Related Experiment Video
Updated: Mar 22, 2026
A Protocol for Explant Cultures of IDH1-mutant Diffuse Low-grade Gliomas
Published on: May 9, 2025
IDH1 Mutation Increases the Sensitivity of Glioma Organoids to Radiotherapy and Targeted Therapy
Chenxuan Yang1,2, Meilin Li3, Dongdong Wu2
1Medical School of Chinese PLA, Beijing, 100853, China.
Introduction:
In gliomas, the Isocitrate Dehydrogenase (IDH1) mutation is a key molecular marker that can distinguish subtypes. IDH1-mutant gliomas grow more slowly, are sensitive to radiotherapy and chemotherapy, and have a better prognosis. However, there are still individual differences in treatment responses and prognosis among patients with this subtype, making traditional treatments difficult to adapt. Organoid models can simulate the characteristics of IDH1-mutant tumors and provide important support for research on their personalized treatment.
Materials And Methods:
Glioma tissues were dissociated, and cells were resuspended in Matrigel before being seeded into culture dishes for IDH1-mutant organoid culture. Organoids at different culture stages were subjected to morphological observation and viability confirmation via live-dead staining. The growth curves of two-dimensional (2D) and three-dimensional (3D) cells were measured by adding CellTiter-Glo (CTG). Hematoxylin and Eosin (H&E) staining and immunohistochemistry were performed on cultured organoids and original tumor tissue sections to validate organoid phenotypes, while immunofluorescence assays confirmed the IDH1 mutation status. Drug sensitivity tests were conducted using temozolomide and vorasidenib on IDH1-mutant and IDHwildtype glioma organoids. Relative luminescence Units (RLU) were measured via CTG at various time points. Organoids were irradiated with 8 Gy γ-rays at day 0 and day 4, and cell viability was measured.
Results:
Immunohistochemical (IHC) results of the constructed organoids were consistent with those of the original tumor tissues, and immunofluorescence confirmed the presence of IDH1 mutations in the organoids. No significant difference in sensitivity to temozolomide was observed between IDH1-mutant and wild-type organoids, whereas IDH1-mutant organoids showed significantly higher sensitivity to vorasidenib than IDH-wildtype organoids. After 4 days of γ-ray irradiation, the cell viability of IDH1-mutant glioma organoids decreased significantly, while no significant change was observed in IDH-wildtype organoids.
Discussion:
This study generated patient-derived IDH1-mutant glioma organoids, validated their value as an individualized precision drug-testing platform by comparing drug sensitivity and radiotherapy responses with IDH-wildtype organoids, and found that IDH1-mutant organoids were more sensitive to radiotherapy and vorasidenib, advancing glioma precision treatment.
Conclusion:
By constructing an IDH1-mutated glioma organoid model and integrating accurate molecular diagnostic information, it is expected to improve treatment efficacy and promote the development of personalized glioma medicine.
Insights
Patient-derived glioma organoids carrying the Isocitrate Dehydrogenase (IDH1) mutation show increased sensitivity to radiotherapy and the drug vorasidenib. This finding supports the development of personalized treatments for IDH1-mutant gliomas.
Area of Science:
- Oncology
- Genetics
- Biotechnology
Background:
- The Isocitrate Dehydrogenase (IDH1) mutation is a critical biomarker in gliomas, differentiating subtypes with distinct prognoses and treatment responses.
- IDH1-mutant gliomas exhibit slower growth and better sensitivity to standard therapies, yet individual variability necessitates personalized treatment approaches.
- Organoid models offer a promising platform for simulating IDH1-mutant tumors and advancing personalized medicine research.
Purpose of the Study:
- To develop and validate patient-derived IDH1-mutant glioma organoids as a model for personalized treatment strategies.
- To assess the drug sensitivity and radiotherapy response of IDH1-mutant glioma organoids compared to IDH-wildtype counterparts.
- To establish an individualized precision drug-testing platform for glioma research.
Main Methods:
- Dissociation of glioma tissues and culture of cells in Matrigel to form 3D organoids.
- Morphological observation, live-dead staining, and CellTiter-Glo assays for viability and growth curve analysis.
- Immunohistochemistry, immunofluorescence, and drug sensitivity testing with temozolomide and vorasidenib, alongside gamma-ray irradiation experiments.
Main Results:
- Constructed organoids accurately reflected the phenotypes of original tumor tissues, with confirmed IDH1 mutations.
- IDH1-mutant organoids demonstrated significantly higher sensitivity to vorasidenib and gamma-ray irradiation compared to IDH-wildtype organoids.
- No significant difference in temozolomide sensitivity was observed between IDH1-mutant and IDH-wildtype organoids.
Conclusions:
- Patient-derived IDH1-mutant glioma organoids serve as a valuable platform for individualized precision drug testing.
- The organoid model validates differential sensitivity to radiotherapy and vorasidenib in IDH1-mutant gliomas, advancing precision treatment.
- Integrating IDH1-mutated glioma organoid models with molecular diagnostics is key to improving treatment efficacy and personalized glioma medicine.
