Dynamics of Gene and Isoform Regulation during EMT and tumor progression

EMT 和肿瘤进展过程中基因和亚型调控的动态

基本信息

批准号：
8852097
负责人：
CHRISTOPHER B BURGE
金额：
$ 69.39万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8852097
关键词：
3&apos Untranslated Regions Address Affect Aggressive behavior Algorithms Alternative Splicing Amino Acid Sequence Apoptosis Behavioral Bioinformatics Biological Biological Assay Biological Markers Breast Cancer Model Breast Epithelial Cells Cancer Patient Cell model Cells Cellular Morphology Code Complex Cultured Cells Cytoskeleton Data Data Analyses Data Set Development Distant Elements Epithelial Epithelial Cells Equipment and supply inventories Event Gene Expression Gene Expression Profile Gene Targeting Genes Genetic Transcription Goals Health Human Image Immunofluorescence Immunologic In Vitro Intercellular Junctions Life Malignant - descriptor Malignant Epithelial Cell Malignant Neoplasms Mediating Mesenchymal Messenger RNA Metastatic breast cancer MicroRNAs Modeling Mouse Mammary Tumor Virus Mus Neoplasm Metastasis Nucleotides Pathway interactions Peptide Sequence Determination Phenotype Phosphorylation Physiology Play Primary Neoplasm Process Production Property Protein Isoforms Proteins RNA RNA Binding RNA Splicing RNA-Binding Proteins Regulation Reporter Research Resistance Role Seeds Series Set protein Signal Pathway Signal Transduction Site Snails Spliced Genes Stem cells Structure Testing Therapeutic Tissues Transgenes Transgenic Mice Untranslated Regions Work cancer cell cell behavior cell motility cohort design epithelial to mesenchymal transition extracellular gene function in vivo insight mRNA Stability neoplastic cell novel programs protein expression protein protein interaction response transcription factor transcriptome sequencing tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): The epithelial-mesenchymal transition (EMT) is a complex cell-biological program that operates during the progression of carcinoma cells to high-grade malignancy, conferring on these cells many of the attributes associated with aggressive tumors, including the ability to disseminate to distant sites and to seed metastatic colonies. This program is orchestrated by a series of pleiotropically acting master transcription factors (EMT- TFs) that organize the complex changes in gene expression causing the replacement of a large cohort of epithelial cell proteins with those associated with the mesenchymal cell state. A major, critical level of control required for expression of the aggressive mesenchymal state is poorly understood however: the precursors of many of the mRNAs whose expression changes during the EMT also undergo alternative splicing (AS) that confer on resulting mature, processed mRNAs altered properties, including changes in stability, protein-coding information, and responsiveness to microRNA-mediated inhibition. The current fragmentary insights into the effects of AS on the execution of the EMT program make it impossible to form a reasonably complete understanding of how this critical cell-biological program is effected. The proposed research will begin by enumerating the hundreds of AS events that occur in response to several alternative mechanisms of inducing an EMT program both in cultured cells and in a living tissue. Having done so, bioinformatics algorithms will be employed to determine the sequences adjacent to involved splice sites. Thereafter, using the known nucleotide-recognizing properties of the large array of already-characterized RNA- binding, splice-regulating proteins, predictions will be made by these algorithms about the identities of the splice-regulators that are likely to b responsible for the observed large-scale shifts in AS occurring during passage through an EMT. This experimental strategy should yield the identities of key regulators of AS that are likely to b as important functionally as the EMT-TFs in executing the EMT program. Experimental tests designed to functionally test the candidacies of these AS factors will be performed. These tests will gauge whether the forced or blocked expression of these factors affect execution of critical components of the EMT program, and whether, as predicted, such imposed changes in AS factor expression affect the production of key EMT-associated proteins, i.e., proteins that play key roles in the expression of the epithelial versus mesenchymal cell phenotypes observed during malignant progression. This work also has the potential to identify novel biomarkers of the EMT program that are applicable, for example, for the detections of stem cells in a variety of epithelial tissues.

描述（由申请人提供）：上皮间质转化（EMT）是一个复杂的细胞生物学程序，在癌细胞发展为高度恶性肿瘤的过程中发挥作用，赋予这些细胞许多与侵袭性肿瘤相关的属性，包括传播到远处并播种转移集落的能力。这该程序由一系列多效性主转录因子（EMT-TF）精心策划，这些因子组织基因表达的复杂变化，导致大量上皮细胞蛋白被与间充质细胞状态相关的蛋白取代。然而，人们对侵袭性间充质状态表达所需的主要、关键控制水平知之甚少：许多在 EMT 期间表达发生变化的 mRNA 的前体也会经历选择性剪接 (AS)，从而赋予最终的成熟、加工过的 mRNA 改变的特性，包括稳定性、蛋白质编码信息以及对 microRNA 介导的抑制反应的变化。目前对 AS 对 EMT 程序执行影响的零碎见解使得不可能对这一关键的细胞生物学程序如何影响形成相当完整的理解。拟议的研究将首先列举数百个 AS 事件，这些事件是对在培养细胞和活体组织中诱导 EMT 程序的几种替代机制作出反应而发生的。完成此操作后，将采用生物信息学算法来确定与所涉及的剪接位点相邻的序列。此后，利用大量已经表征的RNA结合、剪接调节蛋白的已知核苷酸识别特性，这些算法将做出关于剪接调节器的身份的预测，这些剪接调节器可能对观察到的结果负责。在经历 EMT 期间发生 AS 的大规模转变。该实验策略应该能够确定 AS 关键调节因子的身份，这些调节因子在执行 EMT 程序时可能与 EMT-TF 具有同样重要的功能。将进行旨在从功能上测试这些 AS 因素的候选资格的实验测试。这些测试将衡量这些因子的强制或阻断表达是否会影响 EMT 程序关键组成部分的执行，以及 AS 因子表达的这种强加变化是否会影响关键 EMT 相关蛋白的产生，即，在恶性进展期间观察到的上皮细胞与间充质细胞表型的表达中发挥关键作用。这项工作还有可能识别 EMT 程序的新型生物标志物，例如，可用于检测各种上皮组织中的干细胞。