Studying the migration of FGFR3-TACC3 fusion-positive glioblastoma cells in a 3D collagen hydrogel matrix
Epitagama Liyana Arachchige, Navoda Navanjalee Dilhara (2025)
2025
Bioteknologian ja biolääketieteen tekniikan maisteriohjelma - Master's Programme in Biotechnology and Biomedical Engineering
Lääketieteen ja terveysteknologian tiedekunta - Faculty of Medicine and Health Technology
Hyväksymispäivämäärä
2025-07-28
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tuni-202507277812
https://urn.fi/URN:NBN:fi:tuni-202507277812
Tiivistelmä
Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer in adults. Its highly invasive nature makes treatment challenging. Among the genetic alterations associated with GBM, the FGFR3-TACC3 gene fusion has recently been identified as a recurrent oncogenic driver that promotes tumour progression. However, its precise functional role in tumour cell migration, particularly within physiologically relevant 3D environments, remains partially unresolved.
This thesis aimed to investigate the impact of FGFR3-TACC3 fusion on glioblastoma cell migration within a three-dimensional (3D) collagen hydrogel model under different culture conditions. SNB19-derived glioblastoma cell lines overexpressing FGFR3-TACC3, wild-type FGFR3, or control vectors were cultured as spheroids in ultra-low attachment (ULA) plates and embedded in a collagen hydrogel matrix to assess their migration behaviour. Over seven days, spheroid morphology, growth, and migration were monitored using bright-field imaging under a nutrient-rich condition and two distinct nutrient-deprived environments followed by stimulation with fibroblast growth factor (FGF).
The results showed that all SNB19-derived GBM cell lines formed viable spheroids and exhibited consistent migratory patterns within the 3D collagen matrix. FGFR3-TACC3 fusion-positive cells exhibited significantly enhanced migration compared to control and wild-type FGFR3 (WT-FGFR3) overexpressing cells, as revealed by increased radial dispersion.
In conclusion, the present study established and characterized a functional and physiologically relevant 3D in vitro model to study FGFR3-TACC3 fusion-driven invasion in glioblastoma. Further work may be required to optimise this 3D model, for instance, by incorporating patient-derived cells, co-culturing with immune or stromal cells, or integrating hypoxic gradients, to facilitate a deeper exploration of the molecular mechanisms governing the enhanced aggressiveness of FGFR3-TACC3 fusion-positive glioblastomas. Interestingly, this study paves the way for future research on targeted therapies to combat the invasive potential of FGFR3-TACC3 fusion-positive glioblastoma cells.
This thesis aimed to investigate the impact of FGFR3-TACC3 fusion on glioblastoma cell migration within a three-dimensional (3D) collagen hydrogel model under different culture conditions. SNB19-derived glioblastoma cell lines overexpressing FGFR3-TACC3, wild-type FGFR3, or control vectors were cultured as spheroids in ultra-low attachment (ULA) plates and embedded in a collagen hydrogel matrix to assess their migration behaviour. Over seven days, spheroid morphology, growth, and migration were monitored using bright-field imaging under a nutrient-rich condition and two distinct nutrient-deprived environments followed by stimulation with fibroblast growth factor (FGF).
The results showed that all SNB19-derived GBM cell lines formed viable spheroids and exhibited consistent migratory patterns within the 3D collagen matrix. FGFR3-TACC3 fusion-positive cells exhibited significantly enhanced migration compared to control and wild-type FGFR3 (WT-FGFR3) overexpressing cells, as revealed by increased radial dispersion.
In conclusion, the present study established and characterized a functional and physiologically relevant 3D in vitro model to study FGFR3-TACC3 fusion-driven invasion in glioblastoma. Further work may be required to optimise this 3D model, for instance, by incorporating patient-derived cells, co-culturing with immune or stromal cells, or integrating hypoxic gradients, to facilitate a deeper exploration of the molecular mechanisms governing the enhanced aggressiveness of FGFR3-TACC3 fusion-positive glioblastomas. Interestingly, this study paves the way for future research on targeted therapies to combat the invasive potential of FGFR3-TACC3 fusion-positive glioblastoma cells.