Biomolecular Markers for Safe Minimization of Immunosuppression

总体目标是在我们已经取得的进展的基础上再接再厉，并利用我们最近的开创性进展 基因表达谱技术研究进展。 具体目标1：对肾移植受者的尿液进行广泛、公正、深入的测序 接受监测和/或原因活组织检查并发现新的mRNA和miRNA签名 急性排斥反应(AR)、亚临床排斥反应(SAR)和慢性移植物肾病的诊断和预后 (CAN)(SA.IA)，并开发了用于表征移植肾体内免疫状态的新参数 受体及其免疫抑制治疗的充分性(SA。IB)。 特定目的2：测试先前发现的ISS标准化的CD3E的3基因特征和 在连续的尿样中检测IP-10mRNA，可以预测AR和 香港特别行政区(SA.2a)；测试先前发现的波形蛋白、NKCC2、E-钙粘附素的4基因签名 在连续的尿样中检测到的18S rRNA预测了随后的CAN(SA)的发展。 2b)；并测试由miR-21，-21*，-30a-3p，-100；-142-3p/5p，-192，-194，- 200b，-222；和-223，在连续的尿样中测量，预测AR的后续发展， Sar和Can(SA.2C)。具体目标3：测试先前发现的签名和 在连续的尿样中测量的miRNA组与诱导治疗的类型(T 细胞耗尽抗体诱导与抗IL-2受体单抗诱导)和维持。 免疫抑制治疗(他克莫司与不使用他克莫司)。发现信使RNA和microRNA 通过测序将AR、SAR和CAN(SA.1)，并以顺序样本(SA. 2)，将被并入统计分析以确定：(1)先前识别的签名是否 可以进一步优化以提高其诊断和预后效用；以及(2)以前的 识别的特征可以进一步提炼，以提高其跟踪诱导治疗影响的能力 维持治疗对肾移植受者免疫状态的影响。 相关性(请参阅说明)： 过量的免疫抑制药物与感染、恶性肿瘤和心血管疾病有关 器官移植受者代谢异常，而免疫抑制不足与 同种异体排斥反应。因此，优化免疫抑制是器官移植的主要目标。我们 建议开发基于基因的预后和诊断测试来表征体内免疫状态 用于监测肾移植受者的免疫抑制治疗的情况。

The overall gear is to build on the progress we have made and to leverage our recent ground-breaking advances in gene expression profiling technologies. SPECIFIC AIM 1: To perform broad, unbiased, deep sequencing of urine from kidney graft recipients undergoing surveillance and/or for-cause biopsies and discover new mRNA and miRNA signatures diagnostic and prognostic of acute rejection (AR), subclinical AR (SAR) and chronic allograft nephropathy (CAN) (SA. IA) and develop novel parameters for characterizing the in-vivo immune status of kidney graft recipients and the adequacy of their immunosuppressive therapy (SA. IB). SPECIFIC AIM 2: To test whether a previously discovered 3-gene signature of ISS-normalized C D 3 E and IP-10 mRNA, measured in sequential urine specimens, predicts the subsequent development of AR and SAR (SA. 2A); to test whether a previously discovered 4-gene signature of vimentin, NKCC2, E-cadherin and 18S rRNA, measured in sequential urine specimens, predicts the subsequent development of CAN (SA. 2B); and to test whether a miRNA panel comprised of miR-21, -21*, -30a-3p, -100; -142-3p/5p, -192, -194, - 200b, -222; and -223, measured in sequential urine specimens, predicts the subsequent development of AR, SAR and CAN (SA. 2C). SPECIFIC AIM 3: To test whether the previously discovered signatures and the miRNA panel, measured in sequential urine specimens, are associated with the type of induction therapy (T cell depleting antibody induction vs. anti-IL-2 receptor mABs induction) and with maintenance . immunosuppressive therapy (tacrolimus vs. no tacrolimus). Messenger RNAs and microRNAs, discovered by sequencing to be associated with AR, SAR and CAN (SA. 1) and measured in sequential samples (SA. 2), will be incorporated in statistical analyses to determine: (1) whether the previously identified signatures can be further optimized to improve their diagnostic and prognostic utility; and (2) whether the previously identified signatures can be further refined to improve their ability to track the influence of induction therapy and maintenance therapy on the immune status of kidney graft recipients. RELEVANCE (See instructions): An excess of immunosuppressive drugs is associated with infections, malignancy and cardiovascular and metabolic aberrations in organ graft recipients whereas inadequate immunosuppression is associated with allograft rejection. Thus, immunosuppression optimization is a major goal in organ transplantation. We propose to develop gene-based prognostic and diagnostic tests for characterizing the in-vivo immune status of kidney graft recipients and for monitoring the adenuacv of their immunosuppressive therapy.