Gliomas are the most common primary brain malignancies in adults, classified by aggressiveness into low-grade gliomas (LGG, grades II–III) and high-grade gliomas (HGG, grade IV). Glioblastoma (GBM), a grade IV IDH1 wild-type astrocytoma, is the most prevalent and aggressive form, with an incidence of 3.22 cases per 100,000 people. GBM’s high inter- and intra-tumoral heterogeneity contributes to its poor prognosis, with a median overall survival of 15 months and a 5-year survival rate of ~5%. Despite this, the standard of care (SOC)—maximal surgical resection, radiotherapy, and temozolomide (TMZ) chemotherapy—has seen little advancement. LGG research faces challenges due to low proliferation rates and tumor heterogeneity, limiting conventional preclinical models. In this context, patient derived in vitro models like organoids provide a promising platform for personalized medicine.

In this study, we generated patient-derived organoids from over 50 glioma patients, characterized them based on histology, microstructure, expression patterns, and growth, and evaluated their response to SOC therapies. Subsequently, RNA-seq data was used for de novo drug discovery via the DiSCoVER platform, with candidate compounds validated in vitro.

Organoid establishment was successful in most cases, with a higher success rate for GBM-derived organoids compared to LGG. Organoids proliferated and mimicked parental tissue in histology, microstructure, and expression patterns. However, biobanking proved unsatisfactory. Screening of common chemotherapeutic compounds revealed that SOC TMZ and other commonly used treatments had little effect on viability, whereas gefitinib showed a notable impact. Our de novo in silico approach identified alectinib, ruxolitinib, and dabrafenib as promising candidates, which were confirmed to significantly reduce viability, impair migration, and alter key molecular pathways.

In conclusion, while patient-derived glioma organoids present limitations in proliferative capacities and biobanking efficiency that must be addressed for clinical implementation, they remain an invaluable preclinical tool. As demonstrated in this work, the combination of organoids with bioinformatics tools like DiSCoVER establishes a potent platform for testing novel therapeutic strategies in the context of untreatable cancers such as gliomas.