Phospholipase D1 is a regulator of platelet-mediated inflammation

Glycoprotein (GP)Ib is not only required for stable thrombus formation but for platelet-mediated inflammatory responses. Phospholipase (PL)D1 is essential for GPIb-dependent aggregate formation under high shear conditions while nothing is known about PLD1-induced regulation of GPIb in platelet-mediated inflammation and the underlying mechanisms. This study aimed to investigate the relevance of PLD1 for platelet-mediated endothelial and leukocyte recruitment and activation in vitro and in vivo. Pld1^-/- platelets showed strongly reduced adhesion to TNFα stimulated endothelial cells (ECs) under high shear conditions ex vivo. Normal cytoskeletal reorganization of Pld1^-/- platelets but reduced integrin activation after adhesion to inflamed ECs confirmed that defective integrin activation is responsible for reduced platelet adhesion to ECs. This, together with significantly reduced CD40L expression on platelets led to reduced chemotactic and adhesive properties of ECs in vitro. Under flow conditions, recruitment of leukocytes to collagen-adherent platelets was reduced. Under inflammatory conditions in vivo, reduced platelet and leukocyte recruitment and arrest to the injured carotid artery was observed in Pld1^-/- mice. In a second in vivo model of venous thrombosis, platelet adhesion to activated endothelial cells was reduced while leukocyte recruitment was attenuated in PLD1 deficient mice. Mechanistically, PLD1 modulates PLCγ2 phosphorylation and integrin activation via Src kinases without affecting vWF binding to GPIb. Thus, PLD1 is important for GPIb-induced inflammatory processes of platelets and might be a promising target to reduce platelet-mediated inflammation.