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

项目总结 酒精使用障碍(AUD)是一种非常常见和严重的疾病，与 许多躯体疾病、早期死亡、个人和人际关系困难以及直接和间接 给社会带来的经济代价。虽然有几种有效的药物治疗AUD，但他们的 有限的效力助长了澳元对社会的持续负担。做得更好的主要障碍 AUD的治疗方法是缺乏对相关基本分子机制的了解 酒精对大脑的急性和慢性影响。我们实验室正在进行的研究表明，基因 而对乙二酸酶1(GLO1)的药理操作可以减少小鼠饮酒 还有老鼠。GLO1是一种进化上保守的酶，它代谢甲基乙醛(MG)，后者是一种非 糖酵解的副产物，因此存在于所有动物细胞中。GLO1的转基因过表达 降低脑组织中的MG(GLO1‘S底物)。反过来，直接给药MG，基因敲除 Glo1的抑制或GLO1的药物抑制可增加脑组织中MG的浓度。我们展示了MG是 GABA-A受体的选择性激动剂。最近，我们发现GLO1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。 在这项补充申请中，我们延长了由家长R01资助的研究，以进行 新的实验路线。具体地说，我们正在探索将乙醇直接转化为 进入MG的时间刻度与乙醇的急性影响一致。这种令人兴奋的可能性提高了 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信号转导。我们将通过测量不同的MG水平来测试这种可能性 注射乙醇后的时间点。在随后的实验中，我们将探索是否有可能 乙醇直接转化为MG(与间接改变MG相反)，方法是 一种稳定标记的乙醇，其中的氢原子已被重氢取代。我们将使用质量- 规范来评估MG水平，这将使我们能够区分未标记的MG和带有一个或多个标记的MG 重氢原子。这些研究有可能极大地提高我们对 乙醇的作用。还描述了指导计划；主要目标是为学员进入 博士学位课程。

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.