title: Identification of phosphorylase kinase as a novel therapeutic target through high-throughput screening for anti-angiogenesis compounds in zebrafish.
abstract: Angiogenesis is essential for development and tumor progression. With the aim of identifying new compound inhibitors of the angiogenesis process, we used an established enhanced green fluorescent protein-transgenic zebrafish line to develop an automated assay that enables high-throughput screening of compound libraries in a whole-organism setting. Using this system, we have identified novel kinase inhibitor compounds that show anti-angiogenic properties in both zebrafish in-vivo system and in human endothelial cell in-vitro angiogenesis models. Furthermore, we have determined the kinase target of these compounds and have identified and validated a previously uncharacterized involvement of phosphorylase kinase subunit G1 (PhKG1) in angiogenesis in vivo. In addition, we have found that PhKG1 is upregulated in human tumor samples and that aberrations in gene copy number of PhK subunits are a common feature of human tumors. Our results provide a novel insight into the angiogenesis process, as well as identify new potential targets for anti-angiogenic therapies.
authors.affiliation: Center for Regenerative Medicine in Barcelona, Barcelona, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Camus
authors.forename: C
authors.initials: C
authors.lastname: Quevedo
authors.forename: S
authors.initials: S
authors.lastname: Menéndez
authors.forename: I
authors.initials: I
authors.lastname: Paramonov
authors.forename: P F W
authors.initials: PF
authors.lastname: Stouten
authors.forename: R A J
authors.initials: RA
authors.lastname: Janssen
authors.forename: S
authors.initials: S
authors.lastname: Rueb
authors.forename: S
authors.initials: S
authors.lastname: He
authors.forename: B E
authors.initials: BE
authors.lastname: Snaar-Jagalska
authors.forename: L
authors.initials: L
authors.lastname: Laricchia-Robbio
authors.forename: J C
authors.initials: JC
authors.lastname: Izpisua Belmonte
mesh_terms: D020533:Angiogenesis Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D042783:Endothelial Cells / Q000187:drug effects; D018628:Gene Dosage; D057166:High-Throughput Screening Assays; D006801:Humans; D058990:Molecular Targeted Therapy; D009369:Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D010764:Phosphorylase Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics; D015854:Up-Regulation; D015027:Zebrafish
chemical_list: D020533:Angiogenesis Inhibitors; D010764:Phosphorylase Kinase