Targeting Pyruvate Kinase M2: A novel strategy to combat thrombo-inflammation

项目总结 心血管疾病(CVD)和中风夺走的生命比所有形式的癌症加起来还要多，并导致 巨大的健康和经济负担(美国每年3160亿美元)。当前的战略是 在高危患者中预防急性冠脉综合征和缺血性中风依赖于抗血小板药物(例如 阿司匹林和P2Y12抑制剂)，这些药物在三分之一的患者中没有转化为临床疗效。更有力的反- 血小板制剂，如糖蛋白IIbIIIa抑制剂(如阿昔单抗)与出血有关 并发症多，不宜长期使用。因为心血管疾病和中风的特点是血栓形成 和炎症，理想的药物应该是一种在没有重大影响的情况下抑制血栓炎症反应的药物。 出血和激活炎症消解程序导致巨噬细胞从M1极化 (促炎)到M2(抗炎)表型。为了实现这一目标，我们正在探索一种创新的 通过操纵活化血小板中的有氧糖酵解抑制血栓炎症的策略和 白细胞。我们的方法是瞄准有氧糖酵解的关键调节酶--丙酮酸激酶M2 (PKM2)。这种方法利用了最近的发现，就像大多数正常细胞一样，静息的血小板 白细胞主要依赖氧化磷酸化来产生ATP，而激活的血小板和 白细胞表现出高水平的有氧糖酵解(在存在的情况下将葡萄糖转化为乳酸 氧气)。值得注意的是，最近的证据表明，PKM2在单核细胞中高度表达， 冠状动脉疾病患者的巨噬细胞，以及M1巨噬细胞极化的驱动因素。利用 我们产生了新的突变的血小板特异性PKM2缺陷株和髓系特异性PKM2缺陷株 初步数据表明，PKM2在调节血栓炎症中发挥了作用。这项研究的目标是 计划是进一步了解PKM2如何调节血小板和白细胞功能，并确定 靶向二聚体PKM2将抑制高脂血症小鼠模型的血栓炎症。为了促进 这一创新和高额奖励计划的成功，我们将利用互补的遗传和 药理学方法和最新的活体显微镜，以及最新的卒中治疗 学术行业圆桌会议(STAIR)临床前指南。我们有所有的工具，包括试剂和 最先进的活体显微镜和动物模型，以实现我们的目标。我有前提条件 我的业绩记录证明了我的经验，它显示出高生产率和上升轨迹 血栓炎症领域。我召集了一群基础科学家和临床医生，他们的专业知识 将有助于指导拟议的研究从长凳到临床。该项目具有重要的临床意义。 由于对能量代谢及其对血小板和白细胞功能的影响有了明确的认识 功能可以识别血栓形成和炎症共同的新治疗靶点，这可能 改善包括急性缺血性中风在内的血栓炎症性疾病高危患者的预后。

Project summary Cardiovascular disease (CVD) and stroke claim more lives than all forms of cancer combined and result in an immense health and economic burden (>$316 billion annually in the United States). Current strategies to prevent acute coronary syndromes and ischemic stroke in at risk patients rely on anti-platelet drugs (e.g. aspirin and P2Y12 inhibitors), which do not translate into clinical efficacy in 1/3rd of patients. More potent anti- platelet agents such as Glycoprotein IIbIIIa inhibitors (e.g. abciximab) are associated with bleeding complications and are not suitable for long-term use. Since CVD and stroke are characterized by thrombosis and inflammation, an ideal drug would be one that inhibits thrombo-inflammatory responses without major bleeding and activates inflammation resolution programs leading to polarization of macrophages from an M1 (pro-inflammatory) to an M2 (anti-inflammatory) phenotype. To accomplish this, we are exploring an innovative strategy to inhibit thrombo-inflammation by manipulating aerobic glycolysis in activated platelets and leukocytes. Our approach will be to target the key regulatory enzyme of aerobic glycolysis, pyruvate kinase M2 (PKM2). This approach takes advantage of the recent discovery that, like most normal cells, resting platelets and leukocytes rely primarily on oxidative phosphorylation to generate ATP, whereas activated platelets and leukocytes exhibit a high level of aerobic glycolysis (conversion of glucose to lactate in the presence of oxygen). Notably, recent evidence indicates that PKM2 is highly expressed in the monocytes and macrophages from patients with coronary artery disease, and a driver of M1 macrophage polarization. Utilizing novel mutant platelet-specific PKM2 deficient and myeloid-specific PKM2 deficient strains, we have generated preliminary data that suggests a role for PKM2 in modulating thrombo-inflammation. The goals of this research program are to further understand how PKM2 regulates platelet and leukocyte function and to determine if targeting dimeric PKM2 will inhibit thrombo-inflammation in a murine model of hyperlipidemia. To promote the success of this innovative and high reward program, we will utilize complementary genetic and pharmacological approaches and state-of-the art intravital microscopy, and follow updated Stroke Therapy Academic Industry Roundtable (STAIR) pre-clinical guidelines. We have all the tools, including reagents and state-of-the art intravital microscopy and animal models, to accomplish our goals. I have the prerequisite experience as evidenced by my track record, which has shown high productivity and an upward trajectory in the field of thrombo-inflammation. I have assembled a group of basic scientists and clinicians whose expertise will help guide the proposed research from bench to clinic. This project has significant clinical implications since a clear understanding of energy metabolism and its functional consequences on platelet and leukocyte function could identify novel therapeutic targets common to both thrombosis and inflammation, which may improve outcomes in patients at high risk for thrombo-inflammatory disorders including acute ischemic stroke.