Non-canonical Wnt/Dishevelled signaling and cancer cell malignancy

非典型 Wnt/混乱信号传导与癌细胞恶性肿瘤

• 批准号: 7898651
• 负责人: ANTHONY M.C. BROWN
• 金额: $ 31.92万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-08 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7898651
• 关键词: Abbreviations; Binding; Biochemical; Biological Assay; Breast Cancer Cell; Cell Polarity; Cells; Chimeric Proteins; Conditioned Culture Media; Confocal Microscopy; Cysteine-Rich Domain; Data; Development; Dissection; Disseminated Malignant Neoplasm; Dominant-Negative Mutation; Doxycycline; Dsh protein; Ductal; Embryo; Estrogen Receptors; Event; Goals; Green Fluorescent Proteins; Histones; Human; Image; Immunoprecipitation; LDL-Receptor Related Protein 1; Liquid Chromatography; MAPK8 gene; Malignant - descriptor; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Maps; Mass Spectrum Analysis; Mediating; Medical; Migration Assay; Molecular; Monoclonal Antibodies; Morphogenesis; Mouse Mammary Tumor Virus; Movement; Mus; Neoplasm Metastasis; Nuclear Import; Nuclear Translocation; Pathway Analysis; Pathway interactions; Phosphorylation; Phosphorylation Site; Physiological; Play; Property; Proteins; Proteomics; Reporter; Research; Research Personnel; Resistance; Role; Signal Pathway; Signal Transduction; Staging; System; Testing; Tetracyclines; Therapeutic; Time; Transgenes; Transgenic Mice; Transgenic Organisms; Variant; Venus; Wnt proteins; Work; Xenopus; cancer cell; cell motility; detector; homologous recombination; human SFRP4 protein; in vitro Assay; malignant breast neoplasm; malignant phenotype; monolayer; mouse model; mutant; outcome forecast; overexpression; programs; red fluorescent protein; research study; response; retroviral transduction; rho; tandem mass spectrometry; tumor; tumor initiation; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): The overall objectives of this research are to investigate mechanisms of non-canonical Wnt signaling acting via Dishevelled (Dvl) protein, and to test the hypothesis that this pathway regulates cancer cell invasion and is relevant to malignant breast cancer. The canonical Wnt/?-catenin pathway contributes to the initiation of many cancers but recent evidence indicates that ?-catenin-independent, or non-canonical, Wnt signaling is relevant to later stages of malignant progression. This pathway also plays crucial roles in cell polarity and morphogenesis. Wnt-induced phosphorylation of Dvl provides a reliable assay for non-canonical Wnt signaling and a starting point for molecular dissection of the non-canonical pathway and its functions in malignancy. The functional significance of Dvl phosphorylation in Wnt signaling will be investigated by testing the properties of Dvl mutants that are resistant to Wnt-induced phosphorylation. We will also test the hypothesis that non-canonical Wnt signaling inhibits the invasion of ER-negative breast cancer cells. This will be achieved by in vitro assays of cell migration and invasion, in conjunction with signaling assays. The role of Dvl phosphorylation in cell migration will be explored, and we will identify downstream components of non-canonical Wnt signaling that are activated in this context. In view of the key role of Dvl protein in mediating and modulating Wnt signaling, a comprehensive analysis of changes in the subcellular distribution of Dvl in response to non-canonical Wnt signals will be performed by confocal microscopy. A proteomics approach will be taken to identify key changes in the proteins with which Dvl interacts as a result of non-canonical Wnt signals. In addition, transgenic mice will be generated in which non-canonical Wnt signaling will be activated in the mammary gland in a regulated manner. Effects of this signaling on mammary development will be determined and the mice will be crossed with existing mouse models of breast cancer to test directly the consequences of non-canonical Wnt signaling in tumor progression towards invasive and metastatic cancer. Since there is extensive evidence of aberrant Wnt expression in cancer, and most Wnt proteins can elicit non-canonical signals, it is likely that this poorly understood pathway of non-canonical Wnt signaling may be of widespread medical significance. Characterization of the molecular mechanisms involved will provide opportunities for manipulating the pathway for therapeutic purposes.

描述（由申请人提供）：本研究的总体目标是研究通过Dishevelled（Dvl）蛋白发挥作用的非经典Wnt信号传导机制，并检验该途径调节癌细胞侵袭并与恶性乳腺癌相关的假设。经典的Wnt/？-连环蛋白通路有助于许多癌症的发生，但最近的证据表明，不依赖于连环蛋白的或非典型的Wnt信号传导与恶性进展的后期阶段有关。该通路在细胞极性和形态发生中也起着至关重要的作用。Wnt诱导的Dvl磷酸化为非经典Wnt信号传导提供了可靠的测定，并为非经典途径及其在恶性肿瘤中的功能的分子解剖提供了起点。将通过测试抵抗Wnt诱导的磷酸化的Dvl突变体的性质来研究Dvl磷酸化在Wnt信号传导中的功能意义。我们还将检验非经典Wnt信号传导抑制ER阴性乳腺癌细胞侵袭的假设。这将通过细胞迁移和侵袭的体外测定以及信号传导测定来实现。Dvl磷酸化在细胞迁移中的作用将被探索，我们将确定在这种情况下激活的非经典Wnt信号传导的下游组件。鉴于Dvl蛋白在介导和调节Wnt信号传导中的关键作用，将通过共聚焦显微镜对响应于非典型Wnt信号的Dvl亚细胞分布的变化进行全面分析。将采用蛋白质组学方法来鉴定Dvl与之相互作用的蛋白质中的关键变化，其作为非典型Wnt信号的结果。此外，将产生转基因小鼠，其中非经典Wnt信号传导将在乳腺中以受调节的方式激活。将确定这种信号传导对乳腺发育的影响，并将小鼠与现有的乳腺癌小鼠模型杂交，以直接测试非经典Wnt信号传导在肿瘤向侵袭性和转移性癌症进展中的后果。由于有大量证据表明癌症中Wnt表达异常，并且大多数Wnt蛋白可以引发非经典信号，因此这种对非经典Wnt信号传导途径知之甚少的途径可能具有广泛的医学意义。所涉及的分子机制的表征将提供操纵用于治疗目的的途径的机会。

项目成果

Wnt5a Signals through DVL1 to Repress Ribosomal DNA Transcription by RNA Polymerase I.

• DOI: 10.1371/journal.pgen.1006217
• 发表时间: 2016-08
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Dass RA;Sarshad AA;Carson BB;Feenstra JM;Kaur A;Obrdlik A;Parks MM;Prakash V;Love DK;Pietras K;Serra R;Blanchard SC;Percipalle P;Brown AM;Vincent CT
• 通讯作者: Vincent CT

Mammary stem cells and cancer: roles of Wnt signaling in plain view.

乳腺干细胞和癌症：Wnt 信号传导的作用一目了然。

• DOI: 10.1186/bcr2631
• 发表时间: 2010
• 期刊: Breast cancer research : BCR
• 作者: Many AM;Brown AM
• 通讯作者: Brown AM