Characterizing the role of TAK1 in breast cancer metastasis

表征 TAK1 在乳腺癌转移中的作用

• 批准号: 8635761
• 负责人: Min Yu
• 金额: $ 19.74万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8635761
• 关键词: Anoikis; Apoptosis; Apoptotic; Biological; Biological Assay; Biology; Blood; Blood Circulation; Breast Cancer Cell; Cancer Patient; Cancer cell line; Cause of Death; Cell Line; Cells; Cessation of life; Clinical Data; Complex; Critiques; Data; Databases; Dependence; Development; Distant; Dominant-Negative Mutation; Doxycycline; Epithelial; Extracellular Matrix; Foundations; Goals; Human; Individual; Investigation; Knowledge; Lead; Link; Luciferases; Lung; MAP3K7 gene; MAPK14 gene; MAPK8 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mesenchymal; Metastatic Lesion; Microfluidic Microchips; Microfluidics; Molecular; Molecular Profiling; Mus; Neoplasm Circulating Cells; Neoplasm Metastasis; Pancreas; Pathway interactions; Phosphotransferases; Play; Population; Process; Property; Relative (related person); Research; Resistance; Role; Signal Pathway; Signal Transduction; Site; Suspension substance; Suspensions; Time; Tumor Cell Invasion; Work; Writing; Xenograft procedure; base; breast cancer diagnosis; cancer type; career; career development; clinical application; cytokine; in vivo; knock-down; malignant breast neoplasm; meetings; metastatic process; migration; mouse model; neoplastic cell; overexpression; pre-clinical; prevent; public health relevance; research study; small hairpin RNA; therapeutic target; tumor; tumor growth; tumor progression

PROJECT SUMMARY/ABSTRACT Majority of cancer-related deaths are caused by metastasis, a complex process initiated by spreading of tumor cells into circulation, where they are referred to as circulating tumor cells (CTCs). One of the limiting steps towards preventing or treating metastasis is our insufficient knowledge of the biological properties of CTCs, the population that contains metastatic precursors. We have previously developed a microfluidic device to isolate CTCs with high efficiency in human and mice. Our work on analyzing pancreatic CTCs have identified that MAP3K7, also called TGF-¿ activated kinase (TAK1), plays an important role in promoting resistance to anoikis - apoptosis induced by detachment from the extracellular matrix, one of the features to facilitate CTCs to survive during transit to distant sites. In addition, our most recent research has demonstrated the evidence of epithelial-mesenchymal transition (EMT) in CTCs of metastatic breast cancer patients and associated signaling pathways, one of which is the well-known EMT inducing cytokine - TGF-¿. As a downstream target of TGF-¿, TAK1 may be involved in the EMT process thus warrant further investigation. My pilot analysis in Oncomine database indicated a significantly higher expression of TAK1 in breast cancers relative to other type of cancers, however, the role of TAK1 is not very well investigated in breast cancers. In preliminary experiments, I also found that breast cancer cells grown in suspension are exquisitely sensitive to TAK1 inhibition. All the prior work lead to the hypothesis that TAK1 may play an important role in promoting breast cancer metastasis. Whereas pancreatic cancer is often metastatic at the time of diagnosis, breast cancer follows a more prolonged course, where the ability to prevent blood borne metastasis may have real clinical application. Thus, the overall goal of the proposed experiments is to understand the TAK1 signaling pathways involved in breast cancer metastasis, including analyzing the features of anoikis resistance, EMT, and its importance in breast cancer metastatic mouse models. I will analyze the role of TAK1 on anoikis resistance in different subtypes of breast cancer cells by introducing different constructs of TAK1 that can either activate TAK1 or abolish TAK1 kinase function. In addition, I will examine the contribution of TAK1 in promoting EMT in breast cancer cells. Finally, I will evaluate the functional consequences of inhibiting TAK1, via a doxycycline-inducible suppression of TAK1, in CTCs and metastases formation in a xenograft breast cancer metastatic mouse model. Completing these goals will lead to a better understanding of this important kinase pathway and provide potential pre-clinical data on a targetable pathway for preventing breast cancer dissemination. The overall career development goal of the proposed work is to enable me to begin a productive, independent research career focusing on understanding the mechanisms of breast cancer metastasis. !

项目摘要/摘要 大多数与癌症相关的死亡是由转移引起的，这是一个复杂的过程，是通过传播引发的 肿瘤细胞进入循环，在那里它们被称为循环肿瘤细胞（CTC）。限制之一 我们对预防或治疗转移的步骤是我们对生物学特性的了解 CTC，包含转移性前体的人群。我们以前已经开发了微流体设备 分离人和小鼠效率高的CTC。我们在分析的胰腺CTC上的工作 确定MAP3K7，也称为TGF-激活激酶（TAK1），在促进中起着重要作用 抗Anoikis的抗性 - 细胞外基质脱离引起的凋亡，这是 促进CTC在转运到遥远位点的过程中生存。此外，我们最近的研究表明 转移性乳腺癌患者CTC中上皮 - 间质转变（EMT）的证据 相关的信号通路，其中之一是众所周知的EMT诱导的细胞因子-TGF-。作为 TGF-的下游靶标，TAK1可能参与EMT过程，因此保证进一步研究。我的 oncomine数据库中的试点分析表明，乳腺癌中TAK1的表达明显更高 但是，相对于其他类型的癌症，在乳腺癌中，TAK1的作用不是很好。 初步实验，我还发现悬浮液中生长的乳腺癌细胞对 tak1抑制。所有先前的工作都导致了以下假设：tak1可能在促进中起重要作用 乳腺癌转移。而胰腺癌在诊断时通常是转移性的，而乳腺癌 癌症遵循更长的过程，预防血液传播转移的能力可能具有真实 临床应用。这是拟议实验的总体目标是了解tak1信号传导 参与乳腺癌转移的途径，包括Anaikis抗性，EMT， 及其在乳腺癌转移性小鼠模型中的重要性。我将分析tak1在Anoikis上的作用 通过引入不同的tak1构建体，可以在不同亚型的乳腺癌细胞中的抗性 激活TAK1或废除TAK1激酶功能。此外，我将研究tak1的贡献 促进乳腺癌细胞中的EMT。最后，我将评估抑制tak1的功能后果， 通过强力霉素诱导的TAK1抑制，在异种移植乳房中的CTC和转移中形成 癌症转移小鼠模型。完成这些目标将使人们更好地理解这一重要的 激酶途径并提供有关预防乳腺癌的可靶向途径的潜在临床前数据 传播。拟议工作的整体职业发展目标是使我开始 生产性独立研究职业，重点是了解乳腺癌的机制 转移。 呢