Project B – Exploring the physical properties of glioblastoma
This project aims to explore the adaptive response of glioblastoma (GBM) cells to variable stiffness with respect to alterations in their cytoskeleton (CSK), ECM and differentiation status, and in relationship with tumour aggressiveness. GBM will be used as one of the models to examine specific biophysical properties, and to develop and validate computer models.
Research in this project involves:
- Study of the composition and physical properties of the ECM in the available GBM models by solid-state NMR (in collaboration with project C) and indentation-type AFM while being cultured on mechano-controlable hydrogels in vitro. Measurement of spatial heterogeneity of stiffness in GBM and correlation with histology will serve as input/reference for modeling of tissue stiffness in project D.
- CSK adaptations to a variable stiff microenvironment will be studied in vitro by determining major actin- and tubulin microfilament components by biochemical assays and various microscopic techniques. The micrographs on the side illustrate that increasing stiffness of hydrogels induces a stretched-out phenotype, including long protrusions that resemble tumor microtubes.
- Similar as described in project A, the metabolic/glycolytic state will be investigated in relation to stiffness, thus allowing for further comparison between leukemic and glioblastoma cells.
- Exploration of the mechanotransduction pathways that underly the adaptive responses, with special attention to YAP/TAZ, using proteomic analyses and YAP-GFP reporters for visualization.