Understanding the immune response after mild TBI: Do endothelial cells play a major role?

了解轻度 TBI 后的免疫反应：内皮细胞发挥主要作用吗？

• 批准号: 10227663
• 负责人: Colleen Bodnar
• 金额: $ 4.16万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10227663
• 关键词: 3-Dimensional; Acute; Address; Affect; Alzheimer' s Disease; Animals; Antibodies; Arteries; Astrocytes; Behavior; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Blood flow; Brain; Brain Diseases; Brain Injuries; Cells; Central Nervous System Diseases; Cerebrovascular Circulation; Chronic; Closed head injuries; Cognitive; Confocal Microscopy; Development; Diffusion; Disease; Eko; Endothelial Cells; Endothelium; Environment; FDA approved; Fluorescence-Activated Cell Sorting; Fluorescent Dyes; Future; Gene Expression Profiling; Genes; Growth Factor; Hemagglutinin; Histology; Hour; Human; Image; Immune; Immune response; Immunohistochemistry; Individual; Inflammation; Inflammatory; Inflammatory Response; Influenza Hemagglutinin; Injury; Interleukin-1; Interleukin-1 Receptors; Knock-out; Label; Link; Location; Mediating; Mediator of activation protein; Messenger RNA; Minority; Modeling; Mus; Neurodegenerative Disorders; Neuroimmune; Operative Surgical Procedures; Oranges; Pathology; Pathway interactions; Patients; Pattern; Pericytes; Physiological; Play; Positioning Attribute; Postdoctoral Fellow; Proteins; RNA; Recovery; Reporter; Role; Scientist; Severities; Signal Transduction; Structure; Techniques; Testing; Therapeutic; Time; Training; Traumatic Brain Injury; United States; Vascular Diseases; Veins; Work; angiogenesis; brain endothelial cell; career; cell cortex; chemokine; cytokine; density; experience; functional outcomes; genetic manipulation; improved; lipophilicity; mild traumatic brain injury; nano-string; neuroinflammation; neurovascular unit; persistent symptom; receptor; reconstruction; red fluorescent protein; response; skills; tomography

PROJECT SUMMARY/ABSTRACT: The vast majority (over 90%) of traumatic brain injuries (TBI) are classified as mild TBI (mTBI). While most individuals recover from mTBI quickly, a significant minority (10-15%) of patients with mTBI experience persistent symptoms – there are no FDA approved treatments for these individuals. Chronic neuroinflammation and vascular disruption following TBI create an unfavorable environment for recovery. More broadly, neuroinflammation and vascular dysfunction are significant contributors to other CNS diseases as well, including Alzheimer’s disease. My preliminary results demonstrate that interleukin-1 (IL-1) signaling through interleukin-1 receptor 1 (IL-1R1) may be as a link between chronic neuroinflammation and vascular pathology and serve as a therapeutic avenue. IL-1, a major pro-inflammatory cytokine, is upregulated following all TBI severities. IL-1 release has been linked with downstream alterations of the vasculature, including changes to the neurovascular unit, blood flow alteration, angiogenesis, and increased release of cytokines, chemokines, and growth factors. We have found that brain endothelial cells express a high number of IL-1R1 receptors. Further, we have demonstrated that mice lacking IL-1R1 show reduced neuroinflammation following experimental mTBI. We hypothesize that following mTBI, brain endothelial cells, specifically through IL- 1R1, act as a mediator of the neuroimmune and vascular responses. To test this hypothesis, we will use a closed head injury model (CHI) to model a mTBI in mice. Further, we will manipulate IL-1R1 using an inducible endothelial cell-specific IL-1R1 knockout (eKO), a global knockout in which IL-1R1 has been restored only in the endothelial cells (eRestore), and an IL-1R1 reporter mouse in which the protein and mRNA are labeled. In aim 1, we will determine the cellular and temporal expression changes of IL-1R1 in the brain following CHI using the reporter mice. Through this aim, I will be able to determine how TBI alters IL-1R1 across time as well as IL-1R1 association with arteries, veins, and capillaries, while also honing my skills in histology techniques. In aim 2, we will delineate endothelial IL-1R1-dependent neuroinflammatory responses following CHI in wildtype, eKO, and eRestore mice. Through this aim, I will get experience in animal surgeries as well as RNA isolation and Nanostring gene expression analysis. Finally, in aim 3, we will investigate the involvement of endothelial IL-1R1 in vascular structure and blood flow changes after TBI. To do this, we will use vessel painting and speckle contrast diffusion correlated tomography (scDCT). If we find support for our hypothesis, we will provide the groundwork for the development of IL-1 therapeutics for the treatment of mTBI, as well as describe a physiological response of the CNS that can be applied to other diseases. Overall, the question outlined in this proposal not only moves the field forward but also provides significant scientific training which will position me as a strong candidate for post-doctoral work and a future career as an independent scientist.

项目摘要/摘要： 绝大多数(超过90%)的创伤性脑损伤(TBI)被归类为轻度脑损伤(MTBI)。虽然大多数人 个人从mTBI中恢复得很快，相当少数(10%-15%)的mTBI患者有过这样的经历 持续性症状--FDA没有批准针对这些人的治疗方法。慢性神经炎 而颅脑损伤后的血管破裂为康复创造了不利的环境。更广泛地说， 神经炎症和血管功能障碍也是其他中枢神经系统疾病的重要因素， 包括阿尔茨海默氏症。我的初步结果表明，白介素1(IL-1)通过 白细胞介素1受体1(IL-1R1)可能是慢性神经炎和血管病变之间的联系 作为一种治疗途径。IL-1是一种主要的促炎细胞因子，在所有脑损伤后上调 严肃性。IL-1的释放与下游血管系统的变化有关，包括 神经血管单位，血流改变，血管生成，细胞因子，趋化因子， 和增长因素。我们发现脑内皮细胞表达大量的IL-1R1受体。 此外，我们还证明，缺乏IL-1R1的小鼠表现出以下神经炎症的减轻 实验性mTBI。我们假设，在mTBI后，脑内皮细胞，特别是通过IL-1- 1R1，作为神经免疫和血管反应的中介。为了检验这一假设，我们将使用 闭合性脑损伤模型(CHI)，建立小鼠脑创伤模型。此外，我们将使用诱导物操纵IL-1R1 内皮细胞特异性IL-1R1基因敲除(EKO)，在这种全球基因敲除中，IL-1R1仅在 内皮细胞(ERestore)，以及标记了蛋白质和mRNA的IL-1R1报告小鼠。在……里面 目的1、测定慢性脑损伤后脑组织IL-1R1的细胞和时间表达变化 使用的是报告老鼠。通过这个目标，我也将能够确定脑损伤是如何随时间改变IL-1R1的 因为IL-1R1与动脉、静脉和毛细血管联系在一起，同时也磨练了我在组织学技术方面的技能。 在目标2中，我们将描绘血管内皮细胞IL-1R1依赖的神经炎性反应 野生型、Eko和eRestore小鼠。通过这个目标，我将获得动物手术和RNA方面的经验 纳米串基因的分离和表达分析。最后，在目标3中，我们将调查 颅脑损伤后血管结构和血流量中内皮细胞IL-1R1的变化要做到这一点，我们将使用容器 绘画和散斑对比扩散相关层析成像(ScDCT)。如果我们发现我们的假设得到了支持， 我们将为IL-1治疗脊髓损伤的治疗药物的开发提供基础，以及 描述一种可应用于其他疾病的中枢神经系统的生理反应。总体而言，这个问题 这份提案中概述的内容不仅推动了该领域的发展，而且还提供了重要的科学培训， 将使我成为博士后工作的有力候选人，并成为未来独立科学家的职业。