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: 10051334
• 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/10051334
• 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; Immunoblotting; 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; Withdrawal; Withdrawal Symptom; Work; brain tissue; chronic pain; 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.

颅脑损伤是退伍军人发病的主要原因之一。轻度颅脑损伤(MTBI) 与药物滥用有关，作为一种共病，阻碍康复和治疗努力 无论是哪种情况。特别令人感兴趣的是退伍军人中阿片类药物滥用的上升趋势。现有证据 表明脑损伤和阿片类药物滥用的分子和神经通路重叠；例如，炎症性 在阿片类药物滥用和小胶质细胞激活中都观察到了介质和增加的神经变性和小胶质细胞激活 MTBI。据我们所知，还没有报道的临床前研究系统地检查了 在神经行为和神经病理方面，mTBI和随后的阿片类药物暴露之间的串扰 以及神经生化后果。这项研究建议使用一个经过验证的反复闭合的啮齿动物模型 头部损伤(RCHI)与长期暴露于芬太尼药物相结合，以接近模拟mTBI的真实条件 以及鸦片类药物的使用。芬太尼是一种高效但易上瘾的止痛药，通常用于治疗慢性疼痛。 这一建议的中心假设是mTBI后持续的阿片类药物暴露将导致特征 神经功能和神经病理改变，不同于mTBI或阿片类药物单独暴露的情况， 这些变化可以通过神经行为、生化和神经蛋白质组学来识别和评估 接近了。这一建议为国际和平研究所的实验室提供了一个新的方向，并有可能改变范式。目标1 将确定芬太尼阿片类药物暴露1个月，然后在r-chi后停药1个月对 认知、痛感、运动和自发的阿片类药物戒断症状。雄性小鼠(C57BL6)将 使用并接受rCHI或假手术(非损伤)。将有四个试验组：(A) 假手术+生理盐水，(B)rCHI+生理盐水，(C)假手术+芬太尼，(D)rCHI+芬太尼暴露。功能性 评估测试包括：与脑损伤相关的认知(空间记忆表现；莫里斯水迷宫)和焦虑- 行为(高架+迷宫)，或阿片类药物使用/戒断测试；与运动活动相关的疼痛敏感性测试 (温水尾巴戒断和口腔福尔马林试验)(在药物暴露和戒断阶段)，以及药物 戒断依赖症状(在芬太尼戒断阶段)。结果将与 仅从rCHI或芬太尼暴露观察。目标2将评估关键的神经病理、神经细胞 大脑三个区域(皮质、海马体和腹侧被盖区)的标志物和阿片受体水平 与脑损伤和/或阿片受体介导的奖赏回路有关。大脑样本将在两个终点进行分析 -芬太尼给药结束时(第1个月)，停药1个月后(第2个月)，患有或 没有rCHI。神经元、轴突、星形胶质、小胶质、突触和白质标记物，以及 三种阿片受体亚型将通过免疫印迹/酶联免疫吸附试验在不同的动物组中进行评估 并通过免疫组织化学组织病理学分析。目标3将确定mTBI的长期后果和 阿片滥用共病相关病理通路/关键驱动因素作为可能的神经治疗靶点 神经蛋白质组学和系统生物学分析。总体而言，这一新的调查路线旨在提供见解 研究mTBI和损伤后阿片类药物滥用之间的神经生物学和神经病理相互作用。结果是 可能指导可能的新的治疗策略来治疗退伍军人的脑损伤和共病的阿片依赖。