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Agavi Stavropoulou-Tatla

Modelling Human Glioblastoma Migration  for studying Cancer Metastasis

Glioblastoma multiforme (GBM) is the most malignant, aggressive and invasive form of central nervous system cancer. Despite improvements in chemotherapeutic agents, radiotherapy, micro-neurosurgical techniques and neuroimaging, the patients’ outcome remains dismal with a median survival of only about 14.6 months [1, 2]. The highly lethal nature of this tumour mainly originates from its highly infiltrative and dispersive nature [3].

Studying GBM cell migration is of particular interest because tumour angiogenesis and metastasis are strongly related to the deregulated motile behaviour of GBM cells [4]. Metastasis is undoubtedly the main cause of cancer mortality as 90% of solid tumours are due to metastatic spreading [5]. A significant limitation in the development of innovative anti-invasive treatments is the lack of powerful in vitro experimental models to predict GBM cell migration in the brain.

The aim of my PhD project is to develop a reliable GBM migration assay that would facilitate the understanding of the mechanisms that drive cell migration, as well as, serve as a prognostic tool for the initial screening of drug efficacy. The project is supervised by Dr Markaki and Dr Watts and benefits from the links between the Department of Engineering and the Department of Clinical Neurosciences, respectively.

Preliminary experiments were conducted using a cell exclusion zone assay. GBM cells were cultured around a barrier until confluency and then the barrier was removed and the cells were allowed to migrate.  An indicative time-sequence of phase contrast images showing the progression of the gap closure, at times t=0 (A), t=10 hours (B) and t=20 hours (C) is presented in Figure 1. From these images it is clear that after the removal of the barrier the cells become progressively motile on the direction perpendicular to the free edge. Some fingering is observed along the borders, formed by very active leading cells that drag other cells behind them as they progress on the surface.

 

Figure 1: Time-sequence of phase contrast images showing the progression of the gap closure, at times t=0 (A), t=10 hours (B) and t=20 hours (C).

This experimental set-up creates wound zones of reproducible sizes and sharp borders, it does not cause damage to cells by mechanical scraping or electric ablation, it maintains the surface chemistry and finally it is standardised. Future plans include extending the assay to 3D and possibly incorporating co-cultures of cancerous and healthy brain cells.

 

References

[1] Stupp, Roger, et al. "Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma." New England Journal of Medicine 352.10 (2005): 987-996.

[2] Rao, Shreyas S., et al. "Toward 3D biomimetic models to understand the behavior of glioblastoma multiforme cells." Tissue Engineering Part B: Reviews 20.4 (2013): 314-327.

[3] Foty, Ramsey A. "Tumor cohesion and glioblastoma cell dispersal." Future Oncology 9.8 (2013): 1121-1132

[4] Valster, Aline, et al. "Cell migration and invasion assays." Methods 37.2 (2005): 208-215.

[5] Kramer, Nina, et al. "In vitro cell migration and invasion assays." Mutation Research/Reviews in Mutation Research 752.1 (2013): 10-24.