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Published on: October 4, 2019
Single-cell and spatial transcriptome analyses reveal MAZ(+) NPC-like clusters as key role contributing to glioma
Qiang Gu1, Kan Wang2, Tingming Lu1
1Clinical Oncology School of Fujian Medical University, Fujian Cancer Hospital, Fuzhou, China.
Background:
Temozolomide (TMZ) is an important chemotherapeutic agent for glioma treatment. However, the emergence of drug resistance impedes its application. Traditional population-level studies are limited in elucidating resistance mechanisms. Advances in single-cell and spatial transcriptomics technologies provide feasible resolution for studying the cellular composition and dynamics of tumors. In this study, we investigated the heterogeneity of gliomas associated with TMZ resistance at the single-cell and spatial transcriptome levels to identify the resistance mechanisms and potential therapeutic strategies.
Methods:
Single cell sequencing technology was utilized to identify the cellular clusters of gliomas. Drug perturbation analysis and cellular propensity analysis revealed key cluster responding to TMZ. Enrichment analysis was preformed to explore the function of clusters. Transcription factor activity analysis revealed key transcription factors contributing to tumor resistance. Spatial transcriptome data and bulk RNA-seq data validates the role of key transcription factors. Downstream targets of key transcription factors were predicted and validated using gene regulation assays. Drug sensitivity analyses were used to seek viable strategies to overcome drug resistance.
Results:
Glioma cells from before and after temozolomide treatment samples were classified into six clusters: NPC-like cluster, OPC-like cluster, MES-like cluster, AC-like cluster, OC, and Neuron. NPC-like clusters exhibited strong stemness and DNA repair capacity. The activity of MAZ in NPC-like cluster was significantly enhanced after TMZ treatment. The proportion of MAZ( +)_NPC-like cluster was higher in TMZ treated samples. Patients with high proportion of MAZ( +)_NPC-like cluster had poorer survival. Upregulation of MAZ is able to enhance drug resistance in glioma cells, but this phenomenon disappeared when FoxM1 expression was further silenced. The combination of paclitaxel and Trametinib is a promising strategy to overcome resistance.
Conclusions:
NPC-like cluster is prevalent in recurrent and drug-resistant gliomas. MAZ transcription factors are critical regulators that promote the development of drug resistance in NPC-like clusters by enhancing the capacity of DNA repair and stemness. Patients with high proportions of MAZ( +)_NPC-like clusters have poor TMZ sensitivity and prognosis. MAZ enhances stemness and drug resistance in glioma cells by upregulating FOXM1 expression. The combination of paclitaxel and paclitaxel is a promising therapeutic strategy for treating gliomas and overcoming drug resistance.
Insights
Temozolomide resistance in glioma is linked to NPC-like cells with high MAZ activity, which enhances DNA repair and stemness. Silencing MAZ or combining paclitaxel and Trametinib may overcome this resistance.
Area of Science:
- Neuro-oncology
- Cancer Genomics
- Molecular Biology
Background:
- Temozolomide (TMZ) is a key chemotherapy for glioma, but drug resistance limits its effectiveness.
- Single-cell and spatial transcriptomics offer new insights into tumor heterogeneity and resistance mechanisms.
Purpose of the Study:
- Investigate glioma heterogeneity in TMZ resistance using single-cell and spatial transcriptomics.
- Identify mechanisms of TMZ resistance and potential therapeutic strategies.
Main Methods:
- Single-cell sequencing to classify glioma cell clusters.
- Drug perturbation and cellular propensity analyses to identify TMZ-responsive clusters.
- Enrichment and transcription factor activity analyses to uncover resistance drivers.
- Spatial transcriptomics and bulk RNA-seq for validation.
- Gene regulation assays and drug sensitivity analyses for target validation and strategy development.
Main Results:
- Six glioma cell clusters identified: NPC-like, OPC-like, MES-like, AC-like, OC, and Neuron.
- NPC-like clusters showed high stemness and DNA repair capacity.
- MAZ transcription factor activity and proportion increased in NPC-like clusters post-TMZ treatment.
- High MAZ activity in NPC-like clusters correlated with poorer survival and enhanced TMZ resistance.
- MAZ-driven resistance was dependent on FOXM1 expression.
- Combination of paclitaxel and Trametinib showed promise against resistance.
Conclusions:
- NPC-like clusters are associated with recurrent and drug-resistant gliomas.
- MAZ transcription factors are critical regulators of TMZ resistance in NPC-like cells, boosting DNA repair and stemness.
- High MAZ activity in NPC-like clusters predicts poor TMZ sensitivity and prognosis.
- MAZ upregulates FOXM1 to enhance glioma stemness and drug resistance.
- Combined paclitaxel and Trametinib therapy is a potential strategy to overcome TMZ resistance.
