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
Jones, David, Dr.
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.
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
Address
Im Neuenheimer Feld 400
69120 Heidelberg
Themen
Research
- Focus A
- A01: Targeting tumor cell networks to overcome primary and adaptive resistance in glioblastoma
- A02: Development of a specific combination therapy for histone H3-mutant pediatric glioblastoma
- A03: Deciphering resistance against targeted treatments
- A04: Evolution of IDH mutant gliomas during malignant progression
- A05: Predictive biomarkers for MGMT-promoter-methylated glioblastoma
- A06: Resistance mechanisms of glioblastoma against alkylating agents and radiotherapy
Research
- Focus B
- B01: Mechanisms of response and resistance to checkpoint blockade in gliomas
- B02: DNA mis-match repair regulates immune checkpoint blockade therapy in glioblastoma
- B03: Targeting immunosuppressive programs in isocitrate dehydrogenase mutant gliomas
- B04: Impact of myeloid cells on the adaptive immune response in IDH1-mutant glioblastomas
- B05: Dissecting the response of glioblastoma and its tumor microenvironment to focused high-dose radiotherapy
Research
- Focus C
- C01: Comprehensive preclinical pharmacology testing of drugs used for glioblastoma treatment in children and adults
- C02: Radiomics, radiogenomics and deep-learning in neurooncology
- C03: Imaging immune signatures of glioma response and resistance towards immunotherapy
- C04: Identification and spatial mapping of metabolic resistance factors by MALDI mass spectrometry imaging
- C05: Overcoming glioma radio-resistance with particle therapy