COORDINATE REGULATION BY MAP3K4 OF EPIGENETIC MODIFIERS CONTROLLING EMT

MAP3K4 对控制 EMT 的表观遗传修饰子的协调调控

基本信息

批准号：
9328101
负责人：
Amy N Abell
金额：
$ 27.55万
依托单位：
UNIVERSITY OF MEMPHIS
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-25 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9328101
关键词：
Acetylation Acetyltransferase Actins Active Sites Adherence Apical Back Biological Biological Assay Biological Markers Biological Process Breast Cancer Cell Cancer Control Cancer Patient Cause of Death Cell Surface Proteins Cells ChIP-seq Characteristics Chromatin Cytoskeleton Data Deacetylase Deacetylation Decidual Cell Reactions Defect Development Disease Drug Targeting Epigenetic Process Epithelial Epithelial Cells Event Fibrosis GALNT3 gene Gene Expression Gene Expression Profile Gene Expression Regulation Genes Goals HDAC6 gene Histone Acetylation Histone H2A Histone H2B Human In Vitro Individual Inflammation Injection of therapeutic agent Kidney Liver Lung Lysine Malignant Neoplasms Mediating Mesenchymal Molecular Morphology Mus Mutation Neoplasm Metastasis Overlapping Genes Pathologic Pathology Phenotype Phosphotransferases Physiological Processes Process Property Proteins Publishing Regulation Resistance Role Signal Pathway Signal Transduction Stem cells System Tissues Work base blastocyst cancer cell cell motility conventional therapy epigenetic regulation epithelial to mesenchymal transition human disease in vivo in vivo Model inhibitor/antagonist innovation knock-down malignant breast neoplasm new therapeutic target novel novel strategies overexpression prevent programs promoter public health relevance small hairpin RNA stemness targeted treatment therapy resistant tissue regeneration transcription factor transcriptome sequencing trophoblast

项目摘要

DESCRIPTION (provided by applicant): Epithelial to mesenchymal transition (EMT) is a developmental program in which non-motile epithelial cells with tight cell-cell adherence convert into individual, motile mesenchymal cells. Importantly, EMT is reversible through a mesenchymal to epithelial transition (MET) converting mesenchymal cells back to an epithelial state. The EMT program is essential for normal processes during development and for tissue regeneration. EMT is reactivated in human diseases including tissue fibrosis in the kidney, liver, and lung and in cancer metastasis, making EMT a key target for drug therapy. Targeted mutation of the active site lysine of the kinase MAP3K4 in the mouse (KI4) results in severe developmental defects due to disrupted EMT. Epithelial trophoblast stem (TS) cells from kinase-inactive MAP3K4 (KI4) mice (TSKI4 cells) are uniquely paused in the intermediate stages of EMT, expressing both epithelial and mesenchymal characteristics while maintaining stemness. Induction of EMT in TSKI4 cells is due in part to the loss of MAP3K4/CBP mediated acetylation of histone H2B on the promoters of genes controlling the epithelial phenotype. We have recently discovered that loss of MAP3K4 activity increases the expression and activity of another chromatin remodeler, HDAC6. The goal of this project is to define the molecular network regulated by MAP3K4 controlling EMT. This network includes signaling pathways leading to the epigenetic regulation of genes important for EMT. Our approach is based on both our published work and new findings showing that MAP3K4 coordinates EMT by activating CBP and inhibiting HDAC6 on the promoters of genes important for EMT. We predict that genes in this MAP3K4 controlled network are critical to EMT-related pathologies. Our rationale is based on our findings of overlapping gene expression signatures between our TSKI4 stem cells and claudin low breast cancer cells that both display characteristics of stemness and EMT. Cancers with these characteristics frequently are both metastatic and display resistance to therapy. Protein networks that regulate the EMT transition represent potential targets for therapy with the rationale that reversal of EMT would restore sensitivity towards therapy. Our preliminary work using this innovative system has successfully identified new genes in breast cancer that control EMT/MET. Aim1 uses RNA-seq and ChIP-seq to identify a MAP3K4/CBP/HDAC6/H2BK5Ac dependent network that controls EMT. Aim 2 is a mechanistic study of the role of MAP3K4 and HDAC6 in EMT and the impact on the KI4 cellular and organismal phenotypes. The third aim targets GALNT3, a MAP3K4/CBP/HDAC6/H2BK5Ac co- regulated gene, defining the mechanisms by which GALNT3 controls EMT. Together, the three aims of this proposal will define the signaling mechanisms by which MAP3K4 coordinates the cellular phenotype through the co-regulation of the chromatin modifiers CBP and HDAC6. Further, this proposal will identify and characterize novel genes regulating epithelial/mesenchymal states, leading to the discovery of new biomarkers and drug targets for the identification and treatment of EMT related pathologies like fibrosis and metastasis.

描述（由申请人提供）：上皮至间质转化（EMT）是一种发育程序，其中具有紧密细胞间粘附的非活动上皮细胞转化为个体、活动间充质细胞。重要的是，EMT通过间充质至上皮转化是可逆的。 (MET) 将间充质细胞转化回上皮状态，对于发育过程和组织再生的正常过程至关重要。 EMT 在人类疾病中重新激活，包括肾脏、肝脏和肺部的组织纤维化以及癌症转移，这使得 EMT 成为药物治疗的关键靶点，小鼠中激酶 MAP3K4 的活性位点赖氨酸 (KI4) 的靶向突变可导致癌症。来自激酶失活的 MAP3K4 (KI4) 小鼠（TSKI4 细胞）的上皮滋养层干细胞 (TSKI4 细胞) 被破坏，导致严重的发育缺陷。 TSKI4 细胞中 EMT 的诱导部分是由于控制上皮表型的基因启动子上 MAP3K4/CBP 介导的组蛋白 H2B 乙酰化的丧失。发现 MAP3K4 活性的丧失会增加另一种染色质重塑因子 HDAC6 的表达和活性。该项目的目标是定义分子网络。该网络包括导致对 EMT 重要的基因进行表观遗传调控的信号通路。我们的方法基于我们已发表的工作和新发现，表明 MAP3K4 通过激活 CBP 并抑制基因启动子上的 HDAC6 来协调 EMT。我们预测该 MAP3K4 控制网络中的基因对于 EMT 相关病理学至关重要。我们的基本原理是基于我们对我们之间重叠基因表达特征的发现。 TSKI4干细胞和claudin低乳腺癌细胞均表现出干性和EMT特征，具有这些常见特征的癌症均具有转移性并表现出对治疗的抵抗性，调节EMT转变的蛋白质网络代表了潜在的治疗靶标，其基本原理是逆转EMT。 EMT 将恢复对治疗的敏感性。我们使用这一创新系统的初步工作已成功识别出乳腺癌中控制 EMT/MET 的新基因，Aim1 使用 RNA-seq 和 ChIP-seq 来识别。控制 EMT 的 MAP3K4/CBP/HDAC6/H2BK5Ac 依赖性网络是对 MAP3K4 和 HDAC6 在 EMT 中的作用以及对 KI4 细胞和有机体表型的影响的机制研究。 HDAC6/H2BK5Ac 共同调控基因，定义了 GALNT3 控制 EMT 的机制。该提案的三个目标将定义 MAP3K4 通过染色质修饰剂 CBP 和 HDAC6 的共同调节来协调细胞表型的信号传导机制。此外，该提案还将鉴定和表征调节上皮/间质状态的新基因，从而导致这一发现。用于识别和治疗 EMT 相关病理（如纤维化和转移）的新生物标志物和药物靶点。