title: Identification of RAS-mitogen-activated protein kinase signaling pathway modulators in an ERF1 redistribution screen.
abstract: The RAS-mitogen-activated protein kinase (MAPK) signaling pathway has a central role in regulating the proliferation and survival of both normal and tumor cells. This pathway has been 1 focus area for the development of anticancer drugs, resulting in several compounds, primarily kinase inhibitors, in clinical testing. The authors have undertaken a cell-based, high-throughput screen using a novel ERF1 Redistribution assay to identify compounds that modulate the signaling pathway. The hit compounds were subsequently tested for activity in a functional cell proliferation assay designed to selectively detect compounds inhibiting the proliferation of MAPK pathway-dependent cancer cells. The authors report the identification of 2 cell membrane-permeable compounds that exhibit activity in the ERF1 Redistribution assay and selectively inhibit proliferation of MAPK pathway-dependent malignant melanoma cells at similar potencies (IC(50)=< 5 microM). These compounds have drug-like structures and are negative in RAF, MEK, and ERK in vitro kinase assays. Drugs belonging to these compound classes may prove useful for treating cancers caused by excessive MAPK pathway signaling. The results also show that cell-based, high-content Redistribution screens can detect compounds with different modes of action and reveal novel targets in a pathway known to be disease relevant.
authors.affiliation: BioImage A/S, Copenhagen, Denmark.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Grånäs
authors.forename: Betina Kerstin
authors.initials: BK
authors.lastname: Lundholt
authors.forename: Frosty
authors.initials: F
authors.lastname: Loechel
authors.forename: Hans-Christian
authors.initials: HC
authors.lastname: Pedersen
authors.forename: Sara Petersen
authors.initials: SP
authors.lastname: Bjørn
authors.forename: Viggo
authors.initials: V
authors.lastname: Linde
authors.forename: Christian
authors.initials: C
authors.lastname: Krogh-Jensen
authors.forename: Eva-Maria Damsgaard
authors.initials: EM
authors.lastname: Nielsen
authors.forename: Morten
authors.initials: M
authors.lastname: Praestegaard
authors.forename: Søren Jensby
authors.initials: SJ
authors.lastname: Nielsen
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D020935:MAP Kinase Signaling System / Q000187:drug effects* / Q000235:genetics; D015394:Molecular Structure; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000235:genetics; D014162:Transfection
chemical_list: D004268:DNA-Binding Proteins; C107465:ERF protein, human; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D014157:Transcription Factors