Proteomic biosignatures of withdrawal from cocaine in rhesus monkeys

恒河猴戒断可卡因的蛋白质组生物特征

• 批准号: 7762606
• 负责人: SCOTT Edwards HEMBY
• 金额: $ 21.45万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7762606
• 关键词: AMPA Receptors; Acute; Autopsy; Behavior; Behavioral; Biochemical; Biochemical Pathway; Biological Assay; Biological Markers; Biological Markers; Blood; Blood specimen; Brain; Brain region; Chronic; Clinic; Cocaine; Cocaine Abuse; Cocaine Dependence; Collection; Complex; DLG4 gene; Dependence; Development; Diagnosis; Diagnostic; Disease; Drug usage; Funding; Gel; Genetic Markers; Glutamate Receptor; Goals; Hair; High Pressure Liquid Chromatography; Human; Immunoprecipitation; Individual; Intake; Label; Laboratory Study; Lead; Liquid substance; Macaca mulatta; Maps; Measures; Mediating; Medical; Medicine; Membrane; Methods; Modeling; Monitor; Monkeys; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; NR1 gene; National Institute of Drug Abuse; Neurobiology; Neurons; Nucleus Accumbens; Pathway Analysis; Patient Monitoring; Pattern; Peripheral; Pharmaceutical Preparations; Plasma; Plasma Proteins; Predisposition; Principal Component Analysis; Procedures; Protein Analysis; Proteins; Proteomics; Public Health; Relapse; Research; Resources; Rewards; Rodent; Sampling; Self Administration; Self-Administered; Set protein; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Substance abuse problem; Synapses; Therapeutic Intervention; Time; Tissues; United States; Up-Regulation; Urine; Validation; Ventral Striatum; Western Blotting; Withdrawal; addiction; base; biosignature; brain tissue; cocaine exposure; cocaine overdose; cocaine use; craving; prognostic; protein expression; protein protein interaction; public health relevance; receptor; response; tool

DESCRIPTION (provided by applicant): Cocaine abuse and dependence in the United States continues to be a major public health concern yet significant obstacles remain for the appropriate diagnosis and treatment of individuals suffering from addictive disorders. Research efforts have focused primarily on the development and refinement of toxicological assays for detecting cocaine as a measure of recent cocaine use and on the identification of genetic markers indicative of a predisposition to cocaine abuse. While both approaches are useful in their own right, a continuing unmet medical need is the identification of peripheral biological markers that can assist with the objective diagnosis, identification of subsets of patients, and monitor progression and response to treatment. Moreover, biomarkers could also be used as indicators of neurobiological changes associated with chronic cocaine use. To this end, we will use our well developed track record in proteomics research relevant to cocaine abuse to identify and correlate plasma biomarkers with biochemical changes occurring in reward relevant brain regions during withdrawal from chronic cocaine self-administration in rhesus monkeys. These studies will leverage the scientific expertise of Drs. Porrino and Nader and the resources of their ongoing NIDA funded study (R01DA009085). In Aim 1, plasma samples will be collected longitudinally during a 100 day period of cocaine self- administration through a thirty day period of withdrawal. Samples will be analyzed using unbiased complimentary proteomic methods (2D-DIGE and iTRAQ/MDLC with MALDI ToF/ToF analysis) and appropriate validation methods. Multivariate statistical analysis approaches will be used to delineate a finite set of proteins with the greatest degree of discriminability between intake and withdrawal and between acute and chronic withdrawal. Studies in the Aim 2 will explore the cocaine induced dysregulation of NMDA and AMPA receptors and associated proteins in the ventral striatum at one and thirty days of withdrawal from cocaine. In addition, we will use a targeted proteomic approach to evaluate the effects of cocaine withdrawal on the NR1 subunit and associated complex in membrane fractions from the ventral striatum using iTRAQ /MDLC with MALDI ToF/ToF analysis. The expression of peripheral biomarkers identified in Aim 1 will be correlated with neurobiological changes identified in Aim 2. We anticipate these strategies will provide a preliminary biosignature of cocaine use/withdrawal and correlated neuronal dysregulation that ultimately can be used as a diagnostic and prognostic tool in the treatment of cocaine addiction. PUBLIC HEALTH RELEVANCE: A continuing need in addiction medicine is the identification of peripheral biological markers that are indicative of prior chronic drug use and are correlated with changes in the brain that mediate reward, craving and relapse. Using a rhesus monkey model of cocaine self-administration withdrawal, we propose to define a proteomic biosignature in plasma that is correlated with neurobiological changes occurring in response to cocaine withdrawal. The proposed studies are intended to delineate a preliminary set of peripheral biomarkers that can be further evaluated in the rhesus monkey model and in human behavioral laboratory studies before implementing as potential diagnostic and prognostic tool at the clinic.

描述（由申请人提供）：在美国，可卡因滥用和依赖仍然是一个主要的公共卫生问题，但对患有成瘾性疾病的个体进行适当的诊断和治疗仍然存在重大障碍。研究工作主要集中于开发和完善用于检测可卡因的毒理学测定法，作为近期可卡因使用的衡量标准，以及鉴定表明可卡因滥用倾向的遗传标记。虽然这两种方法本身都是有用的，但持续未满足的医疗需求是识别外周生物标志物，这些标志物可以帮助客观诊断、识别患者亚群以及监测进展和治疗反应。此外，生物标志物还可以用作与长期使用可卡因相关的神经生物学变化的指标。为此，我们将利用我们在与可卡因滥用相关的蛋白质组学研究方面的良好记录，来识别血浆生物标志物，并将其与恒河猴长期可卡因自我给药戒断期间奖励相关大脑区域发生的生化变化联系起来。这些研究将利用博士的科学专业知识。 Porrino 和 Nader 以及他们正在进行的 NIDA 资助研究 (R01DA009085) 的资源。在目标 1 中，将在 100 天的可卡因自我给药期间到 30 天的戒断期间纵向收集血浆样本。将使用无偏倚的互补蛋白质组学方法（2D-DIGE 和 iTRAQ/MDLC 以及 MALDI ToF/ToF 分析）和适当的验证方法对样品进行分析。将使用多变量统计分析方法来描述一组有限的蛋白质，在摄入和戒断之间以及急性和慢性戒断之间具有最大程度的区分性。 Aim 2 的研究将探讨可卡因戒断后 1 天和 30 天时可卡因引起的 NMDA 和 AMPA 受体以及腹侧纹状体中相关蛋白的失调。此外，我们将使用 iTRAQ /MDLC 和 MALDI ToF/ToF 分析，采用靶向蛋白质组学方法来评估可卡因戒断对腹侧纹状体膜组分中 NR1 亚基和相关复合物的影响。目标 1 中确定的外周生物标志物的表达将与目标 2 中确定的神经生物学变化相关。我们预计这些策略将提供可卡因使用/戒断和相关神经元失调的初步生物特征，最终可用作可卡因成瘾治疗的诊断和预后工具。 公共健康相关性：成瘾医学的持续需求是识别外周生物标志物，这些生物标志物表明先前长期吸毒，并与介导奖赏、渴望和复发的大脑变化相关。使用可卡因自我戒断的恒河猴模型，我们建议定义血浆中的蛋白质组生物特征，该生物特征与可卡因戒断反应中发生的神经生物学变化相关。拟议的研究旨在描绘一组初步的外周生物标志物，这些标志物可以在恒河猴模型和人类行为实验室研究中进一步评估，然后在临床上作为潜在的诊断和预后工具实施。