Traumatic stress and binge drinking as risk factors for excessive alcohol intake

创伤后应激障碍（PTSD）被归类为创伤或压力相关的障碍，证据表明， 证实了创伤后应激障碍和酒精使用障碍（AUD）之间的联系。尽管 创伤后应激障碍发病率的异质性，创伤后应激障碍和AUD的高共病率在退伍军人中报道， 人口此外，与PTSD/AUD共病的退伍军人的症状加重负相关， 影响康复预后和有效的治疗策略。我们和其他人发现， 作为PTSD动物模型的捕食者气味应激（PS）显著增加了焦虑相关行为 以及随后啮齿动物摄入酒精（乙醇）。最重要的是，我们观察到这些 行为反应，如在人类PTSD/AUD患者中观察到的。此外，既往酗酒（BD） 在应激“敏感”小鼠中，间歇性PS后， 具体地，“PS敏感”雄性和雌性C57 BL/6 J（B6）小鼠表现出乙醇摄入量增加> 70%（> 20%）。 2.5 g/kg），而乙醇摄入量在“弹性”小鼠中没有变化。B6小鼠中的其他研究 确定BD单独在细胞核中的信号通路中产生性差异变化 乙醇摄入量增加后，BD和PS产生了性别差异， 前额叶皮层和海马中与应激轴和神经类固醇合成相关的蛋白质（NAC不 在这项研究中发现）。结合单独的行为和表观遗传适应的证据 潜在的压力“敏感性”和“弹性”，我们假设：（1）不同的行为和分子 适应赋予了BD和PS对后期乙醇的增强作用的“敏感性”和“弹性” 饮酒，（2）BD诱导DNA甲基化（DNAm）信号和相关的基因变化 表达改变了对PS的适应性;（3）潜在的机制存在性别差异 在B6小鼠中。目标1研究将确定是否“敏感性”与“弹性”PS增强饮酒后， 既往BD与焦虑相关行为、心率（HR）和/或乙醇奖励改变相关， 这些关系中是否存在性别差异。将对四组进行试验：第1组（BD+4周23 h 间歇性PS的乙醇饮用;分为“敏感”和“弹性”亚组）;组2 （仅间歇性PS）;第3组（未经治疗）;第4组（仅BD）。PS后饮用后（第1组），最终PS （第2组）或同等时间点（第3组和第4组），HR和焦虑相关行为将在一个 研究和条件性位置偏好的研究将在另一项研究中进行测试。AIM 2将识别表观遗传信号 和NAC中的相关基因表达谱，赋予PS增强的“敏感性”和“弹性”。 在BD之前喝酒。分组与目标1相同，但在最终BD后，PS后对小鼠实施安乐死 最后一个PS我们预测，全基因组DNA分析将确定差异甲基化 胞嘧啶和区域映射到新的和大多数不同的，但一些共享的基因，调控区， 男性和女性的网络，将与“敏感性”和“弹性”PS增强 喝酒DNAm和基因表达关联分析将与行为结果整合， 目标1为功能分析确定目标。目的3将确认所选基因的功能作用（目的 2)重要的“敏感性”和“弹性”PS增强饮酒和相关行为。蛋白水平 将通过蛋白质印迹进行确认。药理学研究将操纵候选分子， 确定对PS增强的饮酒、BD、焦虑和HR的影响，从鉴定的3种分子开始 在目前的融资。阐明赋予“敏感性”的生物和分子机制， BD和创伤性应激反应后对乙醇饮用量增加的“弹性”可以指导 AUD和PTSD的药物治疗策略，男性和女性可能存在差异 并代表了有效治疗退伍军人的重要重点领域。

Post-traumatic stress disorder (PTSD) is classified as a trauma- or stressor-related disorder, and evidence confirms an association between PTSD and the development of an alcohol use disorder (AUD). Despite heterogeneity in the incidence of PTSD, high comorbidity of PTSD and AUD are reported in the veteran population. Furthermore, the exacerbation of symptoms in veterans with comorbid PTSD/AUD negatively influences recovery prognosis and effective therapeutic strategies. We and others found that exposure to predator odor stress (PS), used as an animal model of PTSD, significantly increases anxiety-related behaviors and subsequent alcohol (ethanol) intake in rodents. Most importantly, we observed heterogeneity in these behavioral responses, as is observed in human PTSD/AUD patients. Additionally, prior binge drinking (BD) produced a greater enhancement in ethanol drinking following intermittent PS in stress “sensitive” mice. Specifically, “PS-sensitive” male and female C57BL/6J (B6) mice exhibited > 70% increase in ethanol intake (> 2.5 g/kg over baseline), while ethanol intake was unchanged in “resilient” mice. Additional studies in B6 mice determined that BD alone produced sexually divergent changes in signaling pathways in the nucleus accumbens (NAC), while the enhanced ethanol intake following BD and PS produced sex differences in proteins related to the stress axis and neurosteroid synthesis in prefrontal cortex and hippocampus (NAC not examined in that study). In conjunction with evidence for separate behavioral and epigenetic adaptations underlying stress “sensitivity” and “resilience”, we hypothesize that: (1) distinct behavioral and molecular adaptations confer “sensitivity” and “resilience” to the enhancing effect of BD and PS on later ethanol drinking, (2) BD induces DNA methylation (DNAm) signals and associated changes in gene expression that alter adaptability to PS, and (3) sex differences exist in the underlying mechanisms in B6 mice. Aim 1 studies will determine whether “sensitivity” vs “resilience” to PS-enhanced drinking after prior BD is associated with altered anxiety-related behavior, heart rate (HR), and/or ethanol reward and whether sex differences exist in these relationships. Four groups will be tested: Group 1 (BD+4 weeks 23 h ethanol drinking with intermittent PS; divided into “sensitive” and “resilient” subgroups); Group 2 (intermittent PS only); Group 3 (naïve); Group 4 (BD only). After the post-PS drinking (Group 1), final PS (Group 2), or equivalent time point (Groups 3 & 4), HR and anxiety-related behavior will be assessed in one study and conditioned place preference will be tested in a separate study. Aim 2 will identify epigenetic signals and linked gene expression profiles in the NAC that confer “sensitivity” and “resilience” to PS-enhanced drinking after prior BD. Groups will be as in Aim 1, but mice will be euthanized following final BD, post-PS drinking, or final PS. We predict that the genome-wide DNAm analysis will identify differentially methylated cytosines and regions mapping to novel and mostly distinct but some shared genes, regulatory regions, and networks in males and females that will be associated with “sensitivity” and “resilience” to PS-enhanced drinking. DNAm and gene expression association analysis will be integrated with behavioral outcomes from Aim 1 to pinpoint targets for functional analysis. Aim 3 will confirm the functional role of selected genes (Aim 2) important for “sensitivity” and “resilience” to PS-enhanced drinking and associated behaviors. Protein levels will be confirmed by Western blots. Pharmacological studies will manipulate candidate molecules and determine the effect on PS-enhanced drinking, BD, anxiety, and HR, beginning with 3 molecules identified during current funding. Elucidating biological and molecular mechanisms that confer “sensitivity” and “resilience” to escalated ethanol drinking following BD and traumatic stress can guide pharmacological treatment strategies for AUD and PTSD, which likely differ in males and females and represent important areas of focus in the effective treatment of our veterans.