Lower urinary tract symptoms (LUTS) is common in aging males. Disease etiology is largely unknown but likely includes inflammation and age-related changes in steroid hormones. Diagnosis is currently based on subjective symptom scores, and mainstay treatments can be ineffective and bothersome. Biomarker discovery efforts could facilitate objective diagnostic criteria for personalized medicine and new potential druggable pathways. To identify urine metabolite markers specific to hormone-induced bladder outlet obstruction, we applied our custom synthesized multiplex isobaric tags to monitor the development of bladder outlet obstruction across time in an experimental mouse model of LUTS. Mouse urine samples were collected before treatment and after 2, 4, and 8 weeks of steroid hormone treatment and subsequently analyzed by nanoflow ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry. Accurate and high-throughput quantification of amine-containing metabolites was achieved by 12-plex DiLeu isobaric labeling. Metandem, a novel online software tool for large-scale isobaric labeling-based metabolomics, was used for identification and relative quantification of labeled metabolites. A total of 59 amine-containing metabolites were identified and quantified, 9 of which were changed significantly by the hormone treatment. Metabolic pathway analyses showed that three metabolic pathways were potentially disrupted. Among them, the arginine and proline metabolism pathway was significantly dysregulated both in this model and in a prior analysis of LUTS patient samples. Proline and citrulline were significantly changed in both samples and serve as attractive candidate biomarkers. The 12-plex DiLeu isobaric labeling with Metandem data processing presents an accessible and efficient workflow for an amine-containing metabolome study in biological specimens.

中文翻译：

---

使用自定义 12-Plex 等压 DiLeu 标记进行尿胺代谢组学表征。

下尿路症状 (LUTS) 在老年男性中很常见。疾病病因在很大程度上是未知的，但可能包括炎症和与年龄相关的类固醇激素变化。目前诊断基于主观症状评分，主要治疗可能无效且麻烦。生物标志物发现工作可以促进个性化医疗和新的潜在药物途径的客观诊断标准。为了识别特定于激素诱导的膀胱出口梗阻的尿液代谢物标记物，我们应用我们定制的合成多重同量异位标记来监测 LUTS 实验小鼠模型中随时间变化的膀胱出口梗阻的发展。在治疗前和治疗后 2、4、和 8 周的类固醇激素治疗，随后通过纳流超高效液相色谱与串联质谱联用进行分析。通过 12 重 DiLeu 等压标记实现了含胺代谢物的准确和高通量定量。Metandem 是一种用于大规模等压标记代谢组学的新型在线软件工具，用于标记代谢物的鉴定和相对定量。共鉴定并量化了 59 种含胺代谢物，其中 9 种因激素处理而发生显着变化。代谢途径分析表明，三种代谢途径可能被破坏。其中，精氨酸和脯氨酸代谢途径在该模型和先前对 LUTS 患者样本的分析中均显着失调。脯氨酸和瓜氨酸在两个样品中都发生了显着变化，可作为有吸引力的候选生物标志物。使用 Metandem 数据处理的 12 重 DiLeu 同量异位标记为生物样本中的含胺代谢组研究提供了一种易于访问且高效的工作流程。