Lipid Receptor GPR31 (G-Protein-Coupled Receptor 31) Regulates Platelet Reactivity and Thrombosis Without Affecting Hemostasis.
Lipid Receptor GPR31 (G-Protein-Coupled Receptor 31) Regulates Platelet Reactivity and Thrombosis Without Affecting Hemostasis.
复制标题
DOI：
10.1161/atvbaha.120.315154
复制
发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Covic L
中科院分区：
文献类型：
--
作者：
Van Doren L;Nguyen N;Garzia C;Fletcher EK;Stevenson R;Jaramillo D;Kuliopulos A;Covic L
12-LOX（12-脂氧合酶）产生许多生物活性脂质，包括参与炎症和血小板反应性的 12(S)-HETE。 GPR31（G 蛋白偶联受体 31）是 12(S)-HETE 的拟议受体；然而，尚不清楚 12(S)-HETE-GPR31 信号轴是否有助于增强或抑制血小板活性。利用 pepducin 技术和生化方法，我们使用人血小板和小鼠颈动脉损伤模型提供证据，证明 12(S)-HETE-GPR31 通过 Gi 发出信号，增强 PAR（蛋白酶激活受体）-4 介导的血小板活化和动脉血栓形成。 12(S)-HETE 通过 GPR31 抑制 AC（腺苷酸环化酶）活性，导致 Rap1（Ras 相关蛋白 1）和 p38 激活以及较低但可检测到的钙通量，但不会诱导血小板聚集。 GPR31 的第三个细胞内 (i3) 环衍生的 pepducin GPR310（G 蛋白偶联受体 310）在人和小鼠血小板中显着抑制血小板聚集，以响应凝血酶、胶原蛋白和 PAR4 激动剂 AYPGKF，但在人血小板中相对较少地抑制 PAR1 激动剂 SFLLRN。 GPR310 治疗通过延长闭塞时间，对氯化铁引起的小鼠颈动脉损伤提供了 80% 的显着保护（P=0.0018），且对尾部出血没有任何影响。 GPR310 减弱了 PAR4 介导的致密颗粒分泌和钙通量。与这些结果一致，GPR310 抑制 12(S)-HETE 介导和 PAR4 介导的 Rap1-GTP 和 RASA3 易位至质膜，并减弱 PAR4-Akt 和 ERK 激活。 GPR310 导致凝血酶介导的人血小板聚集发生右移，与抑制 Gi 偶联 P2Y12 受体的效果相当。免疫共沉淀研究表明，GPR31 和 PAR4 在重组系统中形成异二聚体复合物。 12-LOX 产品 12(S)-HETE 刺激 GPR31-Gi 信号通路，从而增强人类血小板和小鼠模型中的凝血酶-PAR4 血小板活化和动脉血栓形成。抑制这种生物活性脂质途径（例如 GPR31 pepducin 拮抗剂）可以提供针对血小板聚集和动脉血栓形成的有益保护作用，同时对止血的影响最小。
12-LOX (12-lipoxygenase) produces a number of bioactive lipids including 12(S)-HETE that are involved in inflammation and platelet reactivity. The GPR31 (G-protein–coupled receptor 31) is the proposed receptor of 12(S)-HETE; however, it is not known whether the 12(S)-HETE-GPR31 signaling axis serves to enhance or inhibit platelet activity. Using pepducin technology and biochemical approaches, we provide evidence that 12(S)-HETE-GPR31 signals through Gi to enhance PAR (protease-activated receptor)-4–mediated platelet activation and arterial thrombosis using both human platelets and mouse carotid artery injury models. 12(S)-HETE suppressed AC (adenylyl cyclase) activity through GPR31 and resulted in Rap1 (Ras-related protein 1) and p38 activation and low but detectable calcium flux but did not induce platelet aggregation. A GPR31 third intracellular (i3) loop–derived pepducin, GPR310 (G-protein–coupled receptor 310), significantly inhibited platelet aggregation in response to thrombin, collagen, and PAR4 agonist, AYPGKF, in human and mouse platelets but relative sparing of PAR1 agonist SFLLRN in human platelets. GPR310 treatment gave a highly significant 80% protection (P=0.0018) against ferric chloride–induced carotid artery injury in mice by extending occlusion time, without any effect on tail bleeding. PAR4-mediated dense granule secretion and calcium flux were both attenuated by GPR310. Consistent with these results, GPR310 inhibited 12(S)-HETE–mediated and PAR4-mediated Rap1-GTP and RASA3 translocation to the plasma membrane and attenuated PAR4-Akt and ERK activation. GPR310 caused a right shift in thrombin-mediated human platelet aggregation, comparable to the effects of inhibition of the Gi-coupled P2Y12 receptor. Co-immunoprecipitation studies revealed that GPR31 and PAR4 form a heterodimeric complex in recombinant systems. The 12-LOX product 12(S)-HETE stimulates GPR31-Gi–signaling pathways, which enhance thrombin-PAR4 platelet activation and arterial thrombosis in human platelets and mouse models. Suppression of this bioactive lipid pathway, as exemplified by a GPR31 pepducin antagonist, may provide beneficial protective effects against platelet aggregation and arterial thrombosis with minimal effect on hemostasis.