Characterization of a novel function-blocking antibody targeted against the platelet P2Y1 receptor. Academic Article uri icon

abstract

  • OBJECTIVE: Platelet hyperactivity is associated with vascular disease and contributes to the genesis of thrombotic disorders. ADP plays an important role in platelet activation and activates platelets through 2 G-protein-coupled receptors, the Gq-coupled P2Y1 receptor (P2Y1R), and the Gi-coupled P2Y12 receptor. Although the involvement of the P2Y1R in thrombogenesis is well established, there are no antagonists that are currently available for clinical use. APPROACH AND RESULTS: Our goal is to determine whether a novel antibody targeting the ligand-binding domain, ie, second extracellular loop (EL2) of the P2Y1R (EL2Ab) could inhibit platelet function and protect against thrombogenesis. Our results revealed that the EL2Ab does indeed inhibit ADP-induced platelet aggregation, in a dose-dependent manner. Furthermore, EL2Ab was found to inhibit integrin GPIIb-IIIa activation, dense and α granule secretion, and phosphatidylserine exposure. These inhibitory effects translated into protection against thrombus formation, as evident by a prolonged time for occlusion in a FeCl3-induced thrombosis model, but this was accompanied by a prolonged tail bleeding time. We also observed a dose-dependent displacement of the radiolabeled P2Y1R antagonist [(3)H]MRS2500 from its ligand-binding site by EL2Ab. CONCLUSIONS: Collectively, our findings demonstrate that EL2Ab binds to and exhibits P2Y1R-dependent function-blocking activity in the context of platelets. These results add further evidence for a role of the P2Y1R in thrombosis and validate the concept that targeting it is a relevant alternative or complement to current antiplatelet strategies.

published proceedings

  • Arterioscler Thromb Vasc Biol

altmetric score

  • 0.5

author list (cited authors)

  • Karim, Z. A., Vemana, H. P., Alshbool, F. Z., Lin, O. A., Alshehri, A. M., Javaherizadeh, P., Paez Espinosa, E. V., & Khasawneh, F. T

citation count

  • 13

complete list of authors

  • Karim, Zubair A||Vemana, Hari Priya||Alshbool, Fatima Z||Lin, Olivia A||Alshehri, Abdullah M||Javaherizadeh, Payam||Paez Espinosa, Enma V||Khasawneh, Fadi T

publication date

  • March 2015