Department of Endoscopy Center and Endoscopy Research Institute, Zhongshan Hospital, Fudan University, Shanghai, China.
Department of Endoscopy, Shanghai Collaborative Innovation Center, Shanghai, China.
Clin Transl Med. 2023 Mar;13(3):e1203. doi: 10.1002/ctm2.1203.
The early diagnosis and intervention of oesophageal squamous cell carcinoma (ESCC) are particularly important because of the lack of effective therapies and poor prognosis. Comprehensive research on early ESCC at the single-cell level is rare due to the need for fresh and high-quality specimens obtained from ESD. This study aims to systematically describe the cellular atlas of human intramucosal ESCC.
Five paired samples of intramucosal ESCC, para-ESCC oesophageal tissues from endoscopically resected specimens and peripheral blood mononuclear cells were adopted for scRNA-seq analysis. Computational pipeline scMetabolism was applied to quantify the metabolic diversity of single cells.
A total of 164 715 cells were profiled. Epithelial cells exhibited high intra-tumoural heterogeneity and two evolutionary trajectories during ESCC tumorigenesis initiated from proliferative cells, and then through an intermediate state, to two different terminal states of normally differentiated epithelial cells or malignant cells, respectively. The abundance of CD8 T s, Tregs and PD1 CD4 T cells suggested an exhausted and suppressive immune microenvironment. Several genes in immune cells, such as CXCL13, CXCR5 and PADI4, were identified as new biomarkers for poor prognosis. A new subcluster of malignant cells associated with metastasis and angiogenesis that appeared at an early stage compared with progressive ESCC was also identified in this study. Intercellular interaction analysis based on ligand-receptor pairs revealed the subcluster of malignant cells interacting with CAFs via the MDK-NCL pathway, which was verified by cell proliferation assay and IHC. This indicates that the interaction may be an important hallmark in the early change of tumour microenvironment and serves as a sign of CAF activation to stimulate downstream pathways for facilitating tumour invasion.
This study demonstrates the changes of cell subsets and transcriptional levels in human intramucosal ESCC, which may provide unique insights into the development of novel biomarkers and potential intervention strategies.
由于缺乏有效的治疗方法和预后不良，食管鳞状细胞癌（ESCC）的早期诊断和干预尤为重要。由于需要从 ESD 获得新鲜且高质量的标本，因此在单细胞水平上对早期 ESCC 进行全面研究很少。本研究旨在系统描述人黏膜内 ESCC 的细胞图谱。
采用 5 对黏膜内 ESCC、内镜切除标本的癌旁食管组织和外周血单核细胞进行 scRNA-seq 分析。应用计算流程 scMetabolism 来量化单细胞的代谢多样性。
共分析了 164715 个细胞。上皮细胞表现出高度的肿瘤内异质性，在 ESCC 肿瘤发生过程中有两个进化轨迹，从增殖细胞开始，然后通过中间状态，分别向两种不同的终末状态发展，即正常分化的上皮细胞或恶性细胞。CD8 T 细胞、Tregs 和 PD1 CD4 T 细胞的丰度表明存在衰竭和抑制性的免疫微环境。在免疫细胞中，如 CXCL13、CXCR5 和 PADI4 等几个基因被确定为预后不良的新生物标志物。与进展性 ESCC 相比，在早期就出现了与转移和血管生成相关的恶性细胞的新亚群。本研究还发现了一种与转移和血管生成相关的恶性细胞亚群，与进展性 ESCC 相比，该亚群在早期出现。基于配体-受体对的细胞间相互作用分析表明，恶性细胞亚群通过 MDK-NCL 途径与 CAFs 相互作用，这通过细胞增殖测定和 IHC 得到验证。这表明这种相互作用可能是肿瘤微环境早期变化的一个重要特征，是 CAF 激活的标志，可刺激下游通路促进肿瘤侵袭。
本研究展示了人黏膜内 ESCC 中细胞亚群和转录水平的变化，这可能为开发新的生物标志物和潜在的干预策略提供独特的见解。
