Transcriptomics and candidate gene/protein expression studies have indicated several biological processes modulated by methylphenidate (MPH), widely used in attention-deficit/hyperactivity disorder (ADHD) treatment. However, the lack of a differential proteomic profiling of MPH treatment limits the understanding of the most relevant mechanisms by which MPH exerts its pharmacological effects at the molecular level. Therefore, our aim is to investigate the MPH-induced proteomic alterations using an experimental design integrated with a pharmacogenomic analysis in a translational perspective. Proteomic analysis was performed using the cortices of Wistar-Kyoto rats, which were treated by gavage with MPH (2 mg/kg) or saline for two weeks (n = 6/group). After functional enrichment analysis of the differentially expressed proteins (DEP) in rats, the significant biological pathways were tested for association with MPH response in adults with ADHD (n = 189) using genome-wide data. Following MPH treatment in rats, 98 DEPs were found (P < 0.05 and FC < -1.0 or > 1.0). The functional enrichment analysis of the DEPs revealed 18 significant biological pathways (gene-sets) modulated by MPH, including some with recognized biological plausibility, such as those related to synaptic transmission. The pharmacogenomic analysis in the clinical sample evaluating these pathways revealed nominal associations for gene-sets related to neurotransmitter release and GABA transmission. Our results, which integrate proteomics and pharmacogenomics, revealed putative molecular effects of MPH on several biological processes, including oxidative stress, cellular respiration, and metabolism, and extended the results involving synaptic transmission pathways to a clinical sample. These findings shed light on the molecular signatures of MPH effects and possible biological sources of treatment response variability.

中文翻译：

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哌醋甲酯治疗反应的综合蛋白质组学和药物基因组学分析。

转录组学和候选基因/蛋白质表达研究表明，哌醋甲酯（MPH）调节了几种生物学过程，广泛用于注意力缺陷/多动症（ADHD）治疗。但是，缺乏MPH治疗的差异蛋白质组学分析限制了对MPH在分子水平上发挥其药理作用的最相关机制的了解。因此，我们的目的是使用实验设计与药物基因组学分析相结合，从翻译的角度研究MPH诱导的蛋白质组学变化。使用Wistar-Kyoto大鼠皮层进行蛋白质组学分析，用MPH（2 mg / kg）或生理盐水灌胃治疗两周（n = 6 /组）。在对大鼠差异表达蛋白（DEP）进行功能富集分析后，使用全基因组数据测试了成人多动症（n = 189）成人中重要的生物学途径与MPH反应的相关性。在大鼠进行MPH治疗后，发现了98个DEP（P <0.05和FC <-1.0或> 1.0）。DEPs的功能富集分析揭示了由MPH调节的18条重要的生物途径（基因集），包括一些具有公认的生物似然性的途径，例如与突触传递有关的那些途径。临床样品中的药物基因组学分析评估了这些途径，揭示了与神经递质释放和GABA传递相关的基因组的名义关联。我们的结果结合了蛋白质组学和药物基因组学，揭示了MPH在几种生物过程中的推定分子作用，包括氧化应激，细胞呼吸和新陈代谢，并将涉及突触传递途径的结果扩展到临床样品。这些发现揭示了MPH效应的分子特征以及治疗反应变异性的可能生物学来源。