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

海湾战争退伍军人疾病（GWVI）的病理生理学仍然知之甚少，治疗方法缺乏- ing.包括认知障碍、注意力缺陷、抑郁和焦虑在内的神经系统症状是最常见的 GWVI患者的投诉。GWVI可能是暴露于旨在保护军事人员的药物的结果 人员免受化学攻击和昆虫的伤害。这些药物包括：1）溴化吡啶斯的明（PB），2）氯菊酯 (PER)和3）二乙基甲苯酰胺（DEET）。虽然这些药物被认为是安全的剂量管理， GW人员，据推测，他们的组合加上战争的压力可能有康- 协同地贡献以产生GWVI。我们已经使用了基于这一假设的GWVI动物模型 通过将小鼠暴露于相关剂量的PB，PER，DEET和压力，发现焦虑，脑神经炎症， 胆碱能、γ-氨基丁酸能和神经营养因子异常是这种疾病的潜在暴露后标志物。 有些变化是性二态性的，表明GWVI的病理生理学和对潜在的 男性和女性的治疗可能不同。我们的数据提供了神经炎症过程的证据， GWVI模型动物的脑，其特征在于海马中的星形胶质细胞和小胶质细胞活化 这与模型中看到的焦虑和记忆异常水平的增加有关， 与GWVI患者异常免疫功能和慢性炎症的报告有关。所以我们 我建议使用我们的GWVI小鼠模型来检验GWVI可以通过靶向治疗来改善的假设 使用芬戈莫德治疗神经炎症，芬戈莫德是一种靶向鞘氨醇-1-磷酸受体的药物， 复发缓解型多发性硬化症（RRMS）的治疗。在RRMS中，它阻止淋巴细胞的浸润 进入CNS，促进髓鞘再生，并对星形胶质细胞发挥神经保护作用。我们建议遏止- 探讨芬戈莫德改善延迟性和持续性GWVI相关的行为、细胞和 雄性和雌性小鼠的分子CNS异常。根据我们的初步数据， 芬戈莫德的剂量将使我们能够区分药物的外周与中枢作用。我们将评估 a）使用特定动物范例评估焦虑、运动、记忆的行为，和 抑郁症; B）用细胞因子组的多重蛋白质组学免疫测定测量的神经炎症， 通过表达Iba 1和GFAP的细胞的免疫组织化学测定; c）BDNF水平和信号传导， ELISA和免疫组织化学测定; d）通过蛋白质印迹法测定胆碱能和GABA能功能的指数; e） 通过全血细胞计数和血清细胞因子组测定来测量外周炎症。本研究 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， 结合认知和情感行为评估，推进GWVI的治疗策略。

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.