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

项目摘要 酒精使用障碍（AUD）给我们的社会带来了巨大的负担。除了它们 心理和社会代价，据估计，澳元在这一年花费了美国经济2490亿美元 仅2010年。虽然已经有几种有效的行为和药物治疗方法， 迫切需要通过新机制发挥作用的新药物疗法。 我们最近发现，酶Glycosidase 1（GLO 1）的抑制剂可以减少自发性 小鼠饮用乙醇（McMurray等人，2017 a）。GLO 1是一种细胞溶质酶， 甲基乙二醛（MG）。MG是糖酵解的非酶促副产物，因此存在于所有 细胞因此，GLO 1活性与MG浓度呈负相关。我们之前已经证明， GLO 1的转基因过表达增加小鼠的焦虑样行为并降低MG 大脑中的浓度。反过来，我们表明，直接管理MG，遗传 Glo 1敲低或使用小分子抑制剂抑制Glo 1均降低焦虑样 行为和增加大脑中的MG浓度。我们发现即使是更高剂量的MG 运动抑制，共济失调和体温过低;综合这些数据表明，MG可能 通过GABA-A受体起作用。事实上，使用膜片钳程序，我们发现MG是一种 GABA-A受体的竞争性部分激动剂。我们还表明，MG是高度选择性的： 不激活GABA-B受体、其他配体门控离子通道或电压门控离子通道。更 最近，我们发现GLO 1抑制具有抗抑郁样作用，提示GLO 1 抑制剂可以治疗焦虑和抑郁，这两种疾病都与AUD共病。鉴于 GABA-A信号在乙醇作用中的重要性，我们推测GLO 1和MG可能 也调节乙醇相关的行为，这使我们研究GLO 1对乙醇的影响 喝酒这些研究表明，GLO 1的抑制减少了乙醇饮用，这是 本申请中提出的实验的基本原理。 我们建议进行研究，旨在了解为什么抑制GLO 1会减少自愿性 酒精饮料在目标1中，我们将使用颅内自刺激（ICSS）程序来检查 GLO 1的遗传和药理学操作对享乐和 小鼠对乙醇的快感缺乏反应。在目标2中，我们将使用条件位置偏好（CPP） 和条件性味觉厌恶（CTA）来研究GLO 1对味觉偏爱和厌恶的影响。 乙醇在小鼠体内最后，在目标3中，我们将使用慢性间歇性乙醇（CIE）程序， 检查Glo 1抑制剂对急性乙醇戒断、强迫样反应和对酒精依赖的影响。 乙醇和恢复乙醇寻求行为后，长期禁欲大鼠。

Project Summary Alcohol use disorders (AUD) place an enormous burden on our society. In addition to their psychological and social toll, it is estimated that AUDs cost the US economy $249 billion in the year 2010 alone. Although there are already several effective behavioral and pharmacological treatments, there is an urgent need for new pharmacotherapies that act via novel mechanisms. We have recently shown that inhibitors of the enzyme Glyoxalase 1 (GLO1) reduce voluntary ethanol drinking in mice (McMurray, et al 2017a). GLO1 is a cytosolic enzyme that metabolizes methylglyoxal (MG). MG is a non-enzymatic side product of glycolysis and is therefore present in all cells. Thus, GLO1 activity is inversely related to MG concentration. We have previously shown that transgenic overexpression of GLO1 increases anxiety-like behavior in mice and decreases MG concentrations in the brain. Reciprocally, we showed that direct administration of MG, genetic knockdown of Glo1 or inhibition of GLO1 using a small molecule inhibitor all decrease anxiety-like behavior and increase MG concentrations in brain. We found that even higher doses of MG produced locomotor depression, ataxia and hypothermia; taken together these data suggested that MG might be acting through GABA-A receptors. Indeed, using a patch clamp procedure, we found that MG is a competitive partial agonist at GABA-A receptors. We also showed that MG is highly selective: it does not activate GABA-B receptors, other ligand gated ion channels or voltage gated ion channels. More recently, we have shown that GLO1 inhibition has antidepressant-like effects, suggesting that GLO1 inhibitors might treat anxiety and depression, both of which are comorbid with AUD. Given the importance of GABA-A signaling in the effects of ethanol, we speculated that GLO1 and MG might also modulate ethanol-related behaviors, which led us to study the effects of GLO1 on ethanol drinking. Those studies showed that inhibition of GLO1 decreased ethanol drinking, which is the rationale for the experiments proposed in this application. We are proposing studies aimed at understanding why inhibition of GLO1 reduces voluntary ethanol drinking. In Aim 1 we will use the intracranial self-stimulation (ICSS) procedure to examine the acute and chronic effects of genetic and pharmacological manipulations of GLO1 on hedonic and anhedonic responses to ethanol in mice. In Aim 2 we will use conditioned place preference (CPP) and conditioned taste aversion (CTA) to study the effects of GLO1 on preference and aversion for ethanol in mice. Finally, in Aim 3, we will use the chronic intermittent ethanol (CIE) procedure to examine the effects of Glo1 inhibitors on acute ethanol withdrawal, compulsive-like responding for ethanol and reinstatement of ethanol seeking behavior after protracted abstinence in rats.