Role of long non-coding RNAs in sarcoma pathogenesis

项目摘要 尤文肉瘤是一种罕见的肿瘤，发生在儿童和年轻人。在这一过程中， 疾病最常见的是易位EWS/FLI。涉及ETS转录的类似易位 因素发生在其他恶性肿瘤，包括前列腺癌，扩大了我们的潜在相关性， 问题研究先前的工作已经确定，EWS/FLI可以通过抑制一些染色质重编程来导致染色质重编程。 增强子，同时通过直接结合激活其他增强子。我们自己的数据表明，EWS/FLI还 上调lncRNA的表达。该提案的主要目标是阐明 尤文肉瘤中的lncRNAs我们的初步数据表明，两个lncRNA，EWSAT 1和EWSAT 2，可能是 参与尤文肉瘤的致癌作用。这些RNA中的任何一种的敲低都会强烈降低 体外和体内尤文瘤的增殖。此外，RNAseq分析表明，这两个 lncRNA抑制大量基因，也已知被EWS/FLI抑制。我们的主要目标是 该建议旨在阐明这些lncRNA通过何种分子功能来促进Ewing 肉瘤。我们将集中研究这些lncRNA如何导致转录抑制。 在目标1中，我们将阐明EWSAT 1和2的表达如何改变细胞中关键的蛋白质-蛋白质相互作用。 尤因细胞。具体来说，我们将检验EWSAT 1或2表达改变EWS/FLI的PPI的假设， EWS或hnRNPK。在目标2中，我们将检验EWSAT 1或2通过调节以下能力发挥作用的假设： EWS/FLI与特异性增强子结合。ChIPseq和ChIRPseq将被用于确定这些lncRNA如何在细胞内表达。 与染色质相互作用以及它们如何改变组蛋白和其他染色质调节剂的结合。在目标3中，我们 使用Ewing患者来源的EWSAT 1/2基因，阐明EWSAT 1/2缺失或过表达的功能后果。 异种移植物和细胞系。

PROJECT SUMMARY Ewing sarcoma is a rare tumor that occurs in children and young adults. The driving oncogenic event in this disease is most commonly the translocation EWS/FLI. Similar translocations involving ETS transcription factors occur in other malignancies, including prostate cancer, broadening the potential relevance of our studies. Prior work has determined that EWS/FLI can lead to chromatin reprogramming by repressing some enhancers while activating others through direct binding. Our own data indicates that EWS/FLI also upregulates expression of lncRNAs. The primary goal of this proposal is to elucidate the molecular function of lncRNAs in Ewing sarcoma. Our preliminary data indicates that two lncRNAs, EWSAT1 and EWSAT2, may be involved in the oncogenic effects of Ewing sarcoma. Knock-down of either of these RNAs strongly decreases proliferation of Ewing tumors in vitro and in vivo. Furthermore, RNAseq analysis indicates that both of these lncRNAs repress a significant number of genes also known to be repressed by EWS/FLI. Our primary goal in this proposal is to elucidate the molecular function through which these lncRNAs act to promote Ewing sarcoma. We will focus our studies on determining how these lncRNAs lead to transcriptional repression. In Aim 1, we will elucidate define how expression of EWSAT1 and 2 alters key protein-protein interactions in Ewing cells. Specifically, we will test the hypothesis that EWSAT1 or 2 expression alters PPIs for EWS/FLI, EWS or hnRNPK. In Aim 2, we will test the hypothesis that EWSAT1 or 2 function by modulating the ability of EWS/FLI to bind to specific enhancers. ChIPseq and ChIRPseq will be used to determine how these lncRNAs interact with chromatin and how they alter binding of histones and other chromatin regulators. In Aim 3, we will elucidate the functional consequences of EWSAT1/2 loss or overexpression using Ewing patient derived xenografts and cell lines.