Regulation of alternative splicing during epithelial-mesenchymal transition

上皮间质转化过程中选择性剪接的调节

• 批准号: 10364651
• 负责人: Chonghui Cheng
• 金额: $ 40万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10364651
• 关键词: Address; Alternative Splicing; Bioinformatics; Biological; CD44 gene; Cells; Cellular biology; Development; Developmental Process; Disease; Epithelial; Fibrosis; Genes; Goals; Mammals; Mesenchymal; Messenger RNA; Molecular; Neoplasm Metastasis; Phenotype; Phosphotransferases; Protein Isoforms; RNA; RNA Splicing; RNA-Binding Proteins; Regulation; Research; Signal Transduction; System; Time; Tissues; Transcript; Work; causal variant; cell motility; experimental analysis; insight; mRNA Precursor; programs

ABSTRACT Alternative splicing of pre-mRNA enables a gene to give rise to multiple distinct mRNA transcripts, yielding protein isoforms with different, even opposing, functions. The long-term goal of our research is to understand the molecular mechanisms and function of alternative splicing on biological activities. The focus of this proposal is to investigate the regulation of alternative splicing in epithelial-mesenchymal transition (EMT). EMT is an essential developmental process that allows cells to change from a tightly packed cobble-stone-like epithelial cellular state to a motile and spindle-shaped mesenchymal cellular state. When abnormally activated, EMT promotes many types of diseases, including tissue fibrosis and cancer metastasis. Through working at the intersection of RNA splicing and cell biology, my lab has made several important findings connecting RNA regulation and EMT. Our work revealed, for the first time, that splice isoform switching of the CD44 gene causally controls EMT. By manipulating CD44 alternative splicing, we were able to convert cells between the epithelial and mesenchymal states. We have also identified the RNA-binding protein (RBP) hnRNPM as a critical splicing factor that promotes EMT through the regulation of alternative splicing. These results suggest that RNA splicing regulation could serve as an important mechanism that provides cellular plasticity. By shifting the programs of alternative splicing, cells are able to convert between the epithelial and mesenchymal cellular states. This capacity of reversing phenotypes is important for normal developmental EMT, as well as for cancer metastasis. Major new discoveries are necessary to fully understand this phenomenon and its underlying mechanisms. Our proposed research program is focused on (1) determining whether splice isoform switching acts as a prevalent mechanism that drives EMT or, if it is largely a phenomenon of byproducts; (2) understanding how RBPs precisely control alternative splicing during EMT; and (3) dissecting how signaling cascades elicit signals to RBPs and trigger alternative splicing changes during EMT. We have made major efforts in the past several years to build up experimental systems and gain expertise to help carry out our proposed studies. These efforts include large-scale RNA profiling of alternative splicing in multiple EMT systems, bioinformatics and experimental analysis of RBPs, as well as our recently completed kinase screen for EMT-associated alternative splicing alterations. Accomplishing the proposed work will provide new insights into our understanding of the regulatory mechanisms of alternative splicing, thus contributing to biological relevance in normal development and diseases.

抽象的 前MRNA的替代剪接使基因能够产生多个不同的mRNA转录本， 产生具有不同甚至相反功能的蛋白质同工型。我们研究的长期目标是 了解替代剪接对生物学活性的分子机制和功能。重点 该建议是研究上皮间质转变（EMT）中替代剪接的调节。 EMT是一个必不可少的发育过程，它允许细胞从类似于鹅卵石的紧密堆满的变化 上皮细胞状态至运动和纺锤形间充质细胞状态。当异常激活时， EMT促进许多类型的疾病，包括组织纤维化和癌症转移。通过在 RNA剪接和细胞生物学的交集，我的实验室已经做出了几个重要发现，将RNA连接起来 调节和EMT。我们的工作首次揭示了CD44基因的剪接同工型切换 控制EMT。通过操纵CD44替代剪接，我们能够在上皮之间转换细胞 和间充质状态。我们还确定了RNA结合蛋白（RBP）HNRNPM作为临界剪接 通过调节替代剪接来促进EMT的因素。这些结果表明RNA剪接 调节可以作为提供细胞可塑性的重要机制。通过转移程序 替代剪接，细胞能够在上皮和间质细胞状态之间转换。这 逆转表型的能力对于正常发育EMT以及癌症转移很重要。 为了充分理解这种现象及其基本机制，必不可少的新发现是必要的。我们的 拟议的研究计划的重点是（1）确定剪接同工型转换是否充当普遍存在 驱动EMT或在很大程度上是副产品现象的机制； （2）了解RBP EMT期间精确控制替代剪接； （3）剖析信号级联向RBP引起信号 并触发EMT期间的替代剪接变化。在过去的几年中，我们已经做出了重大努力 建立实验系统并获得专业知识，以帮助我们进行我们建议的研究。这些努力包括 多个EMT系统，生物信息学和实验性的替代剪接的大规模RNA分析 RBP的分析以及我们最近完成的激酶屏幕，用于EMT相关替代剪接 改变。完成拟议的工作将为我们对监管的理解提供新的见解 替代剪接的机制，从而有助于正常发育和疾病中的生物学相关性。