Leveraging biomarkers for personalized treatment of alcohol use disorder comorbid with PTSD

总结 血液生物标志物核心（BBC）由Silvia Fossati博士领导。对于项目1，所有人的血液和大脑样本 将在处死时收集小鼠以定量：1）经受所有酒精的动物中的分子生物标志物和/或 用两种剂量的托吡酯（TPM）或溶媒治疗后的PTSD样模型，以阐明 TPM作用，和2）用于酒精+PTSD动物模型与酒精+PTSD动物模型的神经生物学比较的生物标志物。 单独的PTSD样动物模型、单独的酒精模型、幼稚对照动物和有弹性的动物。项目 2，将在2个时间点（之前）从所有入组的随机临床试验（RCT）参与者中采集血液 随机化和在第12周），并处理以确定分子生物标志物、治疗性免疫缺陷的预测因子、免疫抑制剂和免疫抑制剂。 疗效和GRIK 1基因型，遵循我们小组开发的血液生物标志物分析最佳实践， 纽约大学科恩退伍军人中心使用Simoa和ELISA检测，我们将确定血液 以下分子的浓度：1）AUD、PTSD中兴奋性和抑制性氨基酸的改变 和PTSD+AUD和TPM（谷氨酸和GABA）的假设作用机制的中心; 2） 调节下丘脑-垂体-肾上腺（HPA）轴反应的肽和激素（促肾上腺皮质激素 释放因子（CRF）、皮质酮/皮质醇、神经肽Y和脑源性神经营养因子（BDNF））; 3)涉及神经炎症的生物标志物（IL 6、IL 10、IL 1 α、TNFα和胶质细胞酸性蛋白）;以及4） 细胞凋亡、氧化应激和线粒体功能障碍标志物（细胞色素C）。跨项目集成 将通过两个设计要素和三个分析策略来实现。集成设计元素包括： 1)项目1中接受酒精和/或创伤后应激障碍样模型的小鼠将被分层并随机分配至 项目2中的托吡酯/赋形剂和PTSD+AUD临床试验参与者将分层并随机分配至 托吡酯/安慰剂，创建平行设计以推进托吡酯作用机制的发现， 治疗反应的预测因子;和2）项目1中小鼠的所有血液生物标志物将在临床试验中确定。 项目2的试验参与者（皮质醇将取代皮质酮）。综合分析策略包括：1） 项目2中托吡酯/安慰剂治疗后的血液生物标志物水平将与血浆生物标志物相关 项目1中托吡酯/赋形剂治疗后酒精+PTSD模型中的水平; 2）血液生物标志物值 在项目2的临床试验参与者中获得的数据将与他们的项目2临床数据和fMRI相关， 来源于项目3的TMS/EEG和MRS标记物;以及3）基因表达和蛋白质表达标记物 在项目1中确定的小鼠PFC、杏仁核、扣带回、VTA和杏仁核中， 在项目3的临床试验参与者的相同脑区中获得的回路标记。此外，因为我们 将能够在人类中收集比小鼠更高的血量，我们将储存血浆，血清，全血， 用于未来系统生物学分析的DNA和RNA，包括基因组学、转录组学、DNA甲基化， 代谢组学和蛋白质组学。

Summary The Blood Biomarkers Core (BBC) is led by Dr. Silvia Fossati. For Project 1, blood and brain samples from all mice will be collected at sacrifice to quantify: 1) molecular biomarkers in animals subjected to all alcohol and/or PTSD-like models after treatment with two doses of topiramate (TPM) or vehicle to clarify the mechanisms of TPM action, and 2) biomarkers for the neurobiological comparison of animal models of alcohol+PTSD with PTSD-like animal models alone, alcohol models alone, naïve control animals, and resilient animals. For Project 2, blood will be collected from all enrolled randomized clinical trial (RCT) participants at 2 time points (before randomization and at week 12), and processed to determine molecular biomarkers, predictors of therapeutic efficacy and GRIK1 genotype, following best practices for blood biomarker analysis developed by our group at the NYU Cohen Veterans Center. Using Simoa and ELISA assays, we will determine the blood concentration of the following molecules: 1) excitatory and inhibitory amino acids altered in AUD, PTSD and PTSD+AUD and central to the hypothesized mechanisms of action of TPM (gluatamate and GABA); 2) peptides and hormones regulating responses of the hypothalamic–pituitary–adrenal (HPA) axis (corticotrophin releasing factor (CRF), corticosterone/cortisol, neuropeptide Y, and brain-derived neurotrophic factor (BDNF)); 3) biomarkers implicated in neuroinflammation (IL6, IL10, IL1α, TNFα, and glial fibrillary acidic protein); and 4) apoptosis, oxidative stress and mitochondrial dysfunction markers (cytochrome C). Integration across projects will be achieved by two design elements and three analytic strategies. Integrative Design Elements include: 1) mice subjected to alcohol and/or PTSD-like models in Project 1 will be stratified and randomized to topiramate/vehicle and PTSD+AUD clinical trial participants in Project 2 will be stratified and randomized to topiramate/placebo, creating parallel designs to advance discovery of mechanisms of topiramate action and predictors of treatment response; and 2) all blood biomarkers in mice in Project 1 will be ascertained in clinical trial participants for Project 2 (cortisol will replace corticosterone). Integrative Analytic Strategies include: 1) blood biomarkers levels after topiramate/placebo treatment in Project 2 will be related to plasma biomarkers levels in alcohol+PTSD models after topiramate/vehicle treatment in Project 1; 2) blood biomarker values obtained in clinical trial participants in Project 2 will be related to their Project 2 clinical data and to fMRI, TMS/EEG and MRS markers derived from Project 3; and 3) gene expression and protein expression markers ascertained in PFC, amygdala, cingulate, insula, VTA and accumbens in mice in Project 1 will be related to circuit markers obtained in the same brain regions in clinical trial participants in Project 3. Further, because we will be able to collect a higher blood volume in humans than mice, we will bank plasma, serum, whole blood, DNA, and RNA for future systems biology analyses, including genomics, transcriptomics, DNA methylation, metabolomics and proteomics.