Large-scale CRISPRi screens link metabolic stress to glioblastoma chemoresistance
Xing Li1,2, Wansong Zhang1,2, Yitong Fang1,2
1School of Medicine, Southern University of Science and Technology, Shenzhen, 518055, Guangdong Province, China.
Background:
Glioblastoma (GBM) patients frequently develop resistance to temozolomide (TMZ), the standard chemotherapy. While targeting cancer metabolism shows promise, the relationship between metabolic perturbation and drug resistance remains poorly understood.
Methods:
We performed high-throughput CRISPR interference screens in GBM cells to identify genes modulating TMZ sensitivity. Findings were validated using multiple GBM cell lines, patient-derived glioma stem cells, and clinical data. Molecular mechanisms were investigated through transcriptome analysis, metabolic profiling, and functional assays.
Results:
We identified phosphoglycerate kinase 1 (PGK1) as a key determinant of TMZ sensitivity. Paradoxically, while PGK1 inhibition suppressed tumor growth, it enhanced TMZ resistance by inducing metabolic stress. This activated AMPK and HIF-1α pathways, leading to enhanced DNA damage repair through 53BP1. PGK1 expression levels correlated with TMZ sensitivity across multiple GBM models and patient samples.
Conclusions:
Our study reveals an unexpected link between metabolic stress and chemoresistance, demonstrating how metabolic adaptation can promote therapeutic resistance. These findings caution against single-agent metabolic targeting and suggest PGK1 as a potential biomarker for TMZ response in GBM.
Insights
Targeting cancer metabolism in glioblastoma (GBM) can paradoxically increase resistance to temozolomide (TMZ). Inhibiting phosphoglycerate kinase 1 (PGK1) boosts DNA repair, highlighting PGK1 as a potential biomarker for GBM treatment response.
Area of Science:
- Oncology
- Cancer Metabolism
- Molecular Biology
Background:
- Glioblastoma (GBM) frequently develops resistance to temozolomide (TMZ), the standard chemotherapy.
- The interplay between metabolic alterations and drug resistance in GBM is not well understood.
Purpose of the Study:
- To identify genes influencing TMZ sensitivity in GBM.
- To elucidate the molecular mechanisms underlying TMZ resistance.
- To explore the role of metabolic pathways in GBM chemoresistance.
Main Methods:
- High-throughput CRISPR interference screens in GBM cells.
- Validation in multiple GBM cell lines, patient-derived glioma stem cells, and clinical data.
- Transcriptome analysis, metabolic profiling, and functional assays to investigate molecular mechanisms.
Main Results:
- Phosphoglycerate kinase 1 (PGK1) was identified as a key determinant of TMZ sensitivity.
- PGK1 inhibition suppressed tumor growth but paradoxically enhanced TMZ resistance by inducing metabolic stress.
- Metabolic stress activated AMPK and HIF-1α pathways, promoting DNA damage repair via 53BP1, with PGK1 levels correlating with TMZ sensitivity.
Conclusions:
- Metabolic adaptation can promote therapeutic resistance in GBM, challenging single-agent metabolic targeting strategies.
- PGK1 plays a critical role in mediating TMZ resistance through metabolic reprogramming and enhanced DNA repair.
- PGK1 may serve as a predictive biomarker for TMZ response in glioblastoma patients.


