Functional and Molecular Heterogeneity in Glioma Stem Cells Derived from Multiregional Sampling

Simple Summary Glioblastoma, the most common and malignant brain tumor, is driven by a population of stem and progenitor cells. We have recently demonstrated that identification of novel drug treatment options by screening glioblastoma stem cell cultures is feasible within a clinical timeframe. We have also identified significant interheterogeneity between glioblastoma stem cell cultures both on the molecular and functional level. Here, we present that glioblastoma stem cell cultures derived from spatially distinct regions representing one entire tumor mass share typical stem cell properties and global gene expression profiles but display considerable differences in somatic mutations and drug sensitivity profiles. However, the heterogeneity within regional glioblastoma stem cell cultures is less complex than what is found between tumors from different patients. Our results suggest that single, focal biopsies used to establish patient-derived GSC cultures may underestimate the total complexity of the glioblastoma stem cell population.Abstract Glioblastoma (GBM) is an aggressive and highly heterogeneous primary brain tumor. Glioma stem cells represent a subpopulation of tumor cells with stem cell traits that are presumed to be the cause of tumor relapse. There exists complex tumor heterogeneity in drug sensitivity patterns between glioma stem cell (GSC) cultures derived from different patients. Here, we describe that heterogeneity also exists between GSC cultures derived from multiple biopsies within a single tumor. From biopsies harvested within spatially distinct regions representing the entire tumor mass, we established seven GSC cultures and compared their stem cell properties, mutations, gene expression profiles, and drug sensitivity patterns against 115 different anticancer drugs. The results were compared to 14 GSC cultures derived from other patients. Between the multiregional-derived GSC cultures, we observed only minor differences in their phenotype, proliferative capacity, and global gene expression. Further, they displayed intratumoral heterogeneity in mutational profiles and sensitivity patterns to anticancer drugs. This heterogeneity, however, did not exceed the extensive heterogeneity found between GSC cultures derived from other GBM patients. Our results suggest that the use of GSC cultures from one single focal biopsy may underestimate the overall complexity of the GSC population and display the importance of including GSC cultures reflecting the entire tumor mass in drug screening strategies.