Proj 1: Imaging Notch Interations with Members of Its Pathway

项目 1：成像缺口与其通路成员的相互作用

基本信息

批准号：
7287030
负责人：
RAPHAEL KOPAN
金额：
$ 24.57万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-28 至 2011-12-11
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7287030
关键词：
Accounting Adult Animals Apoptosis Bioluminescence CADASIL Cancer cell line Cell Line Cell Maintenance Cell Nucleus Cells Collaborations Development Disease Extracellular Domain Generations Genetic Models Genetic Transcription Genomics Goals Human Human Engineering Image Imagery Imaging Device Imaging technology Individual Industry Inherited Invasive Knock-in Mouse Lead Libraries Life Ligand Binding Ligands Luciferases Malignant Neoplasms Mammalian Cell Mediating Methodology Molecular Monitor NF-kappa B Neoplasm Metastasis Notch Signaling Pathway Nuclear Numbers Oncogenes Pathway interactions Patients Pharmaceutical Preparations Process Property Protein Engineering Proteins Reporter Reporting Research Personnel Screening procedure Signal Transduction Small Interfering RNA Staging Stem cells Syndrome System TNFRSF5 gene Technology Therapeutic Intervention Time Tissue Engineering Tissues Tumor Suppressor Genes Tumor Suppressor Proteins Universities Washington Work Xenograft procedure aortic valve base cancer stem cell cell growth cell type cofactor embryonic stem cell high throughput screening in vivo in vivo Cellular and Molecular Imaging Centers inhibitor/antagonist late disease onset medical schools member molecular imaging neoplastic cell notch protein novel paralogous gene programs promoter receptor research study secretase stem tool transmission process tumor tumor growth

项目摘要

Notch signaling is an evolutionary conserved mechanism used by metazoans to direct cell fate decisions, proliferation and apoptosis at all stages of development, including self-renewing adult tissues. Aberrant Notch signaling is implicated in cancer, especially in the emerging field of cancer stem cells. Mammalian cells contain four Notch receptors and five Delta and Jagged cognate ligands; the overall goal of this proposal is to elucidate the significance of context on individual Notch receptor subtype-mediated signals using non-invasive molecular imaging strategies. We propose to develop enabling technologies that will facilitate high throughput screening for agents and gene products that are capable of modulating the Notch signaling pathway in cancer, inherited diseases and facilitate tissue engineering. A comprehensive mechanistic understanding of how to manipulate individual receptors in a context-dependent manner still eludes investigators. We propose to develop a real time imaging system that will go beyond the currently available reporters in providing real time, quantitative accounting of the activation status of individual Notch receptor subtypes. The reporter system we are developing is based on the luciferase complementation imaging technology developed in the ICMIC Molecular Reporter Core at Washington University School of Medicine and enables visualization of the interactions between a specific Notch intracellular domain (NICD) and the common nuclear cofactor RBPjk. The system is versatile; it can be adaptedfor studying the pathway in different cell types and can be easily modified to monitor interaction with other cancer-relevant partners such as components of the NF-kB pathway. While the imaging technology being developed in this application is not directly applicable to imaging in patients as it requires protein engineering, these methodologies will help develop and evaluate novel therapies for Notch-related diseases.

Notch信号传导是后生动物用来指导细胞命运的进化保守机制发育各个阶段的决定、增殖和凋亡，包括自我更新的成体组织。异常的Notch信号传导与癌症有关，尤其是在新兴的癌症干细胞领域。哺乳动物细胞含有四个Notch受体和五个Delta和Jagged同源配体；的总体目标该提案旨在阐明背景对个体Notch受体亚型介导的信号的重要性使用非侵入性分子成像策略。我们建议开发使能技术促进高通量筛选能够调节Notch的试剂和基因产物癌症、遗传性疾病和促进组织工程中的信号通路。全面的对如何以上下文相关的方式操纵个体受体的机械理解仍然存在躲避调查人员。我们建议开发一种实时成像系统，该系统将超越目前的水平可用报告器提供单个 Notch 激活状态的实时、定量统计受体亚型。我们正在开发的报告系统是基于荧光素酶互补华盛顿大学学院 ICMIC 分子报告核心开发的成像技术医学并可实现特定 Notch 细胞内结构域 (NICD) 之间相互作用的可视化和共同的核辅因子 RBPjk。该系统用途广泛；它可以适用于研究途径在不同的细胞类型中，可以轻松修改以监测与其他癌症相关伙伴的相互作用例如 NF-kB 通路的组成部分。虽然成像技术正在开发中应用程序并不直接适用于患者成像，因为它需要蛋白质工程，这些方法将有助于开发和评估Notch相关疾病的新疗法。