Related Experiment Video
Updated: May 13, 2025
Characterization of the Effects of Migrastatic Inhibitors on 3D Tumor Spheroid Invasion by High-resolution Confocal Microscopy
Published on: September 16, 2019
A rhabdomyosarcoma cell migration inhibitor screen using phase-contrast microscopy
Birgitte Bjørnerud1, Jørgen Wesche2, Ellen Margrethe Haugsten1
1Department of Tumor Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway; Centre for Cancer Cell Reprogramming, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway.
Abstract:
Rhabdomyosarcoma (RMS) is a malignant soft tissue tumour occurring in young children. Alterations in fibroblast growth factor receptor 4 (FGFR4) signalling pathways are linked to metastasis and poor prognosis in RMS. A critical trait of metastatic disease is the ability of cancer cells to migrate and invade nearby and distant tissues. To study cell migration, we have developed a computational tool that automatically tracks and measures the velocity of migrating cells in phase contrast images. This tool facilitated an inhibitor screen comprising 462 compounds, analysing nearly 2000 videos and tracking approximately 700 000 RMS-derived cells harbouring a constitutively active FGFR4 (RMS559). The highly selective inhibitor screen targeted various signalling pathways, including receptor tyrosine kinases (RTKs), mitogen-activated protein kinases (MAPK), and phosphoinositide 3-kinases (PI3K). Validation of screen hits revealed that inhibiting proteins in the MAPK and PI3K pathway decreased cell migration and, in some cases, cell viability. Inhibiting FGFR4 resulted in decreased RMS559 cell migration, while, surprisingly, inhibitors targeting Rho-associated protein kinase (ROCK) and focal adhesion kinase (FAK) resulted in increased RMS559 cell migration. Mechanistically, the inhibition of ROCK and FAK resulted in fewer focal adhesions, thereby facilitating increased cell velocity. These findings suggest that targeting specific signalling pathways can effectively modulate migration and survival of metastatic RMS559 cells.
Insights
This study developed a computational tool to screen inhibitors for rhabdomyosarcoma (RMS) cell migration. Targeting MAPK and PI3K pathways decreased migration, while ROCK and FAK inhibition surprisingly increased it by altering cell adhesion.
Area of Science:
- Oncology
- Cell Biology
- Bioinformatics
Background:
- Rhabdomyosarcoma (RMS) is a pediatric soft tissue cancer.
- Fibroblast growth factor receptor 4 (FGFR4) pathway alterations correlate with RMS metastasis and poor prognosis.
- Cancer cell migration and invasion are key features of metastatic disease.
Purpose of the Study:
- To develop a computational tool for analyzing cell migration velocity in phase contrast images.
- To conduct a high-throughput inhibitor screen targeting signaling pathways in RMS cells with active FGFR4.
- To identify novel therapeutic targets for modulating RMS cell migration and survival.
Main Methods:
- Development of an automated computational tool for tracking and measuring cell migration velocity.
- Implementation of a large-scale inhibitor screen (462 compounds) on approximately 700,000 RMS-derived cells (RMS559) with constitutively active FGFR4.
- Validation of inhibitor effects on cell migration and viability, focusing on Receptor Tyrosine Kinases (RTKs), Mitogen-Activated Protein Kinases (MAPK), and Phosphoinositide 3-Kinases (PI3K) pathways.
Main Results:
- Inhibitors of MAPK and PI3K pathways reduced RMS cell migration and, in some cases, cell viability.
- FGFR4 inhibition decreased RMS559 cell migration.
- Unexpectedly, Rho-associated protein kinase (ROCK) and focal adhesion kinase (FAK) inhibitors increased RMS559 cell migration by reducing focal adhesions.
Conclusions:
- Targeting specific signaling pathways can modulate the migration and survival of metastatic RMS cells.
- The developed computational tool is effective for large-scale inhibitor screening.
- ROCK and FAK pathways represent potential targets for modulating RMS cell migration, with complex effects on cell adhesion and velocity.
