Enhancing the Understanding of Ferroptosis Mechanisms in Glioblastoma: An Integrated Approach Utilizing Single-Cell Sequencing and Mendelian Randomization Methods

Background : Ferroptosis, a novel form of regulated cell death, has emerged as a significant research focus due to its involvement in various cancers. This study combines single-cell RNA sequencing with Mendelian randomization (MR) methods to identify genetic factors that are associated with ferroptosis and explore their potential impact on the pathogenesis of glioblastoma (GBM). Methods : MR and multiple validation methods were used to identify ferroptosis-related genes in glioblastoma. Single-cell analysis was performed to evaluate the expression levels of ferroptosis markers across different glioblastoma cell types. Additionally, gene enrichment analysis was conducted to explore gene functions, while survival analysis examined the relationship between gene expression and patient prognosis. Immune cell infiltration analysis was also carried out for genes associated with prognosis. Results : MR and sensitivity analyses identified 9 ferroptosis-related genes that are associated with GBM: SIRT1, KDM3B, VCP, GPT2, PRDX6, CISD2, TP53, FLT3, and FANCD2. Co-localization analysis showed a significant association between the VCP gene and GBM. Single-cell analysis revealed that PRDX6 is highly expressed in tumor tissues. Gene enrichment analysis highlighted the biological processes such as cell metabolism, DNA repair, ubiquitination, and autophagy in the occurrence and progression of GBM. Immune infiltration and survival analyses suggest that CISD2 was related to CD8+ and CD4+ T cell infiltration, and may affect patient prognosis. Conclusions : This study combines MR analysis and single-cell analysis to reveal the crucial role of ferroptosis genes in GBM. These genes influence the occurrence and development of GBM by regulating processes such as metabolism, DNA repair, oxidative stress, and autophagy. Among them, PRDX6 plays an important role in microscopic research, while ubiquitination and autophagy are key drivers of GBM progression. The CISD2 gene may influence patient prognosis by regulating T cell infiltration, thereby promoting immune tolerance mechanisms in tumor cells.