Immunotherapy for esophageal squamous cell carcinoma (ESCC) exhibits notable variability in efficacy. Concurrently, recent research emphasizes circRNAs' impact on the ESCC tumor microenvironment. To further explore the relationship, we leveraged circRNA, microRNA, and mRNA sequence datasets to construct a comprehensive immune-related circRNA–microRNA–mRNA network, revealing competing endogenous RNA (ceRNA) roles in ESCC. The network comprises 16 circular RNAs, 13 microRNAs, and 1,560 mRNAs. Weighted gene co-expression analysis identified immune-related modules, notably cancer-associated fibroblast (CAF) and myeloid-derived suppressor cell modules, correlating significantly with immune and stemness scores. Among them, the CAF module plays a crucial role in extracellular matrix function and effectively discriminates ESCC patients. Four hub collagen family genes within CAF correlated robustly with CAF, macrophage infiltration, and T-cell exclusion. In-house sequencing and RT-qPCR validated their elevated expression. We also identified CAF module-targeting drugs as potential ESCC treatments. In summary, we established an immune-related circRNA–miRNA–mRNA network that not only illuminates ceRNA functionality but also highlights circRNAs' involvement in the CAF through collagen gene targeting. These findings hold promise to predict ESCC immune landscapes and therapy responses, ultimately aiding in more personalized and effective clinical decision-making.

Xin Tang, Qian-Qian Meng, Ye Gao, Chu-Ting Yu, Yan-Rong Zhang, Yan Bian, Jin-Fang Xu, Lei Xin, Wei Wang, Han Lin, Luo-Wei Wang. Gut and Liver 2025;19:198-206. https://doi.org/10.5009/gnl240308

Section Background and Objective Over the past decade, transcription factors (TFs) have emerged as key players in various diseases. Immunotherapy has gained traction as an effective treatment, despite resistance to immune checkpoint inhibitors remains a challenge. The regulatory role of TFs in immunotherapy for esophageal squamous cell carcinoma (ESCC) is poorly understood. This study aims to explore the expression of key TFs, predictive significance, and tumor microenvironment (TME) cell infiltration in ESCC. AbstractSection Methods The expression profiles were obtained from Gene Expression Omnibus (GEO) databases, TF gene data were obtained from the Transcriptional Regulatory Relationships Unraveled by Sentence-based Text mining (TTRUST) database. A protein-protein interaction (PPI) network and enrichment analysis elucidated the molecular processes in ESCC. We investigated the TME cell infiltration landscape in a combined ESCC cohort. The relative abundance of cell infiltration was measured using a single-sample gene-set enrichment analysis (ssGSEA). The diagnostic value of hub TFs for ESCC was investigated using ROC analysis. AbstractSection Results This study found 48 differentially expressed TFs between normal and ESCC tissues and revealed a strong association between EZH2, FOS, KLF4, FOSB, TWIST1, KLF6 and CEBPB. The results suggested that upregulated EZH2, TWIST1, and KLF4 correlated with immunosuppressive cell infiltration (Tregs, MDSCs) and reduced PD-1/PD-L2 checkpoint expression. Downregulated FOSB and CEBPB associated with enhanced NK cell activity. The diagnostic potential of these key transcription factors in ESCC was evaluated through ROC analysis (EZH2, area under the curve [AUC] = 0.95; TWIST1, AUC = 0.85; CEBPB, AUC = 0.84; FOSB, AUC = 0.79; KLF4, AUC = 0.97; FOS, AUC = 0.84; KLF6, AUC = 0.84). Single-cell data confirmed TF expression in tumor cells, macrophages, and T-cell subsets, highlighting their role in TME remodeling. AbstractSection Conclusion The scRNA-seq analysis revealed the expression of hub TFs in different cell clusters. The study investigated the expression of DETFs associated with patient prognosis and TME cell infiltration in ESCC.

INTRODUCTION: Early detection of esophageal squamous cell carcinoma (ESCC) significantly improves patient quality of life and outcomes. We aimed to train and validate a predictive algorithm using methylated DNA markers (MDMs) based on encapsulated sponge cell collection device (CCD) samples. METHODS: CCD samples in the training set (n = 120) were taken from 2 medical centers, and those in the validation set (n = 70) were prospectively collected from another medical center. The case group contained a diverse range of esophageal high-grade lesions, including high-grade intraepithelial neoplasia (HGIN), early ESCC, and advanced ESCC. The algorithms were trained on the training set and tested to evaluate diagnostic performance in a prospective validation set. RESULTS: The MDM model was constructed using logistic regression with 2 MDMs (OTOP2 and KCNA3). The areas under the receiver operating characteristic curves were 0.933 (95% confidence interval [CI] 0.881–0.986) and 0.911 (95% CI 0.841–0.981) in the training and validation sets, respectively. The overall sensitivity was 90.0% at a specificity of 91.4% in the training set. The sensitivity was 90.0%, 95.0%, and 90.0% for HGIN, early ESCC, and advanced ESCC in the training set, respectively. The sensitivity and specificity were 87.5% and 86.7% in the validation set, respectively. The areas under the receiver operating characteristic curves for the complex model that included the 2 MDMs and age were 0.961 (95% CI, 0.920–1.000) and 0.940 (95% CI, 0.889–0.990) in the training and validation sets, respectively. DISCUSSION: The assay of MDMs in CCD samples offers a highly accurate method of predicting HGIN/ESCC, thereby providing the potential for early diagnosis and screening for HGIN/ESCC.

Recently, esophageal cancer has become more common in China. To find a molecular biomarker will provide a handy way to improve precancerous diagnosis and evaluate the state of lymph node metastasis, improving prognosis. The present study aimed to investigate the expression level of hsa-miR-6743-5p in 25 esophageal tissues and to estimate the correlation between clinicopathological features of esophageal squamous cell cancer with miR-6743-5p expression. Quantitative reverse transcription polymerase chain reaction was performed to examine the expression level of miR-6743-5p in 25 pairs of esophageal cancer tissues and adjacent non-cancerous tissues. The correlation between miR-6743-5p level and clinical characteristics was determined. The examined esophageal squamous cell cancer tissues exhibited no statistical difference on miR-6743-5p expression compared to the adjacent non-tumor tissues. miR-6743-5p was positively associated with lymph node metastasis. Downregulation of miR-6743-5p was found in the patients with lymph node metastasis while upregulation of miR-6743-5p was found in those without lymph node metastasis. Our study suggests that the expression of miR-6743-5p is different in different lymph node metastasis statuses. miR-6743-5p expression is downregulated in patients with lymph node metastasis in esophageal cancer.