Targeting Pten - An Upstream, Downstream and Offstream Approach

描述（由申请人提供）：该项目代表了一种高度集成的方法来翻译与PTEN（磷酸酶和张力蛋白同源物）信号通路相关的基础科学发现，该信号通路是人类肿瘤中最常见的失活肿瘤抑制基因之一。这个提议的总体假设是，理解PTEN的调节提供了理解PTEN生物学的新方面的独特机会，以靶向其在人类癌症中频繁失活的后果。为了实现这一目标，该计划项目联合了一组国际知名的合作者，他们将跨越学科界限，为PTEN调控提供新的见解，并为PTEN靶向肿瘤治疗开发新的策略。我们建议一个全面的策略，以涵盖PTEN海滨。我们将首次定义与PTEN通路相关的泛素连接酶、蛋白酶、代谢通量和活性氧。我们还提出了高度集中的努力，在验证新的药物靶点，基于结构的药物设计，并确定分子特征，以表明患者将响应PTEN靶向治疗。在四个核心的综合支持下，这五个项目将共同致力于解决以下PTEN生物学的核心问题：项目1：定义控制PTEN周转的机制，并开发这些途径的拮抗剂，以便PTEN可以在肿瘤中稳定。项目二：确定PTEN如何通过E3连接酶Siah 2调节转移，并使用基于结构的药物设计来开发Siah 2的拮抗剂用于转移性疾病。项目三：定义由PTEN调节的中心碳代谢的方面，并评估这些代谢枢纽是否是PTEN无效肿瘤中的有效药物靶点。项目4：利用基于结构的药物设计开发靶向AKT和Siah 2的新药。项目5：使用新型纳米传感器检测PTEN无效肿瘤的培养物和动物模型中的氧和ROS水平。除管理核心（A）外，还将为siRNA构建体和文库（核心B）、分析人肿瘤细胞系和TMA以获得在拟定研究过程中识别的标志物的分子特征（核心C）以及分析针对项目1-4中在2D、3D培养物和动物模型中研究的每种组分开发的抑制剂（核心D）提供支持。用于监测氧和ROS的纳米传感器的可用性提供了前所未有的机会，以高灵敏度评估培养和动物模型中PTEN信号传导（和本计划项目中开发的抑制剂）ROS和氧水平之间的关系。总的来说，分子生物学，生物化学，代谢组学，基于结构的药物设计和纳米技术的组合提供了一个首屈一指的综合研究，解决关键的肿瘤生物学悬而未决的问题，围绕关键肿瘤抑制基因PTEN的机会。

DESCRIPTION (provided by applicant): This program project represents a highly integrated approach to translate basic science findings pertaining to the PTEN (phosphatase and tensin homolog) signaling pathway which is among the most frequently inactivated tumor suppressor gene in human tumors. The overall hypothesis of this proposal is that understanding regulation of PTEN offers unique opportunities for comprehension of novel aspects of PTEN biology for targeting the consequence of its frequent inactivation in human cancer. To achieve this goal, this Program Project unites an internationally renowned group of collaborators who will cross disciplinary boundaries to provide novel insights into PTEN regulation, and to develop new strategies for PTEN-targeted tumor therapy. We propose a comprehensive strategy for covering the PTEN waterfront. For the first time we will define the ubiquitin ligases, proteases, metabolic fluxes, and reactive oxygen species that are linked to the PTEN pathway. We also propose highly focused efforts in the validation of new drug targets, structure-based drug design, and the identification of molecular signatures to indicate patients that will respond to PTEN-targeted therapy. With the integrated support of the four Cores, the five Projects will work together to address the following central questions in PTEN biology: Project 1: Define the mechanisms that control the turnover of PTEN and develop antagonists of these pathways so that PTEN can be stabilized in tumors. Project 2: Determine how PTEN regulates metastasis through the E3 ligase Siah2 and use structure-based drug design to develop antagonists of Siah2 for metastatic disease. Project 3: Define aspects of central carbon metabolism that are regulated by PTEN, and assess whether these metabolic hubs are valid drug targets in PTEN null tumors. Project 4: Use structure-based drug design to develop novel drugs targeting AKT and Siah2. Project 5: Use novel nanosensors to examine oxygen and ROS levels in cultures and animal models of PTEN null tumors. In addition to the administrative Core (A), support will be provided for siRNA constructs and libraries (Core B), analysis of human tumor cell lines and TMAs for molecular signatures of markers identified in the course of the proposed studies (Core C) and for analysis of inhibitors developed against each of the components studied in projects 1-4 in 2D, 3D cultures and animal model (Core D). The availability of nanosensors for monitoring oxygen and ROS offer unprecedented opportunity to assess at high sensitivity the relation between PTEN signaling (and inhibitors developed in this Program Project) ROS and oxygen levels, in culture and animal models. Overall, the combination of molecular biology, biochemistry, metabolomics, structure based drug design and nanotechnology offers a second to none opportunity for integrated studies that address critical unanswered questions in tumor biology centered around key tumor suppressor gene PTEN.