Peptide Biomarkers of drug exposure: from brain microdialysate to plasma

药物暴露的肽生物标志物：从脑微透析液到血浆

• 批准号: 7762724
• 负责人: Nigel T Maidment
• 金额: $ 30.8万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 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/7762724
• 关键词: Abstinence; Addictive Behavior; Amygdaloid structure; Behavior; Behavioral Model; Biological Markers; Blood specimen; Brain; Brain region; CNS processing; Chronic; Cocaine; Complex; Data; Development; Drug Exposure; Exposure to; Extinction (Psychology); Future; Globus Pallidus; Goals; Home environment; Human; Hypothalamic structure; Individual; Link; Mass Spectrum Analysis; Methods; Microdialysis; Modeling; Morphine; Neuronal Plasticity; Nucleus Accumbens; Peptides; Pharmaceutical Preparations; Phase; Plasma; Potassium; Procedures; Process; Proteome; Rattus; Recording of previous events; Resolution; Sampling; Screening procedure; Self Administration; Serum; Source; Staging; Testing; Tissues; Validation; Ventral Striatum; Withdrawal; addiction; behavior change; conditioning; drug of abuse; extracellular; frontal lobe; interest; preference; public health relevance; research study; response

DESCRIPTION (provided by applicant): The goal of this application is to identify plasma peptide signatures that reflect neuroplasticity in regions of the brain associated with multiple stages of the addictive process, modeled initially using the simple conditioned place preference (CPP) procedure in rats. A focused approach is proposed in which the initial search will begin in the extracellular compartment of brain regions implicated in addictive processes, using microdialysis linked to mass spectrometry (high resolution nanobore LC-MS and data-dependent LC-MS2). We will then search for, and quantify, such identified candidate peptides in plasma in a targeted fashion, using LC-MS3 and/or LC-MRM. The discovery phase of the project will identify arrays of candidate peptides that, in brain regions considered central to addictive behavior, change extracellularly in response to chronic exposure to, and abstinence from, two highly addictive drugs of different pharmacological classes; namely morphine and cocaine. A parallel strategy will focus entirely on readily releasable peptides by constructing a subtractive screen of brain dialysates under potassium depolarization versus basal conditions. In both of these approaches blood plasma samples will be collected simultaneously. Peptides identified in either of these dialysate screens will subsequently be searched for in plasma in a targeted fashion. Peptides that are (a) detectable in both dialysate and plasma, and (b) change in concentration in tandem as a product of drug history or depolarization will be selected for the validation phase. In the validation phase, candidates identified in the discovery phase will be studied in longitudinal experiments using the place preference paradigm to correlate changes in plasma content with multiple phases of the conditioning experiment: drug conditioning, initial testing, incubation (withdrawal), re-testing, extinction and reinstatement. We expect to identify individual peptide signatures in plasma that reflect different stages of the addiction process to the extent that they are modeled by CPP. These experiments will focus on cocaine and morphine, but will be extended to other abused drugs and to other behavioral models (e.g. self-administration) in future experiments beyond the scope of this proposal. PUBLIC HEALTH RELEVANCE: These studies will provide plasma peptide signatures unique to different stages of one simple model of addictive behavior. Once this proof of principle is established the approach could be extended to other more complex addiction models, and ultimately to humans, thereby providing markers of underlying addictive processes. This is a key issue for the development of effective medications to treat addiction to multiple classes of abused substances.

描述（由申请人提供）：本申请的目的是识别反映与成瘾过程多个阶段相关的大脑区域中神经可塑性的等离子体肽特征，最初是使用大鼠中简单条件的位置偏好（CPP）过程建模的。提出了一种集中的方法，即使用与质谱法链接的微透析（高分辨率Nanobore LC-MS和数据依赖性LC-MS2），初始搜索将开始于与成瘾过程有关的大脑区域的细胞外隔室开始。然后，我们将使用LC-MS3和/或LC-MRM以目标方式搜索并量化该鉴定出的候选肽。该项目的发现阶段将确定候选肽的阵列，这些肽在被认为是成瘾行为的核心的大脑区域中，会在响应长期暴露于对不同药理学类别的两种高度上瘾的药物中的慢性暴露而细胞外变化；即吗啡和可卡因。平行策略将通过在去极化钾与基础条件下构建脑透析液的减法筛查，完全集中在易于释放的肽上。在这两种方法中，血浆样品将同时收集。随后将以靶向方式以等离子体的方式搜索这些透析筛网中的任何一个肽。 （a）在透析液和血浆中可检测的肽，以及（b）串联浓度变化作为药物史或去极化的产物，将在验证阶段选择。在验证阶段，将在纵向实验中使用位置偏好范式研究在纵向实验中确定的候选者，以将血浆含量的变化与调节实验的多个阶段相关联：药物调理，初始测试，孵育（戒断），重新测试，灭绝，灭绝和恢复。我们希望在血浆中识别出反映成瘾过程不同阶段的单个肽特征，以通过CPP对它们进行建模。这些实验将集中在可卡因和吗啡上，但将扩展到其他滥用药物以及其他行为模型（例如自我管理）的未来实验，这是该提案范围之外的范围。 公共卫生相关性：这些研究将提供等离子肽特征，这是一种简单的成瘾行为模型的不同阶段所特有的。一旦确定了原则证明，该方法可以扩展到其他更复杂的成瘾模型，并最终扩展到人类，从而提供了基本的成瘾过程的标志。这是开发有效的药物来治疗多种类别滥用物质的有效药物的关键问题。