Urinary Proteome Monitoring for Transplant Injury

移植损伤的尿液蛋白质组监测

• 批准号: 8131911
• 负责人: David G Camp
• 金额: $ 17.97万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-23 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8131911
• 关键词: Acute; Acute Kidney Tubular Necrosis; Acute Renal Failure with Renal Papillary Necrosis; Address; Adopted; Algorithms; Amino Acids; Archives; BK Virus; Bioinformatics; Biological Assay; Biological Markers; Biopsy; Blood; Blood specimen; Camping; Categories; Cations; Child; Childhood; Chronic; Chronic rejection of renal transplant; Clinical; Clinical Data; Clinical Trials Design; Collection; Complement; Creatinine; Cross-Sectional Studies; Data; Data Analyses; Databases; Detection; Development; Diagnosis; Diagnostic; Drug toxicity; End stage renal failure; Environment; Enzyme-Linked Immunosorbent Assay; Equipment and supply inventories; Etiology; Evaluation; Event; Fibrosis; Fourier Transform; Fractionation; Functional disorder; Funding; Gene Proteins; Goals; Graft Survival; Health; Human; Immune; Immunosuppression; Individual; Infection; Injury; Institutional Review Boards; Kidney; Kidney Transplantation; Label; Laboratories; Liquid Chromatography; Mediating; Methods; Molecular Weight; Monitor; Morbidity - disease rate; Nephrotic Syndrome; Organ Transplantation; Outcome; Pacific Northwest; Patients; Pattern; Peptides; Phase; Phenotype; Plasma; Process; Protein Analysis; Proteins; Proteome; Proteomics; Protocols documentation; Publications; Randomized; Reaction; Research Personnel; Resolution; Resources; Risk; Risk Factors; Sampling; Screening procedure; Serum; Staging; Steroids; Technology; Testing; Time; Training; Transplant Recipients; Transplantation; United States National Institutes of Health; Universities; Urine; Validation; Vasculitis; Viral; Viremia; Virus Diseases; allograft rejection; base; candidate validation; clinical application; clinical phenotype; cohort; design; follow-up; functional decline; graft function; improved; kidney allograft; mass spectrometer; non-invasive monitor; pre-clinical; programs; public health relevance; research clinical testing; sample collection; tandem mass spectrometry; translational study; two-dimensional; urinary

DESCRIPTION (provided by applicant): Our goal in this study is to discover highly specific diagnostic non-invasive biomarkers by analyzing a large cohort (n=660) patient urine samples for different kidney transplant injury phenotypes in pediatric renal 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 being an ultrafiltrate of plasma and a direct filtrate of the kidney provides a valuable resource for monitoring graft health and dysfunction. In this proposal, we propose to employ a cutting-edge, comprehensive analysis of the urinary proteome by using recently developed and LC-FT MS/MS-UStags based peptidomics analysis of native urinary peptides and LC-MS/MS based soluble protein analysis of human urine. Availability of state-of-the-art proteomics facility at PNNL and an environment of outstanding bioinformatics with available comprehensive Clinical Database at Stanford will support high-throughput biomarker screening and data analysis for selection of 20 most significant peptides and proteins for differentiating different graft injury phenotypes- AR, CAN, BKV and stable functioning grafts (STA). The selection of the most biologically relevant candidates will be aided by robust integrative proteogenomic data analysis which will use overlapping biopsy microarray data and urine proteomic data from the same patients, 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 SRM on an independent set of 480 urine samples for diagnosis and prediction. The protein/peptide markers will be useful in developing ELISA assays which 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.

描述（由申请人提供）：本研究的目标是通过分析儿科肾移植中不同肾移植损伤表型的大型队列（n=660）患者尿液样本，发现高度特异性的诊断性非侵入性生物标志物。肾移植是儿童终末期肾病的首选治疗方法。移植物存活受到不同损伤的累积的限制，例如急性排斥反应（AR）、慢性同种异体移植物肾病（CAN）和BK病毒感染（BKV），这些损伤通过简单地监测血清肌酐是无法区分的，并且需要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，为监测移植物健康和功能障碍提供了宝贵的资源。在这项提案中，我们建议采用最先进的，全面的分析尿蛋白质组，通过使用最近开发的和LC-FT MS/MS-UStags基于肽组学分析天然尿肽和LC-MS/MS基于可溶性蛋白质分析人尿。PNNL最先进的蛋白质组学设施的可用性和斯坦福大学可用的全面临床数据库的杰出生物信息学环境将支持高通量生物标志物筛选和数据分析，以选择20种最重要的肽和蛋白质，用于区分不同的移植物损伤表型- AR，CAN，BKV和稳定功能移植物（STA）。最具生物学相关性的候选者的选择将通过稳健的整合蛋白基因组数据分析来辅助，该分析将使用在相同时间点收集的来自相同患者的重叠活检微阵列数据和尿蛋白组数据。针对移植物损伤的不同表型的特异性和敏感性非侵入性尿液生物标志物的发现将通过SRM在用于诊断和预测的480个尿液样本的独立集合上进行临床前验证。蛋白质/肽标记物将用于开发ELISA测定，其可用于移植物损伤的临床监测，代替侵入性移植活检。 公共卫生关系：在缺乏有效的非侵入性诊断急性损伤的方法的情况下，肾活检是监测移植肾临床进展的唯一方法。在这项研究中，我们提出了一个全面的搜索生物标志物，可以在以后开发的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志物，这将是移植领域的第一个。