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.

Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Biology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Biology Faculty of Medicine Masaryk University Brno Czech Republic International Clinical Research Center St Anne's University Hospital Brno Czech Republic

Department of Experimental Biology Faculty of Sciences Masaryk University Brno Czech Republic

Department of Internal Medicine Hematology and Oncology Masaryk University Hospital Brno Czech Republic

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$a 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.

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$a Foldynova-Trantirkova, Silvie $u Central European Institute of Technology, Masaryk University, Brno, Czech Republic.

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$a Ghannamova, Barbora El $u Central European Institute of Technology, Masaryk University, Brno, Czech Republic.

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$a Fafilek, Bohumil $u Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.

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$a Varecha, Miroslav $u Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.

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$a Balek, Lukas $u Department of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.

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$a Mayer, Jiri $u Department of Internal Medicine, Hematology and Oncology, Masaryk University Hospital, Brno, Czech Republic.

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$a Krejci, Pavel $u Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.

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