Related Experiment Video
Updated: Jul 7, 2025
Laboratory-Engineered Glioblastoma Organoid Culture and Drug Screening
Published on: January 10, 2025
Patient-derived glioblastoma organoids reflect tumor heterogeneity and treatment sensitivity
Maikel Verduin1, Linde Hoosemans1, Maxime Vanmechelen2,3
1Department of Radiation Oncology (Maastro), GROW School for Oncology and Reproduction, Maastricht University Medical Centre, Maastricht, The Netherlands.
Background:
Treatment resistance and tumor relapse are the primary causes of mortality in glioblastoma (GBM), with intratumoral heterogeneity playing a significant role. Patient-derived cancer organoids have emerged as a promising model capable of recapitulating tumor heterogeneity. Our objective was to develop patient-derived GBM organoids (PGO) to investigate treatment response and resistance.
Methods:
GBM samples were used to generate PGOs and analyzed using whole-exome sequencing (WES) and single-cell karyotype sequencing. PGOs were subjected to temozolomide (TMZ) to assess viability. Bulk RNA sequencing was performed before and after TMZ.
Results:
WES analysis on individual PGOs cultured for 3 time points (1-3 months) showed a high inter-organoid correlation and retention of genetic variants (range 92.3%-97.7%). Most variants were retained in the PGO compared to the tumor (range 58%-90%) and exhibited similar copy number variations. Single-cell karyotype sequencing demonstrated preservation of genetic heterogeneity. Single-cell multiplex immunofluorescence showed maintenance of cellular states. TMZ treatment of PGOs showed a differential response, which largely corresponded with MGMT promoter methylation. Differentially expressed genes before and after TMZ revealed an upregulation of the JNK kinase pathway. Notably, the combination treatment of a JNK kinase inhibitor and TMZ demonstrated a synergistic effect.
Conclusions:
Overall, these findings demonstrate the robustness of PGOs in retaining the genetic and phenotypic heterogeneity in culture and the application of measuring clinically relevant drug responses. These data show that PGOs have the potential to be further developed into avatars for personalized adaptive treatment selection and actionable drug target discovery and as a platform to study GBM biology.
Insights
Patient-derived glioblastoma organoids (PGOs) accurately model tumor heterogeneity and drug responses. Targeting the JNK kinase pathway with temozolomide (TMZ) showed synergistic effects, offering new avenues for glioblastoma treatment.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Glioblastoma (GBM) mortality is driven by treatment resistance and tumor relapse, often due to intratumoral heterogeneity.
- Patient-derived cancer organoids are emerging as a valuable model for studying tumor heterogeneity.
- Developing patient-derived GBM organoids (PGOs) is crucial for investigating treatment response and resistance mechanisms.
Purpose of the Study:
- To develop and validate patient-derived GBM organoids (PGOs) as a model for glioblastoma research.
- To assess the ability of PGOs to recapitulate the genetic and phenotypic heterogeneity of GBM tumors.
- To investigate the response of PGOs to temozolomide (TMZ) treatment and identify potential therapeutic targets.
Main Methods:
- Generation and long-term culture of patient-derived GBM organoids (PGOs).
- Whole-exome sequencing (WES) and single-cell karyotype sequencing for genetic analysis.
- Bulk RNA sequencing and immunofluorescence to assess cellular states and drug response.
- Treatment of PGOs with temozolomide (TMZ) and combination therapy with a JNK kinase inhibitor.
Main Results:
- PGOs demonstrated high genetic variant retention (58%-90%) and preserved copy number variations compared to primary tumors.
- Single-cell sequencing confirmed the maintenance of genetic and cellular heterogeneity within PGOs.
- TMZ treatment elicited differential responses in PGOs, correlating with MGMT promoter methylation.
- Upregulation of the JNK kinase pathway was observed post-TMZ treatment, with synergistic effects noted for combination therapy.
Conclusions:
- PGOs are robust models that retain GBM's genetic and phenotypic heterogeneity.
- PGOs enable the measurement of clinically relevant drug responses.
- PGOs hold potential as avatars for personalized treatment selection, drug target discovery, and studying GBM biology.
