Interplay between Cdh1 and major regulatory pathways in human cancer

描述（由申请人提供）：细胞周期调控缺陷导致基因组不稳定并最终导致癌症发展。E3泛素连接酶APC/Cdh 1是细胞周期进程的主要调节因子，并与DNA损伤修复和肿瘤抑制有关，尽管其确切作用尚不清楚。在这项提案中，我们将探讨Cdh 1与多种主要肿瘤抑制因子和致癌途径的相互作用，这些途径在人类癌症中经常发生改变，以确定Cdh 1的新功能以及调控机制。我们以前报道过Cdh 1靶向Skp 2癌蛋白进行蛋白水解;我们的初步数据表明PI 3 K/Akt通路保护Skp 2免受Cdh 1介导的破坏。在目标1中，我们将利用遗传和生化方法来揭示Akt 1控制Skp 2丰度和细胞定位的潜在分子机制。这些研究将为Skp 2在人类癌症中的过表达提供一种新的机制。它还将推进我们对特定激酶信号级联如何影响APC/Cdh 1复合物控制的蛋白水解的认识，并为开发Akt 1特异性抑制剂作为有效的抗癌药物提供理论基础。在目标2中，我们将定义Cdh 1缺失激活p53和Rb通路的分子机制，以及它们对Cdh 1缺失诱导的早衰的贡献。我们发现，通过控制Claspin破坏，Cdh 1调节Chk 1活性，并进一步影响p53通路，暗示Cdh 1在DNA损伤修复中的作用。Cdh 1对Rb/E2 F1功能有影响。因此，原代人成纤维细胞中Cdh 1的耗竭通过激活p53和Rb途径导致过早衰老的发生。其他肿瘤抑制因子（包括PTEN和VHL）的失活也会诱导过早衰老，这被认为是防止癌症发展的内置故障安全机制。我们在目标2中提出的工作将为肿瘤细胞中较少观察到的Cdh 1丢失提供可能的潜在分子机制，这可能进一步暗示Cdh 1丢失是肿瘤发展的晚期事件。最后，我们的建议探讨了Cdh 1本身是如何监管的。我们的初步数据表明，Mdm 2癌蛋白控制Cdh 1在G1/S晚期的稳定性。这一发现扩展了我们对Cdh 1和p53/Mdm 2通路之间相互作用的理解，并进一步证明了主要细胞周期调节因子的功能相互交织以实现协同效应。我们在目标3中提出的工作将为Cdh 1稳定性控制提供一种新的调节机制，从而揭示Mdm 2致癌功能的另一层机制，并支持使用Mdm 2抑制剂进行癌症治疗。总之，这些研究将显着扩大我们目前的知识Cdh 1的重要功能以外的经典作用，在细胞周期控制，提供深入了解Cdh 1如何整合到网络的主要肿瘤抑制因子（p53和Rb）和癌基因（Akt和Mdm 2）途径，不仅管理细胞周期的进展，但也保持基因组的稳定性，并参与肿瘤抑制。

DESCRIPTION (provided by applicant): Defective cell cycle regulation leads to genomic instability and ultimately cancer development. The E3 ubiquitin ligase APC/Cdh1 is a major regulator of cell cycle progression and has been implicated in DNA damage repair and tumor suppression, although its exact roles remain unclear. In this proposal, we will explore the interaction of Cdh1 with multiple major tumor suppressor and oncogenic pathways frequently altered in human cancers to define the novel functions as well as regulatory mechanisms for Cdh1. We previously reported that Cdh1 targets the Skp2 oncoprotein for proteolysis; our preliminary data showed that the PI3K/Akt pathway protected Skp2 from Cdh1-mediated destruction. In Aim 1, we will utilize both genetic and biochemical approaches to reveal the underlying molecular mechanisms by which Akt1 controls Skp2 abundance and cellular localization. The proposed studies will provide a novel mechanism for Skp2 overexpression in human cancers. It will also advance our knowledge of how specific kinase signaling cascades influence proteolysis governed by the APC/Cdh1 complex, and provide the rationale for developing Akt1-specific inhibitors as potent anti-cancer drugs. In Aim 2, we will define the molecular mechanisms by which loss of Cdh1 activates the p53 and Rb pathways, and their contribution to Cdh1 loss-induced premature senescence. We found that by controlling Claspin destruction, Cdh1 regulates Chk1 activity and further influences the p53 pathway, implicating a role for Cdh1 in DNA damage repair. We also found that Cdh1 could affect Rb/E2F1 function. As a result, depletion of Cdh1 in primary human fibroblasts led to the onset of premature senescence by activating both the p53 and Rb pathways. Inactivation of other tumor suppressors, including PTEN and VHL, also induced premature senescence, which has been proposed as a built-in fail-safe mechanism against cancer development. Our proposed work in Aim 2 will provide the possible underlying molecular mechanism for the less frequently observed Cdh1 loss in tumor cells, which could further imply Cdh1 loss as a late event in tumor development. Lastly, our proposal explores how Cdh1 itself is regulated. Our preliminary data indicated that the Mdm2 oncoprotein controls the stability of Cdh1 in late G1/S phase. This finding extends our understanding of the interplay between the Cdh1 and p53/Mdm2 pathways, and provides further evidence that the functions of major cell cycle regulators are interwoven to achieve synergized effects. Our proposed work in Aim 3 will provide a novel regulatory mechanism for Cdh1 stability control, thus uncovering another layer of mechanism for the oncogenic function of Mdm2 and supporting the use of Mdm2 inhibitors for cancer treatment. Altogether, these studies will significantly expand our current knowledge of the important functions of Cdh1 outside its classic role in cell cycle control by providing insight into how Cdh1 integrates into the network of major tumor suppressor (p53 and Rb) and oncogene (Akt and Mdm2) pathways to not only govern cell cycle progression, but also maintain genomic stability and participate in tumor suppression.