Genetic and Epigenetic Regulation of Addiction Genes

药物成瘾是一种具有强烈遗传成分的复杂疾病，而表观遗传因素的作用 仍然没有解决。我们认为遗传多态性和表观遗传变化之间的相互作用 决定基因表达和可能的mRNA加工，在药物成瘾中起关键作用。大 一部分疑似成瘾易感基因含有CpG岛，其甲基化代表了一种可能的成瘾易感基因。 主要的表观遗传机制。遗传和表观遗传因素都可能导致成瘾易感性 以及由于药物滥用而发生的生理变化。我们将在尸检中研究这些因素 来自迈阿密脑捐赠库的组织，包含约500份可卡因和其他药物样本 滥用者和年龄匹配的对照组。该储存库使相关大脑中的遗传和表观遗传研究成为可能。 涉及成瘾的区域。CpG甲基化可以在基因的两个等位基因之间随机发生 （以总体表达水平表示），或以等位基因选择性方式。后者导致等位基因 表达不平衡（AEI），其代表遗传和 表观遗传顺式作用因子这使我们能够解决几个问题。CpG岛是如何 甲基化在不同的大脑区域是不同的，个体之间的差异是什么？甲基化是否影响 基因表达，交替启动子使用，或选择性剪接？药物滥用的影响是什么 在相关脑区的候选基因CpG岛甲基化？表观遗传因素 有助于临床状态（成瘾）？在这个项目中，我们的目标是含有CpG岛的基因， 涉及成瘾，侧重于生物胺途径，编码合成和分解代谢酶， 囊泡和突触再摄取转运蛋白和受体（MAOA，MAOB，COMT，TH，DAT，NET，VMAT2， DRD2、CHRNA4）。我们已经开发出高通量的工具来测量遗传和表观遗传 对mRNA和蛋白质表达的贡献以及可变剪接。我们的检测是等位基因特异性的 能够评估等位基因表达中的遗传和表观遗传因素， 每个因素的量化影响。这种新的方法，适用于解剖学定义的脑组织， 从吸毒者和控制，有可能产生显着的洞察力的作用和相互作用， 遗传和表观遗传因素之间的联系，并增加了我们对成瘾易感性的理解。

Drug addiction is a complex disorder with a strong genetic component, while the role of epigenetic factors remains unresolved. We propose that the interplay between genetic polymorphisms and epigenetic changes determines gene expression and possibly mRNA processing, serving a critical role in drug addiction. A large portion of suspected addiction susceptibility genes harbors CpG islands, methylation of which represents a main epigenetic mechanism. Both genetic and epigenetic factors likely contribute to addiction susceptibility and physiological changes occurring as a result of substance abuse. We will study these factors in autopsy tissues from the Miami Brain Endowment Bank, containing ~approximately 500 samples from cocaine and other drug abusers and age-matched controls. This repository enables genetic and epigenetic studies in relevant brain regions involved in addiction. CpG methylation can occur randomly between the two allele of a gene (represented in overall expression level), or in an allele-selective fashion. The latter causes an allelic expression imbalance (AEI), which represents a precise and quantitative phenotype for both genetic and epigenetic cis-acting factors. This permits us to address several questions. How does CpG island methylation vary across brain regions, and what is the variability among individuals? Does methylation affect gene expression, alternate promoter usage, or alternative splicing? What is the effect of substance abuse on CpG island methylation in candidate genes, in relevant brain regions? Do epigentic and genetic factors contribute to clinical status (addiction)? In this project, we target genes harboring CpG islands that are implicated in addiction, focusing on biogenic amine pathways, encoding synthetic and catabolic enzymes, vesicular and synaptic reuptake transporters, and receptors (MAOA, MAOB, COMT, TH, DAT, NET, VMAT2, DRD2, CHRNA4). We have developed high-throughput tools for measuring the genetic and epigenetic contribution to mRNA and protein expression, and alternative splicing. Our assays are allele-specific, enabling the evaluation of genetic and epigenetic factors in allelic expression, a powerful tool for assessing the quantitative impact of each factor. This novel approach, applied to anatomically defined brain tissues from drug addicts and controls, has the potential to yield significant insight into the role of and interplay between genetic and epigenetic factors, and add to our understanding of susceptibility to addiction.