Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Reicher, A.
Generation of the FGFR2-BICC1 gene fusion using CRISPR/Cas9 genome editing in immortalized hepatocytes.
[ Diplomarbeit/Master Thesis (UNI) ] University of Graz; 2018. pp.64.
- Autor*innen der Med Uni Graz:
- Reicher Andreas
- Betreuer*innen:
- Pichler Martin
- Altmetrics:
- Abstract:
- Cholangiocarcinoma is a rare cancer with a poor prognosis and limited treatment options. In recent years, gene fusions of the fibroblast growth factor receptor 2 (FGFR2) have been identified in 10-15% of intrahepatic cholangiocarcinomas. In these gene fusions, FGFR2 including its kinase domain is fused to other proteins that provide an oligomerization domain, leading to ligand-independent dimerization and activation of the receptor. Several FGFR2 inhibitors are currently evaluated in clinical trials, but despite the relatively late stage of clinical development, there are no cholangiocarcinoma cell lines available with an endogenous FGFR2 fusion. We therefore used CRISPR/Cas9 to induce the FGFR2-BICC1 gene fusion in immortalized hepatocytes at its endogenous locus on chromosome 10. We successfully established clonal cell lines harboring the FGFR2-BICC1 fusion and detected the expression of the fusion transcript. Interestingly, we could not show activation of oncogenic signaling or promotion of cellular growth in our cells lines. This might mean that other factors such as additional mutations, environmental cues or certain expression levels might be needed for the fusion to have oncogenic effects in the cell lines we have established. Furthermore, we have shown that the plasmid we have created can induce the FGFR2-BICC1 fusion in additional cell lines. Those cell lines will be valuable model systems to study FGFR2 fusions because current models using overexpression of FGFR2 fusions do not recapitulate endogenous expression levels and it is not possible to study the transcriptional regulation of the gene fusion or to study the oncogenic potential of the gene fusion in its natural genomic context.