Enhancer/suppressor screens for angiogenic signaling

血管生成信号传导的增强子/抑制子筛选

基本信息

批准号：
7407545
负责人：
Joanne Chan
金额：
$ 26.28万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2010-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7407545
关键词：
Alkaline Phosphatase Alleles Angiogenesis Inhibitors Animals Antibodies Applications Grants Avastin Blood Circulation Blood Vessels Blood flow Bypass Cancer Patient Cardiovascular system Cell Communication Cell Surface Receptors Cells Chemicals Chromosome Mapping Clinical Trials Cloning Colorectal Cancer Communities Complement Complex Count Data Defect Development Dissection Drosophila genus Drug usage Embryo Enhancers EphB4 Receptor Family Fishes Funding Genes Genetic Genetic Models Genetic Screening Growth Human Knock-out Lead Length Life Life Expectancy Ligands Lipids Lower Organism Mammals Maps Methods Microscope Modeling Mus Mutagenesis Mutation Nature PTK787 Pathway interactions Pharmaceutical Preparations Pharmacologic Substance Phase III Clinical Trials Phenocopy Phenotype Phosphoric Monoester Hydrolases Phosphotransferases Play Process Protein Kinase Proteins Proto-Oncogene Proteins c-akt Rate Receptor Inhibition Receptor Protein-Tyrosine Kinases Receptor Signaling Regulatory Element Resources Role Score Screening procedure Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Molecule Specificity Staging Staining method Stains System Testing Thinking Time Tumor Angiogenesis Up-Regulation Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factors Work Zebrafish angiogenesis cancer cell cancer therapy cell type chemical genetics concept day fly functional loss gene function in vivo inhibitor/antagonist insight kinase inhibitor loss of function loss of function mutation man member mutant null mutation positional cloning receptor receptor function research study small molecule success temperature sensitive mutant therapeutic target tissue culture tumor

项目摘要

DESCRIPTION (provided by applicant): Angiogenesis is a rate-limiting step in the progression of human tumors. Key proteins involved in tumor vessel formation are good targets for therapy, since the forming vasculature is composed of genetically stable cells unlike the malignant cells in the tumor proper. The vascular endothelial growth factor (VEGF) receptors are thought to be master regulators of embryonic and tumor angiogenesis; thus, are currently targets for anti-angiogenic therapy. The transparency of the zebrafish embryo makes it an ideal model for angiogenic studies since blood flow can be easily observed in a living animal. In preliminary studies, a human VEGF receptor inhibitor, PTK787/ZK222584, was used to induce potent inhibition of embryonic vessels in the zebrafish. This chemcial genetic approach allowed an examination of the VEGF receptor signaling pathway as an upregulation of a downstream effector, AKT/PKB, can override the receptor block to provide a robust vessel rescue (Chan et al., 2002). In this grant proposal, chemical inhibition is combined with proven forward genetics to identify critical genes in angiogenic signaling as enhancers or suppressors of the anti-angiogenic phenotype. A pilot screen has been conducted using F2 heterozygous embryos under the influence of PTK787. Angiogenic defects have been confirmed genetically in F3 homozygous recessive embryos. Thus, a large scale screen will allow us to identify physiologically relevant players in the VEGF signaling pathway as additional targets for inhibitor therapy in cancer treatments. The application has two specific aims. Aim 1. To perform an enhancer screen for angiogenic mutations under drug-sensitized VEGF receptor function. Aim 2. To perform a suppressor screen for mutations that can override the drug-induced anti-angiogenics effects in zebrafish embryos.

描述（由申请人提供）：血管生成是人类肿瘤进展中的限速步骤。参与肿瘤血管形成的关键蛋白是治疗的良好靶标，因为形成的血管系统由遗传稳定的细胞组成，与肿瘤本身的恶性细胞不同。血管内皮生长因子 (VEGF) 受体被认为是胚胎和肿瘤血管生成的主要调节因子。因此，它们是目前抗血管生成治疗的目标。斑马鱼胚胎的透明度使其成为血管生成研究的理想模型，因为可以轻松观察活体动物的血流。在初步研究中，使用人类 VEGF 受体抑制剂 PTK787/ZK222584 来诱导斑马鱼胚胎血管的有效抑制。这种化学遗传学方法允许检查 VEGF 受体信号通路，因为下游效应器 AKT/PKB 的上调可以超越受体阻滞，从而提供强大的血管救援（Chan 等，2002）。在这项资助提案中，化学抑制与经过验证的正向遗传学相结合，以确定血管生成信号传导中的关键基因作为抗血管生成表型的增强子或抑制子。在 PTK787 的影响下，使用 F2 杂合胚胎进行了初步筛选。血管生成缺陷已在 F3 纯合隐性胚胎中得到遗传证实。因此，大规模筛选将使我们能够识别 VEGF 信号通路中生理相关的参与者，作为癌症治疗中抑制剂治疗的额外靶点。该应用程序有两个具体目标。目的 1. 在药物敏感的 VEGF 受体功能下进行血管生成突变的增强子筛选。目标 2. 对斑马鱼胚胎中可以克服药物诱导的抗血管生成作用的突变进行抑制筛选。