Biomolecular Markers for Safe Minimization of Immunosuppression

整体齿轮是建立在我们已经取得的进展，并利用我们最近的突破性 基因表达谱技术的进展。 具体目的1：对肾移植受者的尿液进行广泛、无偏倚、深度测序 进行监测和/或有因活检，并发现新的mRNA和miRNA特征 急性排斥反应（AR）、亚临床AR（SAR）和慢性移植物肾病的诊断和预后 (CAN)（SA. IA）并开发用于表征肾移植物的体内免疫状态的新参数 接受者及其免疫抑制治疗的充分性（SA. IB）。 具体目标2：为了测试先前发现的ISS标准化的CD 3 E和CD 3 E的3个基因签名是否是 在连续的尿液标本中测量IP-10 mRNA，可以预测AR的后续发展， SAR（SA. 2A）;以测试先前发现的波形蛋白、NKCC 2、E-钙粘蛋白的4-基因签名是否与先前发现的波形蛋白、NKCC 2、E-钙粘蛋白的4-基因签名相同。 和18 S rRNA，在连续的尿液标本中测量，预测CAN的后续发展（SA. 并测试由miR-21、-21*、-30a-3p、-100、-142-3p/5p、-192、-194、-200、-200、-20 200 b，-222;和-223，在连续的尿标本中测量，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 在连续的尿液样本中测量的miRNA组与诱导治疗的类型相关（T 细胞消耗抗体诱导相对于抗IL-2受体mAb诱导）和维持。 免疫抑制治疗（他克莫司vs.无他克莫司）。发现了信使RNA和微小RNA， 与AR、SAR和CAN（SA）相关联的排序。1)并在连续样品中测量（SA. 2），将纳入统计分析，以确定：（1）是否以前确定的签名 可以进一步优化，以提高其诊断和预后的效用;（2）是否以前的 可以进一步细化识别的特征，以提高其跟踪诱导治疗影响的能力， 以及维持治疗对肾移植受者免疫状态的影响。 相关性（参见说明）： 过量的免疫抑制药物与感染、恶性肿瘤和心血管疾病有关， 器官移植受者的代谢异常，而免疫抑制不足与 同种异体移植排斥反应。因此，免疫抑制优化是器官移植中的主要目标。我们 建议开发基于基因的预后和诊断测试，用于表征体内免疫状态 并用于监测其免疫抑制治疗的效果。

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.