Neuronal basis of OPRM1 A118G polymorphism in alcohol use disorders

酒精使用障碍中 OPRM1 A118G 多态性的神经元基础

• 批准号: 9146155
• 负责人: Apoorva Halikere
• 金额: $ 3.35万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9146155
• 关键词: Acute; Address; Adenine; Affect; Affinity; Agonist; Alcohol abuse; Alcohol consumption; Alcoholism; Alcohols; Alleles; Amino Acid Substitution; Animals; Asparagine; Aspartate; Behavior; Biochemical; Bioinformatics; Brain imaging; Cell Line; Cells; Chronic; D Aspartate; Data; Dependence; Development; Dopamine; Dose; Economics; Electrophysiology (science); Enkephalins; Environmental Risk Factor; Ethnic Origin; Ethnic group; Etiology; European; Exhibits; Genes; Genetic Polymorphism; Genetic Variation; Genetic study; Genotype; Goals; Guanine; Heritability; Human; Immunohistochemistry; Individual; Intoxication; Life; Ligand Binding; Ligands; Mediating; Molecular; Morphology; N-Glycosylation Site; Naltrexone; Neurons; Nicotine Dependence; Opioid; Opioid Receptor; Patients; Pattern; Physiological; Positioning Attribute; Predisposition; Property; Receptor Activation; Receptor Signaling; Recycling; Regulation; Reporting; Research; Risk; Signal Transduction; Single Nucleotide Polymorphism; Societies; Synapses; Synaptic Transmission; System; Techniques; Technology; Testing; Therapeutic Effect; Western Blotting; Work; alcohol exposure; alcohol reinforcement; alcohol response; alcohol reward; alcohol sensitivity; alcohol use disorder; base; brain tissue; effective therapy; excitatory neuron; genetic variant; glycosylation; induced pluripotent stem cell; inhibitory neuron; innovation; insight; mu opioid receptors; nervous system disorder; novel; presynaptic; problem drinker; public health relevance; receptor; receptor expression; receptor function; relating to nervous system; research study; social; stem cell biology; synaptic function; synaptic inhibition; synaptogenesis; trafficking; Acute; Address; Adenine; Affect; Affinity; Agonist; Alcohol abuse; Alcohol consumption; Alcoholism; Alcohols; Alleles; Amino Acid Substitution; Animals; Asparagine; Aspartate; Behavior; Biochemical; Bioinformatics; Brain imaging; Cell Line; Cells; Chronic; D Aspartate; Data; Dependence; Development; Dopamine; Dose; Economics; Electrophysiology (science); Enkephalins; Environmental Risk Factor; Ethnic Origin; Ethnic group; Etiology; European; Exhibits; Genes; Genetic Polymorphism; Genetic Variation; Genetic study; Genotype; Goals; Guanine; Heritability; Human; Immunohistochemistry; Individual; Intoxication; Life; Ligand Binding; Ligands; Mediating; Molecular; Morphology; N-Glycosylation Site; Naltrexone; Neurons; Nicotine Dependence; Opioid; Opioid Receptor; Patients; Pattern; Physiological; Positioning Attribute; Predisposition; Property; Receptor Activation; Receptor Signaling; Recycling; Regulation; Reporting; Research; Risk; Signal Transduction; Single Nucleotide Polymorphism; Societies; Synapses; Synaptic Transmission; System; Techniques; Technology; Testing; Therapeutic Effect; Western Blotting; Work; alcohol exposure; alcohol reinforcement; alcohol response; alcohol reward; alcohol sensitivity; alcohol use disorder; base; brain tissue; effective therapy; excitatory neuron; genetic variant; glycosylation; induced pluripotent stem cell; inhibitory neuron; innovation; insight; mu opioid receptors; nervous system disorder; novel; presynaptic; problem drinker; public health relevance; receptor; receptor expression; receptor function; relating to nervous system; research study; social; stem cell biology; synaptic function; synaptic inhibition; synaptogenesis; trafficking

 DESCRIPTION (provided by applicant): Alcohol use disorders (AUDs) imposing enormous economic and social burdens on society today. AUDs are now understood as a chronic nervous system disorder characterized by a physiological dependence on alcohol beyond voluntary control. While environmental factors contribute to alcohol use, recent genetic studies reveal that the heritability of alcoholism is around 50%, illustrating that common genetic -variation contributes significantly to the etiology of alcoholism. However, the major target mediating alcohol reward signaling and the synaptic mechanisms of alcohol reward are poorly understood. Recent genetic studies have identified alcoholism associated gene variants in patients, one being the single nucleotide polymorphism (SNP) of an adenine to a guanine at position 118 of human OPRM1, a gene encoding the mu opioid receptor (MOR). This results in a nonsynonymous substitution of asparagine (N) at residue 40 to an aspartate (D). Although heterologous expression systems and animal studies suggest altered receptor trafficking and ligand binding affinities, there have been no conclusive studies determining the underlying cellular and molecular mechanisms of the N40D SNP on alcohol reinforcement, especially in a human neuronal context. To better understand the functional consequences of this SNP, the objective of this study is to investigate the interaction of alcohol and opioid signaling in human neurons carrying allelic variants for MOR N40 or D40 at the cellular and synaptic level. Using the novel induced pluripotent stem (iPS) cell and induced neuron (iN) cell techniques, it is possible to elucidate how the N40D gene variant alters alcohol reward signaling in human neurons. Using iPS cell lines from multiple individuals carrying homozygous MOR D40 or N40 alleles, human neurons of excitatory, inhibitory, and dopaminergic subtypes will be derived using the iN cell technology. To characterize the functional impact of this SNP, biochemical and morphological analyses examining changes in MOR expression, localization/trafficking, and degrees of glycosylation will be conducted using western blotting and immunohistochemistry. To give insight into interactions between alcohol exposure and MOR activation, presynaptic and dendritic markers will be analyzed to quantify synapse numbers and dendritic branching in alcohol-naïve, 1-day and 7-day alcohol and/or DAMGO treated iNs. I will also use electrophysiology to elucidate how the MOR N40 or D40 genotype differentially affects the more detailed functional parameters of synaptic transmission in iNs with or without alcohol and/or MOR activation. Our system using iPS cells carrying human gene variants provides the unique opportunity to dissect the interactions of alcohol exposure and MOR activation on synapse functionality in human neurons. As the long-term goal of this project is to understand the effect of human gene variants on alcohol sensitivity and synaptic function, results of this research will likely provide novel information about how to devise therapies for a heterogeneous group of AUDs.

 描述（由适用提供）：当今社会对酒精使用障碍（AUD）施加了巨大的经济和社会燃烧。现在，AUDS被理解为一种慢性神经系统障碍，其特征是对酒精的身体依赖超出自愿控制。虽然环境因素有助于饮酒，但最近的遗传研究表明，酒精中毒的遗传力约为50％，这表明常见的遗传变化对酒精中毒的病因显着贡献。但是，对酒精奖励信号传导和酒精奖励的突触机制的主要目标知之甚少。最近的遗传研究已经确定了患者酒精中毒相关的基因变异，其中一种是腺嘌呤的单个核肽多态性（SNP）与人OPRM1的位置118的鸟嘌呤，这是一个编码MU阿片类受体（MOR）的基因。这会导致40号天冬氨酸（d）中天冬酰胺（n）的非同义替代。尽管异源表达系统和动物研究表明受体运输和配体结合亲和力改变了，但没有确定N40D SNP的基本细胞和分子机制在人类神经元背景下的基本细胞和分子机制。为了更好地了解该SNP的功能后果，这项研究的目的是研究携带MOR N40或D40在细胞和突触水平上的携带等位基因变异的人类神经元中酒精和阿片类药物信号的相互作用。使用新型诱导的多能茎（IPS）细胞和诱导的神经元（IN）细胞技术，有可能阐明N40D基因变体如何改变人类神经元中的酒精奖励信号传导。使用来自多个个体的IPS细胞系，将使用IN IN细胞技术得出兴奋性，抑制性和多巴胺能亚型的人类兴奋性，抑制性和多巴胺能亚型的神经元。为了表征该SNP的功能影响，将使用蛋白质印迹和免疫组织化学进行生化和形态学分析，研究MOR表达，定位/运输以及糖基化程度的变化。为了深入了解酒精暴露与MOR激活之间的相互作用，将分析突触前和树突状标记，以量化酒精，1天和7天的酒精和/或Damgo处理的INS中的突触数和树突分支。我还将使用电生理学来阐明MOR N40或D40基因型如何不同地影响具有或没有酒精和/或MOR激活的INS中突触传播的更详细的功能参数。我们使用携带人类基因变异的IPS细胞的系统为剖析了酒精暴露和MOR激活在人类神经元中突触功能的相互作用的独特机会。由于该项目的长期目标是了解人类基因变异对酒精敏感性和突触功能的影响，因此这项研究的结果可能会提供有关如何为异构AUDS设计疗法的新信息。