A Novel Pharmacotherapy for Alcoholism: Evaluation of Reward, Aversion, Compulsivity, Withdrawal & Reinstatement

项目摘要 酒精使用障碍（AUD）是一种非常常见和严重的疾病， 许多躯体疾病，过早死亡，个人和人际困难，以及直接和间接的 社会的经济成本。虽然有几种有效的药物治疗AUD， 有限的有效性增加了AUD对社会的持续负担。更好地实现这一目标的主要障碍 AUD的治疗是缺乏对相关基本分子机制的理解， 酒精对大脑的急性和慢性影响我们实验室正在进行的工作表明， 而药理学上对Glycosidase 1（GLO 1）的操作可以减少小鼠的乙醇饮用量 还有老鼠GLO 1是一种进化上保守的酶，代谢甲基乙二醛（MG），这是一个非- 糖酵解的酶副产物，因此存在于所有动物细胞中。GLO 1的转基因过表达 降低大脑中MG（GLO 1的底物）。反过来，直接给予MG，基因敲除 Glo 1或Glo 1的药理学抑制增加脑中MG浓度。我们发现MG是 GABA-A受体的选择性激动剂。最近，我们发现GLO 1和MG调节乙醇 饮酒行为，我们假设这是由于MG对GABA-A受体的影响。 在这份补充申请中，我们正在扩展由母项目R 01资助的研究， 新的实验路线。具体来说，我们正在探索乙醇可能直接转化为 在与乙醇的急性效应一致的时间尺度上转化为MG。这种令人兴奋的可能性提出了 乙醇对GABA-A受体的一些影响可能是由于MG（而不是 乙醇），直接影响GABA-A信号传导。我们将测试这种可能性，通过测量MG水平在不同的 乙醇给药后的时间点。在随后的实验中，我们将探索这种可能性， 乙醇通过给药直接转化为MG（与以间接方式改变MG相反）， 氢原子被氘取代的稳定标记乙醇。我们会用质量- 规范，以评估MG水平，这将使我们能够区分未标记的MG与MG标记的一个或多个 氘原子这些研究有可能极大地提高我们对 乙醇的作用。还介绍了指导计划;主要目标是让受训者做好进入 博士程序.

Project summary Alcohol use disorder (AUD) is an extremely common and serious condition that is associated with numerous somatic diseases, early mortality, personal and interpersonal hardship, and direct and indirect economic costs to society. Although there are several effective pharmacological treatments for AUD, their limited effectiveness contributes to the ongoing burden of AUD on society. A major impediment to better treatments for AUD is the lack of understanding surrounding fundamental molecular mechanisms associated with alcohol’s acute and chronic effects on the brain. Ongoing work in our lab has demonstrated that genetic and pharmacological manipulation of the enzyme Glyoxalase 1 (GLO1) can reduce ethanol drinking in mice and rats. GLO1 is an evolutionarily conserved enzyme that metabolizes methylglyoxal (MG), which is a non- enzymatic side product of glycolysis and is thus present in all animal cells. Transgenic overexpression of GLO1 decreases MG (GLO1’s substrate) in the brain. Reciprocally, direct administration of MG, genetic knockdown of Glo1 or pharmacological inhibition of GLO1 increase MG concentrations in the brain. We showed that MG is a selective agonist at GABA-A receptors. More recently, we discovered that GLO1 and MG modulate ethanol drinking behavior, which we hypothesize is due to MG’s effects at GABA-A receptors. In this supplement request, we are extending on the studies funded by the parent R01 to pursue a new line of experimentation. Specifically, we are exploring the possibility that ethanol may be directly converted into MG on a timescale that is consistent with the acute effects of ethanol. This exciting possibility raises the tantalizing possibility that some of ethanol’s effects on GABA-A receptors could be due to MG (rather than ethanol), directly influencing GABA-A signaling. We will test this possibility by measuring MG levels at various time points following administration of ethanol. In subsequent experiments, we will explore the possibility that ethanol is directly converted into MG (as opposed to altering MG in an indirect manner) by administering stable-labeled ethanol in which the hydrogen atoms have been replaced with deuterium. We will use mass- spec to assess MG levels, which will allow us to distinguish unlabeled MG from MG labeled with one or more deuterium atoms. These studies have the potential to greatly enhance our mechanistic understanding of ethanol’s actions. The mentoring plan is also described; the primary goal is to prepare the trainee to enter a Ph.D. program.