1. Real-time sensing of MAPK signaling in medulloblastoma cells reveals cellular evasion mechanism counteracting dasatinib blockade of ERK activation during invasion
- Author
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Alexandre Gries, Michael A. Grotzer, Anuja Neve, Marc Thomas Schönholzer, Min Ma, Karthiga Santhana Kumar, Martin Baumgartner, Jessica Migliavacca, Elena Alvarez, University of Zurich, and Baumgartner, Martin
- Subjects
0301 basic medicine ,MAPK/ERK pathway ,Cancer Research ,ERK, Extracellular regulated kinase ,Basic fibroblast growth factor ,Dasatinib ,Apoptosis ,Cerebellum slice culture ,SHH, sonic hedgehog ,c-Met, cellular mesenchymal epithelial transition factor ,chemistry.chemical_compound ,0302 clinical medicine ,bFGF, basic fibroblast growth factor ,Epidermal growth factor ,Cell Movement ,hemic and lymphatic diseases ,Tumor Cells, Cultured ,1306 Cancer Research ,Kinase ,Cell migration ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Gene Expression Regulation, Neoplastic ,030220 oncology & carcinogenesis ,Hepatocyte growth factor ,Mitogen-Activated Protein Kinases ,medicine.drug ,Nuclear ERK1/2 activation sensor ,Original article ,FGFR, fibroblast growth factor receptor ,MAP Kinase Signaling System ,610 Medicine & health ,Antineoplastic Agents ,lcsh:RC254-282 ,03 medical and health sciences ,medicine ,Humans ,Neoplasm Invasiveness ,Protein kinase A ,Fluorescent biosensor ,Live cell imaging ,Cerebellar Neoplasms ,Cell Proliferation ,EGF, epidermal growth factor ,MB, medulloblastoma ,SKARS, Synthetic Kinase Activation Relocation Sensor ,030104 developmental biology ,WST, water soluble tetrazolium salt ,chemistry ,FDA, food and drug administration ,10036 Medical Clinic ,MAPK, mitogen-activated protein kinases ,Cancer research ,JNK, c-jun N-terminal kinase ,HGF, hepatocyte growth factor ,Medulloblastoma - Abstract
Highlights • Growth factor signaling causes sustained nuclear ERK1/2 activation. • The SCR and BCR/ABL inhibitor dasatinib blocks ERK1/2 and represses cell invasion. • EGF-stimulated cells may escape dasatinib inhibition of invasion through mesenchymal to amoeboid transition. • Combined inhibition of SRC and Rho-kinase signaling is necessary to completely block EGF-induced invasion., Aberrantly activated kinase signaling pathways drive invasion and dissemination in medulloblastoma (MB). A majority of tumor-promoting kinase signaling pathways feed into the mitogen-activated protein kinase (MAPK) extracellular regulated kinase (ERK1/2) pathway. The activation status of ERK1/2 during invasion of MB cells is not known and its implication in invasion control unclear. We established a synthetic kinase activation relocation sensor (SKARS) for the MAPK ERK1/2 pathway in MB cells for real-time measuring of drug response. We used 3D invasion assays and organotypic cerebellum slice culture to test drug effects in a physiologically relevant tissue environment. We found that hepatocyte growth factor (HGF), epidermal growth factor (EGF), or basic fibroblast growth factor (bFGF) caused rapid nuclear ERK1/2 activation in MB cells, which persisted for several hours. Concomitant treatment with the BCR/ABL kinase inhibitor dasatinib completely repressed nuclear ERK1/2 activity induced by HGF and EGF but not by bFGF. Increased nuclear ERK1/2 activity correlated positively with speed of invasion. Dasatinib blocked ERK-associated invasion in the majority of cells, but we also observed fast-invading cells with low ERK1/2 activity. These ERK1/2-low, fast-moving cells displayed a rounded morphology, while ERK-high fast-moving cells displayed a mesenchymal morphology. Dasatinib effectively blocked EGF-induced proliferation while it only moderately repressed tissue invasion, indicating that a subset of cells may evade invasion repression by dasatinib through non-mesenchymal motility. Thus, growth factor-induced nuclear activation of ERK1/2 is associated with mesenchymal motility and proliferation in MB cells and can be blocked with the BCR/ABL kinase inhibitor dasatinib.
- Published
- 2020