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) 的病理生理学仍然知之甚少，并且缺乏治疗方法。 ing。神经系统症状包括认知障碍、注意力缺陷、抑郁和焦虑是最常见的 GWVI 患者的抱怨。 GWVI 可能是接触旨在保护军人的药物的结果 人员免受化学攻击和昆虫的侵害。其中包括：1) 溴化吡斯的明 (PB)，2) 氯菊酯 (PER)和3)二乙基甲苯酰胺(DEET)。尽管这些药物在给药剂量下被认为是安全的 GW 人员，据推测，他们的结合加上战争的压力可能会导致 协同作用以产生 GWVI。基于这一假设，我们使用了 GWVI 动物模型 通过将小鼠暴露于相应剂量的 PB、PER、DEET 和压力中，发现了焦虑、脑神经炎症、 胆碱能、GABA能和神经营养因子异常是这种疾病的潜在暴露后标志。 一些变化是性别二态性的，表明 GWVI 的病理生理学和对潜在的反应 男性和女性的治疗方法可能不同。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，其特征是海马星形胶质细胞和小胶质细胞激活 这与模型中观察到的焦虑和记忆异常程度增加相关，并且可以 与表明 GWVI 患者免疫功能异常和慢性炎症的报告相关。因此，我们 建议使用我们的 GWVI 小鼠模型来检验通过靶向治疗可以改善 GWVI 的假设 使用芬戈莫德治疗神经炎症，芬戈莫德是一种针对 1-磷酸鞘氨醇受体的药物，用于 复发缓解型多发性硬化症（RRMS）的治疗。在 RRMS 中，它可以防止淋巴细胞浸润 进入中枢神经系统，促进髓鞘再生，并对星形胶质细胞发挥神经保护作用。我们建议阻止—— 探究芬戈莫德在改善延迟性和持续性 GWVI 相关行为、细胞和 雄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 芬戈莫德的剂量将使我们能够区分药物的外周作用和中枢作用。我们将评估 治疗效果：a) 使用特定动物范例评估焦虑、运动、记忆和行为 沮丧; b) 使用细胞因子组的多重蛋白质组免疫测定法测量神经炎症和 通过免疫组织化学测定表达 Iba1 和 GFAP 的细胞； c) BDNF 水平和信号传导 ELISA 和免疫组织化学测定； d) 通过Western blot检测胆碱能和GABA能功能的指数； e) 通过全血细胞计数和血清细胞因子组合测定进行炎症的外周测量。这项研究 将合理的药理学与最先进的神经病理学和神经化学技术相结合，以- 结合认知和情感行为评估，推进 GWVI 的治疗策略。