Targeting the sphingosine-1-phosphate system in a mouse model of Gulf War Veterans' Illness

针对海湾战争退伍军人疾病小鼠模型中的 1-磷酸鞘氨醇系统

• 批准号: 10293531
• 负责人: ALPASLAN DEDEOGLU
• 金额: --
• 依托单位: VA BOSTON HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10293531
• 关键词: Acetylcholinesterase; Acetylcholinesterase Inhibitors; Affect; Affective; Animal Model; Animals; Anti-Inflammatory Agents; Anxiety; Astrocytes; Attentional deficit; Axon; Behavior; Behavior assessment; Behavioral; Biological Assay; Blood; Brain; Brain-Derived Neurotrophic Factor; Breathing; Bromides; C-reactive protein; Cerebral cortex; Chemical Warfare; Chronic; Clinical Trials; Cognition; Cognitive; Complement 3c; Complete Blood Count; Complex; Congenital neurologic anomalies; Data; Disease; Disease Marker; Dose; Drug Design; Drug Targeting; Encephalitis; Enzyme-Linked Immunosorbent Assay; Etiology; Exhibits; Exposure to; Fatigue; Female; Functional disorder; Gender; Glial Fibrillary Acidic Protein; Gulf War veteran; Headache; Hippocampus (Brain); Human Resources; Immunoassay; Impaired cognition; Infiltration; Inflammation; Inflammatory; Insect Repellents; Insecta; Insecticides; Intestines; Link; Locomotion; Longitudinal Studies; Lymphocyte; Lymphocyte Count; Measures; Memory; Mental Depression; Methodology; Microglia; Military Personnel; Modeling; Molecular; Monitor; Motor Activity; Multiple Sclerosis; Mus; Nervous system structure; Neuraxis; Neurologic Symptoms; Neurological Models; Neurons; Oligodendroglia; Pain; Patients; Peripheral; Permethrin; Pharmaceutical Preparations; Pharmacology; Platelet Count measurement; Platelet aggregation; Potassium Channel; Process; Production; Proteomics; Relapsing-Remitting Multiple Sclerosis; Reporting; Research; Research Design; Research Methodology; Research Personnel; Role; Sarin; Serum; Signal Transduction; Skin Abnormalities; Sodium Channel; Sphingosine-1-Phosphate Receptor; Stomach; Stress; Symptoms; System; Techniques; Testing; Therapeutic; Thinking; Thromboxanes; Time; Veterans; War; Western Blotting; Woman; analog; base; blood-brain barrier crossing; cell determination; cholinergic; clinical practice; conditioned fear; cytokine; efficacy evaluation; fear memory; field study; forced swim test; gray matter; imaging study; immune function; immune system function; improved; indexing; innovation; male; men; monocyte; mouse model; multiple sclerosis treatment; nerve agent; nervous system disorder; neurochemistry; neuroinflammation; neurotrophic factor; neutrophil; novel; persistent symptom; preclinical trial; prevent; pyridostigmine; remyelination; response; sexual dimorphism; sphingosine 1-phosphate; success; targeted agent; treatment effect

The pathophysiology of Gulf War Veterans’ Illness (GWVI) remains poorly understood, and treatments are lack- ing. Neurological symptoms including cognitive impairment, attention deficits, depression, and anxiety are a top complaint among GWVI patients. GWVI may be the result of exposure to drugs designed to protect military personnel from a chemical attack and from insects. These include: 1) pyridostigmine bromide (PB), 2) permethrin (PER), and 3) diethyltoluamide (DEET). Although these drugs are considered safe at the doses administered to GW personnel, it has been hypothesized that their combination together with the stress of war may have con- tributed synergistically to generate the GWVI. We have used an animal model of GWVI based on this hypothesis by exposing mice to relevant doses of PB, PER, DEET and stress and found anxiety, brain neuroinflammation, cholinergic, GABAergic and neurotrophic factor abnormalities as latent post-exposure markers of this disease. Some changes were sexually dimorphic indicating that the pathophysiology of GWVI and responses to potential treatments may be different in men and women. Our data provide evidence for a neuroinflammatory process in brains of GWVI-model animals, which is characterized by astrocyte and microglia activation in the hippocampus that correlates with the increased level of anxiety and memory abnormalities seen in the model and can be related to reports indicating aberrant immune function and chronic inflammation in GWVI patients. Therefore, we propose to use our mouse model of GWVI to test the hypothesis that GWVI may be ameliorated by targeting neuroinflammation using fingolimod, a drug that targets the sphingosine-1-phosphate receptor and is used for the treatment of relapsing remitting multiple sclerosis (RRMS). In RRMS it prevents the infiltration of lymphocytes into the CNS, promotes remyelination, and exerts neuroprotective effects on astrocytes. We propose to deter- mine the efficacy of fingolimod on ameliorating the delayed and persistent GWVI-related behavioral, cellular and molecular CNS abnormalities in male and female mice. Based on our preliminary data, the use of a high and low dose of fingolimod will allow us to differentiate the drug’s peripheral versus central action. We will assess the treatment effect on: a) behavior using specific animal paradigms for assessing anxiety, locomotion, memory, and depression; b) neuroinflammation measured with multiplexed proteomic immunoassays of cytokine panels and by immunohistochemical determinations of cells expressing Iba1 and GFAP; c) BDNF levels and signaling by ELISA and immunohistochemical assays; d) indices of cholinergic and GABAergic function by Western blot; e) peripheral measures of inflammation by complete blood count and serum cytokine panel assays. This study incorporates rational pharmacology with state-of-the-art neuropathological and neurochemical techniques to- gether with cognitive and affective behavioral assessment to advance therapeutic strategies for GWVI.

海湾战争退伍军人病（GWVI）的病理生理学仍然很了解，并且缺乏治疗方法 - ing。神经系统症状在内 GWVI患者的投诉。 GWVI可能是暴露于旨在保护军事的药物的结果 化学攻击和昆虫的人员。其中包括：1）吡二吡啶溴（PB），2）苄氯菊酯 （PER）和3）二乙基二酰胺（DEET）。尽管这些药物在给予的剂量时被认为是安全的 黄金周的人员，已经假设他们的结合以及战争的压力可能会引起 以协同作用产生GWVI。我们基于此假设使用了GVI的动物模型 通过将小鼠暴露于相关剂量的PB，DEET和压力下，并发现焦虑，脑神经炎症， 胆碱能，GABA能和神经营养因子异常是该疾病的潜在暴露后标记。 一些变化是性二态性的，表明GWVI的病理生理和对潜在的反应 男性和女人的治疗可能有所不同。我们的数据为神经炎症过程提供了证据 GWVI模型动物的大脑，其特征是海马中的星形胶质细胞和小胶质细胞激活 这与模型中看到的动画和记忆异常的水平增加有关，可以是 与表明GWVI患者中异常免疫功能和慢性炎症的报道有关。因此，我们 建议使用我们的GWVI小鼠模型来测试可以通过靶向来改善GWVI的假设 使用Fingerlimod（一种针对鞘氨醇1-磷酸受体的药物）的神经炎症，用于 复发的治疗可使多发性硬化症（RRMS）。在RRMS中，它可以防止淋巴细胞浸润 进入中枢神经系统，促进再髓并对星形胶质细胞执行神经保护作用。我们建议 - 挖掘指指定效率在改善延迟和持续的GVI相关行为，细胞和细胞的效率 男性和雌性小鼠的分子中枢神经系统异常。根据我们的初步数据，使用高和低 Fingerlimod剂量将使我们能够区分该药物的外围与中央作用。我们将评估 治疗效果：a）使用特定动物范式评估动画，运动，记忆和 沮丧; b）通过多重蛋白质组学的细胞因子面板和 通过对表达IBA1和GFAP的细胞的免疫组织化学测定； c）BDNF级别和信号传导 ELISA和免疫组织化学测定； d）Western印迹的胆碱能和GABA能功能指标； e） 通过全血细胞计数和血清细胞因子面板测定法进行炎症的外围度量。这项研究 将理性药理学与最先进的神经病理学和神经化学技术结合在一起。 通过认知和情感行为评估来推动GWVI的治疗策略。