Protein-RNA interactions in cancer

项目总结/摘要 蛋白质-RNA相互作用是基因调控的重要组成部分，但尚未得到充分研究。rna结合 蛋白质执行与mRNA的产生、剪接、加工和稳定性相关的许多功能 分子。几年前，我们发现癌胚RNA结合蛋白IGF 2BP 3结合到3'端， mRNA的非翻译区，并通过涉及RNA诱导的 沉默复合体在最近的研究中，我们发现IGF 2BP 3也结合在3 '-剪接位点附近， 调节可变剪接。总之，我们的研究结果表明IGF 2BP 3在mRNA调控中具有双重作用。 IGF 2BP 3调节与增殖、迁移和信号传导相关的基因，这些基因在胎儿发育中很重要。 发展-而且在癌症中。与这种基因调控功能一致，IGF 2BP 3在一个细胞中过表达。 广泛的恶性肿瘤，包括急性白血病，并且当高表达时预示着不良预后。 使用新的IGF 2BP 3缺陷的小鼠遗传模型，我们现在发现IGF 2BP 3是必需的。 在体内发展成完全渗透的致命白血病。我们之前的大量工作， 出版的和未出版的，提供了一个机制框架，其功能，和一个坚实的基础， 这种蛋白质在疾病中的重要性。为了充分理解这些蛋白质-RNA相互作用的性质， 了解它们在癌症中的作用，我们提出两个目标。在第一个目标中，我们将仔细描述机制 IGF 2BP 3的潜在功能通过仔细执行的实验的组合（目的1）。第二个目标， 我们将使用一组数据来描述IGF 2BP 3在白血病发生、增殖和维持中的重要性。 精心构建的遗传模型和原代细胞中的基因编辑。接下来，我们将描述这两个 提出的基因调节机制- RNA稳定性和前mRNA剪接-在癌症起始中起作用， 使用反向遗传学、miRNA调节和同种型特异性表达的组合。在一起，这项工作 将导致一个分层的，详细的了解蛋白质-RNA相互作用的机制基础是如何密切 与基因表达失调和细胞恶性转化有关。重要的是，它将为 发展新的诊断和治疗方法的方式在恶性肿瘤的特点是巨大的， 转录组失调由RNA结合蛋白的改变支持。

PROJECT SUMMARY/ ABSTRACT Protein-RNA interactions underlie an important and understudied component of gene regulation. RNA-binding proteins carry out numerous functions relating to the production, splicing, processing, and stability of mRNA molecules. A few years ago, we discovered that the oncofetal RNA binding protein, IGF2BP3, binds to the 3’ untranslated region of mRNA and regulates mRNA stability via a mechanism that involves the RNA-induced silencing complex. In more recent work, we found that IGF2BP3 also binds near 3’-splice sites, and may regulate alternative splicing. Together, our findings suggest dual roles in mRNA regulation for IGF2BP3. IGF2BP3 regulates genes that are related to proliferation, migration, and signaling- which are important in fetal development- but also in cancer. Concordant with this gene regulatory function, IGF2BP3 is overexpressed in a wide range of malignancies, including acute leukemia, and portends a poor prognosis when highly expressed. Using novel, murine genetic models of IGF2BP3 deficiency, we have now discovered that IGF2BP3 is required for the development of a fully-penetrant, lethal leukemia in vivo. Together, our extensive prior work, both published and unpublished, provides a mechanistic framework for its function, and a solid foundation for the importance of this protein in disease. To fully understand the nature of these protein-RNA interactions and to understand their role in cancer, we propose two aims. In the first aim, we will carefully delineate the mechanism underlying IGF2BP3 function by a combination of carefully executed experiments (Aim 1). In the second aim, we will characterize the importance of IGF2BP3 in leukemia initiation, propagation and maintenance using a set of carefully constructed genetic models and gene editing in primary cells. Next, we will characterize how the two proposed gene regulatory mechanisms- RNA stability and pre-mRNA splicing- play roles in cancer initiation, using a combination of reverse genetics, miRNA regulation, and isoform specific expression. Together, this work will lead to a layered, detailed understanding of how mechanistic basis of protein-RNA interactions is intimately connected to gene expression deregulation and the malignant transformation of cells. Importantly, it will pave the way to develop novel diagnostic and therapeutic approaches in malignancies characterized by massive transcriptomic dysregulation underpinned by alterations of RNA-binding proteins.