Platelet-Mediated Neutrophil Extracellular Traps Regulate Ischemic Stroke Injury

血小板介导的中性粒细胞胞外陷阱调节缺血性中风损伤

• 批准号: 10593987
• 负责人: Robert A Campbell
• 金额: $ 53.49万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10593987
• 关键词: 3-Dimensional; Acute; Affect; Animals; Aspirin; Binding; Biological Assay; Blood Platelets; Blood Vessels; Brain; Brain Infarction; Brain Ischemia; Cause of Death; Cerebrovascular Circulation; Chromatin; Clinical; Data; Development; Event; Feedback; Female; Genetic; HMGB1 gene; Human; Impairment; In Vitro; Infarction; Infection; Inflammation; Inflammatory; Injury; Ischemia; Ischemic Stroke; Leukocytes; Location; Longitudinal Studies; Mediating; Microbe; Middle Cerebral Artery Occlusion; Modeling; Molecular; Motor; Mus; Nervous System Physiology; Nervous System Trauma; Neurologic; Neurologic Deficit; Neurological outcome; Neutrophil Activation; Neutrophil Infiltration; Outcome; Pathologic; Patients; Pattern; Persons; Plasminogen; Platelet Activation; Play; Public Health; Reaction; Recovery; Role; Site; Stroke; System; Techniques; Testing; Therapeutic; Thrombectomy; Thrombosis; Thrombus; Tissues; Venous Thrombosis; acute stroke; adverse outcome; antimicrobial; brain tissue; circulating biomarkers; cognitive disability; cognitive function; disability; experience; extracellular; fighting; improved; improved outcome; in vivo; inhibitor; innovation; long term recovery; male; microbicide; mortality; neutrophil; novel; novel therapeutics; pathogen; pharmacologic; physically handicapped; post stroke; recruit; response; scaffold; stroke model; stroke outcome; stroke patient; stroke therapy; thrombolysis; thrombotic; thrombotic complications

Despite advancements, about 700,000 people in the U.S. still experience an ischemic stroke annually. Current treatments for acute ischemic stroke are largely restricted to thrombolysis or thrombectomy, for which many stroke patients are ineligible for. Therefore, developing novel therapies for stroke is a significant public health need. Understanding the cellular mechanisms underlying stroke is critical for the development of new stroke therapies. Neutrophils and platelets are critical regulators of ischemic stroke injury. In humans and mice, platelet-neutrophil aggregates increase after ischemic stroke as well do neutrophil extracellular traps (NETs), a marker of neutrophil activation. NETs are critical during inflammation and infection and are released by neutrophils to trap pathogens. While NETs help fight infection, excessive NET formation can be detrimental to the host by promoting thrombosis. However, the pathological role of NET release has not been studied in ischemic stroke injury. Furthermore, the molecular regulators that trigger NET formation during stroke remain unclear. Finally, if targeting NET release during ischemic stroke injury improves outcomes is completely unknown. Here, we will test the innovative hypothesis that platelets are a primary driver of NET release during ischemic stroke and targeting NET formation with a novel, endogenous NET- inhibitory factor (nNIF), will improve stroke outcomes. We will employ complementary clinical, in vitro, and in vivo approaches, along with state-of-the-art techniques and models to rigorously test this hypothesis. Specific Aim 1 will determine if NETs are released after ischemic stroke and if they are present in the brains of human stroke patients and mice after experimental stroke. Furthermore, we will examine if neutrophils are primed to release NETs in ischemic stroke patients. Specific Aim 2 will establish whether platelet high mobility group box 1 (HMGB1), a danger associated molecular pattern released by platelets after activation, regulates NET formation during ischemic stroke injury. Specific Aim 3 will determine if therapeutic nNIF administration blocks NET formation during experimental stroke and improves acute and long-term stroke outcomes, including motor and neurological function. Successful completion of these aims will 1) determine if NETs are present in ischemic stroke injury including intravascular and extravascular locations; (2) establish whether neutrophils and platelets are primed to participate in NET formation during ischemic stroke, (3) determine whether platelet HMGB1 is a critical regulator of platelet-mediated NETosis during ischemic stroke; and (4) determine if NET inhibition improves stroke outcomes and the therapeutic window associated with pathological NET formation. Data generated in this proposal will significantly increase our understanding of how platelets contribute to pathological NET formation during ischemic stroke and associated neurological injury.

尽管取得了进步，但美国每年仍有约70万人患有缺血性中风。 目前急性缺血性卒中的治疗主要限于溶栓或血栓切除术， 很多中风患者都不适合。因此，开发脑卒中的新疗法是一个重要的公共问题。 健康需要。了解中风的细胞机制对于开发新的 中风治疗中性粒细胞和血小板是缺血性卒中损伤的关键调节因子。在人类和 在小鼠中，缺血性卒中后血小板-中性粒细胞聚集体增加，中性粒细胞胞外陷阱也增加 （NET），中性粒细胞活化的标志物。NET在炎症和感染过程中至关重要， 来捕获病原体。虽然NET有助于对抗感染，但过量的NET形成可能会导致 通过促进血栓形成对宿主有害。然而，NET释放的病理作用并没有被 研究缺血性中风损伤。此外，触发NET形成的分子调节剂， 中风仍不清楚。最后，如果在缺血性卒中损伤期间靶向NET释放可以改善结局， 完全未知在这里，我们将测试创新的假设，即血小板是一个主要的驱动力， 缺血性卒中期间NET的释放和用一种新的内源性NET靶向NET形成， 抑制因子（nNIF），将改善中风的结果。我们将采用互补的临床，体外， 和体内方法，沿着最先进的技术和模型来严格检验这一假设。 特定目标1将确定缺血性中风后是否释放NET以及它们是否存在于大脑中 人类中风患者和实验性中风后的小鼠。此外，我们将检查中性粒细胞是否 在缺血性中风患者中释放NET。具体目标2将确定血小板是否高 迁移率族蛋白1（HMGB 1），血小板活化后释放的危险相关分子模式， 在缺血性中风损伤期间调节NET形成。具体目标3将确定治疗性nNIF 给药阻断了实验性卒中期间NET的形成，并改善了急性和长期卒中 结果，包括运动和神经功能。成功完成这些目标将决定：1） NET存在于缺血性卒中损伤中，包括血管内和血管外位置;（2）建立 中性粒细胞和血小板是否在缺血性卒中期间参与NET形成，（3） 确定血小板HMGB 1是否是缺血性卒中期间血小板介导的NETosis的关键调节因子; 以及（4）确定NET抑制是否改善中风结果以及与以下相关的治疗窗： 病理性NET形成。本提案中产生的数据将大大增加我们对以下方面的了解： 缺血性卒中期间血小板如何促进病理性NET形成及相关神经系统疾病 损伤