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: 7653332
• 负责人: RICHARD G PESTELL
• 金额: $ 32.06万
• 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 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7653332
• 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; ERBB2 gene; Engineering; Epidermal Growth Factor Receptor; Epithelial Cells; Epithelium; Event; Eye; Gene Targeting; Genes; Genetic; Growth; Growth Factor Receptors; Human; Knockout Mice; Maintenance; Mammary Neoplasms; Mammary Tumorigenesis; Mammary gland; Mediating; Mitosis; Modeling; Molecular; Molecular Target; 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; public health relevance; 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）：过度活跃的生长因子受体信号传导在许多抵抗当前治疗的肿瘤中仍然活跃。果蝇Dac基因被克隆为过度活跃的EGFR（eclipse）的显性抑制剂。RDGN遗传网络由腊肠犬（dachshund，Eya）、无眼（eyeless，so）基因组成，调节后生动物的细胞命运决定。我们对2,000多例患者的分析表明，细胞核DACH 1表达的增加与细胞有丝分裂呈负相关，并预测乳腺癌患者生存期延长约40个月。我们已经证明DACH 1抑制癌基因介导的乳腺癌发生，阻断乳腺癌上皮细胞DNA合成，集落形成，基质胶生长和小鼠肿瘤生长。小鼠DACH 1基因缺失导致围产期死亡。为了确定DACH 1抑制乳腺肿瘤发生的分子机制，并确定DACH 1在体内的功能意义，我们将： 目标1。确定DACH 1/Eya抑制乳腺肿瘤细胞增殖的机制。我们将确定乳腺肿瘤生长的DACH 1结合蛋白，我们将描述这些成分在人类乳腺癌和研究等位基因失活的DACH 1。我们将使用ChIP芯片检测确定DACH 1的内源性基因靶点。 目标二。使用Dach 1f 1/f1转基因小鼠确定DACH 1作为乳腺肿瘤发病、进展和转移抑制因子的作用。将在原代培养和转基因小鼠（MMTV-Cre/MMTV-ErbB 2）中使用DACH 1 shRNA和DACH 1fl/fl转基因小鼠分析DACH 1作为乳腺肿瘤发生抑制因子的作用。 目标3.确定DACH 1/Eya抑制细胞迁移的机制。DACH 1抑制细胞迁移和趋化性。使用蛋白质组学方法，DACH 1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。我们将在使用异型乳腺癌移植模型中描述这些因素对乳腺癌体内转移的贡献。这些研究将描述一个全新的肿瘤和转移抑制途径。我们将确定DACH 1抑制乳腺肿瘤细胞增殖和转移的体内机制。 公共卫生相关性：今年在美国，41,000人将死于对目前治疗形式耐药的乳腺癌。为了找到治疗乳腺癌的新方法，我们需要乳腺癌进展的新模型和新的分子靶点。我们已经确定了一种全新的基因途径，可以阻止乳腺癌的生长和传播，当在人类乳腺癌中丢失时，预测结果非常糟糕。我们将确定这种新的基因通路如何阻止乳腺癌细胞的生长和转移，为乳腺癌的新型治疗方法提供基础