Determine the Functional Role Dub3 in Breast Cancer Progression and Metastasis

确定 Dub3 在乳腺癌进展和转移中的功能作用

基本信息

批准号：
8895123
负责人：
Binhua P Zhou
金额：
$ 35.71万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8895123
关键词：
Address Biological Markers Brain Breast Breast Cancer Cell Breast cancer metastasis Cell Line Cells Chromatin Clinical Coupled Data Deubiquitination Development Diagnosis Diagnostic Disease Distant Metastasis Drosophila genus E-Cadherin ERG gene Embryonic Development Environment Enzymes Epithelial Event Exposure to Genes Goals Growth Human Inflammatory Intrinsic factor Investigation Knock-out Knowledge Libraries Location Lung MCF7 cell Malignant Neoplasms Mammary Neoplasms Mass Spectrum Analysis Mesenchymal Metabolic Metastatic breast cancer Modeling Molecular Mus Neoplasm Metastasis Oncologist Pathologist Pharmaceutical Preparations Phase Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Prognostic Marker Property Proteins Public Health Reagent Recording of previous events Recruitment Activity Recurrence Regulation Research Role Sampling Signal Transduction Site Small Interfering RNA Snails Specimen Stromal Neoplasm Structural Biologist Structure T47D Testing Therapeutic Tissues Treatment Efficacy Tumor Cell Invasion Ubiquitination Validation Wound Healing Xenograft procedure aerobic glycolysis base beta-Transducin Repeat-Containing Proteins cancer stem cell cancer therapy cell growth regulation cell motility cytokine epithelial to mesenchymal transition functional plasticity gastrulation in vivo inhibitor/antagonist innovation loss of function macrophage malignant breast neoplasm multidisciplinary neoplastic cell new therapeutic target novel strategies outcome forecast overexpression prevent prognostic value programs promoter public health relevance research study screening success targeted treatment therapy resistant tumor tumor microenvironment tumor progression ubiquitin-protein ligase virtual young woman

项目摘要

DESCRIPTION (provided by applicant): Basal-like breast cancer (BLBC) presents with an aggressive clinical history, development of recurrence, distant metastasis, shorter survival, and usually occurs in young women. BLBC has an activated epithelial-mesenchymal transition (EMT) program, which provides cells with the increased phenotypic and cellular plasticity required during embryonic development, tissue remodeling, wound healing and metastasis. Our long-term goal is to discover the molecular circuitry that governs the functional plasticity of BLBC cells in metastasis and to identify molecules that may serve as druggable targets for treating this deadly disease. In the last nine years, we have systematically studied the function of Snail and the molecular mechanism by which Snail represses E-cadherin expression in BLBC. Our studies clearly indicate that Snail levels are regulated predominantly through protein ubiquitination. High levels of Snail, due to protein stabilization, contribute significantly to the increased invasion, therapeutic resistance, CSC-like properties, and tumor recurrence. However, the mechanism by which Snail escapes ubiquitination and degradation remains unknown; uncovering this mechanism is crucially important for the development of novel approaches to treat BLBC. Recently, we discovered that Dub3 is a bona fide deubiquitinase of Snail and contributes to Snail stabilization in breast cancer. Dub3 is evolutionarily conserved from Drosophila to humans. Dub3-knockout downregulated Snail and disrupted gastrulation (an event that requires EMT) in Drosophila. Elevated expression of Dub3 correlates unequivocally with Snail expression in BLBC samples and cell lines. Overexpression of Dub3 in luminal breast tumor MCF7 and T47D cell lines, which contain no detectable endogenous Dub3 and Snail, induces transformation to a basal-like phenotype; whereas, Dub3-knockdown downregulates Snail and suppresses cell migration. Intriguingly, Dub3 was an early response gene that appeared after exposure to inflammatory cytokines. We hypothesize that Dub3 is the long-sought "missing molecule" that regulates cellular plasticity by controlling the level of Snail durig EMT and metastasis. The objective of this proposal is to characterize the function and regulation of the Dub3-Snail axis and explore the clinical value of Dub3 as a prognostic biomarker and a druggable target for treating metastatic breast cancer. Guided by strong preliminary data, we will test this hypothesis by pursuing three specific aims: (1) to delineate th molecular mechanism by which Dub3 controls EMT; (2) to determine the function and regulation of Dub3 in breast cancer; and (3) to define the function and therapeutic efficacy of Dub3 in vivo. Our proposal is innovative and significant, because it will not only uncover a new paradigm that significantly increases our understanding of the regulation of cellular plasticity during tumor progression and metastasis but also leads to the development of specific Dub3 inhibitors for treating metastatic breast cancer.

描述（由申请人提供）：基底样乳腺癌 (BLBC) 具有侵袭性临床病史、复发、远处转移、生存期较短，并且通常发生于年轻女性，具有激活的上皮间质转化 (EMT)。该计划为细胞提供胚胎发育、组织重塑、伤口愈合和转移过程中所需的增强的表型和细胞可塑性。我们的长期目标是发现控制细胞的分子回路。 BLBC 细胞在转移中的功能可塑性，并确定可作为治疗这种致命疾病的药物靶标的分子。在过去的九年中，我们系统地研究了 Snail 的功能以及 Snail 抑制 E-钙粘蛋白表达的分子机制。我们的研究清楚地表明，Snail 水平主要通过蛋白质泛素化进行调节，由于蛋白质稳定，高水平的 Snail 对然而，Snail 逃避泛素化和降解的机制仍然未知；揭示这种机制对于开发治疗 BLBC 的新方法至关重要。 Dub3 是 Snail 的一种真正的去泛素酶，有助于 Snail 在乳腺癌中的稳定，从果蝇到人类，Dub3 敲除后的表达下调。果蝇中的 Snail 和原肠胚形成中断（需要 EMT 的事件） Dub3 的表达升高与 BLBC 样本和细胞系中 Dub3 的过表达明确相关，这些细胞系中未检测到内源性 Dub3 和 Dub3。 Snail 诱导向基底样表型的转化；而 Dub3 敲除则下调 Snail 并抑制细胞迁移。有趣的是，Dub3 是暴露于炎症细胞因子后出现的早期反应基因，我们研究发现 Dub3 是通过控制 Snail 在 EMT 和转移过程中调节细胞可塑性的“缺失分子”。在强有力的初步指导下，表征 Dub3-Snail 轴的功能和调节，并探索 Dub3 作为预后生物标志物和治疗转移性乳腺癌的药物靶标的临床价值。根据数据，我们将通过三个具体目标来检验这一假设：(1) 描述 Dub3 控制 EMT 的分子机制；(2) 确定 Dub3 在乳腺癌中的功能和调节；(3) 定义其功能。我们的建议是创新且意义重大的，因为它不仅揭示了一个新的范式，显着增加了我们对肿瘤进展和转移过程中细胞可塑性调节的理解，而且还导致了特定 Dub3 抑制剂的开发。用于治疗转移性乳腺癌。