Research > Focus C > Work Packages C06
Summary
In subproject A05E we profiled MGMT-promoter methylated glioblastomas, IDH-wt and identified unusual out-of-frame EGFR fusions and exon deletions in glioblastomas with long-term survival. Using mass-spectrometry based interactomics and phosphoproteomics we aim at elucidating the underlying mechanism. Candidate mechanisms will be validated in functional assays. We will evaluate the possibility to mimic the identified mechanism of signaling modification by specific drugs and test the effectiveness in pre-clinical model systems.
Task 1
Determination of the differential interactome / signalling of wildtype and c-terminally truncated/fused EGFR in glioblastoma cells under basal and stressed conditions
Steps / Workflow
- Establishing cell cultures expressing different EGFR structural variants
- Interaction profiling by mass spectrometry: Affinity purification-MS, cross-linking-MS, proximity-ligation-MS
- Monitoring intracellular localisation and trafficking
- Correlation with cellular fate
Task 2
In vitro validation of candidate mechanisms / interactions in functional experiments
Steps / Workflow
- Systematic perturbation of interactions by CRISPR-CAS9 mediated knock-out of interaction partners
- Identification of those interactions being critical for cellular phenotypes.
Task 3
In vivo validation of mechanisms and evaluation of a pharmacological intervention strategy
Steps / Workflow
- Evaluation of the possibility of mimicking the effects of EGFR structural variants on signalling by available drugs
- Confirming the identified mechanism to be of relevance for the treatment response in an in vivo setting
Task 4
Development of biomarker of GB response to RT
Steps / Workflow
- Integrative omics analysis of tumor evolution under RT
- Optimizing tumor delineation for high precision particle RT
- Longitudinal spatio-temporal tracking of tumor and normal tissue evolution with respect to particle RT dose distribution and quality (biophysical model)
PRINCIPLE INVESTIGATORS
REUSS, DAVID E., PD DR. MED.
Chief Consultant
University Hospital Heidelberg, Department of Neuropathology, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany
VON DEIMLING, ANDREAS, PROF. DR. MED.
Chair / Head of Department
University Hospital Heidelberg, Institute of Pathology, Department of Neuropathology & DKFZ, Clinical Cooperation Unit Neuropathology, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany