Research > Focus A > Work Packages A02
Summary
In this project we will use a combination of high-throughput drug and shRNA screening in vitro and in orthotopic animal models to identify molecular mechanisms underlying response and resistance to epigenetic therapies. Our goal is the preclinical development of a histone H3 mutation- specific combination therapy for H3.3-mutant glioblastoma. The visual abstract below shows the aims and planned tasks of the work package.
Task 1
Epigenetic targeted drug profiling of patient-derived tumor samples
Steps / Workflow
- epigenetic compound screening of all H3 K27M, G34V/R and wt models;
- identification of top hit compounds
- analysis of molecular determinants of response
Task 2
Molecular mechanisms underlying K27M-specific response to epigenetic targeted therapy
Steps / Workflow
- RNA interference screen
- identification of over- and underrepresented shRNA targets
- CRISPR/Cas mediated knock-out of genes of interest in cell culture validation
Task 3
High-content compound combination screen
Steps / Workflow
- cell culture combination screen
- generation of dose response profiles and identification of top hit combinations
- cell culture validation
Task 4
In vivo validation of drug combinations and RNAi target genes in the zebrafish xenotransplant model
Steps / Workflow
- tumor cell inoculation
- medium-throughput hit validation (drug/drug and drug/target gene combinations)
- determination of drug-induced changes in tumor volume
Task 5
Validation of top hits/ combinations in preclinical murine PDX models
Steps / Workflow
- orthotopic tumor cell injection with bioluminescence/MRI imaging
- drug treatment & Kaplan-Meier-analysis
- histologic and molecular characterization
PRINCIPLE INVESTIGATORS
Jones, David, Dr.
Group Leader
Hopp Children’s Cancer Center at the NCT Heidelberg (KiTZ) & German Cancer Research Center (DKFZ), Pediatric Glioma Research Group, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany
WITT, OLAF, PROF. DR. MED.
Director Translational Program
Hopp Children’s Cancer Center at NCT Heidelberg (KiTZ) Department of Pediatric Oncology, Hematology, Immunology and Pulmonology, University Hospital Heidelberg, Im Neuenheimer Feld 430, 69120 Heidelberg, Germany