Defining Biomarkers in Pediatric Renal Transplantation

描述（由申请人提供）：我们建议进行一项全面的机制研究，以建立儿科肾移植中的信息性生物标志物，使用来自SNS 01的患者临床数据和组织（活检和血液）-无类固醇与基于类固醇的免疫抑制的随机临床试验。该提案概述了将来自SNS 01的临床和病理学核心与基因组（cDNA阵列和实时PCR）和免疫病理学（核心活检和组织微阵列）核心整合的策略，后两个核心将由该提案生成。本研究的目的有四个方面。(i)通过DNA微阵列分析，在组织和血液中开发稳定移植、急性排斥（AR）和慢性移植物肾病（CAN）的诊断生物标志物。(ii)检测无类固醇和基于类固醇的免疫抑制在这些移植状态中的差异。（iii）使用实时PCR关联组织和血液中所选高度分化基因的表达，以建立用于移植物监测、AR诊断、分类和AR治疗和结果分层的非侵入性血液标志物。(iv)对核心患者活检和组织微阵列进行免疫组织化学分析，以将基因表达数据与AR、CAN和正常移植后肾脏中的细胞浸润表型、密度和活化相关联，包括无类固醇和基于类固醇的免疫抑制。这些研究有望有助于确定稳定移植和不同形式的肾移植物功能障碍、不同类型的免疫抑制（有或无类固醇）的免疫和细胞机制，并产生用于非侵入性移植物监测以及移植物治疗和风险分层的临床工具。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.