Molecular profiling and immunomodulatory interventions

描述(由申请人提供)：我们建议用基因表达谱来表征与实体器官移植相关的先天免疫和获得性免疫。我们还建议进行干预试验，以解决关键的未得到满足的需求；临床研究以机械性研究为基础。我们的建议受到了我们的单中心研究和不断发展的文献的刺激，即外周血细胞、尿细胞和支气管肺泡灌洗(BAL)细胞的mRNA谱可以诊断同种异体移植状态。我们对术中肾移植活检标本的信使核糖核酸图谱进行鉴定，以预测急性排斥反应和移植肾功能结果，为我们的研究设计提供了依据。我们建议进行多点肾脏特异性和多器官联合研究，以确定是否存在炎症和免疫反应的共同分子特征，并提供一种更好的方法来监测功能和应用旨在改善移植结果的干预措施。 在肾移植受者中，我们将调查：(A)是否可以通过连续的尿细胞信使核糖核酸图谱预测急性排斥反应；(B)基于信使核糖核酸图谱的预防性治疗可以预防急性排斥反应并保留GFR，以及(C)信使核糖核酸图谱可用于指导免疫抑制治疗，例如在稳定的受者中停用钙调神经磷酸酶抑制剂。 在不同的移植器官组中的研究将集中在器官特异性和非特异性损伤途径的分子图谱，在供者在购买之前和在移植后立即在受者中，以及它们对器官功能和同种异体反应的影响。更有针对性的研究将在肺和肝脏环境中进行。在同种异体肺移植中，我们将探讨：(A)供体和受体BAL细胞获得的细胞的mRNA谱是否可以预测不良后果，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着急性排斥反应特征。 在同种异体肝移植中，我们将探讨：(A)由独特的促炎症基因的mRNA谱指示的早期炎症反应是否与丙型肝炎复发相关，以及(B)类固醇避免方案是否下调表现出强烈炎症特征的移植物中的丙型肝炎复发。 这些机械性分析将使用动态(实时)定量聚合酶链式反应分析，并在某些情况下使用基于核酸的微阵列，以及对基因表达部位的免疫组织学分析，预计将产生将应用于移植人群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.