Interpreting the regulatory mechanisms underlying the predisposition to substance use disorders

解释物质使用障碍倾向背后的调节机制

• 批准号: 9764328
• 负责人: Andreas Robert Pfenning
• 金额: $ 47.43万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9764328
• 关键词: Alcohol or Other Drugs use; Animal Model; Biological; Biological Assay; Brain; Brain Diseases; Brain region; CRISPR interference; Code; Complex; Computer Analysis; Data Set; Development; Disease; Drug Targeting; Enhancers; Epigenetic Process; Genes; Genetic; Genetic Variation; Genome; Genomics; Health; Human; Human Genetics; Link; Maps; Measures; Methods; Molecular; Mus; Nucleic Acid Regulatory Sequences; Pathway interactions; Personal Satisfaction; Predisposition; Proteins; Reporter; Research; Research Personnel; Smoking Behavior; Substance Use Disorder; Testing; Untranslated RNA; Work; addiction; cell type; epigenomics; flexibility; genetic variant; genome wide association study; genomic tools; in vivo; infancy; mouse model; neural circuit; nicotine exposure; relating to nervous system; therapy development; tool

Summary/Abstract Substance use disorders have a profound impact on human health and wellbeing. The development of new treatments for these disorders has remained difficult due to the complexity of the neural circuits and the underlying genetic mechanisms. Recent genome-wide association studies have begun to identify a few of loci associated with the predisposition to addiction, with many more loci are implicated with lower confidence. The majority of the genetic variants associated with complex brain disorders, including substance use, are likely to be located in non-coding regulatory regions, particularly enhancers, and not within protein-coding genes. Despite the importance of enhancer regions in the brain, the computational and experimental tools to study their function are still in their infancy. To work towards deciphering this critical biological mechanism underlying substance use disorders, we seek to build a framework to study the function of both human and mouse brain enhancer regions in vivo. First, we will analyze publically available genetic and epigenetic data sets to identify human genetic variation at enhancers regions that is likely to influence substance use predisposition. Next, we will measure the impact of that genetic variation using a high-throughput reporter assay, which has the ability to simultaneously test the activity of thousands of enhancers across the mouse brain in vivo under different conditions. Finally, we will validate those predictions using CRISPR interference on conserved orthologous enhancers in the mouse. In an initial test case, we will apply our methods to interpret a genome-wide association study of smoking behavior using cell type-specific human epigenomics. The key candidates that result from the computational analysis will be screened and validated in a mouse model of nicotine exposure. The result of our research effort will be a map that links high-confidence and low-confidence substance use-associated genetic variation to neural enhancer function. Furthermore, the flexible computational and experimental framework has the potential to study any combination of candidate enhancers and genetic variants in the context of different brain regions, different cell types, and different animal models.

摘要/摘要 药物使用障碍对人类健康和福祉产生深远的影响。发展的发展 由于神经回路的复杂性和 潜在的遗传机制。最近的全基因组关联研究已经开始识别一些基因座 与成瘾的易感性相关，更多的基因座的置信度较低。这 与复杂脑疾病相关的大多数遗传变异（包括使用物质）可能都可能 位于非编码调节区域，尤其是增强子，而不是蛋白质编码基因中。 尽管大脑中增强剂区域的重要性，但研究的计算和​​实验工具 他们的功能仍处于起步阶段。 为了破解这种重要的物质使用障碍的关键生物学机制，我们 寻求建立一个框架来研究体内人和小鼠脑增强剂区域的功能。第一的， 我们将分析公开可用的遗传和表观遗传数据集，以识别人类遗传变异 可能影响物质使用易感性的增强剂区域。接下来，我们将衡量 使用高通量记者测定法的遗传变异，该测定能够同时测试 在不同条件下，体内遍布小鼠大脑的数千个增强子的活性。最后，我们会的 使用CRISPR干扰小鼠中保守的直系同源增强子来验证这些预测。在 初始测试案例，我们将应用我们的方法来解释全基因组的吸烟行为研究 使用细胞类型特异性的人类表观基因组学。由计算分析产生的关键候选人 将在尼古丁暴露的鼠标模型中进行筛选和验证。 我们的研究工作的结果将是一张与高信心和低信心物质联系起来的地图 与神经增强子功能的使用相关遗传变异。此外，灵活的计算和 实验框架有可能研究候选增强子和遗传的任何组合 在不同大脑区域，不同细胞类型和不同动物模型的背景下的变体。