Synergism of c-Met and EGFR pathways: their therapeutic role in lung cancer

c-Met 和 EGFR 通路的协同作用：它们在肺癌中的治疗作用

• 批准号: 8244111
• 负责人: NEELU PURI
• 金额: $ 17.5万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-06 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8244111
• 关键词: Active Sites; Apoptosis; Biological Markers; Cancer Patient; Cell Line; Cell Proliferation; Cells; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Development; Drug resistance; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Formalin; Future; Goals; Grant; Growth and Development function; Hepatocyte Growth Factor; Human; Immunoblotting; In Vitro; Individual; Knowledge; Ligands; Lung Neoplasms; MAP Kinase Gene; Malignant neoplasm of lung; Mass Spectrum Analysis; Mediating; Medical; Microarray Analysis; Mutation; Non-Small-Cell Lung Carcinoma; PI3K/AKT; Paraffin Embedding; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Play; Prevalence; Proteins; Receptor Activation; Receptor Protein-Tyrosine Kinases; Research; Research Design; Resistance; Resistance development; Role; Science; Signal Pathway; Signal Transduction; Signaling Protein; Small Interfering RNA; Technology; Testing; Therapeutic; Tissue Sample; Tumor Tissue; Tumorigenicity; Tyrosine Kinase Inhibitor; Western Blotting; Work; base; cancer cell; cancer therapy; cell motility; clinically relevant; human tissue; inhibitor/antagonist; innovation; insight; meetings; new technology; novel; outcome forecast; overexpression; receptor; receptor function; resistance mechanism; response; small hairpin RNA; small molecule; synergism; therapy resistant; tumor; tumor growth; vector; Active Sites; Apoptosis; Biological Markers; Cancer Patient; Cell Line; Cell Proliferation; Cells; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Development; Drug resistance; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Formalin; Future; Goals; Grant; Growth and Development function; Hepatocyte Growth Factor; Human; Immunoblotting; In Vitro; Individual; Knowledge; Ligands; Lung Neoplasms; MAP Kinase Gene; Malignant neoplasm of lung; Mass Spectrum Analysis; Mediating; Medical; Microarray Analysis; Mutation; Non-Small-Cell Lung Carcinoma; PI3K/AKT; Paraffin Embedding; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Play; Prevalence; Proteins; Receptor Activation; Receptor Protein-Tyrosine Kinases; Research; Research Design; Resistance; Resistance development; Role; Science; Signal Pathway; Signal Transduction; Signaling Protein; Small Interfering RNA; Technology; Testing; Therapeutic; Tissue Sample; Tumor Tissue; Tumorigenicity; Tyrosine Kinase Inhibitor; Western Blotting; Work; base; cancer cell; cancer therapy; cell motility; clinically relevant; human tissue; inhibitor/antagonist; innovation; insight; meetings; new technology; novel; outcome forecast; overexpression; receptor; receptor function; resistance mechanism; response; small hairpin RNA; small molecule; synergism; therapy resistant; tumor; tumor growth; vector

DESCRIPTION (provided by applicant): c-Met and Epidermal Growth Factor Receptor (EGFR) are receptor tyrosine kinases (RTKs) that work together to promote the progression of Non-Small Cell Lung Cancer (NSCLC). Our current studies focus on the use of VeraTag technology to quantitatively, accurately and objectively determine if co- activation, co-overexpression and proximity of c-Met and EGFR can predict patient prognosis. Research over the last decade has enabled the development of several RTK-targeted inhibitors (RTKIs) against both c-Met and EGFR. However, recent clinical trials, targeting individual receptors or combinations, are not always effective due to their acquired resistance. Hence, it is important to determine the mechanism of resistance to EGFR/c-Met RTKIs, which is not defined, by utilizing drug resistant cell lines, developed in our lab. Future studies must focus on the development of new inhibitor combinations which may be targeted together to reduce resistance to EGFR/c-Met based therapies. The specific aims of this project are: Specific Aim 1: Determine the role of EGFR and c-Met expression in lung cancer tumorigenicity and elucidate the role of c-Met/EGFR mediated synergism and resistance in human tumor tissues. A. Determine the co-activation, co-overexpression and proximity of c-Met and EGFR in human lung tumor tissues through VeraTag technology and immunostaining for EGFR and c-Met phosphorylated forms and their ligands. Specific Aim 2: Elucidate the mechanism of c-Met and EGFR resistance. A. Determine if resistance developed to EGFR/c-Met RTKIs alters response to EGF or HGF and increases c- Met stability. B. Identify key downstream signaling proteins associated with acquired resistance to c-Met/EGFR therapies by western blotting/mass spectrometry and validate the efficacy of these proteins using si/shRNA vectors. C. Identify activated kinases mediating resistance to EGFR/c-Met inhibitors using kinase enrichment kits and active site probes. Study Design: To understand how c-Met and EGFR function together, we propose to study EGFR and c-Met co-overexpression, co-activation and proximity in NSCLC tumors using novel VeraTag technology and immunostaining. To study the mechanism of resistance to EGFR and c-Met RTKIs, cell lines resistant to EGFR, c-MET, and EGFR/c-Met combined RTKIs have been developed. Mass spectrometry and microarray analysis will be performed to identify specific proteins that may play an important role in resistance to c-Met and EGFR inhibitors. Using siRNA/shRNA vectors the role of a few novel target proteins that play a role in resistance to RTKIs will be validated. Kinase enrichment kits and active site probes will be used in collaboration with Thermo Fisher Scientific to study kinases which are activated specifically in resistant cells. PUBLIC HEALTH RELEVANCE: Advances in medical science have developed novel therapies that target EGFR and c-Met receptors, important proteins involved in the development and progression of lung cancer. Our studies will focus on determining if co-activation, co-overexpression and/or co-localization of EGFR/c-Met are correlated with prognosis in lung cancer patients. Recent clinical trials targeting individual receptors or combinations are not always effective due to their acquired resistance and hence, utilizing drug resistant cell lines developed in our lab will help us determine how tumors become resistant to these therapies and how resistance can be overcome.

描述（由申请人提供）：C-MET和表皮生长因子受体（EGFR）是受体酪氨酸激酶（RTK），它们共同促进非小细胞肺癌（NSCLC）的进展。我们目前的研究着重于使用Veratag技术来定量，准确和客观地确定C-MET和EGFR的共同激活，共同表达和接近是否可以预测患者的预后。在过去的十年中，研究使得针对C-MET和EGFR的几种RTK靶向抑制剂（RTKI）的发展。但是，最近的临床试验（针对单个受体或组合）并不总是由于获得的抗性而有效。因此，重要的是确定对EGFR/C-MET RTKIS的抗性机制，该机制未通过利用耐药细胞系在我们的实验室中开发。未来的研究必须集中于新抑制剂组合的发展，这些组合可能会共同针对，以降低对基于EGFR/C-MET疗法的耐药性。该项目的具体目的是：具体目标1：确定EGFR和C-MET表达在肺癌肿瘤性中的作用，并阐明C-MET/EGFR介导的协同作用和在人类肿瘤组织中的耐药性的作用。答：通过Veratag技术，确定人类肺肿瘤组织中C-MET和EGFR的共同激活，表达和接近性，以及EGFR和C-MET磷酸化形式及其配体的免疫染色。具体目标2：阐明C-MET和EGFR电阻的机理。答：确定对EGFR/C-MET RTKIS产生的电阻是否改变了对EGF或HGF的反应，并增加了C-MET稳定性。 B.通过Western印迹/质谱法确定与获得C-MET/EGFR疗法获得的抗性相关的钥匙，并使用Si/ShRNA载体验证这些蛋白质的功效。 C.使用激酶富集试剂盒和活性位点探针鉴定激活的激酶，从而介导对EGFR/C-MET抑制剂的抗性。研究设计：为了了解C-MET和EGFR如何共同发挥功能，我们建议使用新型的Veratag技术和免疫染色来研究NSCLC肿瘤中EGFR和C-MET共同表达，共激活和接近性。为了研究对EGFR和C-MET RTKI的抗性的机制，已经开发了对EGFR，C-MET和EGFR/C-MET组合RTKIS的细胞系。将进行质谱和微阵列分析，以识别可能在C-MET和EGFR抑制剂抗性中起重要作用的特定蛋白质。使用siRNA/shRNA向量，将验证一些在抗RTKIS中发挥作用的一些新型靶蛋白的作用。激酶富集试剂盒和活性位点探针将与Thermo Fisher Scientific合作研究在抗性细胞中专门激活的激酶。 公共卫生相关性：医学的进步开发了针对EGFR和C-MET受体的新型疗法，这是涉及肺癌发展和发展的重要蛋白质。我们的研究将着重于确定EGFR/C-MET共同激活，共同表达和/或共定位是否与肺癌患者的预后相关。靶向单个受体或组合的最新临床试验并不总是有效的，因为它们获得了耐药性，因此利用我们实验室中开发的耐药细胞系将有助于我们确定肿瘤如何对这些疗法具有抗性以及如何克服耐药性。