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）： 海湾战争退伍军人疾病（GWVI）的病因和病理生理学仍然知之甚少，缺乏治疗方法。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免受昆虫的药物的结果。这些药物包括：1）乙酰苯乙烯溴化物（PB） - 一种可逆的乙酰胆碱酯酶抑制剂（ACHE） - 可防止神经剂，例如Sarin，例如Sarin，永久性地抑制疼痛； 2）苄氯菊酯（PET） - 一种杀虫剂的作用机理是阻止神经元通道； 3）DEET-驱虫剂。这些药物针对神经系统，尤其是通过抑制ACHE，胆碱能系统。尽管这些药物以给予GW人员的剂量安全，但已经假设它们的组合以及GW部署期间所遇到的压力可能会共同贡献了产生多症状疾病GWVI。这已经在毒素/压力动物模型中进行了相当大的成功进行了测试。最近对GW退伍军人和非部署退伍军人进行的地标研究表明，GWVI的病理生理涉及胆碱能副交感神经系统功能的异常。此外，特征GWVI的认知和睡眠障碍与基础前脑胆碱能神经元（BFCN）的功能障碍一致，其正常活性对于记忆，注意力和睡眠过程中的过程至关重要。数据共同指出了与GW相关的上述药物的暴露并引起了中枢神经系统（CNS）中胆碱能神经元的长期功能障碍的可能性。因此，设计治疗方式可以恢复GWVI患者的胆碱能神经元及其靶标的正常功能。实现这一目标的一种策略是使用支持神经元活力和功能的营养因素。特别是大脑衍生的神经营养事实（BDNF），其通过其受体TRKB发出信号。中央副交感神经元和BFCN Express TRKB。 BDNF增加了BFCN的存活率并升高其在细胞培养中的胆碱能标记表达，因此在体内BFCN的产后成熟是必要的。 BDNF可防止大鼠BFCN中轴切开术引起的变性和胆碱能标记的表达丧失。但是，BDNF不会越过血脑屏障（BBB）。 7,8-二羟基氟氟酮（7,8-DHF）是一种有效而有选择性的TRKB激动剂，很容易进入大脑。 7,8-DHF的有益影响 已在PTSD，PD，AD和RETT综合征的模型中报道了。我们发现7,8-DHF在ALS和AD的小鼠模型中有效。拟议研究的总体目标是检验以下假设：在胆碱能神经元的研究中优化的小鼠GWVI模型中，已知的7,8 -DHF和/或中度运动（MEX）的干预措施（MEX）（MEX）将导致大脑功能的恢复。具体而言，我们将使用CHGFP转基因小鼠系，该小鼠系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的细胞分选（FACS）纯化这些细胞，并通过微观成像技术促进它们的可视化。这些小鼠将与限制应力一起暴露于GWVI-COPDAC相关药物组合（PB/PET/DEET），然后接触到我们的治疗方案（即7,8-DHF和/或MEX），在暴露后或在暴露后立即使用，或在延迟4周后治疗已建立的疾病。 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.此外，我们的研究将有助于我们理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型的动物模型，用于未来用于测试与GWVI患者有关的治疗剂。