Abstract
Background
Glioblastoma multiforme (GBM) represents nearly 50% of all malignant brain tumours. Molecular and genomic diagnostics are beginning to unravel the variation between individual tumours. However, there is growing evidence that cellular heterogeneity exists within a single malignancy. Singe cell analysis has demonstrated the presence of subpopulations corresponding to distinct expression profiles.
Objective and experimental approach
characterisation of the intratumor heterogeneity is essential to understand biological behaviour and therapy response. Through combining a genetically labelled mouse model (rosa26-confetti lineage tracing locus) with genetically engineered GBM model we can label distinct cellular lineages.
Results
For three-dimensional imaging of these fluorescently labelled tumours we have optimised tissue clearing protocols. Fluorescence activated sorting of genetically labelled tumour cells identifies distinct populations within single tumours. With these techniques can now interrogate the spatial organisation of clones across large areas and we can compare distinct tumour lineages.
Outlook
Currently, we are engineering human glioblastoma cell lines with genetic fluorescent labels for lineage tracing. Several genetically characterised human cell lines are available for which novel therapeutic targets have been identified. We will apply our lineage tracing approach to investigate the clonal effects of these tailored therapeutics.