DataSheet1_Development and validation of a pyroptosis-related genes signature for risk stratification in gliomas.docx

Background: Glioma is a highly heterogeneous disease, causing the prognostic prediction a challenge. Pyroptosis, a programmed cell death mediated by gasdermin (GSDM), is characterized by cell swelling and the release of inflammatory factors. Pyroptosis occurs in several types of tumor cells, including gliomas. However, the value of pyroptosis-related genes (PRGs) in the prognosis of glioma remains to be further clarified.Methods: In this study, mRNA expression profiles and clinical data of glioma patients were acquired from TCGA and CGGA databases, and one hundred and eighteen PRGs were obtained from the Molecular Signatures Database and GeneCards. Then, consensus clustering analysis was performed to cluster glioma patients. The least absolute shrinkage and selection operator (LASSO) Cox regression model was used to establish a polygenic signature. Functional verification of the pyroptosis-related gene GSDMD was achieved by gene knockdown and western blotting. Moreover, the immune infiltration status between two different risk groups were analyzed through the “gsva” R package.Results: Our results demonstrated that the majority of PRGs (82.2%) were differentially expressed between lower-grade gliomas (LGG) and glioblastoma (GBM) in the TCGA cohort. In univariate Cox regression analysis, eighty-three PRGs were shown to be associated with overall survival (OS). A five-gene signature was constructed to divide patients into two risk groups. Compared with patients in the low-risk group, patients in the high-risk group had obviously shorter OS (p < 0.001). Also, we found that the high-risk group showed a higher infiltrating score of immune cells and immune-related functions. Risk score was an independent predictor of OS (HR > 1, p < 0.001). Furthermore, knockdown of GSDMD decreased the expression of IL-1β and cleaved caspase-1.Conclusion: Our study constructed a new PRGs signature, which can be used to predict the prognosis of glioma patients. Targeting pyroptosis might serve as a potential therapeutic strategy for glioma.

背景：胶质瘤是一种高度异质性疾病，其预后预测极具挑战性。由气体衍生物（GSDM）介导的细胞焦亡是一种程序性细胞死亡，其特征为细胞肿胀和炎症因子的释放。细胞焦亡发生在多种肿瘤细胞中，包括胶质瘤。然而，细胞焦亡相关基因（PRGs）在胶质瘤预后中的价值尚需进一步明确。方法：在本研究中，我们从TCGA和CGGA数据库中获取了胶质瘤患者的mRNA表达谱和临床数据，并从分子签名数据库和GeneCards中获得了118个PRGs。随后，通过一致性聚类分析对胶质瘤患者进行分组。使用最小绝对收缩和选择算子（LASSO）Cox回归模型建立多基因签名。通过基因敲低和蛋白质印迹技术验证了细胞焦亡相关基因GSDMD的功能。此外，通过“gsva”R包分析了两个不同风险组之间的免疫浸润状态。结果：我们的结果表明，在TCGA队列中，大多数PRGs（82.2%）在低级别胶质瘤（LGG）和胶质母细胞瘤（GBM）之间存在差异表达。在单变量Cox回归分析中，83个PRGs与总生存期（OS）相关。构建了一个包含五个基因的签名，将患者分为两个风险组。与低风险组患者相比，高风险组患者的总生存期明显缩短（p < 0.001）。此外，我们还发现高风险组的免疫细胞浸润评分和免疫相关功能较高。风险评分是OS的独立预测因子（HR > 1，p < 0.001）。此外，GSDMD的敲低降低了IL-1β和裂解的caspase-1的表达。结论：本研究构建了一个新的PRGs签名，可用于预测胶质瘤患者的预后。针对细胞焦亡可能成为胶质瘤的一种潜在治疗策略。