DACH1/Eya Cell-fate determination factor and mammary tumoregenesis

DACH1/Eya 细胞命运决定因子与乳腺肿瘤发生

• 批准号: 8193135
• 负责人: RICHARD G PESTELL
• 金额: $ 31.1万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-20 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8193135
• 关键词: Binding Proteins; Biological Assay; Breast; Breast Cancer Cell; Breast Cancer Treatment; Cancer Patient; Cell Proliferation; Cells; Chemotaxis; Complex; Cyclin D1; DNA biosynthesis; Drosophila genus; Engineering; Epidermal Growth Factor Receptor; Epithelial Cells; Epithelium; Event; Eye; Gene Targeting; Genes; Genetic; Growth; Growth Factor Receptors; Health; Human; Knockout Mice; Maintenance; Mammary Neoplasms; Mammary Tumorigenesis; Mammary gland; Mediating; Mitosis; Modeling; Molecular; Molecular Target; Mouse Mammary Tumor Virus; Mus; Neoplasm Metastasis; Nuclear; Oncogenes; Outcome; Pathway interactions; Patients; Perinatal; Phosphorylation; Physiological; Production; Proteomics; Receptor Signaling; Repression; Resistance; Resolution; Retinal; Role; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Transcription Factor AP-1; Transgenic Mice; Transgenic Organisms; Transplantation; Tumor Suppression; United States; Viral Vector; base; cell growth; cell motility; cytokine; improved; in vivo; inhibitor/antagonist; malignant breast neoplasm; matrigel; migration; neoplastic cell; novel; novel strategies; outcome forecast; promoter; recombinase; small hairpin RNA; tumor; tumor growth; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): Hyperactive growth factor receptor signaling remains active in many tumors that resist current therapies. The drosophila Dac gene was cloned as a dominant inhibitor of the hyperactive EGFR (eclipse). The RDGN genetic network, consisting of the dachshund (dac), eyes absent (eya), eyeless, and sine oculis (so) genes, regulates cell fate determination in metazoans. Our analysis of over 2,000 patients demonstrated increased nuclear DACH1 expression correlated inversely with cellular mitosis and predicted a ~40 month improved breast cancer patient survival. We have shown DACH1 inhibits oncogene-mediated breast oncogenesis, blocking breast cancer epithelial cell DNA synthesis, colony formation, growth in matrigel and tumor growth in mice. Genetic deletion of DACH1 in the mouse results in perinatal lethality. To determine the molecular mechanisms by which DACH1 inhibits mammary tumorigenesis and to determine to functional significance of DACH1 in vivo we will: Aim 1. Determine the mechanisms governing DACH1/Eya inhibition of breast tumor cellular proliferation. We will determine the DACH1 binding proteins governing breast tumor growth we will characterize these components in human breast cancer and investigate allelic inactivation of DACH1. We will define endogenous gene targets of DACH1 using ChIP on chip assays. Aim 2. Determine the role of DACH1 as a suppressor of mammary tumor onset, progression and metastasis using Dach1f1/f1 transgenic mice. The role of DACH1 as a suppressor of mammary tumorigenesis will be analyzed using DACH1 shRNA and Dach1fl/fl transgenic mice in primary culture and transgenic mice (MMTV-Cre/MMTV-ErbB2). Aim 3. Determine the mechanism by which DACH1/Eya inhibits cellular migration. DACH1 inhibits cellular migration and chemotaxis. Using a proteomic approach DACH1 was shown to regulate expression and production of secreted cytokines from breast cancer cells. We will characterize the contribution of these factors to breast cancer metastasis in vivo in using heterotypic breast cancer transplant models. These studies will characterize a completely novel tumor and metastasis suppressor pathway. We will determine the mechanism by which DACH1 inhibits breast tumor cellular proliferation and metastasis in vivo. PUBLIC HEALTH RELEVANCE: This year in the United States 41,000 will die from breast cancer resistant to the current forms of therapy. In order for us to find new approaches to treatment of breast cancer new models of breast cancer progression and new molecular targets are required. We have identified a completely novel pathway of genes that block breast cancer growth and spread that when lost in human breast cancer, predict very bad outcome. We and will determine how this new gene pathway blocks breast cancer cellular growth and metastasis to provide the basis for new types of therapies for breast cancer

描述(由申请人提供)：在许多抵抗当前治疗的肿瘤中，过度活跃的生长因子受体信号仍然活跃。果蝇DAC基因被克隆为高活性EGFR(ECLIPSE)的显性抑制基因。RDGN遗传网络由腊肠(Dac)、眼睛缺失(Eya)、盲目和正弦(Sine Oculis，SO)基因组成，调节后生动物细胞命运的决定。我们对2,000多名患者的分析表明，核DACH1表达增加与细胞有丝分裂呈负相关，并预测乳腺癌患者的生存期可提高约40个月。我们已经证明DACH1抑制癌基因介导的乳腺肿瘤的发生，阻止乳腺癌上皮细胞DNA合成、集落形成、基质生长和小鼠肿瘤生长。小鼠DACH1基因缺失会导致围产期死亡。为了确定DACH1抑制乳腺肿瘤发生的分子机制，并确定DACH1在体内的功能意义，我们将： 目的1.探讨DACH1/EYA抑制乳腺肿瘤细胞增殖的机制。我们将确定控制乳腺肿瘤生长的DACH1结合蛋白，我们将描述这些成分在人类乳腺癌中的特征，并研究DACH1的等位基因失活。我们将使用芯片分析来确定DACH1的内源性基因靶点。 目的2.利用Dach1f1/F1转基因小鼠，研究DACH1对乳腺癌发生、发展和转移的抑制作用。在原代培养和转基因小鼠(MMTV-Cre/MMTV-ErbB2)中，将使用DACH1 shRNA和Dach1fl/fl转基因小鼠来分析DACH1作为乳腺肿瘤发生抑制因子的作用。 目的3.确定DACH1/EYA抑制细胞迁移的机制。DACH1抑制细胞迁移和趋化作用。利用蛋白质组学方法，DACH1被证明可以调节乳腺癌细胞分泌的细胞因子的表达和产生。我们将利用异型乳腺癌移植模型来表征这些因素在体内乳腺癌转移中的作用。这些研究将描述一种全新的肿瘤和转移抑制途径。我们将在体内确定DACH1抑制乳腺肿瘤细胞增殖和转移的机制。 与公共卫生相关：今年，美国将有4.1万人死于对当前治疗形式产生抗药性的乳腺癌。为了找到治疗乳腺癌的新方法，需要新的乳腺癌进展模型和新的分子靶点。我们已经确定了一种全新的阻止乳腺癌生长和扩散的基因途径，当这种基因在人类乳腺癌中丢失时，可以预测非常糟糕的结果。我们将确定这一新的基因通路如何阻断乳腺癌细胞的生长和转移，为乳腺癌的新型治疗提供基础。