Understanding and Eliminating Oncogenic EGFR Signaling in Malignant Glioma

了解和消除恶性胶质瘤中的致癌 EGFR 信号转导

基本信息

批准号：
8320724
负责人：
Alain Charest
金额：
$ 66.01万
依托单位：
TUFTS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8320724
关键词：
Adaptor Signaling Protein Address Animal Model Animals Applications Grants Biological Biology Cancer Biology Categories Chemicals Clinical Complement Complex Coupled Custom Data Detection Development Disease Drug Delivery Systems EGFR Protein Overexpression Educational process of instructing Enzymes Epidermal Growth Factor Receptor Etiology Event Failure Funding Gene Expression Gene Proteins Genes Genetic Genetically Engineered Mouse Glioblastoma Goals Heart Human Individual Industry Knowledge Lead Libraries Life Maintenance Malignant Glioma Malignant neoplasm of brain Mass Spectrum Analysis Measurement Mediating Mediator of activation protein Methodology Methods Modeling Mus Nanotechnology Nature Nodal Oncogenic PTEN gene Pharmacologic Substance Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphoserine Phosphotransferases Phosphotyrosine Play Pre-Clinical Model Public Health RNA Interference RNA library Radiation therapy Reagent Receptor Signaling Reporting Research Resistance Resources Retrospective Studies Role Sampling Screening procedure Signal Pathway Signal Transduction Small Interfering RNA Solid System Testing The Cancer Genome Atlas Therapeutic Therapeutic Agents Therapeutic Intervention Threonine Time Toxic effect Translating Tumor Suppressor Genes Vacuum base cell growth efficacy testing falls forest gene function in vivo interest iron oxide loss of function member migration mouse model nanomaterials neoplastic cell p19ARF pre-clinical prevent programs prospective research study small hairpin RNA success therapeutic target therapy resistant tool tumor

项目摘要

DESCRIPTION (provided by applicant): For decades, we have known that overexpression of the epidermal growth factor receptor (EGFR) is a major component of the etiology of glioblastoma multiforme (GBM), yet little is known regarding the signaling events that emanate from activated EGFR in GBM. It is becoming increasingly clear that signaling pathways are complex and highly dynamic and cannot be studied in a vacuum. This certainly helps define GBM's plasticity and explain failures of targeted therapies for GBMs. It is therefore imperative that we study EGFR signaling events globally and in the context of a relevant animal model that we can genetically manipulate at will. We have developed a genetically engineered mouse model (GEMM) of GBM, based on the most common genetic aberrations observed in human tumors, that is overexpression of EGFR along with loss of function of the p16lnk4a/p19ARF and PTEN tumor suppressor genes. We hypothesize that investigating global EGFR signaling pathways in our model using phosphoproteomic methods will reveal key nodal signaling events that are responsible for tumor cell growth, migration and resistance to therapies. By using our model to study signaling events responsible for these effects, our GEMM of GBM will reveal new and insightful information on the etiology of GBMs. This will be accomplished by fulfilling the following goals: 1) To determine and study network dynamics of phosphotyrosine and phosphoserine/threonine signaling events in GBM tumors from our mouse models using mass spectrometry. 2) To study the effects of targeted therapeutic treatment of our GBMs on global signaling phospho-networks. 3) To systematically eliminate the signaling events usurped by EGFR in our GBM tumor cells using custom-made short hairpin RNA (shRNA) libraries, and determine resulting phenotypes (tumor cell growth, invasion, resistance to targeted, chemo and radiation therapies). 4) To validate those key phosphoevents for mouse GBM biology in human GBM samples. 5) To ascertain toxicity profiles and efficacy spectrum of various nanotechnology platforms for the efficient delivery of small interfering RNA (siRNA) molecules to GBM tumor cells in vivo. This application will establish the groundwork for evaluating specific gene function in the context of RNA interference-mediated therapeutic intervention for GBM in a pre-clinical setting, using various nanoplatforms as delivery tools. The power of the prospective research program proposed herein lies in our ability to manipulate gene expression and perform genetic experiments in live animals. Together, these features commensurably complement the retrospective studies brought forward by The Cancer Genome Atlas by providing a much-needed animal system capable of efficient analysis of gene function.

描述（由申请人提供）：几十年来，我们知道表皮生长因子受体（EGFR）的过表达是胶质母细胞瘤多形（GBM）病因的主要组成部分，但关于从活性激活的信号传导事件中，知之甚少EGFR在GBM中。越来越清楚的是，信号通路是复杂且高度动态的，无法在真空中进行研究。当然，这有助于定义GBM的可塑性并解释针对GBM的靶向疗法的失败。因此，我们必须在全球范围内研究EGFR信号传导事件，并在相关的动物模型的背景下我们可以随意进行基因操纵。我们已经根据人类肿瘤中观察到的最常见的遗传像差开发了GBM的基因工程小鼠模型（GEMM），即EGFR的过表达以及p16LNK4A/P19ARF和PTEN抑制肿瘤基因的功能损失。我们假设使用磷酸化蛋白质组学方法在模型中研究全球EGFR信号通路将揭示导致肿瘤细胞生长，迁移和对疗法抗性的关键淋巴结信号事件。通过使用我们的模型研究负责这些效果的信号事件，我们的GBM宝石将揭示有关GBMS病因的新信息。这将通过实现以下目标来实现：1）使用质谱法的GBM肿瘤中的磷酸酪氨酸和磷酸酪氨酸/苏氨酸信号事件的网络动态。 2）研究我们GBM的靶向治疗治疗对全球信号磷酸化网络的影响。 3）使用定制的短发夹RNA（SHRNA）文库系统地消除EGFR在GBM肿瘤细胞中篡夺的信号传导事件，并确定产生的表型（肿瘤细胞的生长，侵袭，对靶标的，化学疗法和辐射疗法）。 4）验证人类GBM样品中小鼠GBM生物学的关键磷酸事件。 5）确定各种纳米技术平台的毒性谱和功效谱，以有效地递送小型干扰RNA（siRNA）分子到体内GBM肿瘤细胞。本应用将在RNA干扰介导的治疗干预措施中，在临床前的环境中，使用各种纳米板作为递送工具，为GBM的RNA干扰介导的治疗干预措施评估特定基因功能的基础。本文提出的前瞻性研究计划的力量在于我们能够操纵基因表达并在活动物中进行遗传实验的能力。这些特征共同补充了癌症基因组图集通过提供急需的动物系统来有效分析基因功能的回顾性研究。