PROJECT 2: Oncogenic Transformation via the PP2A/YAP/Hippo pathway

项目摘要 本申请寻求雅普/TAZ在癌症中的作用的潜在机制。各种各样的 人类癌症改变了通常与预后相关的雅普/TAZ信号传导。雅普和它的表亲 TAZ是Hippo通路的主要效应物，Hippo通路控制器官大小、细胞运动、增殖和分化。 生存该通路还调节其他重要的信号传导通路（Hedgehog、Notch、TGFβ和Wnt）。 我们将使用鼠多瘤病毒（MuPyV）来探测雅普功能。MuPyV引起广泛的肿瘤。其 研究已经阐明了酪氨酸磷酸化和PI 3激酶在癌症中的作用。它继续指向 重要问题，如PP 2A Aβ亚型在癌症中的作用。最近，我们报道了MuPyV的小T 抗原（ST）直接结合雅普，并通过促进其磷酸化、稳定性和功能来改变其磷酸化、稳定性和功能。 与PP 2A的关系这种相互作用使ST能够阻断几个系统中的分化。广泛 关于MT（MuPyV的致癌基因）的初步数据显示MT也与雅普相关。该关联 对于MT驱动的肿瘤转化是重要的。MT也影响雅普磷酸化，但MT的作用是 我们的第一个目的是研究ST的干细胞样细胞的分化的调节。以来 我们知道ST也与TAZ相互作用，我们将测试它是否有助于表型。这一目标将 包括确定所涉及的转录机制和雅普效应蛋白关联 ST用于实现这些改变。我们的第二个目标将是研究雅普在机器翻译驱动中的作用 转型在这里，我们也将对基因表达和蛋白质-蛋白质关联的变化感兴趣。 在我们的最后一个目标中，我们试图使用RT-PCR检测雅普在MuPyV MT介导的体内转化中的作用。 转基因和基因敲除小鼠。 在这个项目的背景下，我们有一个很好的机会利用病毒的相互作用， 更深入地了解人类肿瘤发生中的Hippo信号。通过比较我们的结果对MuPyV的影响 ST/MT对PP 2A/雅普/TAZ的影响，以及我们的同事与SV 40 ST和默克尔ST以及 在K-Ras驱动的人类癌症中研究雅普的哈恩实验室的研究人员，我们应该能够确定 雅普对细胞信号传导的影响非常重要，极大地增强了我们数据的影响。理想情况下，这些 研究将引导我们实现最终目标，将我们的工作转化为人类癌症治疗，就像我们已经做的那样。 之前针对PI 3激酶进行过。

PROJECT SUMMARY This application seeks the mechanisms underlying the roles of YAP/TAZ in cancer. A wide variety of human cancers have altered YAP/TAZ signaling that is generally connected to prognosis. YAP and its cousin TAZ are the major effectors of Hippo pathway that controls organ size, cell movement, proliferation and survival. This pathway also regulates other important signaling pathways (Hedgehog, Notch, TGFβ and Wnt). We will use murine polyomavirus (MuPyV) to probe YAP function. MuPyV causes a broad range of tumors. Its study has illuminated the roles of tyrosine phosphorylation and PI3 kinase in cancer. It continues to point to important issues, such as the role of PP2A Aβ isoform, in cancer. Recently, we reported that MuPyV's small T antigen (ST) directly binds YAP and alters its phosphorylation, stability, and function by promoting its association with PP2A. This interaction allows ST to block differentiation in several systems. Extensive preliminary data on MT, the oncogene of MuPyV, show that MT also associates with YAP. This association is important for MT driven neoplastic transformation. MT also affects YAP phosphorylation, but MT's effects are different than those of ST. Our first aim is to study ST regulation of the differentiation of stem-like cells. Since we know that ST also interacts with TAZ, we will test whether it contributes to the phenotype. This aim will include a determination of the transcriptional mechanisms involved and the YAP effector protein associations that ST uses to achieve these alterations. Our second aim will be to study the roles of YAP in MT driven transformation. Here too we will be interested in changes in gene expression and protein-protein associations. In our last aim we seek to examine the roles of YAP in MuPyV MT mediated transformation in vivo using transgenic and knockout mice. In the context of this Program Project, we have a great opportunity to exploit the virus interactions to gain a deeper insight into Hippo signaling in human oncogenesis. By comparing our results on the effects of MuPyV ST/MT on PP2A/YAP/TAZ with the results of our colleagues working with SV40 ST and Merkel ST and those of the Hahn lab working on YAP in K-Ras driven human cancers, we should be able to determine which of YAP's effects on cellular signaling are truly important, greatly enhancing the impact of our data. Ideally, these studies will lead us to our ultimate goal of translating our work into human cancer therapy, as we have previously done for PI3 kinase.