title: Epidermal growth factor-like domain 7 is a novel inhibitor of neutrophil adhesion to coronary artery endothelial cells injured by calcineurin inhibition.
abstract: BACKGROUND
  We investigated the effect of epidermal growth factor-like domain 7 (Egfl7) on nuclear factor-κB activation, intercellular adhesion molecule-1 expression, and neutrophil adhesion to human coronary artery endothelial cells after calcineurin-inhibition-induced injury.
  
  
  RESULTS
  Human coronary endothelial cells were incubated with cyclosporine (CyA) 10 μg/mL with or without Egfl7 (100 ng/mL) or the Notch receptor activator Jagged1 (200 ng/mL) for 6 to 48 hours. CyA upregulated nuclear factor-κB (p65) activity (128 ± 2% of control, P<0.001) in nuclear extracts, as determined with a DNA-binding activity ELISA. This activity was inhibited by Egfl7 (86 ± 3% of control; P<0.001 versus CyA alone). Jagged1 blocked Egfl7-induced nuclear factor-κB inhibition (105 ± 4% of control; P<0.05 versus CyA plus Egfl7). CyA upregulated cell-surface intercellular adhesion molecule-1 expression (215 ± 13% of control; P<0.001), as determined by flow cytometry. This expression was suppressed by Egfl7 (148 ± 5%; P<0.001 versus CyA alone). Jagged1 attenuated the intercellular adhesion molecule-1-suppressive effect of Egfl7 when administered with CyA (193 ± 3% versus 148 ± 5%; P<0.01). CyA increased neutrophil adhesion to human coronary endothelial cells (control 20 ± 5%, CyA 37 ± 3%; P<0.001 versus control) in a nonstatic neutrophil adhesion assay. This increase was attenuated by Egfl7 (22 ± 6%; P<0.001 versus CyA alone). Jagged 1 attenuated the effect of Egfl7 on neutrophil adhesion (31±3%; P<0.001 versus Egfl7 plus CyA).
  
  
  CONCLUSIONS
  Our study reveals that Egfl7 is a potent inhibitor of neutrophil adhesion to human coronary endothelial cells subsequent to calcineurin-inhibition-induced injury. Mechanistically, Egfl7 blocked nuclear factor-κB pathway activation and intercellular adhesion molecule-1 expression, which suggests that it may have significant antiinflammatory properties. Because Jagged1 blocked the effect of Egfl7, Notch receptor antagonism may contribute to the mechanism of action of Egfl7.
authors.affiliation: Division of Cardiovascular Surgery, Peter Munk Cardiac Centre, Toronto General Hospital-University Health Network, Toronto, Ontario, Canada.
authors.forename: Mitesh V
authors.initials: MV
authors.lastname: Badiwala
authors.forename: Daipayan
authors.initials: D
authors.lastname: Guha
authors.forename: Laura
authors.initials: L
authors.lastname: Tumiati
authors.forename: Jemy
authors.initials: J
authors.lastname: Joseph
authors.forename: Arash
authors.initials: A
authors.lastname: Ghashghai
authors.forename: Heather J
authors.initials: HJ
authors.lastname: Ross
authors.forename: Diego H
authors.initials: DH
authors.lastname: Delgado
authors.forename: Vivek
authors.initials: V
authors.lastname: Rao
mesh_terms: D019703:Calcineurin / Q000187:drug effects; D065095:Calcineurin Inhibitors; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology*; D016572:Cyclosporine / Q000494:pharmacology; D066255:EGF Family of Proteins; D016228:Endothelial Growth Factors / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D000072100:Jagged-1 Protein; D008565:Membrane Proteins / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects*; D009569:Nitric Oxide / Q000378:metabolism; D051880:Receptors, Notch / Q000819:agonists; D000072099:Serrate-Jagged Proteins; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D065095:Calcineurin Inhibitors; D002135:Calcium-Binding Proteins; D066255:EGF Family of Proteins; C527877:EGFL7 protein, human; D016228:Endothelial Growth Factors; D007166:Immunosuppressive Agents; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D016328:NF-kappa B; D051880:Receptors, Notch; D000072099:Serrate-Jagged Proteins; D018799:Intercellular Adhesion Molecule-1; D009569:Nitric Oxide; D016572:Cyclosporine; D019703:Calcineurin