BACKGROUND Esophageal squamous cell carcinoma (ESCC) is one of the most common upper gastrointestinal malignancies worldwide. Tertiary lymphoid structures (TLS) are tumor-infiltrating immune cells aggregates coupled with stromal cells which are similar to secondary lymphoid organs. The objective of this study is to explore the predictive effects of two common genes associated with TLS models on prognosis and immunotherapy effects in ESCC patients. METHODS Clinical information for ESCC patients in the TCGA(The Cancer Genome Altas) cohort and GSE 53625 were collected. All of the samples were classified as either high score group or low score group based on two TLS signatures, and the association between TLS signatures and survival, clinical indicators, genomic burden, stemness indices analysis, tumor microenvironment and immunotherapy response were performed. Furthermore, the mature TLS was also assessed in ESCC tissue microarray. RESULTS In our study, we quantified the score of TLS_9 and TLS_12, respectively, reflecting the different statuses of TLS (TLS_9 = B and T cells in TLSs; TLS_12 = neogenesis of TLSs). Subsequently, we explored the effect of TLS score on ESCC tumor microenvironment quantified by multiple algorithms. We found that a correlation analysis indicated that TLS_9 and TLS_12 were all positively correlated with CD8+ T cell, NK cells, CD4+ T cells, M1 macrophages and so on. Meanwhile, some cells present a different correlation pattern of TLS_9 and TLS_12, including activated CD4+ memory T cells and Tgd cells. Immune-related analysis revealed that the TLS_12 and TLS_9 scores were all positively correlated with immune dysfunction, yet negatively correlated with immune exclusion. Following this, the biological roles of TLS_9 and TLS_12 scores were investigated. Also, we noticed that the TLS score could significantly affect the CAFs infiltration and be associated with the genomic burden and tumor stemness. In addition, we explored the prognostic value of mature TLS through tissue microarray (TMA). Our result displayed ESCC patients with the presence of mature TLS had a better prognosis than ESCC patients without it. CONCLUSIONS Our study indicated that ESCC patients with the presence of TLS had better outcomes and an inflamed immune microenvironment. In addition, both TLS-9 and TLS-12 gene signatures could be used as potential biomarkers for the immunotherapy of ESCC patients.

中文翻译：

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食管癌患者三级淋巴结构与免疫微环境的综合分析

背景技术食管鳞状细胞癌（ESCC）是全世界最常见的上消化道恶性肿瘤之一。三级淋巴结构（TLS）是肿瘤浸润的免疫细胞聚集体与基质细胞的结合，类似于二级淋巴器官。本研究的目的是探讨与 TLS 模型相关的两个常见基因对 ESCC 患者预后和免疫治疗效果的预测作用。方法 收集 TCGA（癌症基因组 Altas）队列和 GSE 53625 中 ESCC 患者的临床信息。根据两个 TLS 签名将所有样本分为高分组或低分组，并进行 TLS 签名与生存、临床指标、基因组负荷、干性指数分析、肿瘤微环境和免疫治疗反应之间的关联。此外，还在 ESCC 组织微阵列中评估了成熟的 TLS。结果在我们的研究中，我们分别量化了 TLS_9 和 TLS_12 的得分，反映了 TLS 的不同状态（TLS_9 = TLS 中的 B 细胞和 T 细胞；TLS_12 = TLS 的新生）。随后，我们探讨了通过多种算法量化的 TLS 评分对 ESCC 肿瘤微环境的影响。相关性分析发现，TLS_9和TLS_12均与CD8+T细胞、NK细胞、CD4+T细胞、M1巨噬细胞等呈正相关。同时，一些细胞呈现出不同的TLS_9和TLS_12相关模式，包括激活的CD4+记忆T细胞和Tgd细胞。免疫相关分析显示，TLS_12和TLS_9评分均与免疫功能障碍呈正相关，与免疫排斥呈负相关。随后，研究了 TLS_9 和 TLS_12 分数的生物学作用。此外，我们注意到 TLS 评分可以显着影响 CAF 浸润，并与基因组负荷和肿瘤干性相关。此外，我们通过组织微阵列（TMA）探讨了成熟 TLS 的预后价值。我们的结果显示，存在成熟 TLS 的 ESCC 患者比不存在成熟 TLS 的 ESCC 患者预后更好。结论 我们的研究表明，存在 TLS 的 ESCC 患者具有更好的预后和发炎的免疫微环境。此外，TLS-9和TLS-12基因特征均可用作食管鳞癌患者免疫治疗的潜在生物标志物。