Urinary Proteome Monitoring for Transplant Injury

移植损伤的尿液蛋白质组监测

• 批准号: 8549203
• 负责人: David G Camp
• 金额: $ 33.97万
• 依托单位: CALIFORNIA PACIFIC MED CTR RES INSTITUTE
• 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 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8549203
• 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; 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; screening; 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）：我们在这项研究中的目标是通过分析一个大队列（n=660）患者尿液样本，发现儿童肾移植中不同肾移植损伤表型的高度特异性诊断性非侵入性生物标志物。肾移植是儿童终末期肾病的治疗选择。移植物存活受到不同损伤累积的限制，如急性排斥反应（AR）、慢性同种异体移植肾病（CAN）和BK病毒感染（BKV），这些损伤无法通过简单监测血清肌酐来区分，需要侵入性活检来诊断已确定的损伤。尿液作为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。在本提案中，我们建议采用最新开发的基于LC-FT MS/MS- ustags的天然尿肽组学分析和基于LC-MS/MS的人尿可溶性蛋白分析，对尿蛋白质组进行尖端的综合分析。PNNL最先进的蛋白质组学设施的可用性和斯坦福大学具有综合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药物的选择将借助强大的综合蛋白质基因组学数据分析，该数据分析将使用在同一时间点收集的来自同一患者的重叠活检微阵列数据和尿液蛋白质组学数据。针对不同表型移植物损伤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480份独立尿液样本上进行临床前验证，以进行诊断和预测。蛋白质/肽标记将有助于开发ELISA检测方法，可用于临床监测移植物损伤，取代侵入性移植活检。