Development of an Anti-Cancer Approach Through a Novel Pathway of Translational Regulation

摘要 信号转导和转录激活因子3(Signal Transducer and Activator of转录3，STAT3)是一种转录调控因子，在转录过程中起着至关重要的作用 在细胞的恶性转化中，肿瘤生长和转移。STAT3活动或表达式为 在70%的人类肿瘤中上调，从而导致癌症的快速进展，因此它是一种潜在的 癌症治疗中的药理靶点。研究表明，STAT3基因敲除降低了异常 细胞生长。然而，STAT3活性的直接和间接抑制剂并没有提供预期的效果， 迫切需要寻找STAT3抑制的替代策略。STAT3基因异常表达的存在 STAT3在不同类型肿瘤中的表达及其治疗癌症的潜在能力 决定了研究的高度意义。当前项目的中心思想是基于对STAT3的控制 在核糖体合成过程中通过调节新途径的成分来表达 是我们最近发现的。RAPP(异常蛋白生产调节)途径的特异性 调节蛋白质的合成，降解不能正确相互作用的蛋白质的mRNAs 核糖体上的合成。我们的假设是，RAPP翻译调节子可能被用来 控制STAT3的表达，从而抑制细胞增殖和肿瘤生长。它的概念是 以前从未探索过STAT3调控的类型，对STAT3翻译伙伴和 关于它们对STAT3表达的调控，从而展示了该项目的新颖性。我们的预赛 研究表明，RAPP通路参与了STAT3的调控。以下是具体目标 为这个项目设计的：(1)在核糖体上合成STAT3的过程中确定新的翻译伙伴， 以及(2)检验RAPP翻译调节因子可以被调节以降低STAT3的假设 表达与癌细胞增殖的关系。这项研究基于独特的检测和检测技术 利用定点光交联法和IPINCH方法鉴定翻译过程中的蛋白质。我们会 使用候选和公正的方法测试STAT3控制中的RAPP调节器。这些实验还将 使用细胞培养模型和RNA干扰技术或过度生产来验证 STAT3调控中的RAPP成分。在这个项目中，我们将确定其潜在的抗癌特性。 RAPP通过测量癌细胞的增殖和凋亡来进行调控。这项提议的成功将建立在 为未来用动物模型和使用化学调节剂(潜在药物)进行研究奠定基础 开发抗癌新药。潜在地，这项提议将导致概念上的新发展 癌症的药物治疗指南。

ABSTRACT STAT3 (Signal Transducer and Activator of Transcription 3) is a transcriptional regulator that plays crucial role in malignant transformation of the cells, tumor growth and metastasis. STAT3 activity or expression is upregulated in 70% of human tumors contributing to a rapid cancer progression and therefore it is a potential pharmacological target in cancer treatments. It was demonstrated that STAT3 knockdown reduced abnormal cell growth. However, direct and indirect inhibitors of STAT3 activity did not provide expected efficacy, demanding search for alternative strategies for STAT3 inhibition. The presence of STAT3 aberrant expression in different types of tumors and potential ability to treat cancer through decrease of STAT3 expression determines high significance of the study. The central idea of the current project is based on control of STAT3 expression during its synthesis on the ribosome through modulation of components of the novel pathway recently discovered by us. The RAPP (Regulation of Aberrant Protein Production) pathway specifically regulates protein synthesis, degrading mRNAs of the proteins that do not properly interact during their synthesis on the ribosome. Our hypothesis is that the RAPP translational regulators may be used to control STAT3 expression and therefore restrict cell proliferation and tumor growth. The concept of this type of STAT3 regulation was never explored before, little is known about STAT3 translational partners and about their modulation to control STAT3 expression, thus demonstrating novelty of the project. Our preliminary data demonstrate that the RAPP pathway is involved in regulation of STAT3. The following specific aims are designed for this project: (1) identify novel STAT3 translational partners during its synthesis on the ribosome, and (2) test the hypothesis that RAPP translational regulators can be modulated to decrease STAT3 expression and cancer cell proliferation. The study is based on unique technologies for detection and identification of proteins during translation by site-specific photo-crosslinking and iPINCH approaches. We will use candidate and unbiased approaches to test RAPP regulators in STAT3 control. The experiments will also involve cell culture models and RNA interference techniques or overproduction to verify involvement of the RAPP components in STAT3 regulation. In this project we will determine potential anti-cancer properties of the RAPP modulation by measuring cancer cell proliferation and apoptosis. The success of the proposal will build a foundation for the future studies with animal models and use of chemical regulators (potential drugs) to develop new anti-cancer medicine. Potentially, this proposal will lead to development of conceptually new directions in pharmacological treatments of cancer.