肾透明细胞肾癌 (ccRCC) 是最常见的肾癌类型，其致癌机制尚未完全阐明。肿瘤异质性在肿瘤进展中起着关键作用，可以通过应用单细胞RNA测序(scRNA-seq)来较大程度上解析。我们从TCGA和Young等人的研究中获取了批量和单细胞RNA表达谱。使用UMAP、TSNE和Louvain聚类算法进行降维，使用FindAllMarkers函数确定差异表达基因(DEGs)。利用Monocle2进行伪时间序列分析，使用SCENIC分析每个细胞亚群的转录因子。利用一系列WB、CFA、CCK-8和EDU分析验证了MT2A在ccRCC致癌过程中的作用。我们观察到在肿瘤组织中有更高程度的T/NK和B细胞浸润，表明免疫细胞在ccRCC致癌中的作用。转录因子分析揭示了CD8+T细胞中EOMES和ETS1的激活，CAF被分为肌-CAF和间质型CAF，间质型CAF中ATF3、JUND、JUNB、EGR1和XBP1富集明显。通过细胞轨迹分析，我们确定了细胞进化的三个不同阶段，其中State2代表正常肾小管细胞向State1和State3转化，随着CNV得分的升高。功能富集分析表明在肿瘤细胞中干扰素γ和炎症反应途径得到增强。共识聚类算法得到了两个分子亚型，其中2号簇与晚期肿瘤阶段和较多的浸润免疫细胞相关。我们通过Cox和LASSO回归模型确定了17个预后相关基因，并用其构建了一个预后模型，其有效性在多个队列中得到验证。此外，我们还研究了其中一个关键基因MT2A在ccRCC致癌中的作用，发现它调节了恶性细胞的增殖和迁移。我们描绘了ccRCC的详细单细胞景观，特别关注CAF、内皮细胞和肾小管细胞。基于差异表达基因构建的预后模型具有高度稳定性和准确性。MT2A在ccRCC致癌中表现出活跃的参与，调控了恶性细胞的增殖和迁移。© 2023. 作者等人，独家许可给Springer-Verlag GmbH Germany，为Springer Nature的一部分。

Clear-cell renal cell carcinoma (ccRCC) appears as the most common type of kidney cancer, the carcinogenesis of which has not been fully elucidated. Tumor heterogeneity plays a crucial role in cancer progression, which could be largely deciphered by the implement of scRNA-seq. The bulk and single-cell RNA expression profile is obtained from TCGA and study conducted by Young et al. We utilized UMAP, TSNE, and clustering algorithm Louvain for dimensionality reduction and FindAllMarkers function for determining the DEGs. Monocle2 was utilized to perform pseudo-time series analysis. SCENIC was implemented for transcription factor analysis of each cell subgroup. A series of WB, CFA, CCK-8, and EDU analysis was utilized for the validation of the role of MT2A in ccRCC carcinogenesis. We observed higher infiltration of T/NK and B cells in tumorous tissues, indicating the role of immune cells in ccRCC carcinogenesis. Transcription factor analysis revealed the activation of EOMES and ETS1 in CD8 + T cells, while CAFs were divided into myo-CAFs and i-CAFs, with i-CAFs showing distinct enrichment of ATF3, JUND, JUNB, EGR1, and XBP1. Through cell trajectory analysis, we discerned three distinct stages of cellular evolution, where State2 symbolizes normal renal tubular cells that underwent transitions into State1 and State3 as the CNV score ascended. Functional enrichment examination revealed an amplification of interferon gamma and inflammatory response pathways within tumor cells. The consensus clustering algorithm yielded two molecular subtypes, with cluster 2 being associated with advanced tumor stages and an abundance of infiltrated immune cells. We identified 17 prognostic genes through Cox and LASSO regression models and used them to construct a prognostic model, the efficacy of which was verified in multiple cohorts. Furthermore, we investigated the role of MT2A, one of our hub genes, in ccRCC carcinogenesis, and found it to regulate proliferation and migration of malignant cells. We depicted a detailed single-cell landscape of ccRCC, with special focus on CAFs, endothelial cells, and renal tubular cells. A prognostic model of high stability and accuracy was constructed based on the DEGs. MT2A was found to be actively implicated in ccRCC carcinogenesis, regulating proliferation and migration of the malignant cells.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.