The ESR will work, in collaboration with ESR-8 (UCL) and ESR-6 (IRE), with biomarker analysis of brain tissue for biomarker discovery and validation of biomarkers in blood. The ESR will develop and apply assays based on our in situ next generation sequencing (NGS) technology, which allows highly multiplexed biomarker analysis at single cell resolution in sections of formalin fixed paraffin embedded (FFPE) tissues from gliomas. The technique can thus generate biomarkers based on both cellular and molecular profiling. In situ NGS is specific and sensitive enough to record also point mutations. We have previously shown that the technology works well for mRNA profiling in tissue, and that is one approach that will be applied. The ESR will develop the technology further to permit miRNA profiling and protein profiling in FFPE tissue sections. The biomarkers will guide development of assays for the blood-based sensors.