title: High-throughput screening system to identify small molecules that induce internalization and degradation of HER2.
abstract: Overexpression of growth factor receptors in cancers, e.g., human epidermal growth factor receptor 2 (HER2) in ovarian and breast cancers, is associated with aggressiveness. A possible strategy to treat cancers that overexpress those receptors is blockade of receptor signaling by inducing receptor internalization and degradation. In this study, we developed a cell-based high-throughput screening (HTS) system to identify small molecules that induce HER2 internalization by employing our recently developed acidic-pH-activatable probe in combination with protein labeling technology. Our HTS system enabled facile and reliable quantification of HER2 internalization with a Z' factor of 0.66 and a signal-to-noise ratio of 44.6. As proof of concept, we used the system to screen a ∼155,000 small-molecule library and identified three hits that induced HER2 internalization and degradation via at least two distinct mechanisms. This HTS platform should be adaptable to other disease-related receptors in addition to HER2.
authors.affiliation: Department of Neurobiology, Graduate School of Medicine and ‡Open Innovation Center for Drug Discovery, The University of Tokyo , Hongo, Tokyo 113-0033, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Isa
authors.forename: Daisuke
authors.initials: D
authors.lastname: Asanuma
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Namiki
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kumagai
authors.forename: Hirotatsu
authors.initials: H
authors.lastname: Kojima
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Okabe
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nagano
authors.forename: Kenzo
authors.initials: K
authors.lastname: Hirose
mesh_terms: D005260:Female; D057166:High-Throughput Screening Assays; D006801:Humans; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D059748:Proteolysis / Q000187:drug effects*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D018719:Erb-b2 Receptor Tyrosine Kinases; D054852:Small Molecule Libraries; C508053:ERBB2 protein, human