BackgroundCigarette smoking is considered as a major risk factor for esophageal carcinoma (ESCA) patients. Neutrophil activation plays a key role in cancer development and progression. However, the relationship between cigarette smoking and neutrophils in ESCA patients remained unclear.MethodsSingle‐cell RNA sequencing (scRNA‐seq) and bulk RNA sequencing data were obtained from public databases. Uniform manifold approximation and projection (UMAP) was used to perform downscaling and clustering based on scRNA‐seq data. The module genes associated with smoking in ESCA patients were filtered by weighted gene co‐expression network analysis (WGCNA). Using the “AUCell” package, the enrichment of different cell subpopulations and gene collections were assessed. “CellChat” and “CellphoneDB” were used to infer the probability and significance of ligand–receptor interactions between different cell subpopulations.ResultsWGCNA was performed to screened module genes associated with smoking in ESCA patients from MEdarkquosie, MEturquoise, and MEgreenyellow. Next, eight cell clusters were identified, and using the AUCell score, we determined that neutrophil clusters were more active in the gene modules associated with smoking in ESCA patients. Two neutrophil subtypes, Neutrophils 1 and Neutrophils 2, exhibited greater enrichment in inflammatory response regulation, intercellular adhesion, and regulation of T cell activation. Furthermore, we found that neutrophils may pass through AMPT‐(ITGA5 + ITGB1) and ICAM1‐AREG in order to promote the development of ESCA, and that the expression levels of the receptor genes insulin‐degrading enzyme and ITGB1 were significantly and positively correlated with cigarette smoking per day.ConclusionCombining smoking‐related gene modules and scRNA‐seq, the current findings revealed the heterogeneity of neutrophils in ESCA and a tumor‐promoting role of neutrophils in the tumor microenvironment of smoking ESCA patients.

中文翻译：

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使用单细胞 RNA 测序和批量 RNA 测序数据揭示食管癌患者吸烟与中性粒细胞活化之间的相关性

背景吸烟被认为是食管癌（ESCA）患者的主要危险因素。中性粒细胞激活在癌症的发生和进展中起着关键作用。然而，吸烟与 ESCA 患者中性粒细胞之间的关系仍不清楚。方法单细胞 RNA 测序 (scRNA-seq) 和批量 RNA 测序数据来自公共数据库。使用均匀流形逼近和投影（UMAP）基于 scRNA-seq 数据进行降尺度和聚类。通过加权基因共表达网络分析（WGCNA）筛选与 ESCA 患者吸烟相关的模块基因。使用“AUCell”包，评估了不同细胞亚群和基因集合的富集。使用“CellChat”和“CellphoneDB”来推断不同细胞亚群之间配体-受体相互作用的概率和显着性。结果通过WGCNA从MEdarkquosie、MEturquoise和MEgreenyellow中筛选出ESCA患者中与吸烟相关的模块基因。接下来，鉴定了八个细胞簇，并使用 AUCell 评分，我们确定中性粒细胞簇在与 ESCA 患者吸烟相关的基因模块中更加活跃。两种中性粒细胞亚型，中性粒细胞1和中性粒细胞2，在炎症反应调节、细胞间粘附和 T 细胞活化调节方面表现出更大的丰富性。此外，我们发现中性粒细胞可以通过AMPT‐(ITGA5+国贸GB1） 和细胞间黏附分子1‐阿雷格为了促进ESCA的发展，研究受体基因胰岛素降解酶和胰岛素降解酶的表达水平国贸GB1与每天吸烟呈显着正相关。结论结合吸烟相关基因模块和scRNA-seq，目前的研究结果揭示了ESCA中中性粒细胞的异质性以及中性粒细胞在吸烟ESCA患者肿瘤微环境中的促癌作用。