Quantitative Proteomics for Monitoring Renal Transplant Injury
Quantitative Proteomics for Monitoring Renal Transplant Injury
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DOI：
10.1002/prca.201900036
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发表时间：
2020-02-16
影响因子：
2
通讯作者：
Xiao, Kunhong
中科院分区：
文献类型：
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作者：
Song, Lei;Fang, Fei;Xiao, Kunhong
目的本研究旨在开发一种分子诊断平台，以提高对福尔马林固定和石蜡包埋（FFPE）标本的定量蛋白质组学分析的解释。实验设计定量蛋白质组学平台包括1）优化的FFPE蛋白质样品制备方法，2）基于串联质量标签TMT 10-plex的蛋白质组学工作流程，和3）已经开发了揭示差异表达蛋白的系统统计分析管道。然后在小样本集（每个表型5个样本）上测试该平台以揭示可以区分T细胞介导的排斥（TCMR）和多瘤病毒BK肾病（BKPyVN）与健康功能稳定的肾组织（STA）的蛋白质组特征。主成分分析表明，所有三个表型调查良好的分离。蛋白质检测和定量具有高度可重复性：重复比较分析表明，检测到的蛋白质重叠率为71-84%，蛋白质测量的变异系数为定量蛋白质组学可应用于存档的FFPE标本，以区分肾移植损伤的不同原因。
Purpose This study is aimed at developing a molecular diagnostics platform to enhance the interpretation of renal allograft biopsies using quantitative proteomic profiling of formalin-fixed and paraffin-embedded (FFPE) specimens.Experimental Design A quantitative proteomics platform composed of 1) an optimized FFPE protein sample preparation method, 2) a tandem mass tag TMT10-plex-based proteomic workflow, and 3) a systematic statistical analysis pipeline to reveal differentially expressed proteins has been developed. This platform is then tested on a small sample set (five samples per phenotype) to reveal proteomic signatures that can differentiate T-cell mediated rejection (TCMR) and polyomavirus BK nephropathy (BKPyVN) from healthy functionally stable kidney tissue (STA).Results Among 2798 quantified proteins, the expression levels of 740 BKPyVN and 638 TCMR associated proteins are significantly changed compared to STA specimens. Principal component analysis demonstrated good segregation of all three phenotypes investigated. Protein detection and quantitation are highly reproducible: replicate comparative analyses demonstrated 71-84% overlap of detected proteins, and the coefficient of variation for protein measurements is Quantitative proteomics can be applied to archived FFPE specimens to differentiate different causes of renal allograft injury.