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，但几乎没有有效的方法 现有的治疗方法只有有限的好处。放射联合细胞毒治疗仍然是大多数患者的主要治疗方法。 然而，它的治疗指标相对较窄，患者总是遭受与治疗相关的系统性疾病。 毒物。为LUSC开发新的靶向疗法仍然是一个高度优先的问题。其中最 肺癌基因组测序的重要发现是Keap1基因频繁(~30%)的改变。 Nrf2信号通路。NRF2抗氧化信号通路构成了主要的细胞防御 对抗氧化应激的系统。负责管理NRF2活动的几种机制是已知的， 然而，事实证明，对于治疗干预来说，它们在很大程度上是棘手的(例如，转录因子)。近期 研究表明，几种蛋白激酶在功能上影响NRF2，尽管全球评估如何 该近缘体指示NRF2的生物学仍未得到测试。作为最易下药的蛋白质类别之一， 激酶和磷酸酶为NRF2指导的治疗干预提供了有吸引力的靶点。我们的初步数据 已经在NRF2和蛋白激酶之间建立了相互通信，包括上游修饰 和下游效应器。因此，我们假设KINOME包含关键的调节因子 NRF2信号转导通路，为NRF2激活的肺癌提供新的治疗靶点。我们已经组建了一支 独特的多学科研究团队，在LUSC分子信号、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学，以破译 Kinome和NRF2信号，识别新的治疗靶点，并在临床前模型中分析它们。我们的 具体目标包括：(1)识别调节NRF2的激酶；(2)识别NRF2反应 以及(3)在新的NRF2模型中评估Kinome功能 激活的LUSC。该项目通过对LUSC肿瘤样本进行动态组蛋白质组图谱的研究，展示了很强的创新性 和细胞系、高通量化学筛选、功能增减基因筛选及其应用 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的方法。 人支气管上皮细胞和新的基因工程小鼠模型和衍生细胞系。这个 这项工作的结果将揭示在功能上影响NRF2生物学的蛋白激酶，并在这样做的过程中可能导致 LUSC和其他NRF2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。最终，我们建议的研究的成功完成将为我们提供一个路线图。 在这些其他人类恶性肿瘤的靶向治疗发现方面做出类似的努力。

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.