Epidermal growth factor-like domain 7 is a novel inhibitor of neutrophil adhesion to coronary artery endothelial cells injured by calcineurin inhibition

Circulation. 2011 Sep 13;124(11 Suppl):S197-203. doi: 10.1161/CIRCULATIONAHA.110.011734.

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.

Methods and 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcineurin / drug effects
  • Calcineurin Inhibitors*
  • Calcium-Binding Proteins / pharmacology
  • Cell Adhesion / drug effects
  • Cells, Cultured
  • Coronary Vessels / cytology*
  • Cyclosporine / pharmacology
  • EGF Family of Proteins
  • Endothelial Growth Factors / pharmacology*
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / metabolism
  • Humans
  • Immunosuppressive Agents / pharmacology
  • Intercellular Adhesion Molecule-1 / metabolism
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Jagged-1 Protein
  • Membrane Proteins / pharmacology
  • NF-kappa B / metabolism
  • Neutrophils / cytology*
  • Neutrophils / drug effects*
  • Nitric Oxide / metabolism
  • Receptors, Notch / agonists
  • Serrate-Jagged Proteins
  • Signal Transduction / drug effects
  • Signal Transduction / physiology

Substances

  • Calcineurin Inhibitors
  • Calcium-Binding Proteins
  • EGF Family of Proteins
  • EGFL7 protein, human
  • Endothelial Growth Factors
  • Immunosuppressive Agents
  • Intercellular Signaling Peptides and Proteins
  • JAG1 protein, human
  • Jagged-1 Protein
  • Membrane Proteins
  • NF-kappa B
  • Receptors, Notch
  • Serrate-Jagged Proteins
  • Intercellular Adhesion Molecule-1
  • Nitric Oxide
  • Cyclosporine
  • Calcineurin