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. Specifically, during Phase I we propose to: 1) develop novel methods to modulate angiogenesis promoters in the subintestinal veins;2) develop software that will locate the zebrafish, identify the region of interest, move that region to the center of the field of view, zoom in and refocus on the SIVs, and collect the image;and 3) develop automated, post-acquisition image processing software that will identify animals in which the extent of budding from the SIV is either enhanced or reduced. Methods for automated handling of the animals will also be developed during Phase I. During Phase II the methods, software and instrumentation will be fully integrated and a high-throughput screen for angiogenic compounds will be performed.
The Phase I and Phase II programs will develop a fully automated tool for discovery of potential therapeutic compounds that either: promote growth of new blood vessels to prevent or reverse damage to heart tissue;or to inhibit growth of new blood vessels as a treatment for numerous diseases including cancer and wet macular degeneration. 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.
