Fully Automated Angiogenesis Drug Discovery Assay in Zebrafish

斑马鱼全自动血管生成药物发现测定

• 批准号: 8253232
• 负责人: ANTHONY A Ferrante
• 金额: $ 55.21万
• 依托单位: PHYSICAL SCIENCES, INC
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8253232
• 关键词: Affect; Algorithms; Angiogenesis Promoter; Animals; Automation; Biological Assay; Birth; Blood Vessels; Clinic; Clinical Management; Collection; Common Neoplasm; Computer software; Data; Defect; Deformity; Development; Disease; Dose; Embryo; Embryonic Development; Ensure; Environment; Evaluation; FDA approved; Fertilization; Fishes; Fluorescence Microscopy; Government; Green Fluorescent Proteins; Heart; Hemangioma; Hour; Image; Imagery; Inborn Genetic Diseases; Individual; Infant; Laboratories; Larva; Lead; Letters; Libraries; Life; Location; Macular degeneration; Malignant Neoplasms; Medical; Methods; Metric; Microscope; Modeling; Mutation; Patients; Pattern; Persons; Phase; Phenotype; Positioning Attribute; Preclinical Drug Evaluation; Probability; Process; Research; Software Tools; Testing; Therapeutic; Time; Vascular Endothelium; Wisconsin; Zebrafish; angiogenesis; assay development; base; computerized data processing; drug development; drug discovery; high throughput screening; human disease; image processing; improved; infancy; instrumentation; interest; malformation; medical schools; mutant; new growth; novel; physical science; programs; repaired; research study; small molecule; software development; tool

DESCRIPTION (provided by applicant): The zebrafish, Danio rerio, has become widely accepted as a model for human disease. Small molecule screens have been performed to identify modifiers of vascular patterning but this process has been performed at relatively low throughput since the tools for fully automated processing are not yet available. We propose to develop a novel, high-throughput screening assay for angiogenesis promoters. The assay would be based on the use of the SideView" microplate to collect clear images of the vascular pattern in animals that express GFP in their vasculature. During Phase I we: 1) developed novel methods to modulate angiogenesis promoters in the plexus; 2) developed software that locates the zebrafish, identifies the region of interest, and moves that region to the center of the field of view; and 3) developed automated, post-acquisition image processing software that distinguishes wild-type zebrafish embryos from xxx mutant embryos with deformities in the plexus. During Phase II the methods, software and instrumentation will be fully integrated and the assay will be validated in a model screen using the Prestwick Library of FDA- approved compounds. PUBLIC HEALTH RELEVANCE: The Phase I and Phase II programs will develop a fully automated tool for discovery of potential therapeutic compounds that affect developing vasculature, which is relevant to embryonic diseases such as vascular anomalies. In addition, compounds that promote or inhibit growth of new blood vessels as a treatment for numerous diseases including cancer and wet macular degeneration will also be discovered. The assay will be performed in zebrafish larvae, which will result in a higher probability that identified compounds, will proceed through the drug development process.

描述（由申请人提供）：斑马鱼（Danio rerio）已被广泛接受为人类疾病的模型。已经进行了小分子筛选来识别血管图案的修饰剂，但由于尚未提供全自动处理的工具，因此该过程的吞吐量相对较低。我们建议开发一种新型的、高通量的血管生成促进剂筛选方法。该测定将基于使用 SideView" 微孔板来收集在血管系统中表达 GFP 的动物的血管模式的清晰图像。在第一阶段，我们：1) 开发了调节神经丛中血管生成启动子的新方法；2) 开发了定位斑马鱼、识别感兴趣区域并将该区域移动到视野中心的软件；3) 开发了自动化的采集后 图像处理软件，可区分野生型斑马鱼胚胎和神经丛畸形的 xxx 突变胚胎。在第二阶段，方法、软件和仪器将完全集成，并且将使用 FDA 批准的化合物的 Prestwick 库在模型筛选中验证测定方法。 公共健康相关性：第一阶段和第二阶段计划将开发一种全自动工具，用于发现影响发育中脉管系统的潜在治疗化合物，这与血管异常等胚胎疾病相关。此外，还将发现促进或抑制新血管生长的化合物，以治疗包括癌症和湿性黄斑变性在内的多种疾病。该测定将在斑马鱼幼虫中进行，这将导致识别出的化合物更有可能进入药物开发过程。