Phosphoproteomics of Opioid Tolerance

阿片类药物耐受性的磷酸化蛋白质组学

• 批准号: 7509546
• 负责人: George Latimer Wilcox
• 金额: $ 22.65万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-15 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7509546
• 关键词: Absence of pain sensation; Adverse effects; Affinity; Agmatine; Arginine; Brain; Cells; Chronic; Complement; Critical Pathways; Dependence; Development; Dopamine; Event; Future; Glutamates; Immunohistochemistry; Individual; Label; MK801; Maintenance; Measures; Mediator of activation protein; Memantine; Morphine; Mus; N-Methyl-D-Aspartate Receptors; NG-Nitroarginine Methyl Ester; NMDA receptor antagonist; Neuronal Plasticity; Neurons; Nitric Oxide Synthase; Opioid; Opioid Analgesics; Parents; Phosphopeptides; Phosphoproteins; Phosphorylation; Physical Dependence; Process; Production; Protein Isoforms; Proteins; Proteomics; Receptor Signaling; Rodent; Self Administration; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Spinal; Spinal Anesthesia; Spinal Cord; Spinal cord injury; Synaptic plasticity; System; Testing; Time; Western Blotting; cell type; chronic pain; computerized data processing; design; effective therapy; inhibitor/antagonist; novel strategies; prevent; protective effect; public health relevance; small molecule

DESCRIPTION (provided by applicant): Opioids are an effective treatment for chronic pain, but repeated use can result in tolerance and physical dependence. These changes are thought to require activation of NMDA receptors and nitric oxide synthase (NOS), because blockade of either can prevent induction of tolerance and dependence. Both of these processes are blocked by the small molecule agmatine (decarboxylated arginine), an endogenous NMDA-receptor antagonist and NOS inhibitor. Because the NMDA-R/NOS system is considered a likely mediator of agmatine's protective effect, the proposed study will explicitly compare its action with that of NMDA-R antagonists and NOS inhibitors as well as searching for other target proteins or proteins altered as a consequence of its action. The proposed project will apply mass spectral phosphoproteomic analysis to compare the complement of proteins phosphorylated in morphine tolerance with those phosphorylated when agmatine or other NMDA-R/NOS inhibitors protects subjects from tolerance. The project will test the hypothesis that agmatine blocks the development of spinal morphine tolerance through signal transduction pathways distinct from the NMDA-R/NOS cascade. The first aim will compare agmatine effects with those of other NMDA-R/NOS inhibitors. The second aim will add co-administered morphine to the first aim and identify the phosphorylation cascades specific to morphine tolerance using 2D LC- MS/MS on spinal cord extract. The third aim will validate these changes and determine the neuronal or glial localization of the altered phosphopeptides. The results of these studies should significantly extend our mechanistic understanding of agmatine modulation of neuroplasticity in opioid tolerance. Future studies could then focus on targeting the identified phosphorylation cascades in a broad spectrum of CNS maladaptive phenomena including opioid self-administration and tolerance, chronic pain, and spinal cord injury. The Public Health Relevance: Tolerance to opioid analgesics remains a key factor limiting access of chronic pain sufferers to adequate relief. This project will apply phosphoprotein analysis to identify the signaling processes activated or suppressed as chronic morphine tolerance develops in rodent spinal cord. Compounds that block spinal tolerance and synaptic plasticity by antagonizing NMDA receptors or inhibiting nitric oxide synthase will be the primary focus.

描述（由申请人提供）：阿片类药物是慢性疼痛的有效治疗方法，但重复使用可以导致耐受性和身体依赖性。人们认为这些变化需要激活NMDA受体和一氧化氮合酶（NOS），因为封锁都可以防止耐受性和依赖性诱导。这两个过程均被小分子agmatine（脱羧精氨酸）（一种内源性NMDA受体拮抗剂和NOS抑制剂）阻止。由于NMDA-R/NOS系统被认为是苹果丁保护作用的可能中介体，因此拟议的研究将显式将其作用与NMDA-R拮抗剂和NOS抑制剂的作用进行比较，并搜索因其作用而改变的其他靶蛋白或蛋白质。拟议的项目将应用质谱磷酸蛋白质组学分析，以比较吗啡耐受性磷酸化的蛋白质的补体与当agmatine或其他NMDA-R/NOS抑制剂保护受试者免受耐受性时的磷酸化的蛋白质与磷酸化的蛋白质。该项目将检验以下假设，即agmatine通过与NMDA-R/NOS Cascade不同的信号转导途径阻止了脊柱吗啡耐受性的发展。第一个目标将将邻玛汀的效应与其他NMDA-R/NOS抑制剂的作用进行比较。第二个目标将在第一个目标中增加辅助吗啡，并在脊髓提取物上使用2D LC-MS/MS确定磷酸化级联对吗啡耐受性。第三个目标将验证这些变化并确定改变的磷酸肽的神经元或神经胶质定位。这些研究的结果应显着扩展我们对阿片类药物耐受性神经可塑性调节的机械理解。然后，未来的研究可以集中于针对广泛的中枢神经系统不良运动现象（包括阿片类药物自我给药和耐受性，慢性疼痛和脊髓损伤）的鉴定磷酸化级联反应。公共卫生相关性：对阿片类镇痛药的耐受性仍然是限制慢性疼痛患者获得足够缓解的关键因素。该项目将采用磷蛋白分析，以确定随着慢性吗啡耐受性在啮齿动物脊髓中的发展而激活或抑制的信号传导过程。通过拮抗NMDA受体或抑制一氧化氮合酶来阻断脊柱耐受性和突触可塑性的化合物将是主要焦点。