12-HETrE regulation of blood coagulation, hemostasis, and thrombosis

12-HETrE 对凝血、止血和血栓形成的调节

• 批准号: 10599220
• 负责人: MICHAEL Allan HOLINSTAT
• 金额: $ 39万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10599220
• 关键词: Adhesions; Affect; Arachidonate 12-Lipoxygenase; Blood; Blood Circulation; Blood Platelets; Blood Vessels; Blood coagulation; Blood flow; Cell Death; Cells; Coagulation Process; Complex; Coupled; Cyclic AMP-Dependent Protein Kinases; Data; Development; Eicosanoids; Endothelium; Enzymes; Event; Excision; Exhibits; Fatty Acids; G-Protein-Coupled Receptors; Hemorrhage; Hemostatic function; Injury; Lipids; Membrane; Morbidity - disease rate; Myocardial Infarction; Nutrient; Oxygen; Pathologic; Pathway interactions; Persons; Physiological; Platelet Activation; Play; Prevention; Process; Regulation; Resolution; Risk; Role; Seminal; Signal Pathway; Site; Stroke; Therapeutic; Thrombosis; Thrombus; Time; cardiovascular risk factor; in vivo; mortality; next generation; novel; oxidized lipid; platelet function; prevent; thrombolysis; thrombotic; vascular injury; wasting

ABSTRACT Platelets play an essential role in the vessel, maintaining hemostasis and normal blood flow following vascular insult or injury under physiological conditions. While activation of the platelet is essential for adhesion and aggregation to occur at the site of vascular injury, excessive platelet reactivity can lead to the formation of occlusive thrombi, the predominant underlying cause of myocardial infarction and stroke. Current anti-platelet treatments have significantly limited morbidity and mortality due to thrombosis, however they often result in an increased risk of bleeding resulting in a need for novel targets to further decrease platelet reactivity while exhibiting a limited increased risk for bleeding. Our lab and others have provided compelling evidence supporting 12-lipoxygenase (12-LOX), an enzyme highly expressed in the platelet whose primary function is thought to be to produce bioactive oxidized lipids (oxylipins) from the fatty acids embedded in the platelet membrane, as playing an important role in the regulation of platelet activation. The role of newly studied fatty acids in the platelet such as DGLA, DHA, and EPA has yielded strong preliminary data supporting fatty acids and their 12-LOX oxylipins as being important for regulation of platelet function through GPCR and non-GPCR mechanisms. Our lab has identified the first 12-LOX negative regulation pathway in the platelet whereby formation of 12(S)-HETrE induces activation of a Gs-coupled GPCR pathway and activation of PKA resulting in inhibition of platelet activation and clot formation in vivo while not causing an observed increase in bleeding. We propose to investigate the underlying mechanisms by which these 12-LOX oxylipins regulate platelet activity, clotting, and thrombosis while only minimally affecting bleeding. Therefore, we will assess how DGLA and its oxylipin 12(S)-HETrE alters the phospho-proteome through activation of PKA, elucidate the mechanism by which DHA and EPA regulate platelet function and clotting both ex vivo and in vivo through their 12-LOX oxylipins while sparing hemostasis, and assess the mechanism by which the DHA 12- LOX oxylipin 14-HpDHA can be oxidized to form a new class of maresins, Mar1-D3, in order to regulate thrombolysis and clot resolution. Preliminary data suggests the platelet may produce a novel pool for this pro- resolving oxylipins that could play an important role in the underlying mechanism of clot resolution in vivo. Successful completion of this study will for the first time define the mechanism by which 12-LOX oxylipins regulate platelet reactivity in order to both prevent clot formation and resolve pre-existing clots. Understanding these complex signaling pathways represents a seminal advancement in our understanding of how oxylipins like 12(S)-HETrE regulate the blood and will enable for identification of new targets on the platelet for development of the next generation of anti-platelet therapeutics with the added benefit of limited bleeding.

抽象的 血小板在血管中起着至关重要的作用，维持止血和血管正常流动 生理条件下的侮辱或伤害。血小板的激活对于粘附和 在血管损伤部位发生的聚集，血小板反应过多会导致形成 闭塞的血栓，这是心肌梗塞和中风的主要根本原因。当前的抗血小板 治疗因血栓形成而引起的发病率和死亡率显着有限，但是它们通常导致 增加出血风险，导致需要新的靶标，以进一步降低血小板反应性 表现出有限的出血风险增加。我们的实验室和其他实验室提供了令人信服的证据 支持12-脂氧酶（12-lox），一种高度表达的酶，其主要功能为 被认为是从嵌入血小板中的脂肪酸中产生生物活性的氧化脂质（氧蛋白） 膜，是在血小板激活调节中起重要作用。新研究的脂肪的作用 血小板中的酸（例如DGLA，DHA和EPA）产生了强大的初步数据，支持脂肪酸 他们的12-洛克斯氧磷酸对于通过GPCR和非GPCR调节血小板功能很重要 机制。我们的实验室已经确定了血小板中的第一个12-lox负调节途径 12（s）的形成 - 诱导GS耦合GPCR途径的激活和PKA的激活，从而激活 在体内抑制血小板激活和凝块形成，而不会引起观察到的出血增加。 我们建议研究这些12-洛克斯氧蛋白调节的潜在机制 血小板活性，凝结和血栓形成，而仅对出血的影响最小。因此，我们会的 评估DGLA及其Oxylipin 12（s）的光如何通过激活PKA来改变磷酸化的蛋白质。 阐明DHA和EPA调节血小板功能的机制，并在体内和体内凝结 通过其12-洛克斯氧蛋白在节省止血的同时，并评估DHA 12--的机制 Lox oxylipin 14-HPDHA可以氧化以形成一类新的Maresins Mar1-D3，以调节 溶栓和凝块分辨率。初步数据表明，血小板可能会为此产生新的池 解决可能在体内凝块分辨率的基本机制中起重要作用的黄脂素。 这项研究的成功完成将首次定义12-洛克斯氧蛋白的机制 调节血小板反应性，以防止凝块形成并解决先前存在的凝块。理解 这些复杂的信号通路代表了我们对氧基蛋白的理解的开创性进步 像12（s） - 光调节血液，并可以识别血小板上的新靶 下一代抗血小板治疗剂的发展，其额外的出血有限。