The role of Infiltrating Macrophages in Seizure Generation Following CNS Infection

浸润巨噬细胞在中枢神经系统感染后癫痫发作中的作用

• 批准号: 10463765
• 负责人: Ana Beatriz de Paula e Silva
• 金额: $ 12.26万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10463765
• 关键词: Acute; Address; Adult; Affect; Animal Model; Anti-Inflammatory Agents; Attention; Automobile Driving; Biological Assay; Biotinylation; Blood; Brain; Cells; Central Nervous System Infections; Central Nervous System Viral Diseases; Cephalic; Chronic; Data; Development; Disease; Down-Regulation; Encephalitis; Epilepsy; Flow Cytometry; GABA Receptor; Generations; Genetic; Grant; Hippocampus (Brain); Homeostasis; Infection; Infiltration; Inflammation; Inflammatory; Interleukin-10; Interleukin-6; Interleukins; Intervention; Knowledge; Laboratories; Lead; Learning; Measures; Mentors; MicroRNAs; Microglia; Modeling; Motor Seizures; Mus; Myeloid Cells; Neuraxis; Neurons; Pathogenesis; Pathologic; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Pharmacology; Phase; Phenotype; Play; Population; Positioning Attribute; Recombinant Proteins; Recurrence; Refractory; Research; Role; Seizures; Severities; Severity of illness; Signal Pathway; Signal Transduction; Source; Stimulus; TMEV; TNF gene; Techniques; Temporal Lobe Epilepsy; Testing; Time; Tissues; Training; Transforming Growth Factors; Viral; Viral Encephalitis; Virus; Virus Diseases; Work; acquired epilepsy; acute infection; astrogliosis; brain parenchyma; career; cytokine; experimental study; improved; macrophage; mouse model; nervous system disorder; neuroinflammation; neuron loss; new therapeutic target; novel; novel therapeutics; peripheral blood; prevent; receptor; receptor expression; response; skills; therapeutic target; tissue repair; trafficking; transcriptome sequencing

Abstract/Project Summary Temporal lobe epilepsy (TLE) is the most prevalent form of acquired epilepsy. 30% of the patients are refractory to therapy and new treatments are urgently needed. Viral infection of the central nervous system (CNS) is a significant cause of TLE, and, while encephalitis is an important contributor, the mechanisms of how inflammation leads to seizures is unclear. Using the first mouse model of viral-induced TLE, which was developed by our laboratory, we found that induction of acute seizures is associated with blood peripheral macrophage infiltration into the brain and secretion of pro-inflammatory cytokines, primarily IL-6. Through RNA sequencing (RNAseq) of infiltrating macrophages isolated from the brains of TMEV-infected mice at the peak of seizure activity, I found that a population of CNS-infiltrating macrophages expresses high levels of the triggering receptor expressed on myeloid cells-1 (TREM1). Activation of TREM1 leads to the secretion of pro-inflammatory cytokines. Increased TREM1 expression is associated with inflammatory conditions, and inhibition of TREM1 function ameliorates inflammation and disease severity in several animal models of inflammatory diseases. However, the involvement of TREM1 in the development of acute seizures is unknown. In Aim 1 I will determine the time course of TREM1 expression in brain-infiltrating macrophages, and address whether genetic and pharmacological inhibition of TREM1 decreases the development of acute seizures in TMEV-infected mice, via videoEEG. The pro-inflammatory cytokine IL-6, which is mainly produced by brain infiltrating macrophages during TMEV infection, plays a pivotal role in inducing seizures; however how IL-6 does it is unknown. In Aim 2, I will determine whether IL-6 induces downregulation of neuronal GABA receptors, via biotinylation assay, and address which IL-6 pathway (classical or trans-signaling) is involved in acute seizures. Since inflammatory macrophages in the brain are associated with generation of acute seizures in TMEV-infected mice, in Aim 3 I will determine whether skewing macrophages towards an anti-inflammatory phenotype will affect the development of acute seizures, via videoEEG. I will also identify microRNAs in brain-infiltrating macrophages that are driving inflammation during the course of acute seizure generation in TMEV-infected mice. VideoEEG and receptor biotinylation assay are critical techniques I will learn during my mentored phase (Aim1/Aim2a), that will be essential to carry out experiments in Phase II (Aim2b/Aim3). These new skills, combined to my previous training, will put me in a unique position to address crucial questions related to neuroinflammation and epilepsy in my own laboratory using cutting edge approaches, and will substantially impact my research and career. These aims will improve the knowledge of the relationship between inflammation and seizures, and has the great potential of revealing inflammatory macrophages as a novel therapeutic target of neuroinflammation, allowing for the development of novel intervention to prevent and treat the development of acquired epilepsy.

摘要/项目摘要 颞叶癫痫（TLE）是获得癫痫的最普遍的形式。 30％的患者是 迫切需要对治疗的难治和新疗法。中枢神经系统（CNS）的病毒感染 是造成TLE的重要原因，尽管脑炎是重要的因素，但其机制的机制是如何 炎症会导致癫痫发作。使用开发的第一个病毒诱导的TLE的小鼠模型 通过我们的实验室，我们发现急性癫痫发作与血液外周巨噬细胞有关 促炎细胞因子的大脑浸润和分泌，主要是IL-6。 通过从TMEV感染的大脑中分离出的浸润巨噬细胞的RNA测序（RNASEQ） 在癫痫活性峰值的小鼠中，我发现一群CNS浸润巨噬细胞表达很高 在髓样细胞-1上表达的触发受体的水平（TREM1）。 trem1的激活导致 促炎细胞因子的分泌。 Trem1表达增加与炎症有关 条件和抑制TREM1功能可以改善几只动物的炎症和疾病严重程度 炎症疾病的模型。但是，TREM1参与急性癫痫发育的是 未知。在AIM 1中，我将确定在脑渗透巨噬细胞中TREM1表达的时间过程，以及 解决遗传学和药理学抑制是否会降低急性癫痫的发展 在TMEV感染的小鼠中，通过Videoeg。促炎性细胞因子IL-6，主要由大脑产生 TMEV感染期间渗透巨噬细胞在诱发癫痫发作中起关键作用。但是IL-6如何做 这是未知的。在AIM 2中，我将确定IL-6是否诱导神经元GABA受体的下调 生物素化测定法和IL-6途径（经典或跨信号）参与急性癫痫发作的地址。 由于大脑中的炎症性巨噬细胞与受TMEV感染的急性癫痫发作有关 小鼠，在目标3中，我将确定偏向巨噬细胞是否会影响抗炎表型 急性癫痫发作的发展，通过Videoeg。我还将识别脑浸润巨噬细胞中的microRNA 在TMEV感染的小鼠中，在急性癫痫发作过程中驱动炎症。 Videoeeg 受体生物素化测定是我将在我的指导阶段（AIM1/AIM2A）中学到的关键技术， 在II阶段（AIM2B/AIM3）中进行实验至关重要。这些新技能，结合了我以前的 培训将使我处于独特的位置，以解决与神经炎症和癫痫有关的关键问题 在我自己的实验室中，采用尖端方法，将大大影响我的研究和职业。 这些目标将提高对炎症与癫痫之间关系的了解，并具有 揭示炎症性巨噬细胞作为神经炎症的新治疗靶点的巨大潜力， 允许开发新的干预措施，以预防和治疗获得的癫痫的发展。