Mild Traumatic Brain Injury and Opiate Exposure Crosstalk: Neuropathological, Neurobehavioral, and Neuroproteomic Assessments

轻度创伤性脑损伤和阿片类药物暴露串扰：神经病理学、神经行为和神经蛋白质组学评估

• 批准号: 10614983
• 负责人: KEVIN Ka Wang WANG
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-11-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10614983
• 关键词: Addictive Analgesics; Animal Model; Animals; Anxiety; Astrocytes; Axon; Behavior; Behavior assessment; Biochemical; Brain; Brain region; Characteristics; Chronic; Closed head injuries; Cognition; Data; Dependence; Drug Exposure; Enzyme-Linked Immunosorbent Assay; Evaluation; Exposure to; Fentanyl; Formalin Tests; Future; Glial Fibrillary Acidic Protein; Goals; Harvest; Health; Hippocampus (Brain); Histopathology; Human; Inflammation Mediators; Injury; Investigation; Knowledge; Label; Laboratories; Locomotion; Mediating; Methods; Microglia; Molecular; Morbidity - disease rate; Motor Activity; Mus; Nerve Degeneration; Neural Pathways; Neurobiology; Neurologic Deficit; Neurologic Symptoms; Neurons; Opiate Addiction; Opioid; Opioid Receptor; Pathologic; Pathway interactions; Patients; Performance; Pharmaceutical Preparations; Phase; Pilot Projects; Procedures; Proteomics; Rehabilitation therapy; Reporting; Rewards; Rodent Model; Saline; Sampling; Shotguns; Stains; Study models; Substance abuse problem; Symptoms; Synapses; Systems Biology; Tail; Testing; Traumatic Brain Injury; Ventral Tegmental Area; Veterans; Water; Western Blotting; Withdrawal; Withdrawal Symptom; Work; brain tissue; chronic pain management; comorbidity; drug withdrawal; essays; experimental group; improved; insight; interest; male; mild traumatic brain injury; military veteran; morris water maze; mortality; neurobehavioral; neuroproteomics; novel therapeutic intervention; opioid abuse; opioid exposure; opioid use; opioid withdrawal; orofacial; osmotic minipump; pain sensation; pain sensitivity; preclinical study; relating to nervous system; spatial memory; tau-1; therapeutic target; trend; white matter; young adult

Traumatic brain injury (TBI) is a major cause of morbidity among the Veteran population. Mild TBI (mTBI) has been associated with substance abuse as a comorbid condition, hampering rehabilitation and treatment efforts of either condition. Of particular interest is the rising trend of opiate abuse among Veterans. Existing evidence indicates overlapping molecular and neural pathways for TBI and opiate abuse; for example, inflammatory mediators and increased neurodegeneration and microglial activation are observed in both opiate abuse and mTBI. To the best of our knowledge, there are no reported preclinical studies that have systematically examined the crosstalk between mTBI and subsequent opiate exposure in terms of neurobehavioral, neuropathological and neurobiochemical consequences. This study proposes to use a validated rodent model of repeated closed head injury (rCHI) paired with chronic exposure to fentanyl drug to closely simulate real-world conditions of mTBI and opiate usage. Fentanyl is a highly effective but addictive analgesic commonly used for treating chronic pain. The central hypothesis of this proposal is that sustained opiate exposure post-mTBI will induce characteristic neurofunctional and neuropathological changes that differ from those seen with mTBI or opiate exposure alone, and that these changes can be identified and evaluated via neurobehavioral, biochemical and neuroproteomic approaches. This proposal is a new direction for the PI’s laboratory and is potentially paradigm-shifting. Aim 1 will determine the effects of 1 month fentanyl opiate exposure followed by 1 month withdrawal after r-CHI on cognition, pain sensation, motor activity and spontaneous opiate withdrawal symptoms. Male mice (C57BL6) will be used and subjected to rCHI or sham procedure (non-injury).There will be four experimental groups: (a) Sham+saline, (b) rCHI+saline, (c) sham+fentanyl, (d) rCHI-followed by Fentanyl exposure. Functional assessment tests include: TBI-related cognition (spatial memory performance; Morris water maze) and anxiety- behavior (elevated plus maze), or opiate-use/withdrawal tests; locomotive activity-related pain sensitivity tests (warm-water tail withdrawal and Orofacial formalin test) (during drug exposure and withdrawal phases), and drug withdrawal-dependence symptoms (during the fentanyl withdrawal phase). The results will be compared with the observations from rCHI or fentanyl exposure alone. Aim 2 will assess key neuropathological, neural cellular markers and opiate receptor levels in three brain regions (cortex, hippocampus and ventral tegmental area) implicated in TBI and/or opioid receptor-mediated reward circuits. Brain samples will be analyzed at 2 endpoints - at the end of fentanyl administration (month 1), and after 1 month of withdrawal (month 2), in animals with or without rCHI. Neuronal, axonal, astroglial, microglial, synaptic and white matter markers, along with and the three subtypes of opioid receptors will be assessed in separate sets of animals by immunoblotting/ELISA essays and by immunohistochemical-histopathological analyses. Aim 3 will identify long-term mTBI consequences and opiate abuse comorbidity-related pathological pathways/key drivers as putative neurotherapeutic targets using neuroproteomic and System Biology analysis. Collectively, this new line of investigation aims to provide insights into the neurobiological and neuropathological interplay between mTBI and post injury opioid abuse. The results may guide possible new therapeutic strategies to treat TBI and comorbid opioid dependence in Veterans.

创伤性脑损伤（TBI）是退伍军人人口发病率的主要原因。轻度TBI（MTBI）具有 与药物滥用有关，以疾病的疾病，阻碍了康复和治疗工作 任何条件。特别令人感兴趣的是优化退伍军人滥用的趋势的上升。现有证据 指示TBI的分子和神经途径重叠并优化滥用；例如，炎症 在优化滥用和 mtbi。据我们所知，尚无研究的临床前研究 MTBI和随后在神经性神经病理学方面的积极暴露之间的串扰 和神经化学后果。这项研究建议使用经过验证的重复封闭的啮齿动物模型 头部损伤（RCHI）与长期暴露于芬太尼药物相结合，以密切模拟M​​TBI的现实状况 并优化用法。芬太尼是一种高效但额外的镇痛药，通常用于治疗慢性疼痛。 该提议的中心假设是MTBI后持续的操作曝光将引起特征 神经功能和神经病理学变化与MTBI看到或仅优化暴露的神经病理学变化， 并且可以通过神经行为，生化和神经蛋白质组学来识别和评估这些变化 方法。该提议是PI实验室的新方向，并且可能正在转移范式。目标1 将确定1个月芬太尼鸦片暴露的影响，然后在R-CHI后1个月提取 认知，疼痛感，运动活动和赞助症状。雄性小鼠（C57BL6）将 被使用并进行RCHI或假手术（非伤害）。将有四个实验组：（a） 假+盐水，（b）rchi+盐水，（c）假+芬太尼，（d）通过芬太尼暴露遵循的rchi。功能 评估测试包括：与TBI相关的认知（空间记忆表现； Morris Water Maze）和动画 - 行为（升高的迷宫），或优化使用/撤回测试；机车活动相关的疼痛敏感性测试 （温暖的水尾撤离和口头福尔马林测试）（药物暴露和戒断阶段）和药物 戒断依赖性症状（在芬太尼戒断阶段）。结果将与 仅来自Rchi或芬太尼暴露的观察结果。 AIM 2将评估关键的神经病理学，神经细胞 标记并优化三个大脑区域的受体水平（皮层，海马和腹侧对段区域） 在TBI和/或阿片类接收器介导的奖励电路中实施。将在2个端点分析大脑样品 - 在芬太尼管理局结束时（第1个月），撤军1个月后（第2个月）， 没有rchi。神经元，轴突，星形胶质细胞，小胶质细胞，突触和白质标记物以及 通过免疫印迹/ELISA论文，将在单独的动物中评估阿片类药物受体的三个亚型 并通过免疫组织化学的 - 依斯科学分析。 AIM 3将确定长期MTBI的后果，并且 鸦片滥用合并症相关的途径/关键驱动因素作为推定的神经治疗目标 神经蛋白质组学和系统生物学分析。总的来说，这一新的投资旨在提供见解 进入MTBI与损伤后阿片类药物滥用之间的神经生物学和神经病理学相互作用。结果 可以指导可能的新治疗策略来治疗退伍军人中TBI和合并症的阿片类药物依赖性。