Mechanisms of RNA turnover during the epithelial-mesenchymal transition

项目摘要/摘要 细胞改变它们的基因表达图景来改变它们的细胞状态。细胞能够迅速地 细胞状态之间的转换对于生物学中的许多过程--从发育到伤口愈合和 再生，在癌症或纤维化等病理条件下经常出错的过程。因此，有一个 生物医学需要了解推动这些快速细胞转变的基本机制。上皮细胞- 间充质转化(EMT)是一种这样的细胞转化，在人类健康和 疾病。虽然关于促进EMT的转录程序知道得很多，但还有其他的水平 控制影响细胞状态转变的基因表达，进而影响EMT。岗位的作用-- 转录调控，以及它如何驱动和促进一系列EMT状态，并不是很好 明白了。在这里，我们研究转录后调控的作用，重点是转录后的周转， 在急救期间。神经脊细胞经历了严格调控的EMT，并提供了一个易于处理的模型系统，在其中 探讨EMT过程中RNA周转的基本机制。 我们假设，除了前EMT转录激活外，抑制EMT的转录本(抗- EMT)或用于维持先前的蜂窝状态必须被降级以驱动EMT和小区状态转变。 使用神经峰EMT作为模型系统，我们试图通过回答以下问题来检验这一假设 问题：1)反EMT和残存转录的目标是什么；以及2)目标是什么 EMT期间的RNA周转以及特定的RNA周转如何对混合EMT状态做出贡献？回答 这些问题，我们将：1)应用无偏多组学和候选基因相结合的方法来 确定在EMT过程中促进RNA周转的RNA结合蛋白；以及2)应用RNA测序 广泛确定RNA周转目标的方法、如何周转它们的机制以及 这对混合EMT状态有何贡献。 这一建议旨在了解EMT过程中RNA周转的机制。这些研究的结果 研究将极大地促进目前对驱动EMT的基本细胞机制的理解，提供 调节EMT在人类健康和疾病中的新靶点。

Project Summary/Abstract Cells alter their gene expression landscape to change their cellular state. The ability of cells to quickly transit between cell states is essential for many processes in biology—from development to wound healing and regeneration, processes which often go awry in pathological conditions like cancer or fibrosis. Thus, there is a biomedical need to understand the basic mechanisms driving these rapid cellular transitions. The epithelial- mesenchymal transition (EMT) is one such cellular transition that has reiterative roles in human health and disease. While much is known about the transcriptional programs that promote EMT, there are additional levels of gene expression control that impact cell state transitions and by extension EMT. The role of post- transcriptional regulation, and how it drives and contributes to a spectrum of EMT states, is not well understood. Here, we examine the role of post-transcriptional regulation, with emphasis on transcript turnover, during EMT. Neural crest cells undergo a tightly regulated EMT and offer a tractable model system in which to investigate the basic mechanisms of RNA turnover during EMT. We hypothesize that, in addition to pro-EMT transcriptional activation, transcripts that inhibit EMT (anti- EMT) or serve to maintain a previous cellular state must be degraded to drive EMT and cell state transitions. Using neural crest EMT as a model system, we seek to test this hypothesis by answering the following questions: 1) How are anti-EMT and residual transcripts targeted for turnover; and 2) What are the targets of RNA turnover during EMT and how does specific RNA turnover contribute to hybrid EMT states? To answer these questions, we will: 1) Apply a combination of unbiased multi-omic and candidate gene approaches to identify the RNA-binding proteins that promote RNA turnover during EMT; and 2) Apply RNA-sequencing approaches to broadly identify the targets of RNA turnover, the mechanism of how they are turned over, and how this contributes to hybrid EMT states. This Proposal seeks to understand the mechanisms of RNA turnover during EMT. The results of these studies will greatly advance current understanding of the basic cellular mechanisms driving EMT, providing novel targets for modulating EMT in human health and disease.