scholarly journals TRPC6 is the endothelial calcium channel that regulates leukocyte transendothelial migration during the inflammatory response

2015 ◽  
Vol 212 (11) ◽  
pp. 1883-1899 ◽  
Author(s):  
Evan W. Weber ◽  
Fei Han ◽  
Mohammad Tauseef ◽  
Lutz Birnbaumer ◽  
Dolly Mehta ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Inflammatory Response ◽  
Calcium Channel ◽  
Transient Receptor Potential ◽  
Leukocyte Adhesion ◽  
Transendothelial Migration ◽  
Dominant Negative ◽  
Ion Concentration ◽  
Free Calcium ◽  
Leukocyte Transendothelial Migration

Leukocyte transendothelial migration (TEM) is a tightly regulated, multistep process that is critical to the inflammatory response. A transient increase in endothelial cytosolic free calcium ion concentration (↑[Ca2+]i) is required for TEM. However, the mechanism by which endothelial ↑[Ca2+]i regulates TEM and the channels mediating this ↑[Ca2+]i are unknown. Buffering ↑[Ca2+]i in endothelial cells does not affect leukocyte adhesion or locomotion but selectively blocks TEM, suggesting a role for ↑[Ca2+]i specifically for this step. Transient receptor potential canonical 6 (TRPC6), a Ca2+ channel expressed in endothelial cells, colocalizes with platelet/endothelial cell adhesion molecule-1 (PECAM) to surround leukocytes during TEM and clusters when endothelial PECAM is engaged. Expression of dominant-negative TRPC6 or shRNA knockdown in endothelial cells arrests neutrophils apically over the junction, similar to when PECAM is blocked. Selectively activating endothelial TRPC6 rescues TEM during an ongoing PECAM blockade, indicating that TRPC6 functions downstream of PECAM. Furthermore, endothelial TRPC6 is required for trafficking of lateral border recycling compartment membrane, which facilitates TEM. Finally, mice lacking TRPC6 in the nonmyeloid compartment (i.e., endothelium) exhibit a profound defect in neutrophil TEM with no effect on leukocyte trafficking. Our findings identify endothelial TRPC6 as the calcium channel mediating the ↑[Ca2+]i required for TEM at a step downstream of PECAM homophilic interactions.

scholarly journals TRPC6 is the endothelial calcium channel that regulates leukocyte transendothelial migration during the inflammatory response

2015 ◽  
Vol 146 (5) ◽  
pp. 1465OIA59
Author(s):  
Evan W. Weber ◽  
Fei Han ◽  
Mohammad Tauseef ◽  
Lutz Birnbaumer ◽  
Dolly Mehta ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Calcium Channel ◽  
Transendothelial Migration ◽  
Leukocyte Transendothelial Migration

scholarly journals TRPC6 is the endothelial calcium channel that regulates leukocyte transendothelial migration during the inflammatory response

2015 ◽  
Vol 210 (7) ◽  
pp. 2107OIA192
Author(s):  
Evan W. Weber ◽  
Fei Han ◽  
Mohammad Tauseef ◽  
Lutz Birnbaumer ◽  
Dolly Mehta ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Calcium Channel ◽  
Transendothelial Migration ◽  
Leukocyte Transendothelial Migration

scholarly journals MYADM controls endothelial barrier function through ERM-dependent regulation of ICAM-1 expression

2013 ◽  
Vol 24 (4) ◽  
pp. 483-494 ◽  
Author(s):  
Juan F. Aranda ◽  
Natalia Reglero-Real ◽  
Beatriz Marcos-Ramiro ◽  
Ana Ruiz-Sáenz ◽  
Laura Fernández-Martín ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Plasma Membrane ◽  
Inflammatory Response ◽  
Barrier Function ◽  
Leukocyte Adhesion ◽  
Cell Junctions ◽  
Membrane Domains ◽  
Filamentous Actin ◽  
Cytokine Tumor Necrosis Factor ◽  
Factor Α

The endothelium maintains a barrier between blood and tissue that becomes more permeable during inflammation. Membrane rafts are ordered assemblies of cholesterol, glycolipids, and proteins that modulate proinflammatory cell signaling and barrier function. In epithelial cells, the MAL family members MAL, MAL2, and myeloid-associated differentiation marker (MYADM) regulate the function and dynamics of ordered membrane domains. We analyzed the expression of these three proteins in human endothelial cells and found that only MYADM is expressed. MYADM was confined in ordered domains at the plasma membrane, where it partially colocalized with filamentous actin and cell–cell junctions. Small interfering RNA (siRNA)-mediated MYADM knockdown increased permeability, ICAM-1 expression, and leukocyte adhesion, all of which are features of an inflammatory response. Barrier function decrease in MYADM-silenced cells was dependent on ICAM-1 expression. Membrane domains and the underlying actin cytoskeleton can regulate each other and are connected by ezrin, radixin, and moesin (ERM) proteins. In endothelial cells, MYADM knockdown induced ERM activation. Triple-ERM knockdown partially inhibited ICAM-1 increase induced by MYADM siRNA. Importantly, ERM knockdown also reduced ICAM-1 expression in response to the proinflammatory cytokine tumor necrosis factor-α. MYADM therefore regulates the connection between the plasma membrane and the cortical cytoskeleton and so can control the endothelial inflammatory response.

scholarly journals Elucidating the Biomechanics of Leukocyte Transendothelial Migration by Quantitative Imaging

2021 ◽  
Vol 9 ◽  
Author(s):  
Amy B. Schwartz ◽  
Obed A. Campos ◽  
Ernesto Criado-Hidalgo ◽  
Shu Chien ◽  
Juan C. del Álamo ◽  
...  
Keyword(s):  
Vascular Endothelial Cells ◽  
Inflammatory Diseases ◽  
Leukocyte Adhesion ◽  
Decisive Role ◽  
Transendothelial Migration ◽  
Physical Contact ◽  
Cell Junctions ◽  
Force Transmission ◽  
Recent Advances ◽  
Leukocyte Transendothelial Migration

Leukocyte transendothelial migration is crucial for innate immunity and inflammation. Upon tissue damage or infection, leukocytes exit blood vessels by adhering to and probing vascular endothelial cells (VECs), breaching endothelial cell-cell junctions, and transmigrating across the endothelium. Transendothelial migration is a critical rate-limiting step in this process. Thus, leukocytes must quickly identify the most efficient route through VEC monolayers to facilitate a prompt innate immune response. Biomechanics play a decisive role in transendothelial migration, which involves intimate physical contact and force transmission between the leukocytes and the VECs. While quantifying these forces is still challenging, recent advances in imaging, microfabrication, and computation now make it possible to study how cellular forces regulate VEC monolayer integrity, enable efficient pathfinding, and drive leukocyte transmigration. Here we review these recent advances, paying particular attention to leukocyte adhesion to the VEC monolayer, leukocyte probing of endothelial barrier gaps, and transmigration itself. To offer a practical perspective, we will discuss the current views on how biomechanics govern these processes and the force microscopy technologies that have enabled their quantitative analysis, thus contributing to an improved understanding of leukocyte migration in inflammatory diseases.

scholarly journals ICAM-1 Clustering on Endothelial Cells Recruits VCAM-1

2010 ◽  
Vol 2010 ◽  
pp. 1-9 ◽  
Author(s):  
Jaap D. van Buul ◽  
Jos van Rijssel ◽  
Floris P. J. van Alphen ◽  
Anna-Marieke van Stalborch ◽  
Erik P. J. Mul ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Adhesion Molecules ◽  
Lipid Rafts ◽  
Leukocyte Adhesion ◽  
Transendothelial Migration ◽  
Human Endothelial Cells ◽  
Strong Adhesion ◽  
Integrin Ligands

In the initial stages of transendothelial migration, leukocytes use the endothelial integrin ligands ICAM-1 and VCAM-1 for strong adhesion. Upon adhesion of the leukocyte to endothelial ICAM-1, ICAM-1 is clustered and recruited to the adhered leukocyte, promoting strong adhesion. In this study, we provide evidence for the colocalization of VCAM-1 at sites of ICAM-1 clustering. Anti-ICAM-1 antibody-coated beads were used to selectively cluster and recruit ICAM-1 on primary human endothelial cells. In time, co-localization of ICAM-1 and VCAM-1 around the adherent beads was observed. Biochemical pull-down assays showed that ICAM-1 clustering induced its association to VCAM-1, suggesting a physical link between these two adhesion molecules. The association was partly dependent on lipid rafts as well as on F-actin and promoted adhesion. These data show that VCAM-1 can be recruited, in an integrin-independent fashion, to clustered ICAM-1 which may serve to promote ICAM-1-mediated leukocyte adhesion.

scholarly journals Transendothelial Migration of Melanoma Cells Involves N-Cadherin-mediated Adhesion and Activation of the β-Catenin Signaling Pathway

2005 ◽  
Vol 16 (9) ◽  
pp. 4386-4397 ◽  
Author(s):  
Jianfei Qi ◽  
Ning Chen ◽  
Junfu Wang ◽  
Chi-Hung Siu
Keyword(s):  
Endothelial Cells ◽  
Cancer Cells ◽  
Cancer Metastasis ◽  
Transendothelial Migration ◽  
Melanoma Cells ◽  
Dominant Negative ◽  
Vitro Assay ◽  
Multistep Process ◽  
Adhesive Interactions

Cancer metastasis is a multistep process involving many types of cell-cell interactions, but little is known about the adhesive interactions and signaling events during extravasation of cancer cells. Transendothelial migration of cancer cells was investigated using an in vitro assay, in which melanoma cells were seeded on top of a monolayer of endothelial cells. Attachment of melanoma cells on the endothelium induced a twofold increase in N-cadherin expression in melanoma cells and the redistribution of N-cadherin to the heterotypic contacts. Transendothelial migration was inhibited when N-cadherin expression was repressed by antisense RNA, indicating a key role played by N-cadherin. Whereas N-cadherin and β-catenin colocalized in the contact regions between melanoma cells and endothelial cells during the initial stages of attachment, β-catenin disappeared from the heterotypic contacts during transmigration of melanoma cells. Immunolocalization and immunoprecipitation studies indicate that N-cadherin became tyrosine-phosphorylated, resulting in the dissociation of β-catenin from these contact regions. Concomitantly, an increase in the nuclear level of β-catenin occurred in melanoma cells, together with a sixfold increase in β-catenin-dependent transcription. Transendothelial migration was compromised in cells expressing a dominant-negative form of β-catenin, thus supporting a regulatory role of β-catenin signaling in this process.

scholarly journals Sinusoidal Endotheliopathy in Nonalcoholic Steatohepatitis: Therapeutic Implications

2021 ◽  
Author(s):  
Samar H Ibrahim
Keyword(s):  
Endothelial Cells ◽  
Inflammatory Response ◽  
Adhesion Molecules ◽  
Immune Cells ◽  
Immune Cell ◽  
Leukocyte Adhesion ◽  
Low Flow ◽  
Liver Sinusoidal Endothelial Cells ◽  
Nonalcoholic Fatty Liver ◽  
Therapeutic Implications

Liver sinusoidal endothelial cells (LSEC) are distinct subtypes of endothelial cells lining a low flow vascular bed at the interface of the liver parenchyma, and the circulating immune cells and soluble factors. Emerging literature implicates LSEC in the pathogenesis and progression of nonalcoholic fatty liver disease (NAFLD). During the evolution of NAFLD, LSEC dysfunction ensues. LSEC undergo morphological and functional transformation known as "capillarization", as well as a pathogenic increase in surface adhesion molecules expression, referred to in this review as "endotheliopathy". LSEC govern the composition of hepatic immune cell populations in nonalcoholic steatohepatis (NASH) by mediating leukocyte subset adhesion through specific combinations of activated adhesion molecules and secreted chemokines. Moreover, extracellular vesicles released by hepatocyte under lipotoxic stress in NASH act as a catalyst for the inflammatory response and promote immune cell chemotaxis and adhesion. In the current review, we highlight leukocyte adhesion to LSEC as an initiating event in the sterile inflammatory response in NASH. We discuss preclinical studies targeting immune cells adhesion in NASH mouse models, and potential therapeutic anti-inflammatory strategies for human NASH.

Regulated release and functional modulation of junctional adhesion molecule A by disintegrin metalloproteinases

Blood ◽  
2009 ◽  
Vol 113 (19) ◽  
pp. 4799-4809 ◽  
Author(s):  
Rory R. Koenen ◽  
Jessica Pruessmeyer ◽  
Oliver Soehnlein ◽  
Line Fraemohs ◽  
Alma Zernecke ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Adhesion Molecule ◽  
Vascular Inflammation ◽  
Neutrophil Infiltration ◽  
Transendothelial Migration ◽  
Cultured Endothelial Cells ◽  
Leukocyte Transendothelial Migration ◽  
Functional Modulation ◽  
Air Pouch Model

Abstract Junctional adhesion molecule A (JAM-A) is a transmembrane adhesive glycoprotein that participates in the organization of endothelial tight junctions and contributes to leukocyte transendothelial migration. We demonstrate here that cultured endothelial cells not only express a cellular 43-kDa variant of JAM-A but also release considerable amounts of a 33-kDa soluble JAM-A variant. This release is enhanced by treatment with proinflammatory cytokines and is associated with the down-regulation of surface JAM-A. Inhibition experiments, loss/gain-of-function experiments, and cleavage experiments with recombinant proteases indicated that cleavage of JAM-A is mediated predominantly by the disintegrin and metalloproteinase (ADAM) 17 and, to a lesser extent, by ADAM10. Cytokine treatment of mice increased JAM-A serum level and in excised murine aortas increased ADAM10/17 activity correlated with enhanced JAM-A release. Functionally, soluble JAM-A blocked migration of cultured endothelial cells, reduced transendothelial migration of isolated neutrophils in vitro, and decreased neutrophil infiltration in a murine air pouch model by LFA-1– and JAM-A–dependent mechanisms. Therefore, shedding of JAM-A by inflamed vascular endothelium via ADAM17 and ADAM10 may not only generate a biomarker for vascular inflammation but could also be instrumental in controlling JAM-A functions in the molecular zipper guiding transendothelial diapedesis of leukocytes.

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