Targeting FGFR to prevent the outgrowth of metastatic breast cancer

项目摘要/摘要 候选人：作为一名研究生和博士后，我完成了全面的研究项目 关于癌症生长和转移的许多方面。我目前的资金来源是 美国癌症学会的博士后奖学金。多年来，我的生产率一直很高 在过去的六年里，持续出版了影响巨大的手稿。我目前担任的职位是 高级研究助理，凯斯西储大学最高博士后培训级别。 在收到并执行此应用程序的指导阶段之后，我的最终目标是运行 一流大学或研究机构的富有成效和活力的独立研究计划。 环境：凯斯综合癌症中心内的研究和核心设施仅次于 什么都没有。在这个环境中，我将能够在基础、临床前和临床之间无缝过渡 研究。特别是在希曼实验室，我一直并将继续收到非凡的 研究技巧、补助金和手稿准备、演示技巧和研究方面的培训 管理层。此外，通过准备这份申请的过程，我挖掘了重要的 案例综合癌症中心内的智力资源。我已经寻求了建议和指导 来自许多高级研究人员，他们的意见帮助我构建了一个专注的和临床的 相关建议。这种独特的培训将使我在过渡到独立调查员的过程中脱颖而出。 研究：乳腺癌确诊为晚期时五年存活率骤降 转移性疾病。事实上，目前的治疗方法在减少转移性乳腺癌生长方面并不有效。 这表明驱动转移性生长的分子途径发生了根本性的变化 反对原发肿瘤的生长。根据定义，三重阴性乳腺癌(TNBC)缺乏所需的靶点 荷尔蒙(雌激素和孕激素受体)和Her2靶向治疗。在TNBC的背景下， 原发肿瘤中EGFR表达增强与疾病预后降低密切相关， 提示EGFR可作为治疗TNBC的有效靶点。然而，EGFR靶向化疗不起作用。 为TNBC患者提供临床益处。表皮生长因子受体的诊断价值与临床的脱节 EGFRi疗法的失败强烈表明TNBCs获得了替代的促生长信号 晚期肿瘤复发和转移的途径。伴随着疾病复发和 转移是上皮-间充质转化(EMT)的获得性。尽管最近在 阐明了参与细胞从上皮到间充质状态转换的分子角色， 人们对EMT导致的分子信号通路的具体变化知之甚少。 EGFR的配体刺激导致丝裂原活化蛋白激酶(MAPK)的强劲激活 以及信号转导和转录激活因子3(STAT3)通路，但这些信号通路是如何 受EMT的影响仍未被探索。在第一个目标中，我们认为EMT的异常启动促进了 乳腺肿瘤细胞通过引起EGFR的基本开关从原发灶扩散 从增殖到侵袭的功能。我们进一步提出，这种功能的转换需要失去 EGFR介导的STAT3激活。为了解决这一假设，我们将使用遗传学和药理学 调节STAT3表达和/或活性的方法，并评估这些STAT3修饰对 EMT促进乳腺癌生长和转移的能力。此外，我们将采用一种新的双重- 底物生物发光技术实时跟踪原位乳腺肿瘤STAT3活性 生长和转移。作为EMT介导的EGFR功能转换的结果， 在转移过程中，必须上调其他生长因子信号通路，以促进转移性生长。 根据我们的初步数据，在目标2中，我们将解决这样的假设，即成纤维细胞生长上调 因子受体(FGFR)能够取代EGFR作为主要的有丝分裂信号模块。 TNBC转移性生长。这种对生长因子依赖的转变，从EGF到成纤维细胞生长因子，提供了一个看似合理的 复发和转移的TNBC不受EGFR靶向治疗影响的机制。因此，我们将 利用遗传学和药理学方法靶向FGFR，有效抑制复发和转移 TNBC病变。最后，在Aim3中，我们将使用一种新的插入突变技术来筛选潜在的 EGFRi的介体--不敏感。此外，我们还将从功能上表征FGFR的诊断潜力 和Tensin1是我们最近建立的EGFRi不敏感的12基因签名的两个成员。我们将利用 人类乳腺癌标本的已知诊断、治疗和转归，以建立诊断 将确定最有可能对FGFRi治疗有反应的TNBC患者的平台。整体而言，建议的 研究将解决目前关于EMT进化的分子机制方面的知识空白 TNBC促生长信号从EGFR到FGFR，导致EGFR靶向药物在 治疗晚期TNBC。更重要的是，我们的研究将建立一个诊断和治疗平台 抑制FGFR作为TNBC转移性患者有效治疗选择的方案 肿瘤复发。

Project Summary/Abstract Candidate: As a graduate student and postdoctoral fellow I have completed comprehensive research projects concerning numerous aspects of cancer growth and progression to metastasis. I am currently funded by a postdoctoral fellowship from the American Cancer Society. My productivity rate has been very strong for the last six years resulting in consistent publication of high impact manuscripts. I currently hold the position of Senior Research Associate, the highest postdoctoral training rank at Case Western Reserve University. Subsequent to receipt and execution of the mentored phase of this application my ultimate goal is run a productive and dynamic independent research program at a leading university or research institute. Environment: The research and core facilities within the Case Comprehensive Cancer Center are second to none. Within this environment I will be able to seamlessly transition between basic, pre-clinical and clinical research. Within the Schiemann laboratory in particular I have been and will continue to receive extraordinary training in research technique, grant and manuscript preparation, presentation skills, and research management. Furthermore, through the process of preparing this application I have tapped into the significant intellectual resources within the Case Comprehensive Cancer Center. I have sought out advice and guidance from numerous high level investigators whose input has aided me in constructing a focused and clinically relevant proposal. This unique training will set me apart as I transition into an independent investigator. Research: The five-year survival rate of breast cancer drops precipitously when diagnosed as late stage metastatic disease. Indeed, current therapies are ineffective at reducing the growth of metastatic breast cancer suggesting that there is a fundamental change in the molecular pathways that drive metastatic outgrowth as opposed to primary tumor growth. By definition triple negative breast cancers (TNBC) lack the required targets of hormonal (estrogen and progesterone receptors) and Her2-targeted therapies. In the context of TNBC, enhanced expression of EGFR within the primary tumor is strongly linked to decreased disease prognosis, suggesting EGFR as an effective target in treating TNBC. However, EGFR-targeted chemotherapies do not offer clinical benefit to TNBC patients. This disconnect between the diagnostic value of EGFR and the clinical failure of EGFRi therapies strongly suggests that TNBCs acquire alternate growth promoting signaling pathways during late-stage tumor recurrence and metastasis. Concomitant with disease recurrence and metastasis is the acquisition of epithelial-mesenchymal transition (EMT). Despite recent advances in elucidating the molecular players involved in the conversion of cells from an epithelial to mesenchymal state, little is known about the specific changes in molecular signaling pathways that result as a consequence of EMT. Ligand stimulation of EGFR leads to the robust activation of both mitogen activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (Stat3) pathways, but how these signaling pathways are affected by EMT remains unexplored. In the first Aim we propose that aberrant initiation of EMT facilitates mammary tumor cell dissemination from the primary lesion by causing a fundamental switch in EGFR function from proliferation to invasion. We further propose that this switch in function requires the loss of EGFR-mediated Stat3 activation. To address this hypothesis we will use genetic and pharmacological approaches to modulate Stat3 expression and/or activity, and assess the impact of these Stat3 modifications on the ability of EMT to enhance breast cancer growth and metastasis. Furthermore, we will employ a novel dual- substrate bioluminescence technique to track real-time in vivo Stat3 activity as orthotopic mammary tumors grow and metastasize. As a consequence of the EMT-mediated switch in EGFR function, following systemic dissemination, other growth factor signaling pathways must be upregulated to facilitate metastatic outgrowth. Based on our preliminary data, in Aim 2 we will address the hypothesis that upregulation of fibroblast growth factor receptor (FGFR) is capable of replacing EGFR as the dominant mitogenic signaling module during metastatic outgrowth of TNBC. This switch in growth factor dependence from EGF to FGF presents a plausible mechanism by which recurrent and metastatic TNBC is unaffected EGFR-targeted therapies. Therefore, we will use genetic and pharmacological approaches to target FGFR and effectively inhibit recurrent and metastatic TNBC lesions. Finally, in Aim3 we will use a novel insertional mutagenesis technique to screen for potential mediators of EGFRi-insensitivity. In addition we will functionally characterize the diagnostic potential of FGFR and Tensin1 two members of our recently established 12-gene signature of EGFRi-insensitivity. We will utilize human breast cancer specimens of known diagnosis, treatment, and outcome to establish a diagnostic platform that would identify TNBC patients most likely to respond to FGFRi therapy. Overall, the proposed studies will address current gaps in knowledge concerning the molecular mechanisms by which EMT evolves TNBC growth promoting signaling from EGFR to FGFR, leading to the failure of EGFR-targeting agents in treating late-stage TNBC. More importantly, our studies will establish a diagnostic platform and therapeutic protocol for the inhibition of FGFR as an effective treatment option for TNBC patients undergoing metastatic tumor recurrence.