The Role of Protein Kinases in NRF2-driven Lung Squamous Cell Carcinoma

项目摘要 尽管晚期肺鳞状细胞癌（LUSC）患者的预后很差，但很少有有效的 仅存在有限益处的治疗。放射性细胞毒性治疗仍然是大多数患者的主要治疗手段。 然而，它的治疗指数相对较窄，患者总是遭受治疗相关的全身性疾病， 毒性LUSC的新靶向疗法的开发仍然是一个高度优先事项。一个最 肺癌基因组测序的一个重要发现是KEAP 1基因的频繁（约30%）改变。 NRF 2信号通路。NRF 2抗氧化信号通路构成了细胞的主要防御 抗氧化应激系统。已知几种负责控制NRF 2活性的机制， 然而，已经证明它们在很大程度上难以进行治疗干预（例如，转录因子）。最近 研究已经揭示了几种蛋白激酶在功能上影响NRF 2，尽管对如何影响NRF 2的全球评估 kinome指示NRF 2生物学仍然未经测试。作为最容易被药物治疗的蛋白质种类之一， 激酶和磷酸酶为NRF 2导向的治疗干预提供了有吸引力的靶标。我们的初步数据 已经建立了NRF 2和蛋白激酶之间的相互通讯，包括上游修饰物 和下游效应器。因此，我们假设激酶组包含以下关键调控因子： NRF 2信号传导，并为NRF 2活性肺癌提供新的治疗靶点。我们组建了一个 独特的多学科研究团队，在LUSC分子信号传导，癌细胞生物学， 动物和细胞培养模型，蛋白质组学和临床治疗，以破译之间的相互作用， 激酶组和NRF 2信号传导，鉴定新的治疗靶点并在临床前模型中分析它们。我们 具体目标包括：（1）鉴定调节NRF 2的激酶;（2）鉴定NRF 2-应答的 激酶和磷酸酶;（3）NRF 2新模型中的增强激酶组功能 主动发光该项目通过LUSC肿瘤样本的激酶组蛋白质组学分析显示出强大的创新性 和细胞系、高通量化学筛选、获得和丧失功能的遗传筛选及其应用 独特的二维和三维细胞培养模型。我们的实验采用基因靶向转化， 人支气管上皮细胞和新的基因工程小鼠模型和衍生的细胞系。的 这项工作的结果将揭示蛋白激酶在功能上影响NRF 2生物学，这样做可能会导致 LUSC和其他NRF 2活性肿瘤的新有效治疗方法，包括头颈癌，膀胱癌， 癌症和卵巢癌。最后，我们建议的研究若能顺利完成， 在这些其他人类恶性肿瘤的靶向治疗发现方面的类似努力。

Project Summary Despite the dismal prognosis for patients with advanced lung squamous cell cancer (LUSC), few effective treatments of only limited benefit exist. Cytotoxic therapy with radiation remains the mainstay for most patients. However, it carries a relatively narrow therapeutic index and patients invariably suffer treatment-related systemic toxicities. The development of new targeted therapies for LUSC remains a high priority. One of the most significant discoveries from lung cancer genome sequencing is the frequent (~30%) alterations of the KEAP1- NRF2 signaling pathway. The NRF2 antioxidant signaling pathway constitutes the primary cellular defense system against oxidative stress. Several mechanisms responsible for governing NRF2 activity are known, however they have proven largely intractable for therapeutic intervention (eg. transcription factors). Recent studies have revealed that several protein kinases functionally impact NRF2, although a global evaluation of how the kinome instructs NRF2 biology remains untested. Being among the most druggable of protein classes, kinases and phosphatases offer attractive targets for NRF2-directed treatment intervention. Our preliminary data have established reciprocal communication between NRF2 and protein kinases, including upstream modifiers and downstream effectors. Therefore, we hypothesize that the kinome encompasses key regulators of NRF2 signaling and holds novel therapeutic targets for NRF2-active lung cancer. We have assembled a unique multidisciplinary team of investigators with experience in LUSC molecular signaling, cancer cell biology, animal and cell culture models, proteomics, and clinical therapeutics to decipher the interactions between the kinome and NRF2 signaling, identify novel therapeutic targets and analyze them in pre-clinical models. Our specific aims include: (1) IDENTIFY KINASES THAT REGULATE NRF2; (2) IDENTIFY NRF2-RESPONSIVE KINASES AND PHOSPHATASES; and (3) EVALUATE KINOME FUNCTION IN NOVEL MODELS OF NRF2 ACTIVE LUSC. This project shows strong innovation through kinome proteomic profiling of LUSC tumor samples and cell lines, high-throughput chemical screens, gain- and loss-of-function genetic screens, and the application of unique 2- and 3-dimensional cell culture models. Our experiments employ gene targeted-transformation of human bronchial epithelial cells and novel genetically engineered mouse models and derived cell lines. The results of this work will reveal protein kinases that functionally impact NRF2 biology, and in doing so may lead to new effective treatments for LUSC and other NRF2-active tumors, including head and neck cancer, bladder cancer, and ovarian cancer. Ultimately, the successful completion of our proposed studies will provide a roadmap for similar efforts on targeted therapeutic discovery in these other human malignancies.