Combinatorial Peptide Ligand Library and two dimensional electrophoresis: New frontiers in the study of peritoneal dialysis effluent in pediatric patients
Combinatorial Peptide Ligand Library and two dimensional electrophoresis: New frontiers in the study of peritoneal dialysis effluent in pediatric patients
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DOI：
10.1016/j.jprot.2015.01.003
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发表时间：
2015-02-26
影响因子：
3.3
通讯作者：
Verrina, Enrico
中科院分区：
文献类型：
--
作者：
Bruschi, Maurizio;Candiano, Giovanni;Verrina, Enrico
腹膜透析流出物（PDE）是腹膜透析（PD）溶液与腹膜（PM）密切接触产生的液体，是一种可用于寻找PM功能或损伤生物标志物的现成材料。我们的实验室已经开发了一种用于PDE的深入蛋白质组学表征的方法，该方法涉及组合肽配体库（CPLL）以减少PDE中蛋白质浓度的动态范围，然后进行双向电泳（2-DE）。在本研究中，我们应用该方法分析了从19名使用葡萄糖PD溶液进行自动腹膜透析（APD）的儿科患者中获得的PDE蛋白质组。这种蛋白质组学方法的组合使用突出了平均700个新蛋白质。PDE蛋白质组谱的差异与APD治疗的持续时间有关。特别是在长期APD患者中，我们观察到intelectin-1增加，凝溶胶蛋白减少。通过用氧化或促纤维化刺激物体外处理间皮细胞也观察到这些变化，这支持了这些蛋白质变化的生物学作用。为了阐明所观察到的差异的生物学意义，我们的进一步研究将包括PDE蛋白质组的纵向评价。生物学意义通过组合肽配体库（CPLL）和双向电泳的联合使用，对儿科患者腹膜透析流出液（PDE）进行深入的蛋白质组学表征，允许检测1788个点，其中的相关部分（724）先前在未用CPLL处理的样品中未检测到。在接受长期自动腹膜透析的患者中，这种蛋白质组学方法可以识别29种潜在的生物标志物，这些生物标志物可能有助于识别患有亚临床炎症和/或腹膜纤维化的患者，从而相应地调整透析治疗。(C)2015爱思唯尔B. V.保留所有权利。
Peritoneal dialysis effluent (PDE) is a fluid resulting from the close contact of peritoneal dialysis (PD) solutions with the peritoneal membrane (PM) and represents a readily available material for the search of biomarkers of PM function or damage. Our laboratory has developed a method for the in-depth proteomic characterization of PDE, which involves Combinatorial Peptides Ligand Library (CPLL) to reduce the dynamic range of protein concentration in PDE, followed by two-dimensional electrophoresis (2-DE). In this study we applied this method to the analysis of PDE proteome obtained from 19 pediatric patients on automated peritoneal dialysis (APD) with glucose-based PD solutions. The combined use of this proteomic approach highlighted a mean of 700 new proteins. Differences in PDE proteome profile were observed in relation with the duration of APD treatment. In particular, in patients on long-term APD, we observed an increase of intelectin-1, and a decrease of gelsolin. These changes were also observed by in vitro treatment of mesothelial cells with oxidative or pro-fibrotic stimulus which supported the biological role of these proteins' changes. In order to clarify the biological meaning of the observed differences, further step of our study will consist of the longitudinal evaluation of PDE proteome.Biological significanceThe in-depth proteomic characterization of peritoneal dialysis effluent (PDE) in pediatric patients by the combined use of Combinatorial Peptide Ligand Library (CPLL) and two dimensional electrophoresis allowed to detect 1788 spots, a relevant part (724) of which were previously undetected in sample untreated with CPLL. In patients on long-term automated peritoneal dialysis, this proteomic approach allowed to identify 29 potential biomarkers that could be of help to identity patients with subclinical inflammation and/or developing peritoneal membrane fibrosis, thus adapting dialysis treatment accordingly. (C) 2015 Elsevier B.V. All rights reserved.