Progesterone-Induced Gene Expression Changes and Risk of Relapse to Cocaine Use

描述（由申请人提供）：这项初步研究将持续2年，将评估黄体酮治疗诱导的基因表达变化与压力诱导的渴望、负面情绪和生理学的实验室测量之间的关系，这些测量先前被证明可以预测可卡因成瘾者早期复发的风险。该项目的长期目标是开发新的分子工具，可用于预测和监测对可卡因依赖的特定治疗的反应。初步研究表明，男性和女性在对可卡因的主观和生理反应以及与复发风险相关的压力或药物线索刺激方面存在差异。男性和女性对可卡因成瘾的各种药理学治疗的临床反应也不同。性腺类固醇水平的差异可能是部分或全部这些差异的基础。最近，一些小规模的实验室调查和临床试验研究了黄体酮作为可卡因成瘾的可能治疗方法。动物模型研究表明，黄体酮受体在雌雄动物的大脑中广泛表达，并调节大量基因的表达。此外，黄体酮已被证明可以激活神经元中的ERK/CREB/bcl-2和Akt第二信使信号通路。在几个实验模型中，这些作用被认为是黄体酮神经保护作用的基础。这些途径的激活可能与应激诱导的基因表达模式的作用相反，这可能介导可卡因依赖个体复发的风险增加。在这项研究中，我们将招募（n=60； 30男30女）寻求治疗的可卡因依赖受试者，他们目前正在被招募参加一项由nida资助的关于黄体酮对压力和线索诱导的可卡因渴望和复发易感性的影响的研究。在这项研究中，受试者被随机分配接受为期五天的黄体酮（400毫克/天）或安慰剂治疗，并接受简短的引导意象治疗，包括个性化的压力、药物提示或中性放松的情况（每次治疗一个意象条件）。在拟议的研究中，我们将从治疗前和黄体酮或安慰剂治疗第5天收集的血液中分离RNA。基因表达水平将通过使用Illumina Sentrix Beadchip （Human-6v2）阵列（与我们初步研究中使用的平台相同）的微阵列杂交来评估。基因表达水平将与药物渴望、焦虑和情绪评级，以及行为困扰反应、心率、血压和每次治疗期间评估的唾液皮质醇测量值相关。选择感兴趣的基因的差异表达将使用定量实时反转录聚合酶链反应（qRT-PCR）确认。具体目的包括：(1)比较黄体酮诱导的可卡因依赖男女外周血基因表达的变化；(2)识别特定的黄体酮调控转录本，其表达与应激或药物线索相关刺激的主观反应相关。

DESCRIPTION (provided by applicant): This preliminary study, which will be of 2 years duration, will assess the relationship between changes in gene-expression induced by treatment with progesterone and laboratory measures of stress-induced craving, negative emotion, and physiology previously shown to predict risk for early relapse in cocaine addicted subjects. The long-term goal of this project is to develop novel molecular tools that can be used to predict and monitor the response to specific treatments for cocaine dependence. Preliminary studies have shown that men and women differ in their subjective and physiological responses to cocaine, as well as stress or drug-cue related stimuli associated with risk for relapse. Men and women also differ in their clinical response to various proposed pharmacological treatments for cocaine addiction. Differences in the level of gonadal steroids may underlie some or all of these differences. Recently, a number of small scale laboratory investigations and clinical trials have investigated progesterone as a possible treatment for cocaine addiction. Studies in animal models have shown that progesterone receptors are widely expressed in the brain and regulate the expression of a large number of genes in the brains of both female and male animals. In addition, progesterone has been shown to activate both the ERK/CREB/bcl-2 and Akt second messenger signaling pathways in neurons. These effects have been proposed to underlie the neuroprotective actions of progesterone in several experimental models. Activation of these pathways may oppose the actions of stress-induced patterns of gene expression, which could mediate increased risk for relapse in cocaine dependent individuals. In this study, we will recruit (n=60; 30 male and 30 female) treatment-seeking cocaine-dependent subjects who are currently being recruited to take part in a NIDA-funded study of progesterone effects on stress and cue- induced cocaine craving and relapse susceptibility. In that study, subjects are randomly assigned to receive either progesterone (400mg/day) or placebo for a period of five days and undergo brief guided imagery sessions involving personalized stressful, drug-cue or neutral-relaxing situations (one imagery condition per session). In the proposed study, we will isolate RNA from blood collected prior to treatment and on day 5 of treatment with either progesterone or placebo. Gene-expression levels will be assessed by microarray hybridization using Illumina Sentrix Beadchip (Human-6v2) arrays (the same platform used in our preliminary studies). Gene-expression levels will be correlated with drug craving, anxiety and emotion ratings, as well as behavioral distress responses, heart rate, blood pressure, and salivary cortisol measures assessed during each session. Differential expression of selected genes of interested will be confirmed using Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Specific aims include: (1) comparing progesterone-induced changes in peripheral blood gene-expression in cocaine-dependent men and women; and (2) identifying specific progesterone-regulated transcripts whose expression is correlated with the subjective response to stress or drug-cue related stimuli. PUBLIC HEALTH RELEVANCE: These studies will allow us to better understand the molecular basis for individual differences in the response to progesterone in cocaine dependent men and women. Better understanding of these differences will allow us to identify those individuals (both men and women) most likely to benefit from treatment with progesterone. In addition, better understanding of the mechanism of action of progesterone at the genomic level may help to guide the development of novel hormonally based pharmacotherapies for the treatment of cocaine dependence.