Endothelial Cell Cortactin Coordinates Intercellular Adhesion Molecule-1 Clustering and Actin Cytoskeleton Remodeling during Polymorphonuclear Leukocyte Adhesion and Transmigration

Yang, Lin; Kowalski, Jennifer R.; Yacono, Patrick W.; Bajmoczi, Milan; Shaw, Sunil K.; Froio, Richard M.; Golan, David E.; Thomas, Sheila M.; Luscinskas, Francis W.

doi:10.4049/jimmunol.177.9.6440

Cited by 105 publications

(108 citation statements)

References 38 publications

(59 reference statements)

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“…It was also shown that cortactin phosphorylation is required for leukocyte-mediated cytoskeletal remodeling as well as clustering of ICAM-1 (43). During, TEM, ICAM-1 clustering is required for numerous signaling processes in EC, thus Src-cortactin signaling may be the initiating event upstream of other ICAM-1-dependent signals (43). In line with this, expression of VE-cadherin Y658F or Y731F mutants had no effect on the distribution or recruitment of ICAM-1 to sites of leukocyte adhesion (data not shown) strengthening the idea that the ICAM-Src-cortactin pathway functions upstream of VE-cadherin phosphorylation.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…Yang et al (22,43) have demonstrated a requirement for Src-mediated phosphorylation of cortactin specifically in paracellular leukocyte TEM. It was also shown that cortactin phosphorylation is required for leukocyte-mediated cytoskeletal remodeling as well as clustering of ICAM-1 (43). During, TEM, ICAM-1 clustering is required for numerous signaling processes in EC, thus Src-cortactin signaling may be the initiating event upstream of other ICAM-1-dependent signals (43).…”

Section: Discussionmentioning

confidence: 99%

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ICAM-1-Mediated, Src- and Pyk2-Dependent Vascular Endothelial Cadherin Tyrosine Phosphorylation Is Required for Leukocyte Transendothelial Migration

Allingham

Buul

Burridge

2007

The Journal of Immunology

292

304

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Leukocyte transendothelial migration (TEM) has been modeled as a multistep process beginning with rolling adhesion, followed by firm adhesion, and ending with either transcellular or paracellular passage of the leukocyte across the endothelial monolayer. In the case of paracellular TEM, endothelial cell (EC) junctions are transiently disassembled to allow passage of leukocytes. Numerous lines of evidence demonstrate that tyrosine phosphorylation of adherens junction proteins, such as vascular endothelial cadherin (VE-cadherin) and β-catenin, correlates with the disassembly of junctions. However, the role of tyrosine phosphorylation in the regulation of junctions during leukocyte TEM is not completely understood. Using human leukocytes and EC, we show that ICAM-1 engagement leads to activation of two tyrosine kinases, Src and Pyk2. Using phospho-specific Abs, we show that engagement of ICAM-1 induces phosphorylation of VE-cadherin on tyrosines 658 and 731, which correspond to the p120-catenin and β-catenin binding sites, respectively. These phosphorylation events require the activity of both Src and Pyk2. We find that inhibition of endothelial Src with PP2 or SU6656 blocks neutrophil transmigration (71.1 ± 3.8% and 48.6 ± 3.8% reduction, respectively), whereas inhibition of endothelial Pyk2 also results in decreased neutrophil transmigration (25.5 ± 6.0% reduction). Moreover, overexpression of the nonphosphorylatable Y658F or Y731F mutants of VE-cadherin impairs transmigration of neutrophils compared with overexpression of wild-type VE-cadherin (32.7 ± 7.1% and 38.8 ± 6.5% reduction, respectively). Our results demonstrate that engagement of ICAM-1 by leukocytes results in tyrosine phosphorylation of VE-cadherin, which is required for efficient neutrophil TEM.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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ICAM-1-Mediated, Src- and Pyk2-Dependent Vascular Endothelial Cadherin Tyrosine Phosphorylation Is Required for Leukocyte Transendothelial Migration

Allingham

Buul

Burridge

2007

The Journal of Immunology

292

304

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“…In the absence of PECAM-1 recycling, leukocyte transmigration is arrested [10]. Cortactin-mediated rearrangement of the actin cytoskeleton [7] and intermediate filaments [11] is also critical for successful transmigration. Overall, these results reveal that endothelial cells are active participants in transmigration.…”

Section: Introductionmentioning

confidence: 99%

“…For example, the junctional protein vascular endothelial cadherin (VE-cadherin) is phosphorylated and redistributes during neutrophil transmigration [7][8][9]. This results in the formation of transient gaps that allow for the passage of neutrophils [9].…”

Section: Introductionmentioning

confidence: 99%

Endothelial paxillin and focal adhesion kinase (FAK) play a critical role in neutrophil transmigration

et al. 2012

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CanadaDuring an inflammatory response, endothelial cells undergo morphological changes to allow for the passage of neutrophils from the blood vessel to the site of injury or infection. Although endothelial cell junctions and the cytoskeleton undergo reorganization during inflammation, little is known about another class of cellular structures, the focal adhesions. In this study, we examined several focal adhesion proteins during an inflammatory response. We found that there was selective loss of paxillin and focal adhesion kinase (FAK) from focal adhesions in proximity to transmigrating neutrophils; in contrast the levels of the focal adhesion proteins b1-integrin and vinculin were unaffected. Paxillin was lost from focal adhesions during neutrophil transmigration both under static and flow conditions. Down-regulating endothelial paxillin with siRNA blocked neutrophil transmigration while having no effect on rolling or adhesion. As paxillin dynamics are regulated partly by FAK, the role of FAK in neutrophil transmigration was examined using two complementary methods. siRNA was used to down-regulate total FAK protein while dominant-negative, kinase-deficient FAK was expressed to block FAK signaling. Disruption of the FAK protein or FAK signaling decreased neutrophil transmigration. Collectively, these findings reveal a novel role for endothelial focal adhesion proteins paxillin and FAK in regulating neutrophil transmigration. IntroductionNeutrophils are essential mediators of host defense. During inflammation, neutrophils leave the bloodstream and traffic into the tissue in a well-described series of steps [1,2]. The molecular interactions governing the initial stages of tethering, rolling and firm adhesion have been well studied [1,2], whereas a clear picture of the latter steps underlying the passage of the leukocytes through the blood vessel has been slower to emerge [3][4][5].During transmigration endothelial cells undergo rapid changes in their structural organization [5,6]. For example, the junctional protein vascular endothelial cadherin (VE-cadherin) is phosphorylated and redistributes during neutrophil transmigration [7][8][9]. This results in the formation of transient gaps that 436allow for the passage of neutrophils [9]. Platelet endothelial cell adhesion molecule-1 (PECAM-1), another junctional protein, is regulated by the action of the kinesins and requires an intact microtubule cytoskeleton to enable PECAM-1 recycling from membrane compartments to the lateral junctions [10]. In the absence of PECAM-1 recycling, leukocyte transmigration is arrested [10]. Cortactin-mediated rearrangement of the actin cytoskeleton [7] and intermediate filaments [11] is also critical for successful transmigration. Overall, these results reveal that endothelial cells are active participants in transmigration.The importance of cytoskeletal and junctional proteins during leukocyte transmigration led us to ask whether another class of cellular structures, the focal adhesions, also play a role during this process. Focal a...

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Septin and actin contributions to endothelial cell–cell junctions and monolayer integrity

et al. 2022

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Septins in endothelial cells (ECs) have important roles supporting the integrity of the endothelial monolayer. Cell–cell junctions in EC monolayers are highly dynamic, with continuous retractions and protrusions. Depletion of septins in ECs leads to disruption of cell–cell junctions, which are composed of VE‐cadherin and other junctional proteins. In EC monolayers, septins are concentrated at the plasma membrane at sites of cell–cell contact, in curved‐ and scallop‐shaped patterns. These membrane‐associated septin accumulations are located in regions of positive membrane curvature, and those regions are often associated with and immediately adjacent to actin‐rich protrusions with negative membrane curvature. EC septins associate directly with plasma membrane lipids, based on findings with site‐specific mutations of septins in ECs, which is consistent with biochemical and cell biological studies in other systems. Loss of septins leads to disruption of the EC monolayer, and gaps form between cells. The number and breadth of cell–cell contacts and junctions decreases, and the number and frequency of retractions, ruffles, and protrusions at cell edges also decreases. In addition, loss of septins leads to decreased amounts of F‐actin at the cortical membrane, along with increased amounts of F‐actin in stress fibers of the cytoplasm. Endothelial monolayer disruption from loss of septins is also associated with decreased transendothelial electric resistance (TEER) and increased levels of transendothelial migration (TEM) by immune and cancer cells, owing to the gaps in the monolayer. A current working model is that assembly of septin filaments at regions of positive membrane curvature contributes to a mechanical footing or base for actin‐based protrusive forces generated at adjoining regions of the membrane. Specific molecular interactions between the septin and actin components of the cytoskeleton may also be important contributors. Regulators of actin assembly may promote and support the assembly of septin filaments at the membrane, as part of a molecular feedback loop between the assembly of septin and actin filaments.

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Endothelial Cell Cortactin Coordinates Intercellular Adhesion Molecule-1 Clustering and Actin Cytoskeleton Remodeling during Polymorphonuclear Leukocyte Adhesion and Transmigration

Cited by 105 publications

References 38 publications

ICAM-1-Mediated, Src- and Pyk2-Dependent Vascular Endothelial Cadherin Tyrosine Phosphorylation Is Required for Leukocyte Transendothelial Migration

ICAM-1-Mediated, Src- and Pyk2-Dependent Vascular Endothelial Cadherin Tyrosine Phosphorylation Is Required for Leukocyte Transendothelial Migration

Endothelial paxillin and focal adhesion kinase (FAK) play a critical role in neutrophil transmigration

Septin and actin contributions to endothelial cell–cell junctions and monolayer integrity

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