Development of a computer-assisted high-throughput screening platform for anti-angiogenic testing.
Angiogenesis is widely accepted to play a central role in tumor growth and spreading. Therefore, anti-angiogenic agents appear as promising tools in cancer therapy, and great efforts have been devoted to their identification in recent years. Despite fast progress in this area, a quantitative and objective system for rapid anti-angiogenic testing is not currently available. In this work, we describe a new platform for the large-scale screening of agents with potential anti-angiogenic activity. This involved the miniaturization of an in vitro two-dimensional assay model of vascular morphogenesis to run in a high-density format (384- or 1536-well microplates) and the development of a specific digital image-analysis system for the automated and quantitative evaluation of biological activity. To test the efficacy and reproducibility of the platform, we used a wide variety of well-characterized angiogenesis inhibitors, including antibodies against components of the extracellular matrix, peptides, and chemicals. The system proved to be a reproducible, polyvalent, and powerful tool for massive screening of agents with potential application in cancer therapy. Ongoing developmental work, directed toward optimizing this system for use with a fully automated high-throughput screening platform, should accelerate research into the mechanisms of tumor angiogenesis and the development of drugs that target the essential tumor vasculature.
Sanz
Laura
L
Department of Immunology, Hospital Universitario Clínica Puerta de Hierro, Madrid, Spain.
Pascual
Mario
M
Muñoz
Adolfo
A
González
Miguel A
MA
Salvador
Carlos H
CH
Alvarez-Vallina
Luis
L
D001331:Automation; D002460:Cell Line; D003094:Collagen / Q000494:pharmacology; D003201:Computers; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D004730:Endothelium, Vascular / Q000166:cytology / Q000473:pathology*; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008833:Microcirculation / Q000166:cytology; D009389:Neovascularization, Pathologic; D011509:Proteoglycans / Q000494:pharmacology; D015203:Reproducibility of Results; D012984:Software
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen