title: Soluble VE-cadherin is involved in endothelial barrier breakdown in systemic inflammation and sepsis.
abstract: OBJECTIVE
  Microvascular endothelial barrier breakdown in sepsis precedes organ failure and death in patients. We tested the hypothesis that the formation of endothelium-derived soluble vascular endothelial (VE)-cadherin fragments (sVE-cadherin) is involved in inflammation-induced endothelial barrier disruption.
  
  
  RESULTS
  Incubation of human dermal microvascular endothelial cells (HDMEC) with tumour necrosis factor-α (TNF-α) and bacterial lipopolysaccharide (LPS) led to endothelial barrier disruption which correlated with significantly increased sVE-cadherin at a size of ∼90 kDa in cell culture supernatants. Inhibition of the VE-cadherin-cleaving disintegrin and metalloproteinase ADAM10 using GI254023X attenuated inflammation-induced formation of sVE-cadherin and endothelial barrier disruption, suggesting ADAM10-mediated shedding as a mechanism underlying sVE-cadherin release. Formation of VE-cadherin fragments at 90 and 110 kDa was observed when recombinant VE-cadherin (rVE-cadherin) was digested with recombinant ADAM10. Mass spectrometry of the VE-cadherin fragments showed that they originated from cleavage of the extracelluar domain and thereby several cleavage sites of ADAM10 were identified. Atomic force microscopy measurements demonstrated that cell culture supernatants containing sVE-cadherin and application of rVE-cadherin blocked VE-cadherin binding. Accordingly rVE-cadherin dose-dependently led to loss of endothelial barrier functions in HDMEC monolayers. Finally, in patients suffering from severe sepsis or septic shock with clinical signs of a microvascular leackage, serum levels of sVE-cadherin were significantly increased.
  
  
  CONCLUSIONS
  Taken together, formation of sVE-cadherin is associated and contributes to inflammation-induced breakdown of endothelial barrier functions by inhibition of VE-cadherin binding. The underlying mechanism of VE-cadherin cleavage involves ADAM10 and appears to be of clinical relevance since sVE-cadherin was augmented in patients with severe sepsis.
authors.affiliation: Department of General, Visceral, Vascular and Paediatric Surgery (Department of Surgery I), University of Wuerzburg, Oberduerrbacherstraße 6, Wuerzburg D-97080, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Flemming
authors.affiliation: Department of General, Visceral, Vascular and Paediatric Surgery (Department of Surgery I), University of Wuerzburg, Oberduerrbacherstraße 6, Wuerzburg D-97080, Germany.
authors.forename: Natalie
authors.initials: N
authors.lastname: Burkard
authors.affiliation: Department of General, Visceral, Vascular and Paediatric Surgery (Department of Surgery I), University of Wuerzburg, Oberduerrbacherstraße 6, Wuerzburg D-97080, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Renschler
authors.affiliation: Institute of Anatomy and Cell Biology, University of Munich, Munich D-80336, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Vielmuth
authors.affiliation: Department of General, Visceral, Vascular and Paediatric Surgery (Department of Surgery I), University of Wuerzburg, Oberduerrbacherstraße 6, Wuerzburg D-97080, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Meir
authors.affiliation: Department of Anaesthesia and Critical Care, University of Wuerzburg, Wuerzburg D-97080, Germany.
authors.forename: Martin Alexander
authors.initials: MA
authors.lastname: Schick
authors.affiliation: Department of Anaesthesia and Critical Care, University of Wuerzburg, Wuerzburg D-97080, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Wunder
authors.affiliation: Department of General, Visceral, Vascular and Paediatric Surgery (Department of Surgery I), University of Wuerzburg, Oberduerrbacherstraße 6, Wuerzburg D-97080, Germany.
authors.forename: Christoph-Thomas
authors.initials: CT
authors.lastname: Germer
authors.affiliation: Institute of Anatomy and Cell Biology, University of Munich, Munich D-80336, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Spindler
authors.affiliation: Institute of Anatomy and Cell Biology, University of Munich, Munich D-80336, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Waschke
authors.affiliation: Department of General, Visceral, Vascular and Paediatric Surgery (Department of Surgery I), University of Wuerzburg, Oberduerrbacherstraße 6, Wuerzburg D-97080, Germany Schlegel_N@ukw.de.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Schlegel
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors / Q000502:physiology; D000072197:ADAM10 Protein; D000368:Aged; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000502:physiology; D015703:Antigens, CD / Q000032:analysis / Q000502:physiology*; D015820:Cadherins / Q000032:analysis / Q000502:physiology*; D002478:Cells, Cultured; D004151:Dipeptides / Q000494:pharmacology; D042783:Endothelial Cells / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007249:Inflammation / Q000503:physiopathology*; D008297:Male; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000502:physiology; D008875:Middle Aged; D010539:Permeability; D058988:Phosphodiesterase 4 Inhibitors / Q000494:pharmacology; D018805:Sepsis / Q000503:physiopathology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D000099152:Cadherin 5
chemical_list: D051722:ADAM Proteins; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases; D015703:Antigens, CD; D015820:Cadherins; D004151:Dipeptides; D006877:Hydroxamic Acids; D008565:Membrane Proteins; D058988:Phosphodiesterase 4 Inhibitors; D014409:Tumor Necrosis Factor-alpha; D000099152:Cadherin 5; C555398:3-(formylhydroxyamino)-2-(3-phenyl-1-propyl)butanoic acid (2,2-dimethyl-1-methylcarbamoyl-1-propyl)amide; C491236:ADAM10 protein, human
keywords: ADAM10; Endothelial barrier; Inflammation; Metalloproteinases; VE-cadherin