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

基本信息

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
项目类别：
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

项目摘要

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），它们共同作用促进非小细胞肺癌（NSCLC）的进展。我们目前的研究重点是利用VeraTag技术定量、准确、客观地确定c-Met和EGFR的共激活、共过表达和接近是否可以预测患者预后。过去十年的研究已经开发出多种针对 c-Met 和 EGFR 的 RTK 靶向抑制剂 (RTKI)。然而，最近针对单个受体或组合的临床试验由于其获得性耐药性并不总是有效。因此，通过利用我们实验室开发的耐药细胞系来确定对 EGFR/c-Met RTKI 的耐药机制（尚未定义）非常重要。未来的研究必须集中于开发新的抑制剂组合，这些组合可以共同降低对基于 EGFR/c-Met 的疗法的耐药性。该项目的具体目标是：具体目标1：确定EGFR和c-Met表达在肺癌致瘤性中的作用，阐明c-Met/EGFR介导的人体肿瘤组织中的协同和耐药作用。 A. 通过 VeraTag 技术和 EGFR 和 c-Met 磷酸化形式及其配体的免疫染色，确定人肺肿瘤组织中 c-Met 和 EGFR 的共激活、共过表达和接近性。具体目标2：阐明c-Met和EGFR耐药的机制。 A. 确定对 EGFR/c-Met RTKI 产生的耐药性是否会改变对 EGF 或 HGF 的反应并增加 c-Met 稳定性。 B. 通过蛋白质印迹/质谱鉴定与 c-Met/EGFR 疗法获得性耐药相关的关键下游信号蛋白，并使用 si/shRNA 载体验证这些蛋白的功效。 C. 使用激酶富集试剂盒和活性位点探针鉴定介导 EGFR/c-Met 抑制剂耐药性的活化激酶。研究设计：为了了解 c-Met 和 EGFR 如何共同发挥作用，我们建议使用新型 VeraTag 技术和免疫染色来研究 NSCLC 肿瘤中 EGFR 和 c-Met 的共过表达、共激活和邻近性。为了研究对 EGFR 和 c-Met RTKI 的耐药机制，开发了对 EGFR、c-MET 和 EGFR/c-Met 联合 RTKI 耐药的细胞系。将进行质谱和微阵列分析，以确定可能在 c-Met 和 EGFR 抑制剂耐药性中发挥重要作用的特定蛋白质。使用 siRNA/shRNA 载体，将验证一些在 RTKI 抗性中发挥作用的新型靶蛋白的作用。将与 Thermo Fisher Scientific 合作使用激酶富集试剂盒和活性位点探针来研究在耐药细胞中特异性激活的激酶。公共健康相关性：医学科学的进步已经开发出针对 EGFR 和 c-Met 受体的新疗法，这些受体是参与肺癌发生和进展的重要蛋白质。我们的研究将重点确定 EGFR/c-Met 的共激活、共过表达和/或共定位是否与肺癌患者的预后相关。最近针对单个受体或组合的临床试验由于其获得性耐药性并不总是有效，因此，利用我们实验室开发的耐药细胞系将帮助我们确定肿瘤如何对这些疗法产生耐药性以及如何克服耐药性。