Novel Molecular Functions of WEE1 in Esophageal Adenocarcinoma

项目摘要/摘要（30行） 食管腺癌（EAC）的发病率急剧增加， 在美国的三十年。大约80%的患者在晚期（III期）被诊断出 或IV），5年生存率低于5%。EAC的治疗选择有限，而且往往无效 因为缺乏基于生物学的靶向治疗。根据美国癌症协会，吸烟是 这是EAC的一个主要和重要的危险因素。我们新的初步数据表明，WEE 1表达水平 在EAC细胞中，吸烟和WEE 1的频繁异常过表达诱导了令人惊讶的细胞溶质 在EAC组织样品和体外细胞模型中的定位。该提案的长期目标是评估 WEE 1在EAC中的分子、生物学和治疗潜力。目标，这是一个桥梁， 长期目标是研究激活吸烟-WEE 1-STAT 3信号传导的分子机制 途径以及靶向WEE 1在人EAC中的疗效和临床结果。核心假设是， 吸烟诱导的WEE 1异常过表达为癌细胞提供了有效的促存活优势 通过激活STAT 3信号通路。靶向WEE 1-STAT 3轴可能是一种新的治疗方法 在EAC的方法。这一假设主要是由国际原子能机构进行的初步研究得出的。 申请人的工作。为了检验这一假设，将实现三个具体目标。目标1。调查机制 吸烟诱导EAC中的WEE 1。目标二。确定WEE 1-STAT 3信号转导的分子功能 轴线目标3。探讨WEE 1-STAT 3轴在EAC中的临床意义。本提案的创新之处 包括以下创新假设：1）吸烟诱导人EAC中胞质WEE 1过表达 激活STAT 3信号传导，2）靶向WEE 1以改善EAC结果的新治疗方法， 和3）利用遗传工程化的tet诱导细胞模型，慢病毒表达， CRISPR基因敲低系统、患者来源的异种移植物（PDX）和利用该系统的SynergySeq分析 NIH Library of Integrated Network-Based Cellular Signatures（LINCS）项目资源。这项建议是 重要的，可以有积极的影响，不仅揭示了一个新的分子WEE 1-STAT 3信号轴 还因为其平移分量。翻译研究包括1）识别潜在的 使用RNA-seq在3种最佳与3种最差WEE 1抑制剂（MK 1775）PDX中进行治疗反应的生物标志物 缓解者和2）确定MK 1775作为单药或联合用药的潜在治疗疗效 FDA批准的抗癌药物在临床前环境中通过复杂的计算疗法预测 方法使用SynergySeq.整合的分子，功能和翻译的方法，在这方面， 这些建议有望为EAC的生物学和分子基础提供新的理解。结果 可以为循证治疗奠定基础，并迅速转化为改善治疗， EAC患者的预后。

Project summary/abstract (30 lines) A sharp increase in the incidence of esophageal adenocarcinoma (EAC) has been documented over the past three decades in the United States. Approximately 80% of patients are diagnosed at advanced stages (III or IV) with a five-year survival rate less than 5%. Therapeutic options for EAC are limited and often ineffective due to the lack of biology based targeted therapeutics. According to the American Cancer Society, smoking is a major and important risk factor of EAC. Our novel preliminary data demonstrate that WEE1 expression level is induced by smoking in EAC cells and a frequent aberrant overexpression of WEE1 with surprising cytosolic localization in EAC tissue samples and in vitro cell models. The long-term goal of this proposal is to evaluate the molecular, biological, and therapeutic potential of WEE1 in EAC. The objective, which is a bridge to the long-term goal, is to investigate molecular mechanisms by which activate smoking-WEE1-STAT3 signaling pathway and the efficacy and clinical outcome of targeting WEE1 in human EAC. The central hypothesis is that smoking induced aberrant overexpression of WEE1 provides potent pro-survival advantage to cancer cells through activation of STAT3 signaling pathway. Targeting the WEE1-STAT3 axis could be a novel therapeutic approach in EAC. This hypothesis has been generated primarily from the preliminary studies conducted by the applicant’s work. Three specific aims will be achieved to test this hypothesis. Aim 1. Investigate mechanisms by which smoking induces WEE1 in EAC. Aim 2. Determine the molecular functions of WEE1-STAT3 signaling axis. Aim 3. Investigate the clinical significance of WEE1-STAT3 axis in EAC. The innovations in this proposal include innovative hypothesis that 1) smoking induced cytosolic WEE1 overexpression in human EAC activates STAT3 signaling, 2) a novel therapeutic approach targeting WEE1 for improving outcome of EAC, and 3) experimental design that utilizes genetically engineered tet-inducible cell models, lenti-viral expression, CRISPR gene knockdown systems, patient-derived xenografts (PDXs) and SynergySeq analysis utilizing the NIH Library of Integrated Network-Based Cellular Signatures (LINCS) project resources. This proposal is significant and can have a positive impact not only by revealing a novel molecular WEE1-STAT3 signaling axis but also because of its translational components. The translational studies include 1) identify potential biomarkers for therapeutic response using RNA-seq in 3 best versus 3 worst WEE1 inhibitor (MK1775) PDX responders and 2) determine the potential therapeutic efficacy of MK1775 as a single agent or in combination with FDA approved anti-cancer drugs in pre-clinical settings predicted by complex computational therapeutics approach using SynergySeq. The integrated molecular, functional, and translational approaches in this proposal are expected to provide a novel understanding of the biology and molecular basis of EAC. The results can lay the foundation for evidence-based therapy and swiftly be translated for improving therapy and prognosis in EAC patients.