title: One reporter for in-cell activity profiling of majority of protein kinase oncogenes.
abstract: In-cell profiling enables the evaluation of receptor tyrosine activity in a complex environment of regulatory networks that affect signal initiation, propagation and feedback. We used FGF-receptor signaling to identifyEGR1as a locus that strongly responds to the activation of a majority of the recognized protein kinase oncogenes, including 30 receptor tyrosine kinases and 154 of their disease-associated mutants. TheEGR1promoter was engineered to enhancetrans-activation capacity and optimized for simple screening assays with luciferase or fluorescent reporters. The efficacy of the developed, fully synthetic reporters was demonstrated by the identification of novel targets for two clinically used tyrosine kinase inhibitors, nilotinib and osimertinib. A universal reporter system for in-cell protein kinase profiling will facilitate repurposing of existing anti-cancer drugs and identification of novel inhibitors in high-throughput screening studies.
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Gudernova
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Silvie
authors.initials: S
authors.lastname: Foldynova-Trantirkova
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Barbora El
authors.initials: BE
authors.lastname: Ghannamova
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Bohumil
authors.initials: B
authors.lastname: Fafilek
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Varecha
authors.affiliation: Department of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.
authors.forename: Lukas
authors.initials: L
authors.lastname: Balek
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Hruba
authors.affiliation: Department of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Jonatova
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Jelinkova
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Kunova Bosakova
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Lukas
authors.initials: L
authors.lastname: Trantirek
authors.affiliation: Department of Internal Medicine, Hematology and Oncology, Masaryk University Hospital, Brno, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Mayer
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.|International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.
authors.forename: Pavel
authors.identifier: 0000-0003-0618-9134
authors.initials: P
authors.lastname: Krejci
mesh_terms: D000818:Animals; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods*; D006801:Humans; D000069416:Intravital Microscopy; D051379:Mice; D015513:Oncogene Proteins / Q000032:analysis*; D061848:Optical Imaging; D011494:Protein Kinases / Q000032:analysis*
chemical_list: D015513:Oncogene Proteins; D011494:Protein Kinases
keywords: activity; cancer biology; cell biology; human; in cell; mouse; profiling; protein kinase; receptor tyrosine kinase; reporter