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: 10590459
• 负责人: MICHAEL Allan HOLINSTAT
• 金额: $ 8.23万
• 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 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10590459
• 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; GTP-Binding Protein alpha Subunits, Gs; Hemorrhage; Hemostatic function; Injury; LOX gene; Lead; Lipids; Membrane; Morbidity - disease rate; Myocardial Infarction; Nutrient; Oxides; Oxygen; Pathologic; Pathway interactions; Persons; Physiological; Platelet Activation; Play; Prevention; Process; Proteome; 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.

摘要 血小板在血管中起着至关重要的作用，维持血管后的止血和正常血流 生理条件下的侮辱或伤害。虽然血小板的活化对于粘附和粘附是必不可少的， 当血小板聚集发生在血管损伤部位时，过度的血小板反应性可导致 闭塞性血栓是心肌梗死和中风的主要潜在原因。当前抗血小板药物 治疗显著限制了由于血栓形成引起的发病率和死亡率，然而，它们经常导致 出血风险增加，导致需要新的靶点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-脂氧合酶（12-LOX），一种在血小板中高度表达的酶，其主要功能是 被认为是从嵌入血小板的脂肪酸中产生生物活性氧化脂质（氧化脂质 膜，因为在血小板活化的调节中起重要作用。新研究的脂肪酸的作用 血小板中的DGLA、DHA和EPA等脂肪酸已经产生了强有力的初步数据，支持脂肪酸 以及它们的12-LOX氧脂素对于通过GPCR和非GPCR调节血小板功能是重要的 机制等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）-HETrE的形成诱导Gs-偶联的GPCR途径的活化和PKA的活化，导致 抑制体内血小板活化和凝块形成，同时不引起观察到的出血增加。 我们建议研究这些12-LOX氧化脂质调节的潜在机制， 血小板活性、凝血和血栓形成，而仅最小程度地影响出血。所以我们会 评估DGLA及其oxlipin 12（S）-HETrE如何通过PKA的活化改变磷酸化蛋白质组， 阐明DHA和EPA调节血小板功能和体内外凝血的机制 通过他们的12-LOX氧化脂质，同时保留止血，并评估DHA 12- LOX oxylipin 14-HpDHA可被氧化形成一类新的maresins，Mar1-D3，以调节 血栓溶解和凝块消退。初步数据表明，血小板可能会产生一个新的池，这种亲， 分解氧脂素，其可在体内血块分解的潜在机制中发挥重要作用。 这项研究的成功完成将首次确定12-LOX氧脂素 调节血小板反应性，以防止凝块形成并消除预先存在的凝块。理解 这些复杂的信号通路代表了我们对氧脂素 如12（S）-HETrE调节血液，并将能够识别血小板上的新靶点， 开发下一代抗血小板治疗药物，并增加限制出血的益处。