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？

基本信息

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
项目类别：
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&apos 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

项目摘要

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：脑外伤（TBI）的绝大多数（超过90％）被归类为轻度TBI（MTBI）。虽然大多数个体很快从MTBI中恢复过来，MTBI经验的大量少数（10-15％）持续的症状 - 这些人没有FDA批准的治疗方法。慢性神经炎症 TBI后的血管破坏创造了一个不利的恢复环境。更广泛地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病。我的初步结果表明，白介素-1（IL-1）通过白介素-1受体1（IL-1R1）可能是慢性神经炎症与血管病理学之间的联系并充当治疗大道。 IL-1是一种主要的促炎细胞因子，在所有TBI之后都进行了更新严重性。 IL-1释放与脉管系统的下游变化有关，包括更改神经血管单元，血流改变，血管生成以及细胞因子，趋化因子，释放增加和增长因素。我们发现脑内皮细胞表达大量的IL-1R1受体。此外，我们已经证明缺乏IL-1R1的小鼠显示出降低的神经炎症实验MTBI。我们假设遵循MTBI，脑内皮细胞，特别是通过IL- 1R1充当神经免疫和血管反应的介体。为了检验这一假设，我们将使用闭合的头部损伤模型（CHI）为小鼠的MTBI建模。此外，我们将使用诱导内皮细胞特异性IL-1R1敲除（EKO），这是一个全球敲除，其中IL-1R1仅恢复内皮细胞（Erestore）和IL-1R1报告基因小鼠，其中蛋白质和mRNA被标记。 AIM 1，我们将在CHI后确定IL-1R1的细胞和临时表达变化使用记者小鼠。通过这个目标，我将能够确定TBI也如何改变IL-1R1 作为IL-1R1与动脉，静脉和毛细血管的关联，同时还磨练了我在组织学技术方面的技能。在AIM 2中，我们将描述CHI之后的内皮IL-1R1依赖性神经炎症反应 Wildtype，Eko和Erestore小鼠。通过这个目标，我将获得动物手术和RNA的经验分离和纳米基因表达分析。最后，在AIM 3中，我们将研究 TBI后血管结构和血流变化的内皮IL-1R1。为此，我们将使用船只绘画和斑点对比扩散相关断层扫描（SCDCT）。如果我们找到对假设的支持，我们将为开发IL-1治疗的基础，以治疗MTBI以及描述可以应用于其他疾病的中枢神经系统的物理反应。总体而言，问题该提案中概述不仅会向前推进该领域，而且还提供了重大的科学培训我将把我作为博士后工作的有力候选人，并且是独立科学家的未来职业。