Grhl2-regulation of Developing Ectoderm as a Model to Identify Suppressors of EMT

Grhl2-发育外胚层的调控作为识别 EMT 抑制因子的模型

• 批准号: 8712104
• 负责人: Heather Joy Ray
• 金额: $ 3.06万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8712104
• 关键词: Affect; Automobile Driving; Binding Sites; Bioinformatics; Biological Assay; Breast Cancer Cell; Breast Cancer Model; CCL4 gene; Cancer cell line; Cell Line; Cell model; Cells; Characteristics; Clinical; Development; Distal; Distant; Dorsal; E-Cadherin; Ectoderm; Ectoderm Cell; Embryo; Embryonic Development; Ensure; Epithelial; Epithelium; Family; Gene Activation; Gene Expression Regulation; Genes; Head; Homologous Gene; Human; Image; In Situ Hybridization; In Vitro; Knockout Mice; Knowledge; Laminin; Life; Luciferases; Malignant Neoplasms; Mesenchymal; Modeling; Molecular; Morphology; Mus; Nature; Neoplasm Metastasis; Neural Crest; Neural Crest Cell; Neural Tube Closure; Neural Tube Defects; Neural tube; Neuroectoderm; Outcome; Play; Process; Promoter Regions; Proteins; RNA Sequences; RNA Splicing; Regulation; Reporter; Role; Site; System; TRIM Motif; Testing; Time; Tissues; Travel; Work; cancer cell; cell behavior; cell type; epithelial to mesenchymal transition; epitheliasin; gene function; genetic regulatory protein; in vivo; loss of function; malignant breast neoplasm; migratory population; mouse model; new therapeutic target; novel; public health relevance; recombinase; relating to nervous system; therapeutic target; tool; transcription factor; transcriptome sequencing; tumor

DESCRIPTION (provided by applicant): The epithelial-to-mesenchymal (EMT) transition plays an essential role during embryonic development. Studies have also found that EMT can be involved in the promotion of various epithelial cancers. Recent evidence supports the idea that the EMT process between development and cancer is similar at the molecular level. Therefore, using an in vivo developmental system to study how EMT is regulated may identify genes that are relevant targets for treatment of human cancers. During embryonic neural tube closure, neural crest cells undergo an EMT as they delaminate out of the neuroectoderm to migrate to distant sites. At the same time, the neighboring non-neural ectoderm (NNE) cells must remain tightly associated within the same epithelium in order for neural tube closure to proceed correctly. Much is known about the regulatory mechanisms driving neural crest EMT, however less is known about how the non-neural ectoderm enforces its epithelial fate. The transcription factor Graineyhead like 2 (Grhl2) is expressed within the NNE cells during neural tube closure and may act as a suppressor of EMT via its direct downstream targets. In fact, GRHL2 has recently been shown to act in this manner in a breast cancer model. Isolation of NNE from embryos at the time of neural tube closure followed by high throughput RNA-sequencing led to the identification of several genes that may be involved in EMT suppression in this tissue. In this project, a loss of function Grhl2 mouse model (Grhl21Nisw/1Nisw) will be used to explore these genes for a potential role within a Grhl2-regulated network of EMT suppression in the NNE of the mouse embryo. Further assessment of these genes will show how loss of gene function affects the dynamics of NNE cell behavior during neural tube closure. Genes that are found to suppress EMT in the developing neural tube will then be further investigated in a breast cancer model to determine if they affect both in vitro EMT as well as in vivo metastasis. Through this study, a further understanding of a Grhl2- regulated gene network of EMT suppression will be gained and targets may be identified that could impact the progression of epithelial cancers.

描述（由申请人提供）：上皮细胞向间充质细胞（EMT）转化在胚胎发育过程中发挥重要作用。研究还发现，EMT可参与促进各种上皮癌。最近的证据支持这样一种观点，即发育和癌症之间的EMT过程在分子水平上是相似的。因此，使用体内发育系统来研究EMT是如何调节的，可以鉴定作为治疗人类癌症的相关靶点的基因。在胚胎神经管闭合期间，神经嵴细胞经历EMT，因为它们从神经外胚层分层迁移到远处。同时，相邻的非神经外胚层（NNE）细胞必须在同一上皮内保持紧密结合，以使神经管闭合正确进行。关于驱动神经嵴EMT的调节机制知之甚多，然而关于非神经外胚层如何执行其上皮命运知之甚少。转录因子Graineyhead like 2（Grhl 2）在神经管闭合期间在NNE细胞内表达，并且可以通过其直接下游靶标作为EMT的抑制剂。事实上，GRHL 2最近已被证明在乳腺癌模型中以这种方式起作用。在神经管闭合时从胚胎中分离NNE，然后进行高通量RNA测序，鉴定出可能参与该组织中EMT抑制的几个基因。在这 项目中，将使用功能缺失的Grhl 2小鼠模型（Grhl 21 Nisw/1 Nisw）来探索这些基因在小鼠胚胎的NNE中的EMT抑制的Grhl 2调节网络内的潜在作用。对这些基因的进一步评估将显示基因功能的丧失如何影响神经管闭合期间NNE细胞行为的动力学。在发育中的神经管中发现抑制EMT的基因将在乳腺癌模型中进一步研究，以确定它们是否影响体外EMT和体内转移。通过这项研究，将获得对EMT抑制的Grhl 2调节基因网络的进一步理解，并可能确定可能影响上皮癌进展的靶点。