■肿瘤微环境的异质性显著影响肝细胞癌(HCC)患者的预后,通过配体-受体复合物的细胞通讯起着核心作用。
■我们使用CellChat对10个HCC组织进行了单细胞转录组学分析,以鉴定参与恶性HCC细胞通讯的配体-受体基因。利用来自TCGA肝癌(TCGA-LIHC)和肝癌的RNA-Seq数据-RIKEN,JP(LIRI-JP)同伙,我们采用Cox回归分析筛选预后相关基因.通过无监督聚类和差异基因表达分析构建预后风险模型。随后,建立了涉及肿瘤细胞和巨噬细胞的共培养系统。一系列的实验,包括Transwell分析,免疫荧光染色,免疫沉淀,流式细胞术,和免疫组织化学,进行以阐明HCC细胞通过CCL16-CCR1轴募集巨噬细胞的机制。
■单细胞分析揭示了恶性HCC细胞与巨噬细胞之间的显着相互作用,鉴定76个相关的配体受体基因。根据八种预后相关的配体受体基因的表达模式,将患者分为三种亚型。预后最差的亚型表现为T细胞相关免疫细胞浸润减少,免疫检查点基因的下调,和增加M2样肿瘤相关巨噬细胞评分。体外实验证实了CCL16-CCR1轴在肿瘤相关巨噬细胞的募集和M2极化中的关键作用。临床样本显示CCL16蛋白表达水平与晚期之间存在显著关联,淋巴结转移,和远处转移。免疫组织化学和免疫荧光染色进一步证实了CCL16与CCR1、CD68、CD206之间的相关性,以及CD68+CCR1+巨噬细胞浸润。
■我们的研究确定了分子亚型,预后模型,和基于配体-受体相互作用的恶性肝癌细胞通讯的免疫微环境特征。此外,我们揭示了HCC细胞通过CCL16-CCR1轴募集M2样肿瘤相关巨噬细胞的促瘤作用.
The heterogeneity of the tumor microenvironment significantly influences the prognosis of hepatocellular carcinoma (HCC) patients, with cell communication through ligand-receptor complexes playing a central role.
We conducted single-cell transcriptomic analysis on ten HCC tissues to identify ligand-receptor genes involved in malignant HCC cell communication using CellChat. Leveraging RNA-Seq data from the TCGA Liver Cancer (TCGA-LIHC) and Liver Cancer - RIKEN, JP (LIRI-JP) cohorts, we employed Cox regression analysis to screen for prognosis-related genes. Prognostic risk models were constructed through unsupervised clustering and differential gene expression analysis. Subsequently, a co-culture system involving tumor cells and macrophages was established. A series of experiments, including Transwell assays, immunofluorescence staining, immunoprecipitation, flow cytometry, and immunohistochemistry, were conducted to elucidate the mechanism through which HCC cells recruit macrophages via the
CCL16-CCR1 axis.
Single-cell analysis unveiled significant interactions between malignant HCC cells and macrophages, identifying 76 related ligand-receptor genes. Patients were classified into three subtypes based on the expression patterns of eight prognosis-related ligand-receptor genes. The subtype with the worst prognosis exhibited reduced infiltration of T cell-related immune cells, downregulation of immune checkpoint genes, and increased M2-like tumor-associated macrophage scores. In vitro experiments confirmed the pivotal role of the
CCL16-CCR1 axis in the recruitment and M2 polarization of tumor-associated macrophages. Clinical samples demonstrated a significant association between
CCL16 protein expression levels and advanced stage, lymph node metastasis, and distant metastasis. Immunohistochemistry and immunofluorescence staining further confirmed the correlation between
CCL16 and CCR1, CD68, and CD206, as well as CD68+CCR1+ macrophage infiltration.
Our study identified molecular subtypes, a prognostic model, and immune microenvironment features based on ligand-receptor interactions in malignant HCC cell communication. Moreover, we revealed the pro-tumorigenic role of HCC cells in recruiting M2-like tumor-associated macrophages through the
CCL16-CCR1 axis.