ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signalling

Chongxiu Sun, MacK H. Wu, Mingzhang Guo, Mark L. Day, Eugene S Lee, Sarah Y. Yuan

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

AimsEndothelial barrier dysfunction is a key event in the pathogenesis of vascular diseases associated with inflammation. ADAM (a disintegrin and metalloprotease) 15 has been shown to contribute to the development of vascular inflammation. However, its role in regulating endothelial barrier function is unknown. The aim of this study was to examine the effect of ADAM15 on endothelial permeability and its underlying mechanisms.Methods and resultsBy measuring albumin transendothelial flux and transendothelial electric resistance in cultured human umbilical vein endothelial cell monolayers, we found that depletion of ADAM15 expression via siRNA decreased endothelial permeability and attenuated thrombin-induced barrier dysfunction. In contrast, endothelial cells overexpressing either wild-type or catalytically dead mutant ADAM15 displayed a higher basal permeability and augmented hyperpermeability in response to thrombin. In addition, ADAM15 knockdown inhibited whereas ADAM15 overexpression promoted neutrophil transendothelial migration. Further molecular assays revealed that ADAM15 did not cleave vascular endothelial-cadherin or cause its degradation. However, overexpression of ADAM15 promoted extracellular signal-regulated kinase (ERK)1/2 phosphorylation in both non-stimulated and thrombin-stimulated endothelial cells in a protease activity-independent manner. Pharmacological inhibition of Src kinase or ERK activation reversed ADAM15-induced hyperpermeability and neutrophil transmigration.ConclusionThe data provide evidence for a novel function of ADAM15 in regulating endothelial barrier properties. The mechanisms of ADAM15-induced hyperpermeability involve Src/ERK1/2 signalling independent of junction molecule shedding.

Original languageEnglish (US)
Pages (from-to)348-355
Number of pages8
JournalCardiovascular Research
Volume87
Issue number2
DOIs
StatePublished - 2010

Fingerprint

Thrombin
Permeability
Neutrophils
Endothelial Cells
Inflammation
Transendothelial and Transepithelial Migration
Disintegrins
Mitogen-Activated Protein Kinase 3
src-Family Kinases
Mitogen-Activated Protein Kinase 1
Extracellular Signal-Regulated MAP Kinases
Human Umbilical Vein Endothelial Cells
Metalloproteases
Electric Impedance
Vascular Diseases
Small Interfering RNA
Blood Vessels
Albumins
Peptide Hydrolases
Phosphorylation

Keywords

  • Endothelial permeability
  • Metalloproteinase
  • Signal transduction
  • Vascular inflammation

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)
  • Physiology
  • Medicine(all)

Cite this

ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signalling. / Sun, Chongxiu; Wu, MacK H.; Guo, Mingzhang; Day, Mark L.; Lee, Eugene S; Yuan, Sarah Y.

In: Cardiovascular Research, Vol. 87, No. 2, 2010, p. 348-355.

Research output: Contribution to journalArticle

Sun, Chongxiu ; Wu, MacK H. ; Guo, Mingzhang ; Day, Mark L. ; Lee, Eugene S ; Yuan, Sarah Y. / ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signalling. In: Cardiovascular Research. 2010 ; Vol. 87, No. 2. pp. 348-355.
@article{cbfe873c51f54d11a5c192035d9f6b36,
title = "ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signalling",
abstract = "AimsEndothelial barrier dysfunction is a key event in the pathogenesis of vascular diseases associated with inflammation. ADAM (a disintegrin and metalloprotease) 15 has been shown to contribute to the development of vascular inflammation. However, its role in regulating endothelial barrier function is unknown. The aim of this study was to examine the effect of ADAM15 on endothelial permeability and its underlying mechanisms.Methods and resultsBy measuring albumin transendothelial flux and transendothelial electric resistance in cultured human umbilical vein endothelial cell monolayers, we found that depletion of ADAM15 expression via siRNA decreased endothelial permeability and attenuated thrombin-induced barrier dysfunction. In contrast, endothelial cells overexpressing either wild-type or catalytically dead mutant ADAM15 displayed a higher basal permeability and augmented hyperpermeability in response to thrombin. In addition, ADAM15 knockdown inhibited whereas ADAM15 overexpression promoted neutrophil transendothelial migration. Further molecular assays revealed that ADAM15 did not cleave vascular endothelial-cadherin or cause its degradation. However, overexpression of ADAM15 promoted extracellular signal-regulated kinase (ERK)1/2 phosphorylation in both non-stimulated and thrombin-stimulated endothelial cells in a protease activity-independent manner. Pharmacological inhibition of Src kinase or ERK activation reversed ADAM15-induced hyperpermeability and neutrophil transmigration.ConclusionThe data provide evidence for a novel function of ADAM15 in regulating endothelial barrier properties. The mechanisms of ADAM15-induced hyperpermeability involve Src/ERK1/2 signalling independent of junction molecule shedding.",
keywords = "Endothelial permeability, Metalloproteinase, Signal transduction, Vascular inflammation",
author = "Chongxiu Sun and Wu, {MacK H.} and Mingzhang Guo and Day, {Mark L.} and Lee, {Eugene S} and Yuan, {Sarah Y.}",
year = "2010",
doi = "10.1093/cvr/cvq060",
language = "English (US)",
volume = "87",
pages = "348--355",
journal = "Cardiovascular Research",
issn = "0008-6363",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signalling

AU - Sun, Chongxiu

AU - Wu, MacK H.

AU - Guo, Mingzhang

AU - Day, Mark L.

AU - Lee, Eugene S

AU - Yuan, Sarah Y.

PY - 2010

Y1 - 2010

N2 - AimsEndothelial barrier dysfunction is a key event in the pathogenesis of vascular diseases associated with inflammation. ADAM (a disintegrin and metalloprotease) 15 has been shown to contribute to the development of vascular inflammation. However, its role in regulating endothelial barrier function is unknown. The aim of this study was to examine the effect of ADAM15 on endothelial permeability and its underlying mechanisms.Methods and resultsBy measuring albumin transendothelial flux and transendothelial electric resistance in cultured human umbilical vein endothelial cell monolayers, we found that depletion of ADAM15 expression via siRNA decreased endothelial permeability and attenuated thrombin-induced barrier dysfunction. In contrast, endothelial cells overexpressing either wild-type or catalytically dead mutant ADAM15 displayed a higher basal permeability and augmented hyperpermeability in response to thrombin. In addition, ADAM15 knockdown inhibited whereas ADAM15 overexpression promoted neutrophil transendothelial migration. Further molecular assays revealed that ADAM15 did not cleave vascular endothelial-cadherin or cause its degradation. However, overexpression of ADAM15 promoted extracellular signal-regulated kinase (ERK)1/2 phosphorylation in both non-stimulated and thrombin-stimulated endothelial cells in a protease activity-independent manner. Pharmacological inhibition of Src kinase or ERK activation reversed ADAM15-induced hyperpermeability and neutrophil transmigration.ConclusionThe data provide evidence for a novel function of ADAM15 in regulating endothelial barrier properties. The mechanisms of ADAM15-induced hyperpermeability involve Src/ERK1/2 signalling independent of junction molecule shedding.

AB - AimsEndothelial barrier dysfunction is a key event in the pathogenesis of vascular diseases associated with inflammation. ADAM (a disintegrin and metalloprotease) 15 has been shown to contribute to the development of vascular inflammation. However, its role in regulating endothelial barrier function is unknown. The aim of this study was to examine the effect of ADAM15 on endothelial permeability and its underlying mechanisms.Methods and resultsBy measuring albumin transendothelial flux and transendothelial electric resistance in cultured human umbilical vein endothelial cell monolayers, we found that depletion of ADAM15 expression via siRNA decreased endothelial permeability and attenuated thrombin-induced barrier dysfunction. In contrast, endothelial cells overexpressing either wild-type or catalytically dead mutant ADAM15 displayed a higher basal permeability and augmented hyperpermeability in response to thrombin. In addition, ADAM15 knockdown inhibited whereas ADAM15 overexpression promoted neutrophil transendothelial migration. Further molecular assays revealed that ADAM15 did not cleave vascular endothelial-cadherin or cause its degradation. However, overexpression of ADAM15 promoted extracellular signal-regulated kinase (ERK)1/2 phosphorylation in both non-stimulated and thrombin-stimulated endothelial cells in a protease activity-independent manner. Pharmacological inhibition of Src kinase or ERK activation reversed ADAM15-induced hyperpermeability and neutrophil transmigration.ConclusionThe data provide evidence for a novel function of ADAM15 in regulating endothelial barrier properties. The mechanisms of ADAM15-induced hyperpermeability involve Src/ERK1/2 signalling independent of junction molecule shedding.

KW - Endothelial permeability

KW - Metalloproteinase

KW - Signal transduction

KW - Vascular inflammation

UR - http://www.scopus.com/inward/record.url?scp=77954317110&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77954317110&partnerID=8YFLogxK

U2 - 10.1093/cvr/cvq060

DO - 10.1093/cvr/cvq060

M3 - Article

C2 - 20189953

AN - SCOPUS:77954317110

VL - 87

SP - 348

EP - 355

JO - Cardiovascular Research

JF - Cardiovascular Research

SN - 0008-6363

IS - 2

ER -