Molecular Fingerprint of Cocaine Abuse Analysis

描述(申请人提供)：美国的可卡因滥用是一种 主要的公共卫生问题。可卡因的使用会导致特异性神经元的激活 大脑中的回路，最著名的是中脑边缘多巴胺神经元。(续) 使用时，这些神经元会发生神经适应性变化，从而导致更多 长期使用这种药物。在以下方面取得了重大进展： 阐明神经适应性变化背后的生化机制， 然而，可卡因对人死后基因转录的调节 组织受到的关注要少得多。第一个目标是比较 腹侧被盖区(VTA)与腹侧被盖区(VTA)的局部基因表达 外侧实质黑质(L-SN)和伏隔核(NAc)与背侧 可卡因过量患者死后组织中d-CP的检测 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相关 VTA和NAC中编码多巴胺和信号转导相关蛋白 分别与L-SN和d-CP进行比较。在第二个目标中，我们将 通过比较来检测离散神经元群体中的基因表达 VTA和1-SN内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关系。为此，我们预测， 编码多巴胺和信号的基因表达优先改变 VTA内多巴胺神经元信号转导相关蛋白的比较 L-SN中的可卡因过量受害者。这个目标是基于这样一个想法，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的证据 涉及多个转录本和详细的表达谱 人类滥用可卡因。最终目标是评估蛋白质水平的变化和 与基因表达谱相关的功能，特别是 多巴胺相关和信号转导相关的蛋白质。该应用程序将 利用人类死后组织和最先进的区域和单一 神经元表达和cDNA阵列方法学提供了第一个广泛的 可卡因成瘾的表达谱。改变表达的特征 数千个基因的模式将提供潜在的全景 可卡因强化的分子基础与神经适应性改变 在这些神经元中与长期使用有关。此外，识别 差异表达基因可能为基因治疗提供新靶点 药物治疗的发展和/或现有药物的提炼 药物疗法。

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.