Rare Disease & LCM Laboratories, Daiichi Sankyo Co., Ltd, 1-2-58, Hiromachi, Shinagawa-ku, Tokyo, 140-8710, Japan.
Sci Rep. 2018 Jul 2;8(1):9955. doi: 10.1038/s41598-018-28181-8.
Vasodilator-stimulated phosphoprotein (VASP) is a member of actin regulatory proteins implicated in platelet adhesion. In addition, phosphorylation of VASP is utilised for the assessment of platelet reactivity in patients treated with P2Y receptor antagonists, a class of antiplatelet agents. However, the role of VASP in platelet aggregation, thrombogenesis, haemostasis, and the antiplatelet effect of P2Y receptor antagonists remains unclear. We investigated these effects using heterozygous and homozygous VASP knockout rats generated with a CRISPR/Cas9 system. Baseline characteristics, such as haematology and other biochemical parameters, were comparable among the genotypes. In vitro platelet aggregation stimulated by adenosine diphosphate (ADP) or collagen, P-selectin expression of rat platelets treated with ADP, and in vivo thrombocytopenia induced by collagen were also comparable among the genotypes. In addition, in vivo thrombogenesis in a ferric chloride-induced arterial thrombosis model and bleeding time were also comparable among the genotypes. Furthermore, the in vitro antiplatelet effect of prasugrel, a third-generation P2Y receptor antagonist, was unaffected by VASP knockout. Although phosphorylated VASP is still an important surrogate marker specific for P2Y antagonists, our findings demonstrate that VASP is not a major mediator of platelet aggregation, thrombogenesis, haemostasis, and the antiplatelet effect of prasugrel in rats.
血管扩张刺激磷蛋白（VASP）是一种肌动蛋白调节蛋白，与血小板黏附有关。此外，VASP 的磷酸化被用于评估接受 P2Y 受体拮抗剂治疗的患者的血小板反应性，P2Y 受体拮抗剂是一类抗血小板药物。然而，VASP 在血小板聚集、血栓形成、止血和 P2Y 受体拮抗剂的抗血小板作用中的作用尚不清楚。我们使用 CRISPR/Cas9 系统生成的杂合子和纯合子 VASP 敲除大鼠来研究这些作用。基因型之间的基线特征，如血液学和其他生化参数，是可比的。体外由二磷酸腺苷（ADP）或胶原蛋白刺激的血小板聚集、用 ADP 处理的大鼠血小板上的 P-选择素表达以及胶原蛋白诱导的体内血小板减少在基因型之间也是可比的。此外，在三氯化铁诱导的动脉血栓形成模型和出血时间中的体内血栓形成在基因型之间也是可比的。此外，第三代 P2Y 受体拮抗剂普拉格雷的体外抗血小板作用不受 VASP 敲除的影响。尽管磷酸化的 VASP 仍然是 P2Y 拮抗剂的一个重要替代标志物，但我们的研究结果表明，VASP 不是大鼠血小板聚集、血栓形成、止血和普拉格雷抗血小板作用的主要介质。
