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(申请人提供)：酒精中毒在美国是一个严重的健康和社会经济问题。了解酒精如何产生奖励，刺激进一步消费，并最终导致成瘾，对于设计有效的酒精使用障碍(AUD)治疗方法是必要的。突触传递介导了所有这些行为，然而，关于酒精对人类神经元突触传递的影响，人们知之甚少。单核苷酸多态(SNP)rs1799971(OPRM1 A118G)在阿片受体(MOR)上产生非同义氨基酸替换，其中天冬酰胺40(MOR N40)被天冬氨酸(MOR D40)取代，并与特定种族的AUDS相关。重要的是，非选择性MOR拮抗剂纳曲酮对患有MOR D40的酗酒者有很强的治疗作用。我们从7名携带MORN40或MORD40纯合子的受试者中产生了人类神经元。我们的初步数据表明，携带D40的人神经元在被DAMGO([D-Ala2，NME-Phe4，Gly-ol5]-Enkephalin)激活MOR后表现出缺陷的再敏化，提示MORS的缺陷运输。支持这一点的是，生物信息学分析和人类MOR N40D的小鼠模型表明，D40破坏了MOR上的一个N-糖基化位点。然而，MOR蛋白转运缺陷影响乙醇和阿片类药物相互作用的机制尚不清楚。尤其重要的是揭示N40D MOR变异体在其天然神经元环境中的潜在功能的分子机制，因为先前在异源系统中进行的研究揭示了不确定和令人困惑的结果。此外，还提出了一种特定物种的MORS运输机制。这项建议的目的是研究酒精和阿片类药物信号对携带N40和D40MOR等位基因变体的小鼠和人类神经元的影响，重点关注可能构成行为基础的突触机制。中心假说是，缺陷的D40MOR转运导致酒精和阿片类药物之间的相互作用对奖赏神经回路中突触功能的影响发生了改变。我们将首先研究酒精对携带这些基因变体的人类神经元组成的明确神经回路中突触功能的影响。接下来，我们将使用人类N40D的小鼠模型，研究中脑腹侧被盖区(VTA)的奖赏神经回路中的突触机制与MOR功能和乙醇的关系。这项拟议的研究具有创新性，因为我们将结合干细胞生物学、最先进的突触生理学和新的微制造技术的最新发展，直接探讨酒精和阿片信号对携带OPRM1基因变体的小鼠和人类神经网络突触功能的影响。我们期望揭开MOR N40D变种的物种和细胞类型特异性的机制，这可能为理解AUDS提供新的信息。