Inhibition of the plasma SCUBE1, a novel platelet adhesive protein, protects mice against thrombosis

Arterioscler Thromb Vasc Biol. 2014 Jul;34(7):1390-8. doi: 10.1161/ATVBAHA.114.303779. Epub 2014 May 15.

Abstract

Objective: Signal peptide-CUB-EGF domain-containing protein 1 (SCUBE1), a secreted and surface-exposed glycoprotein on activated platelets, promotes platelet-platelet interaction and supports platelet-matrix adhesion. Its plasma level is a biomarker of platelet activation in acute thrombotic diseases. However, the exact roles of plasma SCUBE1 in vivo remain undefined.

Approach and results: We generated new mutant (Δ) mice lacking the soluble but retaining the membrane-bound form of SCUBE1. Plasma SCUBE1-depleted Δ/Δ mice showed normal hematologic and coagulant features and expression of major platelet receptors, but Δ/Δ platelet-rich plasma showed impaired platelet aggregation in response to ADP and collagen treatment. The addition of purified recombinant SCUBE1 protein restored the aggregation of platelets in Δ/Δ platelet-rich plasma and further enhanced platelet aggregation in +/+ platelet-rich plasma. Plasma deficiency of SCUBE1 diminished arterial thrombosis in mice and protected against lethal thromboembolism induced by collagen-epinephrine treatment. Last, antibodies directed against the epidermal growth factor-like repeats of SCUBE1, which are involved in trans-homophilic protein-protein interactions, protected mice against fatal thromboembolism without causing bleeding in vivo.

Conclusions: We conclude that plasma SCUBE1 participates in platelet aggregation by bridging adjacent activated platelets in thrombosis. Blockade of soluble SCUBE1 might represent a novel antithrombotic strategy.

Keywords: antibodies, neutralizing; blood platelets; mice, knockout; platelet aggregation; thrombosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / pharmacology
  • Blood Coagulation* / drug effects
  • Blood Platelets / drug effects
  • Blood Platelets / metabolism*
  • Calcium-Binding Proteins
  • Cell Shape
  • Disease Models, Animal
  • Fibrinolytic Agents / pharmacology
  • Intercellular Signaling Peptides and Proteins / blood*
  • Intercellular Signaling Peptides and Proteins / chemistry
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Mutant Strains
  • Mutation
  • Platelet Aggregation* / drug effects
  • Protein Structure, Tertiary
  • Pulmonary Embolism / blood
  • Pulmonary Embolism / genetics
  • Pulmonary Embolism / prevention & control*
  • Signal Transduction
  • Thrombosis / blood
  • Thrombosis / genetics
  • Thrombosis / prevention & control*
  • Time Factors

Substances

  • Antibodies, Monoclonal
  • Calcium-Binding Proteins
  • Fibrinolytic Agents
  • Intercellular Signaling Peptides and Proteins
  • Scube1 protein, mouse