Center for Clinical Pharmacy, Cancer Center, Department of Pharmacy, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Hangzhou, China; Key Laboratory of Endocrine Gland Diseases of Zhejiang Province, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Hangzhou, China.
International Medical Department, The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.
Biochim Biophys Acta Gen Subj. 2023 Sep;1867(9):130420. doi: 10.1016/j.bbagen.2023.130420. Epub 2023 Jul 9.
In this study, we integrated single-cell RNA sequencing (scRNA-seq) data to investigate cell heterogeneity and utilized MSigDB and CIBERSORTx to explore the pathways of major cell types and the relationships between different cell subtypes. Subsequently, we explored the correlation of cell subtypes with survival and used Gene Set Enrichment Analysis (GSEA) analyses to assess the pathways associated with the infiltration of specific cell subtypes. Finally, multiplex immunohistochemistry in tissue microarray cohort were performed to validate differences in protein level and their correlation with survival.
iCCA presented a unique immune ecosystem, with increased proportions of Epi (epithelial)-SPP1-2, Epi-S100P-1, Epi-DN (double negative for SPP1 and S100P expression)-1, Epi-DN-2, Epi-DP (double positive for SPP1 and S100P expression)-1, Plasma B-3, Plasma B-2, B-HSPA1A-1, B-HSPA1A-2 cells, and decreased proportions of B-MS4A1. High level of Epi-DN-2, Epi-SPP1-1, Epi-SPP1-2, B-MS4A1, and low level of Epi-DB-1, Epi-S100P-1, and Epi-S100P-2 was significantly associated with longer overall survival (OS), and high level of B-MS4A1_Low_Epi-DN-2_Low was associated with the shortest OS. Moreover, the results of MsigDB and GSEA suggest that bile acid metabolism is a crucial process in iCCA. Finally, we found that S100P+, SPP1+, SPP1 + S100P+, and MS4A1-SPP1 + S100P+ were highly expressed, whereas MS4A1 was lowly expressed in iCCA, and patients with high level of S100P+, SPP1 + S100P+, and MS4A1-SPP1 + S100P+ exhibited shorter survival.
We identified the cell heterogeneity of iCCA, found that iCCA is a unique immune ecosystem with many cell subtypes, and showed that the novel cell subtypes of SPP1 + S100P+ and MS4A1-SPP1 + S100P+ were key subpopulations in iCCA.
在这项研究中，我们整合了单细胞 RNA 测序 (scRNA-seq) 数据，以研究细胞异质性，并利用 MSigDB 和 CIBERSORTx 来探索主要细胞类型的途径以及不同细胞亚型之间的关系。随后，我们探索了细胞亚型与生存的相关性，并使用基因集富集分析 (GSEA) 分析来评估与特定细胞亚型浸润相关的途径。最后，在组织微阵列队列中进行了多重免疫组织化学验证，以验证蛋白水平的差异及其与生存的相关性。
iCCA 呈现出独特的免疫生态系统，Epi (上皮)-SPP1-2、Epi-S100P-1、Epi-DN (SPP1 和 S100P 表达均为阴性)-1、Epi-DN-2、Epi-DP (SPP1 和 S100P 表达均为阳性)-1、Plasma B-3、Plasma B-2、B-HSPA1A-1、B-HSPA1A-2 细胞的比例增加，而 B-MS4A1 的比例减少。Epi-DN-2、Epi-SPP1-1、Epi-SPP1-2、B-MS4A1 高水平和 Epi-DB-1、Epi-S100P-1、Epi-S100P-2 低水平与总生存期 (OS) 延长显著相关，而 B-MS4A1_Low_Epi-DN-2_Low 高水平与 OS 最短相关。此外，MsigDB 和 GSEA 的结果表明，胆汁酸代谢是 iCCA 中的一个关键过程。最后，我们发现 S100P+、SPP1+、SPP1+S100P+和 MS4A1-SPP1+S100P+在 iCCA 中高表达，而 MS4A1 低表达，高水平的 S100P+、SPP1+S100P+和 MS4A1-SPP1+S100P+患者的生存时间更短。
我们确定了 iCCA 的细胞异质性，发现 iCCA 是一个具有多种细胞亚型的独特免疫生态系统，并表明 SPP1+S100P+和 MS4A1-SPP1+S100P+等新型细胞亚型是 iCCA 的关键亚群。
