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

抗体介导的排斥反应中 microRNA 对基因表达的转录后调控

• 批准号: 10522285
• 负责人: Robert L Fairchild
• 金额: $ 70.11万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522285
• 关键词: Address; Affect; Allografting; Antibodies; Calcineurin inhibitor; Cells; Chronic; Clinical; Communities; Complement; Complex; Data; Disease; Event; Fibrosis; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Graft Rejection; Human; Injury; Injury to Kidney; Kidney; Kidney Failure; Kidney Transplantation; Knowledge; Lead; Maps; Mediating; Messenger RNA; MicroRNAs; Mus; Pathology; Pathway interactions; Play; Post-Transcriptional Regulation; Post-Translational Regulation; Process; Proteins; RNA; RNA-Binding Proteins; RNA-Induced Silencing Complex; Regulation; Reporting; Resources; Role; Sampling; Signal Pathway; Testing; Toxic effect; Transplantation; Untranslated RNA; allograft rejection; antibody-mediated rejection; base; clinically relevant; clinically translatable; crosslink; differential expression; digital; insight; integrin-linked kinase; interest; kidney allograft; kidney cell; member; miRNA expression profiling; mouse model; novel; prediction algorithm; prevent; renal damage; response; response to injury

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.

慢性抗体介导的排斥反应（AMR）是肾同种异体移植排斥反应的主要原因。然而 尽管其临床重要性，但对抗体反应如何的综合理解和 补体介导的攻击是否受移植肾调节尚未确定。 我们知识上的差距至少部分是由于对反应的不完全理解造成的 由肾脏产生，导致基因调节，促进或预防损伤。微小RNA (miRNA) 是一类在转录后调节基因表达的小非编码 RNA。 miRNA 在调节肾损伤中发挥着重要作用。然而，miRNA 的研究 排斥反应和肾损伤很大程度上是基于对总细胞 miRNA 的分析，这些 miRNA 是 疾病期间差异表达。这种方法是有问题的，因为它没有提供 有关这些 miRNA 靶向的 mRNA 的信息。为了解决这个问题，我们询问是否 可以分离 miRNA 和它们在 RNA 诱导沉默中靶向的 mRNA 通过分离与 RNA 结合蛋白 (RBP) AGO2 交联的 RNA 形成复合物 (RISC)。使用 通过这种方法，我们定义了第一个移植相关肾损伤的 miRNA-mRNA 相互作用图谱。 这些原理验证研究表明，在 miRNA-mRNA 靶组内，有可能 定义的 miRNA 及其靶向的 mRNA 在细胞中经历独特的变化 受伤。通路富集分析表明 RISC 复合体靶标中存在 miRNA mRNA 编码可能导致损伤的途径中的蛋白质。基于这些研究，我们 假设 miRNA-mRNA 靶组可用于识别基因通路 为 AMR 做出贡献。为了验证这一假设，我们将使用临床相关的小鼠模型来 确定 AMR 的 miRNA-mRNA 图谱，并使用目标组阐明的信息来 检查 miRNA 调控下的基因通路。我们检查我们的临床相关性 通过检查人类肾移植中是否发生类似的变化来得出结论。这些研究 将为驱动与 AMR 相关的病理过程提供独特的见解，这些信息 可用于区分 AMR 与其他类型的伤害，并为 移植和更广泛的科学界。