Molecular Fingerprint of Cocaine Abuse Analysis

描述（由申请人提供）：在美国，可卡因滥用是一种 重大公共卫生问题。使用避孕药会激活特定的 大脑中的神经回路，尤其是中脑边缘多巴胺神经元。与持续 使用时，神经适应性变化发生在这些神经元中， 长期使用药物。在以下方面取得了重大进展： 阐明了神经适应性变化的生化机制， 然而，可卡因对人类死后基因转录的调节 组织受到的关注相当少。第一个目标将比较 腹侧被盖区（VTA）与 外侧黑质（l-SN）和延髓核（NAc）与背侧 可卡因过量受害者死后组织中的尾壳核（d-CP）， 年龄匹配的非药物对照组根据初步数据，我们预测慢性 可卡因的使用优先与基因表达的改变有关， 编码腹侧被盖区和NAc中的多巴胺和信号转导相关蛋白 分别与l-SN和d-CP进行比较。第二个目标，我们将 通过比较来检查离散神经元群体中的基因表达， VTA和1-SN中酪氨酸羟化酶免疫阳性神经元的分布 可卡因过量受害者和对照组之间的差异为此，我们预测 优先改变编码多巴胺和信号的基因的表达 在腹侧被盖区的多巴胺神经元中的转导相关蛋白与 可卡因过量的受害者中的l-SN这一目标是基于这样的想法，即协调 离散神经元群体中几个基因的失调与 可卡因滥用这些研究的结果将提供相关证据， 多个转录本的参与和详细的表达谱， 人类可卡因滥用最终目的是评估蛋白质水平的变化， 与基因表达谱相关的功能，特别是 多巴胺相关蛋白和信号转导相关蛋白。应用程序将 利用人类死后组织和最先进的区域和单个 神经元表达和cDNA阵列方法，以提供第一个广泛的 可卡因成瘾的表达谱。改变表达的表征 数千个基因的模式将提供一个潜在的全景图， 可卡因强化的分子基础和神经适应性变化 在这些神经元中与长期使用有关。此外，识别 差异表达的基因可能提供新的靶点， 药物开发和/或现有药物的改进 药物治疗

DESCRIPTION (provided by applicant): Cocaine abuse in the United States is a major public health concern. Cocaine use leads to activation of specific circuits in the brain, most notably mesolimbic dopamine neurons. With continued use, neuroadaptive changes occur in these neurons which lead to even more protracted use of the drug. Significant progress has been made in the elucidation of the biochemical mechanisms underlying the neuroadaptive changes, however, the regulation of gene transcription by cocaine in human post-mortem tissue has received considerably less attention. The first aim will compare regional gene expression between the ventral tegmental area (VTA) versus lateral substantial nigra (l-SN) and nucleus accumbens (NAc) versus dorsal caudate-putamen (d-CP) in post-mortem tissue from cocaine overdose victims and age-matched, non-drug controls. Based on preliminary data, we predict chronic cocaine use is preferentially associated with altered expression of genes encoding dopamine- and signal transduction-related proteins in the VTA and NAc compared with the l-SN and d-CP, respectively. In the second aim, we will examine the gene expression in a discrete neuronal population by comparing profiles of tyrosine hydroxylase immuno-positive neurons in the VTA and 1-SN between cocaine overdose victims and controls. To this end, we predict preferentially altered expression of genes encoding dopamine- and signal transduction-related proteins in dopamine neurons in the VTA compared with the l-SN in cocaine overdose victims. This aim is based on the idea that coordinate dysregulation of several genes in discrete neuronal populations is linked to cocaine abuse. Results from these studies will provide correlative evidence of the involvement of multiple transcripts and a detailed expression profile of human cocaine abuse. The final aim is to evaluate changes in protein levels and function associated with the expression profile of genes, in particular dopamine related and signal transduction-related proteins. The application will utilize human post-mortem tissue and state-of-the-art regional and single neuron expression and cDNA array methodologies to provide the first extensive expression profile of cocaine addiction. Characterization of altered expression patterns for thousands of genes will provide a panoramic view of the potential molecular underpinnings of cocaine reinforcement and the neuroadaptive changes in these neurons associated with long-term use. In addition, identification of differentially expressed genes may provide novel targets for the pharmacotherapeutic development and/or the refinement of existent pharmacotherapies.