A New Workflow for Proteomic Analysis of Urinary Exosomes and Assessment in Cystinuria Patients
A New Workflow for Proteomic Analysis of Urinary Exosomes and Assessment in Cystinuria Patients
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DOI：
10.1021/pr501003q
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发表时间：
2015-01-01
影响因子：
4.4
通讯作者：
Guerrera, Ida Chiara
中科院分区：
文献类型：
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作者：
Bourderioux, Matthieu;Thao Nguyen-Khoa;Guerrera, Ida Chiara
胱氨酸尿症是一种由胱氨酸转运体基因突变引起的纯肾脏罕见遗传病，以胱氨酸重吸收缺陷导致肾结石为特征。在14%的病例中，患者接受了肾切除术，但鉴于难以预测疾病的发展，识别肾损害标志物将改善对高风险患者的随访。本研究的目的是开发一种可靠的、可重复的、无创的方法，使用高分辨率质谱法对尿外泌体进行蛋白质组学分析。一项针对8名胱氨酸尿病患者和10名对照者进行的临床试点研究突出了165种蛋白，其中38种蛋白上调，这些蛋白将胱氨酸尿病患者与对照者区分开来，并进一步区分严重和中度胱氨酸尿病患者。这些蛋白包括肾损伤标志物、循环蛋白和中性粒细胞标记。在另外6名胱氨酸尿病患者身上，通过免疫印迹分析选择的蛋白质，验证了质谱数据。据我们所知，这是第一个成功的胱氨酸尿蛋白组学研究，揭示了炎症在这种疾病中的潜在作用。我们开发的工作流程适用于研究不同肾脏疾病的尿外泌体，并寻找诊断/预后标志物。数据可通过ProteomeXchange获得，标识符为PXD001430。
Cystinuria is a purely renal, rare genetic disease caused by mutations in cystine transporter genes and characterized by defective cystine reabsorption leading to kidney stones. In 14% of cases, patients undergo nephrectomy, but given the difficulty to predict the evolution of the disease, the identification of markers of kidney damage would improve the follow-up of patients with a higher risk. The aim of the present study is to develop a robust, reproducible, and noninvasive methodology for proteomic analysis of urinary exosomes using high resolution mass spectrometry. A clinical pilot study conducted on eight cystinuria patients versus 10 controls highlighted 165 proteins, of which 38 were up-regulated, that separate cystinuria patients from controls and further discriminate between severe and moderate forms of the disease. These proteins include markers of kidney injury, circulating proteins, and a neutrophil signature. Analysis of selected proteins by immunobloting, performed on six additional cystinuria patients, validated the mass spectrometry data. To our knowledge, this is the first successful proteomic study in cystinuria unmasking the potential role of inflammation in this disease. The workflow we have developed is applicable to investigate urinary exosomes in different renal diseases and to search for diagnostic/prognostic markers. Data are available via ProteomeXchange with identifier PXD001430.