Master Student - Pediatric Leukemia Research

Project
Glycogen synthase kinase-3 (GSK3) has emerged as a critical regulator of multiple cellular processes and potential therapeutic target in various cancers, including acute myeloid leukemia (AML). There are two isoforms of GSK3: α and β, which cannot fully compensate each other. GSK3β plays a complex role in AML, as its inhibition activates the Wnt/β-catenin signaling pathway, a known hallmark of cancer. Previous GSK3 inhibitors showed non-specific inhibition of both α and β isoforms. Recent efforts have focused on designing inhibitors that selectively inactivate one of the two isoforms. Eraglusib, a specific early clinical trial GSK3β inhibitor, has shown promising results across various cancer subtypes. Our own analysis of AML patient data sets has revealed an elevated expression of GSK3β, and our preliminary results are expected to provide promising responses across a broad range of human leukemia cell lines and primary cell-derived murine leukemia models.
However, the molecular mechanisms underlying this regulation in AML remain insufficiently understood and potential combinatorial treatment regimens are missing. We thus aim to decipher post-transcriptional and post-translational mechanisms of GSK3β in pediatric AML. To this end, the project aims to elucidate the effects of GSK3β inhibition in human and murine AML cell lines using pharmacological inhibition with Eraglusib and genetic manipulation via CRISPR-mediated genome editing and RNA-interference. A multi-omics approach -including IP/MS and RNA-sequencing- will be performed to identify protein interactors and deregulated pathways associated with GSK3β inhibition.
Based on the anticipated goals, the project further aims to generate AML cell lines with endogenously fluorescence-tagging of GSK3β with CRISPR-mediated genome editing, followed by high-content CRISPR screening for modulating factors. With the already available CRISPR libraries targeting >1500 RNA-binding proteins, >800 factors of the ubiquitin system, and a drug-repurposing CRISPR library, we aim to probe for both mechanistic and therapeutically relevant targets modulating GSK3β in pediatric AML.
This comprehensive approach aims to uncover both mechanistic insights and therapeutically relevant targets modulating GSK3β in pediatric AML. This project offers an excellent opportunity to contribute to cutting-edge research in pediatric oncology and will be performed in close collaboration with clinicians within the GPOH and an Eraglusib clinical trial currently under preparation. The successful candidate will develop a strong foundation in translational cancer research and gain valuable insights into the development of targeted therapies for childhood leukemia.

Methods
Multi-omics approaches. CRISPR-Cas9-based generation of reporter cell lines. Genotyping and characterization of reporter cells. CRISPR-Cas high-throughput screening (HTS) for putative drug targets altering ARID3A expression. Arrayed HTS validation. Lentiviral cloning, packaging, transduction. Flow cytometry. Viability and cell cycle assays. Protein analysis.

Applications are open for motivated, talented and curious Master students (Molecular Medicine, Molecular Biology, Biotechnology or Biology). Potential candidates with a completed BSc degree in a relevant field and with skills in cell and molecular biology are very welcome to apply. Previous experience in cell culture and cloning is beneficial. We are looking for candidates who are highly self-motivated, creative and organized team players who will join our international and dynamic research team.

We offer an excellent supervision and exciting research in well-equipped research laboratories. Master projects (6-12 months) can start immediately or at any time point, upon mutual agreement.