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

基本信息

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
项目类别：
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

项目摘要

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）：在许多抗电流疗法的肿瘤中，多动活跃生长因子受体信号传导保持活跃。果蝇DAC基因被克隆为多动性EGFR（Eclipse）的主要抑制剂。 RDGN遗传网络由腊肠犬（DAC），眼睛不存在（EYA），无眼和正弦声（SO）基因组成，可调节后生动物中的细胞命运测定。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相关，并预测约40个月改善的乳腺癌患者生存率。我们已经显示DACH1抑制了癌基因介导的乳腺癌发生，阻断了乳腺癌上皮细胞DNA合成，菌落形成，矩阵的生长和小鼠的肿瘤生长。小鼠中DACH1的遗传缺失会导致围产期致死性。为了确定DACH1抑制乳腺肿瘤发生并确定DACH1在体内的功能显着性的分子机制，我们将：目标1。确定乳腺肿瘤细胞增殖的DACH1/EYA抑制的机制。我们将确定有关乳腺肿瘤生长的DACH1结合蛋白，我们将表征人类乳腺癌中的这些成分并研究DACH1的等位基因失活。我们将使用芯片分析中的芯片定义DACH1的内源基因靶标。 AIM 2。使用DACH1F1/F1转基因小鼠确定DACH1作为乳腺肿瘤发作，进展和转移的抑制作用。将使用DACH1 shRNA和DACH1FL/FL转基因小鼠在原代培养和转基因小鼠（MMTV-CRE/MMTV-ERBB2）中分析DACH1作为乳腺肿瘤发生的抑制剂的作用。目标3。确定DACH1/EYA抑制细胞迁移的机制。 DACH1抑制细胞迁移和趋化性。使用蛋白质组学方法DACH1显示可调节乳腺癌细胞的分泌细胞因子的表达和产生。我们将表征这些因素对使用异型乳腺癌移植模型的体内乳腺癌转移的贡献。这些研究将表征一种完全新颖的肿瘤和转移抑制途径。我们将确定DACH1在体内抑制乳腺肿瘤细胞增殖和转移的机制。公共卫生相关性：今年在美国，有41,000名将因抗乳腺癌的抗药性而死亡。为了使我们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标。我们已经确定了一种完全新颖的基因途径，该途径阻断了乳腺癌的生长并传播，当在人类乳腺癌中丢失时，预测结果非常不好。我们并且将确定该新基因途径如何阻止乳腺癌细胞的生长和转移，以为乳腺癌的新疗法提供基础