Synthetic glycopolymers and natural fucoidans cause human platelet aggregation via PEAR1 and GPIbα

Blood Adv. 2019 Feb 12;3(3):275-287. doi: 10.1182/bloodadvances.2018024950.

Abstract

Fucoidans are sulfated fucose-based polysaccharides that activate platelets and have pro- and anticoagulant effects; thus, they may have therapeutic value. In the present study, we show that 2 synthetic sulfated α-l-fucoside-pendant glycopolymers (with average monomeric units of 13 and 329) and natural fucoidans activate human platelets through a Src- and phosphatidylinositol 3-kinase (PI3K)-dependent and Syk-independent signaling cascade downstream of the platelet endothelial aggregation receptor 1 (PEAR1). Synthetic glycopolymers and natural fucoidan stimulate marked phosphorylation of PEAR1 and Akt, but not Syk. Platelet aggregation and Akt phosphorylation induced by natural fucoidan and synthetic glycopolymers are blocked by a monoclonal antibody to PEAR1. Direct binding of sulfated glycopolymers to epidermal like growth factor (EGF)-like repeat 13 of PEAR1 was shown by avidity-based extracellular protein interaction screen technology. In contrast, synthetic glycopolymers and natural fucoidans activate mouse platelets through a Src- and Syk-dependent pathway regulated by C-type lectin-like receptor 2 (CLEC-2) with only a minor role for PEAR1. Mouse platelets lacking the extracellular domain of GPIbα and human platelets treated with GPIbα-blocking antibodies display a reduced aggregation response to synthetic glycopolymers. We found that synthetic sulfated glycopolymers bind directly to GPIbα, substantiating that GPIbα facilitates the interaction of synthetic glycopolymers with CLEC-2 or PEAR1. Our results establish PEAR1 as the major signaling receptor for natural fucose-based polysaccharides and synthetic glycopolymers in human, but not in mouse, platelets. Sulfated α-l-fucoside-pendant glycopolymers are unique tools for further investigation of the physiological role of PEAR1 in platelets and beyond.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biopolymers / pharmacology
  • Calcium / blood
  • Humans
  • Mice
  • Mice, Knockout
  • Platelet Aggregation / drug effects*
  • Platelet Glycoprotein GPIb-IX Complex / metabolism*
  • Polysaccharides / pharmacology*
  • Receptors, Cell Surface / blood*
  • Syk Kinase / blood

Substances

  • Biopolymers
  • PEAR1 protein, human
  • PEAR1 protein, mouse
  • Platelet Glycoprotein GPIb-IX Complex
  • Polysaccharides
  • Receptors, Cell Surface
  • adhesion receptor
  • fucoidan
  • SYK protein, human
  • Syk Kinase
  • Calcium