Novel neurotrophic therapies in an optimized mouse model of GWVI

优化的 GWVI 小鼠模型中的新型神经营养疗法

• 批准号: 8815008
• 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: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8815008
• 关键词: Acetylcholinesterase; Acetylcholinesterase Inhibitors; Agonist; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Animals; Arthralgia; Attention; Autonomic nervous system; Axotomy; Behavior; Behavioral; Behavioral Symptoms; Biology; Blood - brain barrier anatomy; Blood Volume; Brain; Brain Stem; Brain-Derived Neurotrophic Factor; Bromides; Cell Culture Techniques; Cells; Cerebrum; Chemical Warfare; Chronic; Cognitive; Data; Defect; Disease; Dose; Drug Combinations; Drug Design; Drug Targeting; Environment; Etiology; Evaluation; Exercise; Exposure to; Fatigue; Fluorescence-Activated Cell Sorting; Functional disorder; Future; Gene Expression Profiling; Genes; Goals; Green Fluorescent Proteins; Gulf War; Headache; Health; Hippocampus (Brain); Human; Human Resources; Imagery; Imaging Techniques; Impaired cognition; Insect Repellents; Insecta; Insecticides; Intervention; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Measurement; Mediating; Memory; Microscopic; Military Personnel; Modality; Modeling; Moderate Exercise; Mus; Myalgia; Nervous system structure; Neuraxis; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 2; Outcome Measure; Parkinson Disease; Patients; Permethrin; Pharmaceutical Preparations; Physiological; Post-Traumatic Stress Disorders; Rattus; Recovery; Regimen; Reporting; Rett Syndrome; Role; Sarin; Signal Transduction; Skin Abnormalities; Sleep; Sleep disturbances; Sodium Channel; Stress; Symptoms; Synapses; Synaptic plasticity; System; Techniques; Telemetry; Testing; Therapeutic; Toxin; Transgenic Mice; Veterans; basal forebrain cholinergic neurons; behavioral pharmacology; cholinergic; cholinergic neuron; design; gastrointestinal; in vivo; memory process; mouse model; nerve agent; neurochemistry; neurogenesis; novel; nucleus ambiguus; postnatal; prevent; public health relevance; pyridostigmine; receptor; response; restraint stress; septohippocampal; spatial memory; success; therapy design

DESCRIPTION (provided by applicant): The etiology and pathophysiology of Gulf War Veterans' Illness (GWVI) remain poorly understood and treatments are lacking. Most studies suggest that GWVI may be the result of exposure to drugs designed to protect military personnel from a chemical warfare attack and from insects. These drugs include: 1) pyridostigmine bromide (PB) - a reversible inhibitor of acetylcholinesterase (AChE) - that prevents nerve agents, such as sarin, from inhibiting AChE permanently; 2) permethrin (PET) - an insecticide whose mechanism of action is to block neuronal sodium channels; and 3) DEET - an insect repellent. These drugs target the nervous system and in particular, via the inhibition of AChE, the cholinergic system. Although these drugs are safe at the doses given to GW personnel, it has been hypothesized that their combination together with the stress encountered during the GW deployment may have contributed collectively to generate the multi-symptom disease, GWVI. This has been tested in toxin/stress animal models with considerable success. Recent landmark studies performed on GW Veterans and non-deployed Veterans indicate that pathophysiology of GWVI involves abnormalities in the function of the cholinergic parasympathetic system. Moreover, cognitive and sleep disturbances that characterize GWVI are consistent with a dysfunction of the basal forebrain cholinergic neurons (BFCN) whose normal activity is central to the processes of memory, attention and sleep. Together the data point to the possibility that the GW-associated exposure to the above-listed drugs and to stress caused a long-term dysfunction of cholinergic neurons within central nervous system (CNS). Therefore it would be desirable to design treatment modalities that could restore the normal functioning of cholinergic neurons and their targets in patients with GWVI. One strategy to accomplish this goal would be to use trophic factors that support neuronal viability and function. Specifically brain derived neurotrophic facto (BDNF) that signals via its receptor, TrkB. The central parasympathetic neurons and BFCN express TrkB. BDNF increases BFCN survival and elevates their cholinergic marker expression in cell culture and it is necessary for postnatal maturation of BFCN in vivo. BDNF prevents axotomy-induced degeneration and loss of cholinergic marker expression in BFCN in rats. However, BDNF does not cross the blood-brain barrier (BBB). 7,8-dihydroxyflavone (7,8-DHF) is a potent and selective TrkB agonist that readily enters the brain. Beneficial effects of 7,8-DHF have been reported in models of PTSD, PD, AD and Rett syndrome. We found that 7,8-DHF is effective in mouse models of ALS and AD. The overall goal of the proposed studies is to test the hypothesis that administration of 7,8-DHF and/or moderate exercise (MEX) - interventions known to generate a trophic neuronal environment - will cause a recovery of brain function in a mouse model of GWVI optimized for the studies of cholinergic neurons. Specifically, we will use the CHGFP transgenic mouse line that expresses the green fluorescent protein (GFP) exclusively in cholinergic cells. This permits the purification of these cells by fluorescence-activated cell sorting (FACS) and facilitates their visualization with microscopic imaging techniques. These mice will be exposed to the GWVI- associated drug combination (PB/PET/DEET) together with restraint stress, and then to our therapeutic regimens (i.e. 7,8-DHF and/or MEX) that will be employed immediately after the exposure, or after a delay of 4 weeks to treat an established illness. These studies incorporate principles of rational pharmacology and behavioral evaluation combined with state-of- the-art MR imaging and spectroscopy, physiological telemetry as well as neuropathological, neurochemical and gene-analytic techniques to define the therapeutic benefits of a novel neurotrophic compound that crosses the BBB as well as a non-pharmacological treatment modality in a unique GWVI mouse model. In addition, our studies will contribute to our understanding of the basic biology of cholinergic neurons and their role in GWVI and further characterize a novel animal model for future use to test therapeutics relevant to patients with GWVI.

描述（由申请人提供）： 海湾战争退伍军人病（GWVI）的病因和病理生理学仍然知之甚少，缺乏治疗。大多数研究表明，GWVI可能是暴露于旨在保护军事人员免受化学战攻击和昆虫攻击的药物的结果。这些药物包括：1）溴化吡啶斯的明（PB）-一种可逆的乙酰胆碱酯酶（AChE）抑制剂-可防止神经毒剂（如沙林）永久抑制AChE; 2）氯菊酯（PET）-一种杀虫剂，其作用机制是阻断神经元钠通道; 3）避蚊胺（DEET）-一种驱虫剂。这些药物靶向神经系统，特别是通过抑制乙酰胆碱酯酶，胆碱能系统。尽管这些药物在给予GW人员的剂量下是安全的，但据推测，它们与GW部署期间遇到的压力结合在一起可能共同导致了多症状疾病GWVI。这已经在毒素/应激动物模型中进行了测试，取得了相当大的成功。最近对GW退伍军人和未部署退伍军人进行的里程碑式研究表明，GWVI的病理生理学涉及胆碱能副交感神经系统功能异常。此外，认知和睡眠障碍的特点GWVI是一致的功能障碍的基底前脑胆碱能神经元（BFCN）的正常活动是中央的记忆，注意力和睡眠的过程。总之，这些数据表明，与GW相关的上述药物暴露和应激可能导致中枢神经系统（CNS）内胆碱能神经元的长期功能障碍。因此，希望设计能够恢复GWVI患者胆碱能神经元及其靶点正常功能的治疗方式。实现这一目标的一个策略是使用支持神经元活力和功能的营养因子。特别是脑源性神经营养因子（BDNF），通过其受体TrkB发出信号。中枢副交感神经元和BFCN表达TrkB。BDNF可增加BFCN的存活率，并提高其胆碱能标记物的表达，这是BFCN在体内成熟所必需的。BDNF可预防大鼠轴突切断术诱导的BFCN胆碱能标记物表达的退化和丢失然而，BDNF不能穿过血脑屏障（BBB）。7，8-二羟基黄酮（7，8-DHF）是一种有效的选择性TrkB激动剂，易于进入大脑。7，8-DHF的有益效果 已经在PTSD、PD、AD和Rett综合征的模型中报道。我们发现7，8-DHF在ALS和AD小鼠模型中有效。拟议研究的总体目标是检验以下假设：给予7，8-DHF和/或适度运动（MEX）-已知可产生营养神经元环境的干预措施-将导致针对胆碱能神经元研究优化的GWVI小鼠模型中脑功能恢复。具体而言，我们将使用CHGFP转基因小鼠系，其仅在胆碱能细胞中表达绿色荧光蛋白（GFP）。这允许通过荧光激活细胞分选（FACS）纯化这些细胞，并促进它们通过显微成像技术的可视化。这些小鼠将暴露于GWVI相关药物组合（PB/PET/DEET）以及束缚应激，然后暴露于我们的治疗方案（即7，8-DHF和/或MEX），所述治疗方案将在暴露后立即使用，或在延迟4周后使用以治疗已确定的疾病。这些研究结合了合理药理学和行为评价的原则，结合了最先进的MR成像和光谱学、生理遥测以及神经病理学、神经化学和基因分析技术，以确定一种新型神经营养化合物的治疗益处，该化合物可穿过BBB，以及在独特的GWVI小鼠模型中的非药理学治疗方式。此外，我们的研究将有助于我们了解胆碱能神经元的基本生物学及其在GWVI中的作用，并进一步表征一种新的动物模型，供将来用于测试与GWVI患者相关的治疗方法。