Urinary Proteome Monitoring for Transplant Injury

描述(申请人提供)：我们在这项研究中的目标是在一大群(n=580)儿童肾移植不同移植物损伤表型的患者尿样中发现诊断的非侵入性生物标记物。肾移植是儿童终末期肾病的首选治疗方法。移植物的存活受到不同损伤的累积的限制，如急性排斥反应(AR)、慢性移植物肾病(CAN)和BK病毒感染(BKV)，这些损伤无法通过简单地监测血清肌酐来区分，需要进行侵入性活检来诊断已建立的损伤。尿液是血浆的超滤液和肾脏的直接滤液，为监测移植物的健康和功能障碍提供了宝贵的资源。我们建议利用基于LC-MALDI的多肽组分析和基于LC-MS/MS的尿液可溶性蛋白分析来应用尖端的、全面的尿蛋白组分析。卓越的生物信息学环境和全面的临床数据库的可获得性将支持高通量数据分析，选择50种最重要的多肽和蛋白质来区分不同的移植物损伤表型-AR、CAN、BKV和稳定功能移植物(STA)。为了能够选择最具生物学相关性的候选对象，将使用重叠的活检微阵列数据和同一患者在同一时间点收集的尿蛋白组学数据进行稳健的综合蛋白质组数据分析。为移植物损伤的不同表型发现的特定和敏感的非侵入性尿液生物标记物将在一组独立的400个尿样上接受MRM的临床前验证，以进行诊断和预测。这一建议的成果是丰富了~10个可用于临床移植损伤监测的尿蛋白生物标志物，取代了有创性移植活检。 公共卫生相关性： 在缺乏有效的非侵入性诊断急性肾损伤的方法下，肾活检是监测移植肾临床进展的唯一途径。在这项研究中，我们建议对生物标记物进行全面的搜索，这些生物标记物可以在以后开发为损伤特异性临床试验。我们将使用两种最强大的蛋白质组学方法来鉴定这些生物标记物，这在移植领域将是第一次。

DESCRIPTION (provided by applicant): Our goal in this study is to discovery of diagnostic non-invasive biomarkers in a large cohort (n=580) patient urine samples for different graft injury phenotypes in pediatric kidney transplantation. Kidney transplantation is the treatment of choice for children with end stage kidney disease. Graft survival is limited by the accrual of different injuries such as acute rejection (AR), chronic allograft nephropathy (CAN), and viral infection with the BK virus (BKV), which are indistinguishable by simply monitoring the serum creatinine and require an invasive biopsy for diagnosis of established injury. Urine is an ultrafiltrate of plasma and a direct filtrate of the kidney and provides a valuable resource for monitoring graft health and dysfunction. We propose to employ cutting-edge, comprehensive analysis of the urinary proteome by LC-MALDI based peptidomics and LC-MS/MS based soluble protein analysis of human urine. An environment of outstanding bioinformatics and the availability of a comprehensive Clinical Database will support high-throughput data analysis for selection of 50 most significant peptides and proteins for differentiating different graft injury phenotypes- AR, CAN, BKV and stable functioning grafts (STA). To enable selection of the most biologically relevant candidates, robust integrative proteogenomic data analysis will be performed using overlapping biopsy microarray data and urine proteomic data from the same patient, collected at the same time-point. Specific and sensitive non-invasive urinary biomarkers discovery for different phenotypes of graft injury will undergo pre-clinical validation by MRM on an independent set of 400 urine samples for diagnosis and prediction. The deliverable from this proposal is to enrich for ~10 urinary protein biomarkers that can be used for clinical monitoring of graft injury, replacing the invasive transplant biopsy. PUBLIC HEALTH RELEVANCE: In the absence of an effective noninvasive way of diagnosing acute injury, kidney biopsy is the only way to monitor the clinical progress the transplanted kidney. In this study, we propose a comprehensive search for biomarkers that could be later developed as an injury specific clinical test. We are going to use two most powerful proteomic methods to identify these biomarkers which will be first of its kind in the field of transplantation.