Background: TheN6-methyladenosine (m6A) modification of eukaryotic mRNA is the most prevalent of such epigenetic modifications and has recently been identified as a potential player in the pathogenesis and progression of hepatocellular carcinoma (HCC). With the increasing emergence of immunotherapy in the treatment of HCC, we have evaluated the potential of m6A-related genes in predicting overall survival and the therapeutic efficacy of immunotherapy in HCC patients.Methods: We employed transcriptomic data from TCGA-LIHC and GSE76427, comprising a total of 485 HCC patients, as the training set. Based on 23 recognized m6A regulators, we performed clustering analysis on HCC patients. The intersecting differentially expressed genes (DEGs) among subtypes were used in least absolute shrinkage and selection operator (LASSO) Cox and multivariate Cox regression analyses to construct the risk model. For the quantification of a risk model of HCC patients, a risk score was developed and correlated with clinical and immunological parameters. Furthermore, a single-cell transcriptomic atlas was used to analyze the relationship between model genes and immune cell subpopulations. Mechanistic studies included in vitro assays to validate the association between the m6A-related geneANLNand the progression of HCC.Results: Internal (TCGA and GEO) and external validation (ICGC) suggested that an 8-gene risk score provides an accurate and stable prognostic assessment for HCC. Furthermore, the high-risk score, characterized by elevated TP53 mutation frequency, tumor mutation burden (TMB), and tumor stem cell characteristics indicated a poor prognosis. The prognostic signature was associated with immune cell infiltration in HCC. Those patients with a high-risk score had lower immune tolerance with a better prediction of the efficacy of immunotherapy. The risk model helps to assess and predict the response and prognosis of HCC patients to immune checkpoint inhibitors (ICIs). Additionally, single-cell RNA sequencing data revealed that the high-risk group had a higher proportion of T cells and fewer immunosuppressive T cells, potentially correlating with a better response to immunotherapy. Finally, in vitro experiments showed thatANLN, an m6A-related gene, promoted the proliferation and migration of HCC cells.Conclusions: In this study, we identified and validated an m6A gene signature consisting of eight genes that can be used to predict prognosis and immunotherapy efficacy in HCC patients.
背景：真核生物信使RNA的N6-甲基腺苷（m6A）修饰是此类表观遗传修饰中最普遍的一种，最近被确定为肝细胞癌（HCC）发病机制和进展中的潜在参与者。随着免疫疗法在HCC治疗中的应用日益增多，我们评估了m6A相关基因在预测HCC患者总生存期和免疫疗法疗效方面的潜力。 方法：我们采用来自TCGA-LIHC和GSE76427的转录组数据，共包含485名HCC患者，作为训练集。基于23个公认的m6A调控因子，我们对HCC患者进行了聚类分析。利用亚型间交叉的差异表达基因（DEGs），通过最小绝对收缩和选择算子（LASSO）Cox回归及多变量Cox回归分析构建风险模型。为量化HCC患者的风险模型，我们开发了风险评分，并将其与临床和免疫学参数相关联。此外，利用单细胞转录组图谱分析了模型基因与免疫细胞亚群之间的关系。机制研究包括体外实验，以验证m6A相关基因ANLN与HCC进展之间的关联。 结果：内部（TCGA和GEO）和外部验证（ICGC）表明，一个包含8个基因的风险评分能为HCC提供准确且稳定的预后评估。此外，高风险评分以TP53突变频率升高、肿瘤突变负荷（TMB）增加和肿瘤干细胞特征为特点，预示着不良预后。该预后特征与HCC中的免疫细胞浸润相关。高风险评分的患者免疫耐受性较低，对免疫疗法疗效的预测更佳。该风险模型有助于评估和预测HCC患者对免疫检查点抑制剂（ICIs）的反应和预后。此外，单细胞RNA测序数据显示，高风险组T细胞比例较高，免疫抑制性T细胞较少，这可能与对免疫疗法更好的反应相关。最后，体外实验表明，m6A相关基因ANLN促进了HCC细胞的增殖和迁移。 结论：在本研究中，我们鉴定并验证了一个由八个基因组成的m6A基因特征，可用于预测HCC患者的预后和免疫疗法疗效。
Stratification of Hepatocellular Carcinoma UsingN6-Methyladenosine
肿瘤(癌症)患者之家