Role of bradykinin receptor 2 in tPA stroke therapy

缓激肽受体 2 在 tPA 中风治疗中的作用

• 批准号: 9767297
• 负责人: Fabricio Simao
• 金额: $ 21.47万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9767297
• 关键词: Address; Adoptive Transfer; Adverse effects; Affect; Aftercare; Agonist; Alteplase; Area; Blood - brain barrier anatomy; Blood Vessels; Blood specimen; Bone Marrow; Bone Marrow Cells; Bradykinin; Bradykinin Receptor; Brain; Brain hemorrhage; Cell Death; Cells; Cerebral hemisphere hemorrhage; Cerebrum; Chimera organism; Clinical; Coculture Techniques; Collaborations; Data; Development; Endothelial Cells; Endothelium; Endothelium-Dependent Relaxing Factors; Extravasation; FDA approved; Factor XII; Glucose; Goals; Grant; Growth and Development function; Hemorrhage; Hospitals; Human; Immune; Immune Cell Activation; Immune system; Immunobiology; Infarction; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Infusion procedures; Injury; Ischemic Stroke; K-Series Research Career Programs; Laboratories; Link; Mediating; Mentors; Mentorship; Molecular; Mus; Neurosciences; Oxygen; Pathway interactions; Patients; Phagocytosis; Plasma; Plasma Kallikrein; Plasminogen; Process; Prostate-Specific Antigen; Research; Research Personnel; Risk; Role; Series; Signal Transduction; Splenectomy; Stroke; Testing; Therapeutic; Thrombolytic Therapy; Time; Training; Treatment Efficacy; Wild Type Mouse; Writing; acute stroke; brain endothelial cell; career development; cell injury; deprivation; experimental study; improved; in vitro Model; in vitro activity; innovation; macrophage; migration; monocyte; neuron loss; new therapeutic target; novel; novel strategies; paracrine; post stroke; prevent; programs; recruit; skills; stroke therapy

Project Summary The widespread use of tissue plasminogen activator (tPA), the only FDA-approved acute stroke treatment, remains limited by its narrow therapeutic time window and related risks of brain hemorrhage. Bradykinin (BK) is well known for its actions as an endothelial-dependent vasodilator and a powerful proinflammatory agonist. A number of experimental studies have show that BK receptors are upregulated after stroke by mediating inflammatory effects during early stages after stroke, indicating that BK aggravates cerebral injury. In preliminary studies, we found that: 1) BK formation is increased after tPA therapy through plasma kallikrein activation; 2) co-administration of a BK receptor 2 antagonist with tPA treatment after stroke reduces hemorrhage transformation; 3) B2R agonist does not induce endothelial cell death during oxygen and glucose deprivation; 4) splenectomized or macrophage-depleted mice with tPA decreased brain hemorrhage; 5) BK infusion after stroke increases hemorrhage transformation, which was blocked in macrophage-depleted mice. The objective of the proposed studies is to define the mechanisms by which BK affects and activates macrophages after tPA therapy, thus causing brain hemorrhage via proinflammatory processes during vascular injury. Our ultimate goal is to delineate the mechanistic underpinnings of tPA therapy responsible for improving the time window and to render tPA safer and more widely usable in patients with stroke. We propose to test the following specific aims: (i) to determine the contribution of conditional endothelium- and macrophage-specific B2R deletion to tPA-induced hemorrhage transformation; (ii) to characterize the role of BK in activated macrophages on accumulation and recruitment after tPA treatment; (iii) to delineate the molecular mechanisms by which BK regulates macrophage function and its interaction with brain endothelial cells. I will use this mentored career development award (K01) to fulfill a series of training objectives, which build upon my prior skills in neuroscience, but also expand my expertise in ways, which are essential to my transition to independent investigator. The training component of this application will build on my expertise in the immune system by investigating vascular injury involved in brain hemorrhage after tPA therapy. Together with training in molecular immunobiology, this will enable me to establish my own cutting-edge research program in stroke. The career development activities will be mentored by Dr. George King in collaboration with Drs. Magdy Selim, Kazuhide Hawakaya, and Stephan Kissler because of their areas of scientific expertise, and the technical and scientific advice that I stand to gain from our mentoring relationship. My career development activities will be focused on: 1) mentorship and guidance on laboratory management and organization; 2) the development and growth of my independent research program; 3) institutional responsibilities and fulfilling requirements for promotion and expanding my scientific network; 4) grant writing skills.

项目概要 组织纤溶酶原激活剂 (tPA) 的广泛使用是 FDA 批准的唯一急性中风治疗方法， 仍然受到其狭窄的治疗时间窗口和相关脑出血风险的限制。缓激肽 (BK) 是 以其作为内皮依赖性血管扩张剂和强大的促炎激动剂的作用而闻名。一个 大量实验研究表明，BK 受体在中风后通过介导而上调 中风后早期的炎症作用，表明 BK 会加重脑损伤。在 初步研究，我们发现：1）tPA治疗后通过血浆激肽释放酶增加BK形成 激活; 2) 中风后联合使用 BK 受体 2 拮抗剂与 tPA 治疗可减少 出血转化； 3) B2R激动剂在氧和葡萄糖过程中不诱导内皮细胞死亡 剥夺; 4) 脾切除或巨噬细胞耗竭的小鼠使用tPA可减少脑出血； 5) 黑克 中风后输注会增加出血转化，而这种转化在巨噬细胞耗尽的小鼠中被阻断。 拟议研究的目的是确定 BK 影响和激活的机制 tPA 治疗后的巨噬细胞，从而通过血管过程中的促炎症过程引起脑出血 受伤。我们的最终目标是描绘 tPA 治疗的机制基础，负责改善 时间窗口并使 tPA 更安全、更广泛地用于中风患者。我们建议测试 以下具体目标：（i）确定条件内皮细胞和巨噬细胞特异性的贡献 B2R 缺失对 tPA 诱导的出血转化； (ii) 表征 BK 在激活中的作用 tPA 处理后巨噬细胞的积累和募集； (iii) 描绘分子机制 BK 通过其调节巨噬细胞功能及其与脑内皮细胞的相互作用。我会用这个 指导职业发展奖（K01）以实现一系列培训目标，这些目标建立在我之前的基础上 神经科学方面的技能，同时也以多种方式扩展我的专业知识，这对于我过渡到 独立调查员。该应用程序的培训部分将建立在我在免疫方面的专业知识的基础上 通过调查 tPA 治疗后脑出血涉及的血管损伤来建立系统。与训练一起 在分子免疫生物学领域，这将使我能够建立自己的中风前沿研究项目。 职业发展活动将由乔治·金博士与博士合作指导。马格迪·塞利姆, Kazuhide Hawakaya 和 Stephan Kissler 因其在科学专业领域以及技术和 我将从我们的指导关系中获得科学建议。我的职业发展活动将是 重点关注：1）实验室管理和组织方面的辅导和指导； 2）发展与 我的独立研究计划的发展； 3）机构职责及履行要求 促进和扩大我的科学网络； 4）授予写作能力。