Cannabinoid Receptor (CNR1) MicroRNAs and Addictive Disease

大麻素受体 (CNR1) MicroRNA 与成瘾性疾病

• 批准号: 7291044
• 负责人: HENRY M FURNEAUX
• 金额: $ 14.37万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-26 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7291044
• 关键词: 3' Untranslated Regions; Addictive Behavior; Affect; African American; Alcohols; Algorithms; Be++ element; Beryllium; Binding; Candidate Disease Gene; Cannabinoids; Cells; Code; Collaborations; Complex; Computer Simulation; Control Groups; DNA; DNA Insertion Elements; Disease; Down-Regulation; Drug Addiction; Elements; Enzymes; Epigenetic Process; Genes; Genetic; Genetic Polymorphism; Hand; Human; Indium; Individual; Knock-out; Lead; Luciferases; Mediating; Messenger RNA; MicroRNAs; Mus; Mutate; Mutation; Nucleotides; Opiates; Outcome; Pathway interactions; Patients; Population; Predisposition; Process; Proteins; Reagent; Regulation; Reporter; Repression; Response Elements; Role; Sampling; Small RNA; Testing; Variant; addiction; cannabinoid receptor; design; expectation; experience; human DICER1 protein; nervous system disorder; receptor expression; research study; response

DESCRIPTION (provided by applicant): Drug addiction is a complex neurological disorder likely to involve genetic components. For example, studies in mice have shown that elimination of the gene that encodes the receptor for cannabinoids blunts their "addictive response " to opiates and alcohol. Although many genes have been suspected to contribute to addictive behavior, unique human variants in these genes that unequivocally promote susceptibility to drug addiction have not been identified. So far, most attempts to identify such variants have focused on the protein coding segments of the suspect genes. However, it is quite possible that variants that alter the expression of such candidate genes may also render individuals susceptible to addictive disease. Recently, it has become clear that many human genes are regulated by a new epigenetic mechanism, which involves the specific annealing of a small RNA (microRNA) to a target element in the mRNA. In most cases, the annealing of the microRNA leads to a profound decrease in expression of the target mRNA. In our preliminary studies, we have identified a 50 nucleotide element in the 3'UTR of the human cannabinoid receptor mRNA that is likely to be the target of a microRNA. We have observed that insertion of this element into a luciferase reporter significantly reduces its expression. Importantly, a mutation in the target element that is predicted to compromise the binding of microRNAs abrogates the suppressive effect of the target element. Interestingly, this mutation has previously been identified as a rare polymorphism in African American populations. Thus, our overarching hypothesis is that the cannabinoid receptor is regulated by microRNAs and that variants in the cannabinoid receptor element may influence its expression and thus render the individual susceptible to drug addiction. In our first aim, we will confirm and extend our initial observations and identify the microRNA that mediates repression of the cannabinoid receptor. These studies will be aided by our experience in designing and using "Antagomir" reagents. Antagomirs are modified RNAs that can be introduced into cells and can potently down regulate the expression of specific microRNAs. In our second aim, we will examine whether any human variants in the cannabinoid receptor element correlate with susceptibility to addiction. To do this, we have established collaboration with Drs. Kranzler, Covault and Oncken who have access to DNA from control and affected patient populations.

描述（由申请人提供）：药物成瘾是一种复杂的神经系统疾病，可能涉及遗传成分。例如，对小鼠的研究表明，消除编码大麻素受体的基因会减弱它们对阿片类药物和酒精的"成瘾反应"。尽管许多基因被怀疑与成瘾行为有关，但这些基因中明确促进药物成瘾易感性的独特人类变异尚未被确定。到目前为止，大多数识别这种变异的尝试都集中在可疑基因的蛋白质编码片段上。然而，很有可能改变这些候选基因表达的变异也可能使个体易患成瘾性疾病。最近，已经清楚的是，许多人类基因由一种新的表观遗传机制调节，该机制涉及小RNA（microRNA）与mRNA中的靶元件的特异性退火。在大多数情况下，microRNA的退火导致靶mRNA表达的显著降低。在我们的初步研究中，我们已经在人类大麻素受体mRNA的3'UTR中鉴定了一个50个核苷酸的元件，该元件可能是microRNA的靶标。我们已经观察到，将该元件插入荧光素酶报告基因显著降低其表达。重要的是，靶元件中被预测损害microRNA结合的突变消除了靶元件的抑制作用。有趣的是，这种突变以前被确定为非洲裔美国人群体中罕见的多态性。因此，我们的总体假设是，大麻素受体受microRNA的调节，大麻素受体元件的变体可能会影响其表达，从而使个体容易成瘾。在我们的第一个目标中，我们将确认并扩展我们的初步观察结果，并确定介导大麻素受体抑制的microRNA。我们在设计和使用"Antagomir"试剂方面的经验将有助于这些研究。拮抗剂是经过修饰的RNA，可以引入细胞并有效下调特定microRNA的表达。在我们的第二个目标中，我们将研究大麻素受体元件中的任何人类变体是否与成瘾易感性相关。为此，我们与Kranzler、Covault和Oncken博士建立了合作关系，他们可以从对照组和受影响的患者群体中获得DNA。