Deciphering the neural basis of alcohol use disorders using human and mouse neurons

利用人类和小鼠神经元破译酒精使用障碍的神经基础

• 批准号: 9029804
• 负责人: ZHIPING P. PANG
• 金额: $ 34.69万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-05 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9029804
• 关键词: Acute; Address; Affect; Agonist; Alcohol abuse; Alcoholism; Alcohols; Amino Acid Substitution; Asparagine; Aspartate; Behavior; Biochemical; Biochemistry; Bioinformatics; Cell physiology; Chronic; Consumption; Data; Defect; Development; Enkephalins; Ethanol; Ethnic group; Genes; Genetic study; Goals; Health; High Prevalence; Human; Human Genetics; Individual; Label; Mediating; Methods; Microfabrication; Microfluidics; Midbrain structure; Modeling; Molecular; Mus; N-Glycosylation Site; Naltrexone; Nervous System Physiology; Neuraxis; Neurobiology; Neurons; Opioid; Opioid Receptor; Physiology; Receptor Activation; Receptor Signaling; Recycling; Regulation; Research; Rewards; Signal Transduction; Single Nucleotide Polymorphism; Slice; Specific qualifier value; Stem cells; Synapses; Synaptic Transmission; System; Technology; Testing; Therapeutic Effect; Variant; Ventral Tegmental Area; Work; addiction; alcohol effect; alcohol sensitivity; alcohol use disorder; base; cell type; design; dopamine system; dopaminergic neuron; drinking behavior; effective therapy; endogenous opioids; genetic variant; innovation; insight; mouse model; mu opioid receptors; novel; problem drinker; protein transport; public health relevance; receptor; receptor function; relating to nervous system; socioeconomics; stem cell biology; synaptic function; trafficking

 DESCRIPTION (provided by applicant): Alcoholism is a serious health and socioeconomic problem in the U.S. Understanding how alcohol produces reward, motivates further consumption, and eventually leads to addiction is necessary to design effective treatments for alcohol use disorders (AUDs). Synaptic transmission mediates all of these behaviors, however, little is known about the effect of alcohol on synaptic transmission in the context of human neurons. The single nucleotide polymorphism (SNP) rs1799971 (OPRM1 A118G) produces a non-synonymous amino acid substitution in the mu-opioid receptor (MOR), in which Asparagine 40 (MOR N40) is replaced with Aspartate (MOR D40), and is associated with AUDs in specific ethnic groups. Importantly, Naltrexone, a nonselective MOR antagonist, has potent therapeutic effects in alcoholic individuals with MOR D40. We generated human neurons from 7 subjects carrying either homozygous MOR N40 or MOR D40. Our preliminary data suggest that human neurons carrying D40 show defective re-sensitization after MOR activation by DAMGO ([D-Ala2, NMe-Phe4, Gly-ol5]-enkephalin), suggesting defective trafficking of MORs. Supporting this, bioinformatic analyses and mouse models of human MOR N40D suggest that D40 disrupts an N-glycosylation site on MOR. However, the mechanism by which MOR protein trafficking defects affect the interaction between ethanol and opioids is not known. It is particularly important to reveal the molecular mechanisms underlying the function of N40D MOR variants in their native neuronal context because previous studies performed in heterologous systems have revealed inconclusive and confusing results. Moreover, a species-specific trafficking mechanism of MORs has been suggested. The objective of this proposal is to study the impact of alcohol and opioid signaling in both mouse and human neurons carrying both the N40 and D40 MOR allelic variants, focusing on the synaptic mechanisms that likely underlie behavior. The central hypothesis is that defective D40 MOR trafficking results in an altered effect of the interaction between alcohol and opioids on synaptic function in the reward neurocircuitry. We will first examine the effect of alcohol on synaptic function in a defined neurocircuitry composed of human neurons carrying these gene variants. Next, we will use a mouse model of human N40D, and study the synaptic mechanism in the reward neurocircuitry, i.e. the midbrain ventral tegmental area (VTA), in relation to MOR function and ethanol. The proposed research is innovative, because we will combine recent developments in stem cell biology, the state-of-the-art synaptic physiology, and novel microfabrication technologies to directly probe the impact of alcohol and opioid signaling on synaptic function in both mouse and human neuronal networks carrying OPRM1 gene variants. We expect to unravel a species- and cell type-specific mechanism of MOR N40D variants that may provide novel information for understanding AUDs.

 描述（由适用提供）：酒精中毒是美国的严重健康和社会经济问题，了解酒精如何产生奖励，激励进一步的消费以及最终导致成瘾是为了设计有效的酒精使用障碍治疗（AUDS）所必需的。突触传播介导了所有这些行为，但是，对于人类神经元中酒精对合成传播的影响知之甚少。单个核苷酸多态性（SNP）rs1799971（OPRM1 A118G）在MU-Apioid受体（MOR）中产生非同义的氨基酸取代，其中天质40（MOR N40）与天冬氨酸（MOR D40）相关联，并与特定元素相关联。重要的是，非选择性MOR拮抗剂Naltrexone对MOR D40的酒精个体具有潜在的治疗作用。我们从携带纯合MOR N40或MOR D40的7名受试者中产生了人类神经元。我们的初步数据表明，携带D40的人神经元在Damgo（[D-Ala2，NME-Phe4，Gly-Ol5] -enkephalin）激活后表现出有缺陷的重新敏化，这表明对MOR的运输有缺陷。在支持这一点的情况下，人类MOR N40D的生物信息学分析和小鼠模型表明，D40破坏了MOR上的N-糖基化位点。但是，尚不清楚MOR蛋白运输缺陷影响乙醇与阿片类药物之间相互作用的机制。揭示N40D MOR变体在其天然神经元背景下功能功能的基础的分子机制尤其重要，因为先前在异源系统中进行的研究揭示了不确定且令人困惑的结果。此外，MOR的物种特异性运输机制我们将首先研究该提案的目的是研究酒精和阿片类药物信号在携带N40和D40 MOR等位基因变体的小鼠和人类神经元中的影响，重点是侧重于可能行为不足的合成机制。中心假设是，有缺陷的D40 MOR运输会导致酒精与阿片类药物之间相互作用对奖励神经记录中突触功能的影响改变。我们将首先检查酒精对由携带这些基因变异的人神经元组成的定义神经循环中突触功能的影响。接下来，我们将使用人类N40D的小鼠模型，并研究奖励神经循环中的突触机制，即中脑腹侧盖区域（VTA），都与MOR功能和乙醇有关。拟议的研究具有创新性，因为我们将结合干细胞生物学的最新发展，最新的合成生理学以及新型的微生物制作技术，以直接探测酒精和阿片类药物信号对携带OPRM1基因变异体的小鼠和人类神经元网络中突触功能的影响。我们期望揭开MOR N40D变体的物种和细胞类型特异性机制，这些机制可能提供新的信息以理解AUDS。