A novel urinary biomarker protein panel to identify children with ureteropelvic junction obstruction - A pilot study.
A novel urinary biomarker protein panel to identify children with ureteropelvic junction obstruction - A pilot study.
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DOI：
10.1016/j.jpurol.2020.05.163
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发表时间：
2020-08
影响因子：
2
通讯作者：
Shapiro LH
中科院分区：
文献类型：
--
作者：
Devarakonda CKV;Shearier ER;Hu C;Grady J;Balsbaugh JL;Makari JH;Ferrer FA;Shapiro LH
可靠的尿液生物标志物蛋白对于识别输尿管肾盂连接处梗阻（UPJO）的儿童具有极其重要的价值，因为现有的生物标志物蛋白在预测能力方面并不一致。因此，本研究的目的是识别UPJO儿童中一致且可靠的尿液生物标志物蛋白。
为了识别候选生物标志物蛋白，对年龄受限（<2岁）且性别匹配（男性）的对照组（n = 22）和UPJO组（n = 21）尿液样本中的总蛋白进行了质谱分析。优先在UPJO样本中鉴定出的蛋白经过筛选（两步法），并根据其诊断比值比的值进行排序。选择比值比最高的前十种蛋白，并用酶联免疫吸附测定（ELISA）分别进行检测。将每种蛋白的总量根据尿肌酐进行标准化，并确定对照组和UPJO组样本的中位数及四分位间距。此外，还确定了最终选定的5种蛋白组合中位数的倍数变化（UPJO/对照组）。最后，我们计算了对照组样本中5种蛋白各自的平均值 + 3倍标准差（SD）和平均值 + 4倍标准差的值，并将其用作任意截断值来对单个对照组和UPJO组样本进行分类。
在我们筛选过程的第一步，我们在UPJO样本中鉴定出171种在大多数对照组样本（16/22个样本，即72.7%）中未检测到的蛋白。在这171种蛋白中，只有50种蛋白在至少11/21（52.4%）的UPJO样本中被检测到，因此在第二步中被选中。随后，根据比值比对这50种蛋白进行排序，排名前10的蛋白通过ELISA进行验证。10种蛋白中的5种——前列腺素还原酶 - 1、纤维胶凝蛋白 - 2、烟酸核苷酸焦磷酸化酶[羧化]、含免疫球蛋白超家族的富亮氨酸重复蛋白和血管细胞黏附分子 - 1在UPJO样本中的水平较高（蛋白浓度中位数的倍数变化范围为2.9 - 9.4），并成为一组识别梗阻性尿路疾病的生物标志物。最后，这5种蛋白在UPJO样本中的出现频率顺序为PTGR1>FCN2>QPRT>ISLR>VCAM1。
总之，这种独特的筛选策略导致了以前未知的生物标志物蛋白的识别，当对这些蛋白进行集体筛选时，可能可靠地区分梗阻与非梗阻的婴儿，并可能有助于为许多疾病的生物样本识别有信息价值的生物标志物组合。
总结图. PTGR1、FCN2、QPRT、ISLR和VCAM1形成一组UPJO固有尿液生物标志物蛋白。
（A）对照组和UPJO组样本中标准化的PTGR1、FCN2、QPRT、ISLR和VCAM1的量。
（B）标准化的PTGR1、FCN2、QPRT、ISLR和VCAM1量的中位数的倍数变化（UPJO/对照组）。
Reliable urinary biomarker proteins would be invaluable in identifying children with ureteropelvic junction obstruction (UPJO) as the existing biomarker proteins are inconsistent in their predictive ability. Therefore, the aim of this study was to identify consistent and reliable urinary biomarker proteins in children with UPJO. To identify candidate biomarker proteins, total protein from age-restricted (<2 years) and sex-matched (males) control (n=22) and UPJO (n=21) urine samples was analyzed by mass spectrometry. Proteins that were preferentially identified in UPJO samples were selected (2-step process) and ranked according to their diagnostic odds ratio value. The top ten proteins with highest odds ratio values were selected and tested individually by ELISA. The total amount of each protein was normalized to urine creatinine and the median with interquartile ranges for control and UPJO samples was determined. Additionally, fold change (UPJO/Control) of medians of the final panel of 5 proteins was also determined. Finally, we calculated the average + 3(SD) and average + 4(SD) values of each of the 5 proteins in the control samples and used it as an arbitrary cutoff to classify individual control and UPJO samples. In the first step of our selection process, we identified 171 proteins in UPJO samples that were not detected in the majority of the control samples (16/22 samples, or 72.7%). Of the 171 proteins, only 50 proteins were detected in at least 11/21 (52.4%) of the UPJO samples and hence were selected in the second step. Subsequently, these 50 proteins were ranked according to the odds ratio value and the top 10 ranked proteins were validated by ELISA. Five of the 10 proteins – prostaglandin-reductase-1, ficolin-2, nicotinate-nucleotide pyrophosphorylase [carboxylating], immunoglobulin superfamily-containing leucine-rich-repeat-protein and vascular cell adhesion molecule-1 were present at higher levels in the UPJO samples (fold-change of the median protein concentrations ranging from 2.9 – 9.4) and emerged as a panel of biomarkers to identify obstructive uropathy. Finally, the order of prevalence of the 5 proteins in UPJO samples is PTGR1>FCN2>QPRT>ISLR>VCAM1. In summary, this unique screening strategy led to the identification of previously unknown biomarker proteins that when screened collectively, may reliably distinguish between obstructed vs. non-obstructed infants and may prove useful in identifying informative biomarker panels for biological samples from many diseases. Summary Figure. PTGR1, FCN2, QPRT, ISLR and VCAM1 form a panel of UPJO-inherent urinary biomarker proteins. (A) Normalized PTGR1, FCN2, QPRT, ISLR and VCAM1 amounts in control and UPJO samples. (B) Fold change (UPJO/control) of medians of normalized PTGR1, FCN2, QPRT, ISLR and VCAM1 amounts.
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