Mechanisms of RNA turnover during the epithelial-mesenchymal transition

项目总结/摘要 细胞改变其基因表达景观以改变其细胞状态。细胞迅速地 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， 再生，这一过程往往在癌症或纤维化等病理条件下出错。因此，有一个 生物医学需要了解驱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，其在人类健康中具有促进作用， 疾病虽然我们对促进EMT的转录程序了解很多， 影响细胞状态转换和EMT的基因表达控制。后的作用- 转录调控，以及它如何驱动和促进一系列EMT状态， 明白在这里，我们研究转录后调节的作用，重点是转录周转， 在急诊室神经嵴细胞经历严格调控的EMT，并提供了一个易于处理的模型系统， 研究EMT过程中RNA周转的基本机制。 我们假设，除了pro-EMT转录激活，抑制EMT的转录物（抗-EMT）， EMT）或用于维持先前蜂窝状态的服务必须被降级以驱动EMT和蜂窝状态转换。 使用神经嵴EMT作为模型系统，我们试图通过回答以下问题来验证这一假设 问题：1）抗EMT和残留转录本是如何靶向周转的; 2） EMT期间的RNA周转以及特定RNA周转如何促成混合EMT状态？回答 这些问题，我们将：1）应用无偏多组学和候选基因方法的组合， 鉴定在EMT期间促进RNA周转的RNA结合蛋白;和2）应用RNA测序 广泛识别RNA周转目标的方法，它们如何周转的机制，以及 这对混合EMT状态有何影响 该提案旨在了解EMT期间RNA周转的机制。的结果予以 这些研究将极大地推进目前对驱动EMT的基本细胞机制的理解， 调节人类健康和疾病中EMT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.