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

摘要 血液生物标志物核心(BBC)由西尔维亚·福萨蒂博士领导。对于项目1，所有人的血液和大脑样本 小鼠将在牺牲时被收集以量化：1)在所有酒精和/或 两种剂量托吡酯(TPM)或赋形剂治疗后的创伤后应激障碍模型 TPM作用，以及2)酒精+PTSD动物模型的神经生物学比较的生物标志物与 类似创伤后应激障碍的动物模型、单纯的酒精模型、天真的对照动物和有弹性的动物。对于项目 2、将在2个时间点(之前)从所有入选的随机临床试验(RCT)参与者中采集血液 随机化和第12周)，并进行处理以确定分子生物标记物，治疗的预测因素 有效性和GRIK1基因，遵循我们小组开发的血液生物标记物分析的最佳实践，网址为 纽约大学科恩退伍军人中心。使用SIMA和ELISA法，我们将检测血液 以下分子的浓度：1)AUD、PTSD中兴奋性和抑制性氨基酸的变化 和PTSD+AUD，是TPM(谷氨酸和GABA)假想作用机制的核心； 调节下丘脑-垂体-肾上腺(HPA)轴反应的多肽和激素(促肾上腺皮质激素 释放因子、皮质酮/皮质醇、神经肽Y、脑源性神经营养因子)； 3)与神经炎症有关的生物标志物(白介素6、白介素10、白介素1α、肿瘤坏死因子α和胶质纤维酸性蛋白)； 细胞凋亡、氧化应激和线粒体功能障碍标记物(细胞色素C)。跨项目集成 将通过两个设计元素和三个分析策略来实现。一体化设计元素包括： 1)项目1中的酒精和/或类似创伤后应激障碍模型的小鼠将被分层和随机分为 项目2中的托吡酯/Vehicle和PTSD+AUD临床试验参与者将被分层和随机分配到 托吡酯/安慰剂，创造平行设计以促进托吡酯作用机制的发现和 治疗反应的预测因素；以及2)项目1中小鼠的所有血液生物标记物将在临床上确定 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记物水平将与血浆生物标记物相关 项目1中托吡酯/赋形剂治疗后酒精+创伤后应激障碍模型的水平；2)血液生物标志物的值 在项目2的临床试验参与者中获得的将与他们的项目2的临床数据和功能磁共振相关， 来自项目3的TMS/EEG和MRS标记；以及3)基因表达和蛋白质表达标记 在项目1中在小鼠的前额叶、杏仁核、扣带核、岛叶、VTA和伏隔核中确定的将与 在项目3的临床试验参与者中，在相同的大脑区域中获得电路标记。此外，因为我们 将能够在人类体内采集比小鼠更高的血量，我们将储存血浆，血清，全血， DNA和RNA用于未来的系统生物学分析，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.