CRISPR targeting of key enhancers as a new therapeutic strategy in glioblastoma
Most genetic variants that predispose to cancer are located in non-coding regions enriched in putative enhancers, whose systematic rearrangements have a high impact on gene expression in cancer. In this project, we study how reprogramming of regulatory regions and topological changes in 3D-chromatin organisation determine gene dysregulation in glioblastoma. Having identified such functional alterations in key enhancers, the goal is to target them by CRISPR/Cas9-based (epi)genome editing to modulate the expression of their target genes and thus affect the malignancy of the glioblastoma cells.
Glioblastoma is the most lethal and aggressive, but also the most common of all primary brain tumors, for which no cure is available and survival rates have not considerably improved in the last decades. Despite the number of susceptibility loci identified in glioblastoma, a functional understanding of how the non-coding regulatory genome contributes to the pathogenesis is lacking. Studying how reprogramming of regulatory regions and topological changes in 3D chromatin organization determine gene dysregulation in glioblastoma allows us to identify functionally relevant regulatory regions that contribute to tumor heterogeneity and invasiveness. The identified regulatory regions will be first screened by in vitro CRISPR-Cas9 targeting to select the candidates with highest impact on proliferation and invasiveness in human glioblastoma cells. Ultimately, CRISPR-based enhancer targeting will be performed in vivo in mouse models to validate the applicability of the method for therapy. The outcome of this study will provide functional support for the application of enhancer targeting as a new therapeutic strategy in glioblastoma.