Plasma TNFSF10B levels associated with acamprosate treatment response in patients with alcohol use disorder. Plasma TNFSF10B levels associated with acamprosate treatment response in patients with alcohol use disorder

Acamprosate is an anti-craving drug used in alcohol use disorder (AUD) pharmacotherapy. However, only a subset of patients achieves optimal treatment outcomes. The identification of predictive biomarkers of acamprosate treatment response in patients with AUD would be a substantial advance in addiction medicine. We designed this study to use proteomics data as a quantitative biological trait as a step toward identifying inflammatory modulators that might be associated with acamprosate treatment outcomes. The NIAAA-funded Mayo Clinic Center for the Individualized Treatment of Alcoholism study had previously recruited 442 AUD patients who received three months of acamprosate treatment. However, only 267 subjects returned for the 3-month follow-up visit and, as a result, had treatment outcome information available. Baseline alcohol craving intensity was the most significant predictor of acamprosate treatment outcomes. Baseline plasma TNFSF10 concentration was associated with alcohol craving intensity and variation in acamprosate treatment outcomes among AUD patients. We also performed RNA sequencing using baseline peripheral blood mononuclear cells from AUD patients with known acamprosate treatment outcomes which revealed that inflammation-related pathways were highly associated with relapse to alcohol use during the three months of acamprosate treatment. These observations represent an important step toward advancing our understanding of the pathophysiology of AUD and molecular mechanisms associated with acamprosate treatment response. In conclusion, applying omics-based approaches may be a practical approach for identifying biologic markers that could potentially predict alcohol craving intensity and acamprosate treatment response. Overall design: Peripheral blood mononuclear cells (PBMCs) samples were isolated from 15 ml of whole blood before acamprosate treatment using Ficoll density gradient centrifugation. We lysed the cells in Trizol and extracted total RNA using the RNeasy mini kit (Qiagen, Valencia, CA, USA). The RNA integrity numbers (RIN) were between 8.5-9.2 for the PBMC samples (relapse: n=27, non-relapse: n=26). RNA-seq experiments were conducted by GENEWIZ using an Illumina HiSeq 4000 with eight samples in each lane using 100bp paired end index reads. Fastq files containing paired RNA-Seq reads were aligned with STAR (23) against the UCSC human reference genome (hg19). We performed RNA-seq differential expression analysis using the DESeq2 package with default parameters.

阿坎酸（acamprosate）是一种用于酒精使用障碍（alcohol use disorder, AUD）药物治疗的抗渴求药物。然而，仅有部分患者可获得最佳治疗结局。识别酒精使用障碍患者阿坎酸治疗应答的预测生物标志物，将是成瘾医学领域的重大进展。本研究旨在以蛋白质组学数据作为定量生物学性状，逐步探索可能与阿坎酸治疗效果相关的炎症调节因子。
由美国国家酒精滥用与酒精中毒研究所（NIAAA）资助的梅奥诊所酒精成瘾个体化治疗中心研究此前招募了442名接受3个月阿坎酸治疗的酒精使用障碍患者，但仅有267名受试者完成了3个月随访，因此可获取其治疗结局信息。基线酒精渴求强度是阿坎酸治疗结局最显著的预测因子。基线血浆TNFSF10浓度与酒精使用障碍患者的酒精渴求强度及阿坎酸治疗结局的变异度相关。
我们还对已知阿坎酸治疗结局的酒精使用障碍患者的基线外周血单个核细胞进行了RNA测序（RNA-seq），结果显示，在阿坎酸治疗的3个月期间，炎症相关通路与酒精复吸行为高度相关。上述发现为加深我们对酒精使用障碍病理生理学以及阿坎酸治疗应答相关分子机制的理解迈出了重要一步。综上，应用组学相关方法或许是识别可潜在预测酒精渴求强度及阿坎酸治疗应答的生物标志物的可行途径。
总体实验设计：在阿坎酸治疗前，通过Ficoll密度梯度离心法从15ml全血中分离外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）。我们使用Trizol裂解细胞，并采用RNeasy迷你试剂盒（Qiagen，美国加利福尼亚州巴伦西亚）提取总RNA。外周血单个核细胞样本的RNA完整性数（RNA integrity number, RIN）介于8.5至9.2之间（复吸组：n=27，非复吸组：n=26）。RNA测序实验由GENEWIZ公司完成，使用Illumina HiSeq 4000平台，每个泳道搭载8个样本，采用100bp双端索引读长。包含双端RNA测序读段的Fastq文件通过STAR（23）比对至UCSC人类参考基因组（hg19）。我们使用DESeq2软件包以默认参数进行RNA测序差异表达分析。