Impaired Activation of Platelets Lacking Protein Kinase C Theta (PKCΘ) Isoform.

Abstract

Protein kinase C (PKC) has been implicated in platelet functional responses, but the contribution of individual isoforms has not been directly evaluated. PKCΘ is activated by glycoprotein VI (GPVI) and protease-activated receptor (PAR) agonists, but not by ADP. In human platelets, PKCΘ-selective receptor for activated C kinase (RACK) antagonistic peptide inhibited agonist-induced aggregation and secretion. Consistently, in murine platelets lacking PKCΘ, GPVI- or PAR-mediated aggregation and secretion were also impaired. Previously, fibrinogen receptor has been shown to be activated independently by calcium and PKC pathways. In the presence of dimethyl BAPTA, AYPGKF-induced platelet aggregation was inhibited by PKCΘ antagonistic RACK peptides, suggesting a role for this isoform in PKC-dependent fibrinogen receptor activation. In addition, the levels of thromboxane A2 (TXA2) release measured in GPVI and PAR-mediated activation of PKCΘ −/− murine platelets, were significantly lower compared to WT platelets. Moreover, agonist-induced extracellular-signal regulated kinase (ERK) phosphorylation was also significantly decreased in PKCΘ −/− murine platelets, which could be contributing to decreased TXA2 levels. PKCΘ −/− mice displayed unstable thrombus formation and prolonged arterial occlusion in the FeCl3 in vivo thrombosis model versus WT mice. In conclusion, PKCΘ isoform plays a significant role in platelet functional responses downstream of GPVI and PARs.