Post-transcriptional regulation of gene expression by microRNAs in antibody-mediated rejection

慢性抗体介导的排斥反应(AMR)是肾移植排斥反应的主要原因。还没有 尽管它在临床上很重要，但对抗体和 补体介导的攻击是否受移植肾的调控尚未建立。 我们知识上的这种差距至少部分是由于对反应的不完全理解。 由肾脏制造，导致基因调节，促进或防止损伤。MicroRNAs(MiRNAs) 是一类小的非编码RNA，在转录后调节基因表达。 MiRNAs在调节肾脏损伤中发挥重要作用。然而，对miRNAs的研究 排斥反应和肾脏损伤在很大程度上是基于对细胞总miRNA的分析，这些miRNA是 在疾病期间表达的差异。这种方法是有问题的，因为它没有提供 有关这些miRNAs靶向的mRNAs的信息。为了解决这个问题，我们询问了 在RNA诱导的沉默中分离miRNAs及其靶向的mRNAs是可能的 通过分离与RNA结合蛋白(RBP)AGO2交联的RNA来形成复合体(RISC)。vbl.使用 通过这种方法，我们定义了首个移植相关肾损伤的miRNA-mRNA相互作用图谱。 这些原则证明研究表明，在miRNA-mrna靶标体内，有可能 已定义的miRNAs和它们针对的在细胞中经历独特变化的mRNAs 受伤。途径富集化分析表明，miRNAs存在于RISC复合靶中 在可能导致损伤的途径中编码蛋白质的mRNAs。基于这些研究，我们 假设miRNA-mRNA目标组可用于识别 为AMR做出贡献。为了验证这一假设，我们将使用一个临床相关的小鼠模型来 确定AMR的miRNA-mRNA图谱，并使用靶组阐明的信息来 研究miRNAs调控下的基因途径。我们研究了我们的临床相关性 通过检查人类肾脏移植是否发生类似的变化来发现这一点。这些研究 将提供对驱动与AMR相关的病理过程的独特洞察，信息 可用于将AMR与其他类型的损伤区分开来，并为 移植和更广泛的科学界。

Chronic antibody-mediated rejection (AMR) is a major cause of renal allograft rejection. Yet despite its clinical importance an integrated understanding of how responses to antibody and complement mediated attack are regulated by the transplanted kidney has not been established. This gap in our knowledge is due at least in part to an incomplete understanding of responses made by the kidney that result in gene regulation promote or prevent injury. MicroRNAs (miRNAs) are a class of small noncoding RNAs that regulate gene expression post-transcriptionally. miRNAs play an important role in regulating renal injury. However, the study of miRNAs in rejection and renal injury has largely been based on analysis of total cellular miRNAs that are differentially expressed during disease. This approach is problematic because it does not provide information on the mRNAs targeted by these miRNAs. To address this issue, we asked whether it is possible to isolate miRNAs and the mRNAs they are targeting in the RNA-Induced Silencing Complex (RISC) by isolating RNAs cross-linked to the RNA Binding Protein (RBP) AGO2. Using this approach we defined the first miRNA-mRNA interaction map for transplant related renal injury. These proof-of-principle studies revealed that within the miRNA-mRNA targetome it is possible to defined miRNAs and the mRNAs they target that undergo unique changes in cells undergoing injury. Pathway enrichment analysis indicated that miRNAs present in the RISC complex target mRNAs encoding proteins in pathways that may contribute to injury. Based on these studies, we hypothesize that the miRNA-mRNA targetome can be used to identify gene pathways that contribute to AMR. To test this hypothesis, we will use a clinically relevant murine model to determine the miRNA-mRNA map for AMR and use information elucidated by the targetome to examine gene pathways under regulation by miRNAs. We examine the clinical relevance of our findings by examining whether similar changes occur in human kidney transplants. These studies will provide unique insight into process that drive pathology associated with AMR, information that could be used to distinguish AMR from other types of injury, and provides a novel resource to the transplantation and wider scientific community.