Mrgprb2 mediated neuroinflammation after cerebral ischemia

摘要 我即将完成我在约翰霍普金斯医学院神经外科的脑血管研究。 医学，我将从神经外科助理教授开始，专注于脑血管的实践 疾病我正在申请一个指导外科医生科学家职业发展奖（CDA），以获得进一步的 神经科学、免疫学和脑缺血动物模型培训。这将进一步促进我的长期 探索脑缺血潜在的神经免疫机制的职业目标，并希望提供 缺血性卒中患者新研究的基础。 中风病理学的一个主要机制涉及神经炎症， 小胶质细胞和外周白细胞的浸润使神经元损伤和细胞死亡恶化。尽管有大量的预- 尽管临床上进行了努力，但还没有有效的治疗方法来限制中风后神经炎性损伤。桅杆 细胞是组织特异性、长寿命的免疫效应细胞，被认为是第一反应细胞之一， 在免疫监视和激活中，激活先天免疫系统的多种途径。因此， 抑制肥大细胞活化的能力可广泛减弱脑炎症后的多种途径， 中风最近，Mrgprb 2被鉴定为肥大细胞特异性G偶联蛋白受体，用于碱性磷酸酶的诱导。 促分泌素，介导IgE非依赖性激活。我已经初步表明，删除Mrgprb 2的结果， 短暂性大脑中动脉闭塞（tMCAO）后每搏输出量减少， 脑卒中后TNF α和CCL 2的转录和产生，以及减少中性粒细胞募集到 脑缺血这个CDA建议建立在我的药理学研究生训练的细胞机制， 死亡和可卡因成瘾，在神经科学和免疫学的界面走向一个新的方向。我希望 这个CDA将使我能够在神经科学和细胞科学部门接受多学科培训， 分子医学具体培训目标包括：（1）高级神经科学基础技术培训 包括硬脑膜和脑膜显微切割，以及活体动物成像（2）动物模型培训 （3）免疫学教学和实验培训 和神经免疫相互作用（4）在道德和负责任的研究行为的额外培训。 该研究计划旨在解决Mrgprb 2是一种关键的肥大细胞特异性受体的假设 其上调脑缺血后的初始免疫激活反应， Mrgprb 2的生理配体可减少中风后的神经炎症。提案的具体目标 目的是：（1）研究在缺血性卒中中Mrgprb 2调节哪些肥大细胞介质亚群（2） 确定Mrgprb 2在中枢神经系统肥大细胞中的定位和表达模式（3） 脑缺血中Mrgprb 2的内源性配体通过筛选已知的tMCAO后Mrgprb 2激动剂， 沿着Mrgprb 2抑制剂作为中风潜在治疗剂的测试。

ABSTRACT I am finishing my cerebrovascular fellowship in the department of neurosurgery at the Johns Hopkins School of Medicine, where I will start as an assistant professor of neurosurgery with a practice focused on cerebrovascular diseases. I am applying for a mentored surgeon-scientist career development award (CDA) to obtain further training in neuroscience, immunology, and animal models of cerebral ischemia. This will further my long-term career goals of exploring neuro-immunologic mechanisms underlying cerebral ischemia, and hopefully provide a basis for new investigations in patients experiencing ischemic stroke. A major mechanism underlying stroke pathology involves neuroinflammation, where activation of microglia and infiltration of peripheral leukocytes worsen neuronal injury and cell death. Despite substantial pre- clinical efforts, there exists no efficacious therapeutic in limiting post-stroke neuroinflammatory damage. Mast cells are tissue specific, long-lived immunologic effector cells which are thought to be one of the first-responders in immune surveillance and activation, activating multiple pathways of the innate immune system. As such, the ability to suppress mast cell activation may broadly attenuate multiple pathways of cerebral inflammation after stroke. Recently, Mrgprb2 was identified as a mast cell-specific G-coupled protein receptor for basic secretagogues, mediating IgE-independent activation. I have shown preliminarily that deletion of Mrgprb2 results in decreased stroke volume after transient middle cerebral artery occlusion (tMCAO), with decreased transcription and production of TNFa and CCL2 after stroke, and decreased neutrophil recruitment into the ischemic brain. This CDA proposes building upon my pharmacology graduate training in mechanisms of cell death and cocaine addiction, towards a new direction at the interface of neuroscience and immunology. I hope this CDA will allow me to receive multidisciplinary training in the Departments of Neuroscience and Cellular and Molecular Medicine. Specific training goals include: (1) Training in advanced basic neuroscience techniques including dural and meningeal microdissection, and in-vivo live-animal imaging (2) Training in animal models of cerebral ischemia including photo-thrombotic stroke (3) Didactic and experimental training in immunology and neuro-immune interactions (4) Additional training in the ethical and responsible conduct of research. The research plan seeks to address the hypothesis that Mrgprb2 is a critical mast cell specific receptor which upregulates the initial immunologic activation response after cerebral ischemia, and that inhibition of the physiologic ligand for Mrgprb2 may decrease neuroinflammation after stroke. The Specific Aims of the proposal are to: (1) Investigate which subset of mast cell mediators are regulated by Mrgprb2 in ischemic stroke (2) Determine the localization and expression pattern of Mrgprb2 in central nervous system mast cells (3) Find the endogenous ligand for Mrgprb2 in cerebral ischemia by screening of known Mrgprb2 agonists after tMCAO, along with testing of inhibitors of Mrgprb2 as potential therapeutics of stroke.