario, Monica Corada, Fabrizio Orsenigo, Adriana Zanetti, Takamune Takahashi, Giovanna Balconi, Anna Cattelino, Rolf Kemler, Thomas O DanielAbstract:Confluent Endothelial cells respond poorly to the proliferative signals of VEGF. Comparing isogenic Endothelial cells differing for Vascular Endothelial Cadherin (VE-Cadherin) expression only, we found that the presence of this protein attenuates VEGF-induced VEGF receptor (VEGFR) 2 phosphorylation in tyrosine, p44/p42 MAP kinase phosphorylation, and cell proliferation. VE-Cadherin truncated in β-catenin but not p120 binding domain is unable to associate with VEGFR-2 and to induce its inactivation. β-Catenin–null Endothelial cells are not contact inhibited by VE-Cadherin and are still responsive to VEGF, indicating that this protein is required to restrain growth factor signaling. A dominant-negative mutant of high cell density–enhanced PTP 1 (DEP-1)//CD148 as well as reduction of its expression by RNA interference partially restore VEGFR-2 phosphorylation and MAP kinase activation. Overall the data indicate that VE-Cadherin–β-catenin complex participates in contact inhibition of VEGF signaling. Upon stimulation with VEGF, VEGFR-2 associates with the complex and concentrates at cell–cell contacts, where it may be inactivated by junctional phosphatases such as DEP-1. In sparse cells or in VE-Cadherin–null cells, this phenomenon cannot occur and the receptor is fully activated by the growth factor.
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a monoclonal antibody to Vascular Endothelial Cadherin inhibits tumor angiogenesis without side effects on Endothelial permeability
Blood, 2002Co-Authors: Monica Corada, Ferruccio Breviario, Francescafang Liao, Lucia Zanetta, Maria Grazia Lampugnani, Fabrizio Orsenigo, Sergio Bernasconi, Daniel J Hicklin, Peter Bohlen, Elisabetta DejanaAbstract:Vascular Endothelial Cadherin (VE-Cadherin) is an Endothelial-specific, trans-membrane protein that promotes homophilic cell adhesion. Inhibition of VE-Cadherin by the blocking monoclonal antibody (mAb) BV13 inhibited angiogenesis and tumor growth in vivo. However, this effect was accompanied by a marked increase in lung and heart permeability. In the present paper, we characterize a different VE-Cadherin mAb (BV14) that is able to inhibit angiogenesis without affecting Vascular permeability. In vitro studies show that BV14, in contrast to BV13, did not increase paracellular permeability of Endothelial monolayers and did not disrupt VE-Cadherin clusters at junctions. However, both antibodies could inhibit formation of Vascularlike structures in collagen gels and increase migration of Endothelial cells into wounded areas. In vivo, BV14 and BV13 were equally active in inhibiting angiogenesis in the mouse cornea and in reducing the growth of hemangioma and C6 glioma. In contrast to BV13, BV14 did not change Vascular permeability in all the organs tested and at any dose used. BV14 and BV13 bind to VE-Cadherin extracellular repeats EC4 and EC1, respectively. We propose that, in resting vessels, where junctions are stable and well-structured, antibody binding to EC1 but not EC4 disrupts their organization and increases permeability. In contrast, in growing vessels, where Endothelial cells are migrating and junctions are weaker, antibody binding to EC4 may be sufficient to disrupt cell-to-cell adhesion and inhibit assembly of new Vascular structures.
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monoclonal antibodies directed to different regions of Vascular Endothelial Cadherin extracellular domain affect adhesion and clustering of the protein and modulate Endothelial permeability
Blood, 2001Co-Authors: Monica Corada, Ferruccio Breviario, Maria Grazia Lampugnani, Daniel J Hicklin, Peter Bohlen, Fang Liao, Maria Lindgren, Ronald Frank, William A Muller, Elisabetta DejanaAbstract:Vascular Endothelial Cadherin (VE-Cadherin) is an Endothelial cell–specific Cadherin that plays an important role in the control of Vascular organization. Blocking VE-Cadherin antibodies strongly inhibit angiogenesis, and inactivation of VE-Cadherin gene causes embryonic lethality due to a lack of correct organization and remodeling of the vasculature. Hence, inhibitors of VE-Cadherin adhesive properties may constitute a tool to prevent tumor neoVascularization. In this paper, we tested different monoclonal antibodies (mAbs) directed to human VE-Cadherin ectodomain for their functional activity. Three mAbs (Cad 5, BV6, BV9) were able to increase paracellular permeability, inhibit VE-Cadherin reorganization, and block angiogenesis in vitro. These mAbs could also induce Endothelial cell apoptosis in vitro. Two additional mAbs, TEA 1.31 and Hec 1.2, had an intermediate or undetectable activity, respectively, in these assays. Epitope mapping studies show that BV6, BV9, TEA 1.31, and Hec 1.2 bound to a recombinant fragment spanning the extracellular juxtamembrane domains EC3 through EC4. In contrast, Cad 5 bound to the aminoterminal domain EC1. By peptide scanning analysis and competition experiments, we defined the sequences TIDLRY located on EC3 and KVFRVDAETGDVFAI on EC1 as the binding domain of BV6 and Cad 5, respectively. Overall, these results support the concept that VE-Cadherin plays a relevant role on human Endothelial cell properties. Antibodies directed to the extracellular domains EC1 but also EC3-EC4 affect VE-Cadherin adhesion and clustering and alter Endothelial cell permeability, apoptosis, and Vascular structure formation.
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inhibition of cultured cell growth by Vascular Endothelial Cadherin Cadherin 5 ve Cadherin
Journal of Clinical Investigation, 1996Co-Authors: Luis Caveda, Pilar Navarro, M G Lampugnani, F Breviario, Monica Corada, D Gulino, Ines Martinpadura, Elisabetta DejanaAbstract:Endothelial cell proliferation is inhibited by the establishment of cell to cell contacts. Adhesive molecules at junctions could therefore play a role in transferring negative growth signals. The transmembrane protein VE-Cadherin (Vascular Endothelial Cadherin/Cadherin-S) is selectively expressed at intercellular clefts in the endothelium. The intracellular domain interacts with cytoplasmic proteins called catenins that transmit the adhesion signal and contribute to the anchorage of the protein to the actin cytoskeleton. Transfection of VE-Cadherin in both Chinese hamster ovary (CHO) and L929 cells confers inhibition of cell growth. Truncation of VE-Cadherin cytoplasmic region, responsible for linking catenins, does not affect VE-Cadherin adhesive properties but abolishes its effect on cell growth. Seeding human umbilical vein Endothelial cells or VE-Cadherin transfectants on a recombinant VE-Cadherin amino-terminal fragment inhibited their proliferation. These data show that VE-Cadherin homotypic engagement at junctions participates in density dependent inhibition of cell growth. This effect requires both the extracellular adhesive domain and the intracellular catenin binding region of the molecule.
D K James - One of the best experts on this subject based on the ideXlab platform.
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Vascular Endothelial Cadherin and β catenin in human fetoplacental vessels of pregnancies complicated by type 1 diabetes associations with angiogenesis and perturbed barrier function
Diabetologia, 2004Co-Authors: L. Leach, M. O. Babawale, A. Gruchy, C Foster, Terry M. Mayhew, C. Gray, Sam Staton, D K JamesAbstract:Aims/hypothesis Increased angiogenesis of fetoplacental vessels is a feature of pregnancies complicated by Type 1 diabetes mellitus, but the underlying molecular mechanisms are unknown. This investigation tests whether the diabetic maternal environment alters the phenotypic expression of placental Vascular Endothelial Cadherin and β-catenin, which have been implicated as key molecules in barrier formation and angiogenesis in the endothelium.
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Vascular Endothelial Cadherin and ??-catenin in human fetoplacental vessels of pregnancies complicated by Type 1 diabetes: Associations with angiogenesis and perturbed barrier function
Diabetologia, 2004Co-Authors: L. Leach, M. O. Babawale, A. Gruchy, C Foster, Terry M. Mayhew, C. Gray, Sam Staton, D K JamesAbstract:AIMS/HYPOTHESIS: Increased angiogenesis of fetoplacental vessels is a feature of pregnancies complicated by Type 1 diabetes mellitus, but the underlying molecular mechanisms are unknown. This investigation tests whether the diabetic maternal environment alters the phenotypic expression of placental Vascular Endothelial Cadherin and beta-catenin, which have been implicated as key molecules in barrier formation and angiogenesis in the endothelium. METHODS: Term placental microvessels from normal pregnancies (n=8) and from those complicated by Type 1 diabetes (n=8) were perfused with 76-Mr dextran tracers (1 mg/ml) and subjected to immunocytochemistry, immunoblotting and microscopy. Junctional integrity, localisation and phosphorylation were investigated along with total protein levels of Vascular Endothelial Cadherin, beta-catenin and Vascular Endothelial growth factor. Stereological sampling and estimation tools were used to quantify aspects of angiogenesis and Endothelial proliferation. RESULTS: In the Type 1 diabetic placentae, junctional localisations of Vascular Endothelial Cadherin and beta-catenin altered significantly, with more than 50% of microvessels showing complete loss of immunoreactivity and with no overall loss of total protein. Tracer leakage was associated with these vessels. There was a two- to three-fold increase in vessels showing junctional phospho-tyrosine immunoreactivity and hyperphosphorylated beta-catenin. Vascular Endothelial growth factor levels were higher in these placentae. A four-fold increase in Endothelial proliferation was observed, along with an increase in total length of capillaries without any change in luminal diameter. CONCLUSIONS/INTERPRETATION: Molecular perturbations of Vascular Endothelial Cadherin and beta-catenin occur in fetoplacental vessels of pregnancies complicated by Type 1 diabetes. Phosphorylation and loss of these molecules from the adherens junctional domains may be influenced in part by the elevated levels of Vascular Endothelial growth factor in the placenta. Perturbations of the junctional proteins may explain the observed breach in barrier integrity and may contribute to the mechanisms that drive proliferation and increases in capillary length.
