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

• 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
• 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

 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）：上皮细胞向间充质细胞转化（EMT）是一种发育程序，其中具有紧密细胞-细胞粘附的非运动性上皮细胞转化为个体运动性间充质细胞。重要的是，EMT是可逆的，通过间充质上皮转化（MET）将间充质细胞转化回上皮状态。EMT程序对于发育期间的正常过程和组织再生至关重要。EMT在包括肾、肝和肺中的组织纤维化和癌症转移的人类疾病中被重新激活，使得EMT成为药物治疗的关键靶标。在小鼠（KI 4）中激酶MAP 3 K4的活性位点赖氨酸的靶向突变由于EMT被破坏而导致严重的发育缺陷。来自激酶失活的MAP 3 K4（KI 4）小鼠（TSKI 4细胞）的上皮滋养层干细胞（TS）在EMT的中间阶段独特地暂停，表达上皮和间充质特征，同时保持干细胞性。TSKI 4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的MAP 3 K4/CBP介导的组蛋白H2 B乙酰化的丧失。我们最近发现，MAP 3 K4活性的丧失增加了另一种染色质重塑剂HDAC 6的表达和活性。本研究的目的是明确MAP 3 K4调控EMT的分子网络。该网络包括导致对EMT重要的基因的表观遗传调节的信号通路。我们的方法是基于我们发表的工作和新的发现，表明MAP 3 K4通过激活CBP和抑制EMT重要基因启动子上的HDAC 6来协调EMT。我们预测MAP 3 K4控制的网络中的基因对EMT相关的病理至关重要。我们的理由是基于我们的TSKI 4干细胞和claudin低乳腺癌细胞之间重叠基因表达特征的发现，这两种细胞都显示出干性和EMT的特征。具有这些特征的癌症通常是转移性的并且显示出对治疗的抗性。调节EMT转变的蛋白质网络代表了治疗的潜在靶点，其基本原理是EMT的逆转将恢复对治疗的敏感性。我们使用这种创新系统的初步工作已经成功地确定了乳腺癌中控制EMT/MET的新基因。Aim 1使用RNA-seq和ChIP-seq来鉴定控制EMT的MAP 3 K4/CBP/HDAC 6/H2 BK 5Ac依赖性网络。目的2是MAP 3 K4和HDAC 6在EMT中的作用以及对KI 4细胞和生物表型的影响的机制研究。第三个目标靶向GALNT 3，一种MAP 3 K4/CBP/HDAC 6/H2 BK 5Ac共调节基因，定义了GALNT 3控制EMT的机制。总之，该提案的三个目标将定义MAP 3 K4通过染色质修饰剂CBP和HDAC 6的共调节来协调细胞表型的信号传导机制。此外，该提案将鉴定和表征调节上皮/间充质状态的新基因，从而发现用于鉴定和治疗EMT相关病理如纤维化和转移的新生物标志物和药物靶标。