ASK1 a novel regulator of platelet function

心血管疾病是人类的头号杀手。大多数CVD都与 动脉粥样硬化和血栓形成。越来越多的证据表明，血小板是起始物。 动脉粥样硬化和血栓形成。已知血小板中的激动剂刺激可激活 MAPK，已证明它们对体内和体内的血小板活性都很重要。 体外培养。尽管MAPK信号对血小板功能有明显的作用，但 它们调节血小板活性的机制还不完全清楚。我们有 作为MAP3K家族的一员，凋亡信号调节蛋白1(ASK1)是 存在于人和小鼠的血小板中，由生理激动剂激活。我们有 显示Ask1的活性支持血小板聚集和分泌，以及Ask1的消融 在体内血栓形成模型中具有保护作用。因此，我们假设 血小板ASK1是动脉粥样硬化形成、动脉粥样硬化血栓形成和缺血的关键调节因子 心肌梗死和卒中血栓溶解引起的再灌注(I/R)损伤。我们还有 根据已发表的虚拟数据库中的信息确定了几种结构不同的ASK1抑制剂 化学图书馆的屏幕。其中两种化合物显示出了良好的保护效果。 小鼠血栓形成，止血效果最小，通过尾部出血时间和 激光诱导止血模型。R01提案的重点是描绘血小板的作用 ASK1在启动动脉粥样硬化形成、动脉粥样硬化血栓形成、加重I/R损伤中起重要作用。因此， 提出了三个具体目标。《特殊目的1》将检验这样一种假设 ASK1是启动动脉粥样硬化形成的关键。我们将使用高脂血症小鼠模型(APOE-/-小鼠 高脂饲料喂养)，研究消融或抑制血小板Ask1对斑块的影响 队形。《特殊目的2》将检验以下假设：血小板ASK1是 动脉粥样硬化斑块破裂时的血小板活化(动脉粥样硬化血栓形成)。我们将使用 模拟斑块破裂部位血栓形成的创新小鼠模型。特定的 目的3验证血小板ASK1加重I/R损伤的假说。我们 将使用卒中的一过性大脑中动脉闭塞模型(TMCAO)来评估 消融/抑制血小板ASK1对I/R损伤的影响成功完成本建议书 将有助于开发一系列治疗血栓相关疾病的干预措施 例如动脉粥样硬化、心肌梗塞和中风。

Cardiovascular disease (CVD) is the number one killer of mankind. Most CVDs are associated with atherosclerosis and thrombosis. Mounting evidence suggests that platelets are the initiators of both atherosclerosis and thrombosis. Agonist stimulation in platelets is known to activate MAPKs, and it has been shown that they are important for platelet activities both in vivo and in vitro. Despite this evident role of MAPK signaling contributing to platelet functions, the mechanisms through which they regulate platelet activities are not fully understood. We have identified that a member of the MAP3K family, apoptosis signal-regulating kinase (ASK1) is present in both human and murine platelets and is activated by physiological agonists. We have shown that Ask1 activity supports platelet aggregation and secretion, and ablation of Ask1 confers a protective effect in in vivo models of thrombosis. We therefore hypothesize that platelet ASK1 is a key regulator of atherogenesis, atherothrombosis, and ischemia reperfusion (I/R) injury resulting from clot dissolution in MI, and stroke. We have also identified several structurally distinct ASK1 inhibitors based on information from published virtual chemical library screens. Two of these compounds have shown excellent efficacy in protecting mice from thrombosis with minimal effects on hemostasis, as assessed by tail bleeding time and laser-induced hemostasis model. This R01 proposal is focused on delineating the role of platelet ASK1 in initiating atherogenesis, atherothrombosis, and aggravating I/R injury. Accordingly, three Specific Aims have been proposed. Specific Aim 1 will test the hypothesis that platelet ASK1 is key in initiating atherogenesis. We will use a hyperlipidemia mouse model (Apoe-/- mice fed with high-fat diet) to study the effect of ablation or inhibition of platelet Ask1 on plaque formation. Specific Aim 2 will test the hypothesis that platelet ASK1 is a central regulator of platelet activation during atherosclerotic plaque rupture (atherothrombosis). We will use an innovative mouse model to mimic thrombus formation at the site of plaque rupture. Specific Aim 3 will test the hypothesis that platelet ASK1 is responsible for aggravating I/R injury. We will use the transient middle cerebral artery occlusion model (tMCAO) of stroke to assess the effect of ablation/inhibition of platelet ASK1 on I/R injury. Successful completion of this proposal will help to develop a number of therapeutic interventions for thrombosis associated diseases such as atherosclerosis, MI, and stroke.