Effects of Long-Term Morphine Treatment on Opioid Receptor Signaling and Inflammation in the Chronic Post-TBI Period

长期吗啡治疗对慢性 TBI 后阿片受体信号传导和炎症的影响

• 批准号: 10454092
• 负责人: Kelly Bosse
• 金额: --
• 依托单位: JOHN D DINGELL VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454092
• 关键词: Absence of pain sensation; Acute; Address; Affect; Affinity; Agonist; Amygdaloid structure; Analgesics; Area; Astrocytes; Award; Awareness; Behavior; Behavioral; Binding; Biological Assay; Brain; Brain Injuries; Cell Nucleus; Chronic; Clinical; Cognition; Cognitive; Coupled; Data; Dependence; Diagnosis; Dorsal; Dose; Emotional; Enzyme-Linked Immunosorbent Assay; Epidemic; Experimental Designs; Exposure to; Extracellular Signal Regulated Kinases; Female; Flow Cytometry; Foundations; Freedom; Functional disorder; Future; GTP-Binding Proteins; Gene Expression; Guanosine Triphosphate; Hippocampus (Brain); Human Resources; Hyperalgesia; Immunity; Immunofluorescence Immunologic; Impaired cognition; Impairment; Individual; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-1 beta; Label; Ligand Binding; Link; Long-Term Care; Long-Term Effects; Measurement; Measures; Mediating; Mediator of activation protein; Microglia; Mission; Molecular Analysis; Morphine; Mus; Narcotics; Nature; Neuroglia; Neurologic; Neurons; Nociception; Nucleus Accumbens; Opioid; Opioid Receptor; Outcome; Pain; Pain management; Paper; Pathologic; Pathology; Pathway interactions; Patient Care; Pattern; Pharmaceutical Preparations; Pharmacology; Phenotype; Population; Precipitation; Prefrontal Cortex; Process; Property; Proteins; Published Comment; Publishing; Receptor Signaling; Recurrent pain; Regimen; Rehabilitation therapy; Rewards; Risk; Services; Signal Pathway; Signal Transduction; Signaling Protein; Stains; Substance Use Disorder; System; TLR4 gene; Testing; Time; Tissues; Traumatic Brain Injury; Ventral Tegmental Area; Veterans; Withdrawal; addiction; adverse outcome; base; chronic pain management; cytokine; density; effective therapy; glial activation; high risk; immune activation; immune system function; improved; innovation; interest; male; midbrain central gray substance; morphine administration; mu opioid receptors; novel; operation; opioid misuse; opioid use; pain relief; radioligand; receptor; receptor density; receptor expression; recruit; response; service member; side effect; tool

Increasing awareness has been placed in understanding the issues related to rehabilitation of the estimated 330,000 service personnel diagnosed with traumatic brain injury (TBI) since the start of Operation Enduring Freedom and Operation Iraqi Freedom, who suffer from physical, cognitive, emotional dysfunction and recurrent pain. Specifically, opioid misuse is a growing epidemic nationwide, and recent evidence indicates that Veterans who sustain moderate/severe TBI are more likely to receive long-term treatment with opioid- based medications and engage in higher-risk opioid use patterns. These findings highlight the critical need for studies to establish what adaptive changes occur in the opioid system in the injured brain that may influence nociceptive, cognitive and reward/dependence outcomes. Morphine, the prototypical opioid antinociceptive, binds to the mu opioid receptor (MOR) to stimulate G-protein signaling and downstream effectors, such as extracellular signal-regulated kinase (ERK). In addition to this classical signaling pathway, morphine can act through the immunity toll-like like receptor 4 (TLR4) to activate glial cells and stimulate the release of proinflammatory mediators, like interleukin 1 beta (IL-1). While both signaling pathways contribute to morphine-induced phenotypes, evidence indicates immune activation disrupts morphine’s efficacy to promote analgesia, worsens pathological pain and exacerbates dependence and withdrawal. While TBI is recognized to induce long-term effects on immune system function alone, to date, no published papers have explored the combined influence of TBI and opioids on these signaling outcomes, adding to the significance and innovation of the proposed project. The current SPiRE proposal will test the hypothesis that repeated exposure to morphine following TBI will upregulate MOR expression/function and selectively potentiate glial-mediated inflammatory processes upon subsequent re-exposure to morphine in regions associated with reward, cognition and pain. The objectives in this proposal will be accomplished through two Specific Aims with tissue measurements in the nucleus accumbens, ventral tegmental area, prefrontal cortex, periaqueductal gray, dorsal hippocampus and amygdala. Aim 1 will assess longitudinal changes in MOR status as measured by radioligand binding density/affinity as well as gene expression (real-time PCR) and immunofluorescent co- localization on neurons and glia in tissue collected 7 days, 1, 3 and 6 months following TBI/repeated morphine. Aim 2 will characterize differential activation of classical and inflammatory pathways stimulated by morphine re- exposure at long-term timepoints (3 and 6 months) following combined TBI and subchronic morphine. Activity through classical pathways will be evaluated by G-protein (GTPS) activity and extent of phosphorylated ERK immunofluorescence. Inflammatory pathway signaling will be assessed by measuring the extent of glial activation with immunofluorescence, TLR4 microglial staining and flow cytometry, and IL-1 protein quantification with ELISA. The hypothesized finding of a potentiated inflammatory response to morphine following TBI would be consistent with conferring an increased risk of adverse outcomes associated with activation of this pathway, including addiction vulnerability, impaired cognition and hyperalgesia, making these studies highly relevant to the mission for VA patient care and rehabilitation. These data will also provide critical evidence to serve as a foundation for a comprehensive future study to explore the behavioral implications of these signaling outcomes and pharmacologic strategies exploring biased agonism.

提高人们的认识，了解与康复相关的问题 自行动开始以来，估计有 330,000 名军人被诊断患有创伤性脑损伤 (TBI) 持久自由和伊拉克自由行动，遭受身体和情感功能障碍的人 具体而言，阿片类药物滥用在全国范围内日益流行，最近的证据表明。 患有中度/重度 TBI 的退伍军人更有可能接受阿片类药物的长期治疗 这些发现凸显了对阿片类药物的迫切需求。 研究确定受伤大脑中阿片类药物系统发生哪些适应性变化可能会影响 伤害性、认知性和奖励/依赖结果吗啡，典型的阿片类抗镇痛药， 与 mu 阿片受体 (MOR) 结合，刺激 G 蛋白信号传导和下游效应器，例如 除了这种经典的信号传导途径外，吗啡还可以发挥作用。 通过免疫类Toll样受体4（TLR4）激活神经胶质细胞并刺激释放 促炎症介质，如白细胞介素 1 β (IL-1)，而这两种信号通路均有助于促进炎症反应。 吗啡诱导的表型，证据表明免疫激活会破坏吗啡促进的功效 镇痛会加剧病理性疼痛并加剧依赖性和戒断症状，​​而 TBI 被认为会导致这种情况。 仅对免疫系统功能产生长期影响，迄今为止，还没有发表的论文探讨过 TBI 和阿片类药物对这些信号传导结果的综合影响，增加了重要性和创新性 拟议项目的。 目前的 SPiRE 提案将检验以下假设：TBI 后反复接触吗啡 将上调 MOR 表达/功能并选择性增强神经胶质介导的炎症 随后在与奖赏、认知和相关区域再次暴露于吗啡时的过程 该提案中的目标将通过组织的两个具体目标来实现。 伏隔核、腹侧被盖区、前额皮质、导水管周围灰质的测量， 目标 1 将评估 MOR 状态的纵向变化（通过测量）。 放射性配体结合密度/亲和力以及基因表达（实时 PCR）和免疫荧光共 TBI/重复吗啡后 7 天、1、3 和 6 个月收集的组织中神经元和神经胶质细胞的定位。 目标 2 将表征由吗啡重新刺激的经典途径和炎症途径的差异激活。 TBI 和亚慢性吗啡联合活动后的长期暴露（3 个月和 6 个月）。 通过经典途径将通过 G 蛋白 (GTP-S) 活性和磷酸化 ERK 的程度进行评估 免疫荧光将通过测量神经胶质的范围来评估炎症通路信号传导。 通过免疫荧光、TLR4 小胶质细胞染色和流式细胞术以及 IL-1 蛋白进行激活 首次发现吗啡可增强炎症反应。 TBI 后发生不良后果的风险增加是一致的 该通路的激活，包括成瘾脆弱性、认知受损和痛觉过敏，使这些 与退伍军人事务部患者护理和康复任务高度相关的研究这些数据也将提供重要的信息。 证据作为未来全面研究的基础，以探索行为的影响 这些信号转导结果和药理学策略探索了偏向激动作用。