Characterizing the role of TAK1 in 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，由 肿瘤细胞进入循环，在那里它们被称为循环肿瘤细胞(CTCs)。限制之一 预防或治疗转移的步骤是我们对肿瘤的生物学特性了解不足 CTC，包含转移前体细胞的群体。我们之前已经开发了一种微流控设备 目的：在人和小鼠体内高效分离CTCs。我们在分析胰腺CTCs方面的工作有 研究发现，MAP3K7又称转化生长因子激活的蛋白激酶(TAK1)，在细胞周期调控中起重要作用。 抵抗失巢凋亡-从细胞外基质脱离诱导的凋亡，这是 促进CTC在运往遥远地点的过程中存活。此外，我们最新的研究表明， 转移性乳腺癌患者CTC上皮-间充质转化的证据及意义 相关的信号通路，其中之一是众所周知的EMT诱导细胞因子-转化生长因子-β。作为一名 下游靶向的转化生长因子-β、TAK1可能参与了EMT过程，值得进一步研究。我的 Oncomine数据库中的Pilot分析表明TAK1在乳腺癌中的表达显著升高 然而，相对于其他类型的癌症，TAK1在乳腺癌中的作用还没有得到很好的研究。在……里面 初步实验中，我还发现在悬浮液中生长的乳腺癌细胞对 TAK1抑制作用。所有以前的工作都导致了这样的假设：TAK1可能在促进 乳腺癌转移。而胰腺癌在确诊时通常是转移的，而乳腺癌 癌症遵循更长的病程，在这种情况下，预防血液转移的能力可能是真实的 临床应用。因此，所提出的实验的总体目标是理解TAK1信号 参与乳腺癌转移的途径，包括分析失巢耐药的特征，EMT， 以及它在乳腺癌转移小鼠模型中的重要性。我将分析TAK1在失巢症中的作用 通过引入不同结构的TAK1在不同亚型的乳腺癌细胞中产生耐药性 要么激活TAK1，要么取消TAK1激酶功能。此外，我将研究TAK1在 促进乳腺癌细胞的EMT。最后，我将评估抑制TAK1的功能后果， 通过多西环素诱导抑制CTCs中的TAK1和异种乳腺转移瘤的形成 肿瘤转移小鼠模型。完成这些目标将使我们更好地理解 并为预防乳腺癌的靶向途径提供潜在的临床前数据 传播。拟议工作的总体职业发展目标是使我能够开始 专注于了解乳腺癌发病机制的富有成效的独立研究生涯 转移。 好了！

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. !