Molecular profiling and immunomodulatory interventions

描述（由申请人提供）：我们提出基因表达谱用于表征与实体器官移植相关的先天免疫和适应性免疫。我们还提出了干预性试验，将解决关键的未满足的需求，临床研究的基础上的机制研究。我们的建议是由我们的单中心研究和不断发展的文献刺激的，即外周血细胞、尿细胞和支气管肺泡灌洗（BAL）细胞的mRNA谱分析可以诊断同种异体移植物状态。我们对术中肾移植活检标本的mRNA表达谱的鉴定可以预测急性排斥反应和肾移植功能结果，这为我们的研究设计提供了依据。我们建议合作多部位肾脏特异性和多器官研究，以确定是否有共同的分子特征的炎症和免疫反应，这可以提供一个更好的方法来监测功能和应用干预方式，旨在改善移植物的结果。 在肾移植受者中，我们将研究：（a）急性排斥反应是否可以通过尿细胞的连续mRNA谱预测;（B）基于mRNA谱的抢先治疗，预防急性排斥反应并保护GFR;（c）mRNA谱是否可以用于指导免疫抑制管理，例如在稳定的受者中停用钙调磷酸酶抑制剂。 对不同移植器官组的研究将集中在器官特异性和非特异性损伤途径的分子特征分析上，这些损伤途径在供体获得前和移植后立即在受体中，以及它们对器官功能和同种异体反应性的影响。将在肺部和肝脏环境中进行更有针对性的研究。在肺同种异体移植物中，我们将探索：（a）供体和受体BAL细胞获得的细胞的mRNA谱是否预测不良结果，如严重炎症和移植物功能受损的发展，（B）BAL细胞的持续炎症特征是否伴随急性排斥特征。 在同种异体肝移植中，我们将探讨：（a）促炎基因的独特mRNA谱所指示的早期炎症反应是否与HCV复发相关，以及（B）在表达强烈炎症特征的移植物中，避免类固醇治疗方案是否下调HCV复发。 这些机制分析将使用动态（真实的）时间定量PCR分析进行，并且在选择的情况下使用基于核酸的微阵列以及基因表达位点的免疫组织学分析，并且预期产生将应用于移植群体的常规临床护理的数据。

DESCRIPTION (provided by applicant): We propose gene expression profiling for the characterization of innate immunity and adaptive immunity pertinent to solid organ transplantation. We also propose interventional trials that will address critical unmet needs; the clinical studies are underpinned by mechanistic studies. Our proposals were stimulated by our single center studies and evolving literature that mRNA profiling of peripheral blood cells, urinary cells, and bronchoalveolar lavage (BAL) cells are diagnostic of allograft status. Our identification of mRNA profiles of intraoperative renal allograft biopsy specimens that predict acute rejection and renal allograft functional outcome has informed our research design. We propose cooperative multi-site kidney-specific and multi-organ studies to determine whether there is common molecular profile characteristic of inflammatory and immune responses, and which can provide a better method to monitor function and apply interventional modalities aimed at improving graft outcome. In renal allograft recipients, we will investigate whether: (a) acute rejection can be predicted by sequential mRNA profiling of urinary cells; (b) preemptive treatment, based on mRNA profiles, prevents acute rejection and preserves GFR, and (c) mRNA profiles can be used to guide immunosuppression management, such as the withdrawal of calcineurin inhibitors in stable recipients. Studies in diverse groups of transplanted organs will center on the molecular profiling of organ specific and nonspecific injury pathways in the donor prior to procurement and in the recipient immediately after transplantation, and their impact on organ function and alloreactivity. More focused studies will be done in the lung and the liver setting. In lung allografts, we will explore: (a) whether mRNA profiling of cells obtained by donor and recipient BAL cells predict adverse outcomes such as intense inflammation and development of impaired graft function, (b) whether a persistent inflammatory signature of BAL cells is followed by an acute rejection signature. In liver allografts, we will explore: (a) whether an early inflammatory response indicated by a unique mRNA profile of proinflammatory genes is correlated with HCV recurrence, and (b) whether a steroid-avoidance protocol downregulates HCV recurrence in grafts expressing intense inflammatory signature. These mechanistic assays will be carried out with the use of kinetic (real) time quantitative PCR assay, and in select instances with the use of nucleic-acid based microarrays, as well as immunohistologic analysis of the site of gene expression, and are expected to yield data that will be applied in routine clinical care of the transplanted population.