Integration of Single-Cell RNA and Bulk RNA Sequencing Reveals Cellular Heterogeneity and Identifies Survival-Associated Regulatory Networks in Glioblastoma

单细胞RNA和批量RNA测序的整合揭示了胶质母细胞瘤的细胞异质性并鉴定了与生存相关的调控网络

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Abstract

Glioblastoma is a highly aggressive and devastating brain malignancy with dismal prognosis and extremely limited therapeutic options. Identification of prognostic biomarkers and therapeutic targets from multi-omics data is critical for improving patient outcomes. In this study, we investigated the clinical significance of cellular heterogeneity and super-enhancer-driven regulatory networks, which are critically implicated in glioblastoma progression and treatment resistance. We first performed scRNA-seq to dissect tumour microenvironment heterogeneity, identifying 16 distinct cell clusters, including astrocytes, macrophages, and CD8+ T cells. CellChat analysis revealed key intercellular signalling pathways, with astrocytes and macrophages acting as central communication hubs. To integrate bulk RNA sequencing data, we applied the Scissor algorithm to identify survival-associated cell states. By combining single-cell and bulk transcriptomic data, we uncovered 642 survival-related genes, including QKI and RBM47, which robustly predicted patient survival and immunotherapy response. Furthermore, WGCNA analysis identified seven co-expression modules and super enhancer-regulated networks orchestrated by transcription factors (RFX2, RFX4) and hub genes (NEAT1, CFLAR). These networks stratified patients into high- and low-risk groups with significant survival differences. Collectively, our findings elucidate the intricate interplay between cellular heterogeneity and super enhancer-driven gene regulation in glioblastoma, providing a translational framework for targeting oncogenic hubs and modulating microenvironment interactions.

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