Defining Biomarkers in Pediatric Renal Transplantation

描述(申请人提供)：我们建议进行一项全面的机制研究，利用患者的临床数据和SNS01的组织(活检和血液)来建立儿科肾移植中的信息生物标记物。SNS01是一项关于非类固醇和基于类固醇的免疫抑制的随机临床试验。该建议概述了将SNS01的临床和病理核心与基因组(基因芯片和实时聚合酶链式反应)和免疫病理学(核心活组织检查和组织微阵列)核心整合的战略，后两个核心将由该提议产生。这项研究的目的有四个方面。(I)通过DNA微阵列分析开发组织和血液中用于稳定移植、急性排斥(AR)和慢性移植物肾病(CAN)的诊断生物标记物。(Ii)检测这些移植物状态在非类固醇免疫抑制和类固醇免疫抑制方面的差异。(Iii)利用实时定量聚合酶链式反应(Real-Time-PCR)将选定的高分化基因在组织和血液中的表达进行关联，以建立非侵入性血液标记物，用于移植物监测、AR诊断、分类以及AR治疗和结果的分层。(Iv)利用核心患者活检组织和组织微阵列的免疫组织化学分析，将基因表达数据与AR、CAN和移植后正常肾脏中的细胞渗透表型、密度和激活相关联，包括在非类固醇和基于类固醇的免疫抑制中。这些研究有望促进稳定移植和不同形式的肾移植功能障碍、不同类型的免疫抑制(使用和不使用类固醇)的免疫和细胞机制的定义，并为非侵入性移植监测以及移植治疗和风险分层提供临床工具。基因组和组织微阵列数据库将向学术界公开，以促进未来的大规模合作研究。

DESCRIPTION (provided by applicant): We propose to undertake a comprehensive mechanistic study to establish informative biomarkers in pediatric renal transplantation, using patient clinical data and tissue (biopsy and blood) from SNS01- a randomized clinical trial of steroid-free versus steroid-based immunosuppression. This proposal outlines a strategy for integrating clinical and pathology cores from SNS01 with genomic (cDNA arrays and real-time PCR) and immunopathology (core biopsies and tissue microarrays) cores, the latter two cores to be generated by this proposal. The aims of this study are fourfold. (i) To develop diagnostic biomarkers by DNA microarray analysis, in tissue and blood for stable transplants, acute rejection (AR) and chronic allograft nephropathy (CAN). (ii) To detect differences in these graft states in steroid-free and steroid-based immunosuppression. (iii)To correlate the expression of selected highly differentiated genes in tissue and blood, using real-time PCR, to establish non-invasive blood markers for graft surveillance, AR diagnosis, classification, and stratification of AR therapy and outcome. (iv) To use immunohistochemistry analysis on core patient biopsies and tissue microarrays to correlate gene expression data with cellular infiltration phenotypes, density and activation in AR, CAN and normal post-transplant kidneys, in steroid-free and steroid-based immunosuppression. These studies are expected to facilitate the definition of immunological and cellular mechanisms underlying stable transplants and different forms of renal allograft dysfunction, different types of immunosuppression (with and without steroids), and generate clinical tools for non-invasive graft monitoring, as well as graft therapeutic and risk stratification. The genomic and tissue microarray databases will be made publicly available for the academic community to facilitate future large-scale collaborative studies.