Guo Zichao, Feng Haoran, Jia Wenqing, Zhang Tianshuai, Chen Mengdi, Xu Zhuoqing, Zheng Fang, Huang Chenhao, Li Zhiliang, Liu Kun, Zhang Tao, Zhao Ren, Zhang Wei, Cheng Xi
Department of General Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Shanghai Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Int J Surg. 2025 Aug 1;111(8):5088-5104. doi: 10.1097/JS9.0000000000002568. Epub 2025 May 30.
Tertiary lymphoid structures (TLS) are emerging biomarkers for enhanced anti-tumoral immunity and treatment efficacy in colorectal cancer (CRC). This study aimed to develop a CRC-specific TLS signature and evaluate its predictive roles in survival prognosis and therapeutic responses to chemotherapy, and PD-1 blockade.
Transcriptomic data from TCGA database were analyzed to establish a TLS signature. Survival and treatment responses were validated in multiple cohorts, including TCGA cohort, RJ cohort (neoadjuvant FOLFOX, n = 16), CHcohort (neoadjuvant sintilimab, n = 17), and several GEO datasets. Genetic mutations, tumor microenvironment (TME) features, and its spatial interactions were assessed via maftools, Bayesprism, CIBERSORT, IBOR, TIMER 2.0, NicheNet analysis and multiplex immunofluorescence. Tumor-immune cells co-cultured miniPDX/PDO models were used to validate PD-1 blockade in MSS CRC.
In TCGA cohort, patients in TLS_high group exhibited significantly longer overall survival ( P = 0.008) and demonstrated a higher proportion of complete response/partial response (CR/PR) ( P = 0.017). Further analyses on TME revealed that tumor mutation burden was positively correlated with TLS signature scores ( R = 0.31, P < 0.001). High TLS scores depicted a higher proportion of CD8 + T cells, B cells, and dendritic cells, increased activation of anti-tumoral signaling pathways, enhanced cytokines secretion, and higher immune checkpoints expression, including CTLA4, LAG3, and TIM3. Sensitive group in RJ cohort and clinical CR group in CH cohort scored higher of TLS signature ( P < 0.05), and depicted activation of signaling pathways related to TLS formation. Mature TLS and enhanced intercellular receptor-ligand interactions were identified. miniPDX/PDO models confirmed TLS-driven PD-1 blockade sensitivity in MSS CRC.
The CRC-specific TLS signature identifies patients with favorable prognosis and enhanced responses to chemotherapy and PD-1 blockade, including MSS CRC resistant to immune checkpoint blockade. TLS abundance reflects an immunogenic TME, offering a biomarker for personalized treatment strategies.
三级淋巴结构（TLS）正成为结直肠癌（CRC）中增强抗肿瘤免疫和治疗效果的新兴生物标志物。本研究旨在建立一种CRC特异性的TLS特征，并评估其在生存预后以及对化疗和PD-1阻断治疗反应中的预测作用。
分析来自TCGA数据库的转录组数据以建立TLS特征。在多个队列中验证生存情况和治疗反应，包括TCGA队列、RJ队列（新辅助FOLFOX，n = 16）、CH队列（新辅助信迪利单抗，n = 17）以及几个GEO数据集。通过maftools、Bayesprism、CIBERSORT、IBOR、TIMER 2.0、NicheNet分析和多重免疫荧光评估基因突变、肿瘤微环境（TME）特征及其空间相互作用。使用肿瘤-免疫细胞共培养的miniPDX/PDO模型验证微卫星稳定（MSS）型CRC中PD-1阻断的效果。
在TCGA队列中，TLS高分组患者的总生存期显著更长（P = 0.008），且完全缓解/部分缓解（CR/PR）比例更高（P = 0.017）。对TME的进一步分析表明，肿瘤突变负荷与TLS特征评分呈正相关（R = 0.31，P < 0.001）。高TLS评分表明CD8 + T细胞、B细胞和树突状细胞比例更高，抗肿瘤信号通路的激活增加，细胞因子分泌增强，以及免疫检查点表达更高，包括CTLA4、LAG3和TIM3。RJ队列中的敏感组和CH队列中的临床CR组TLS特征评分更高（P < 0.05），并显示出与TLS形成相关的信号通路激活。鉴定出成熟的TLS和增强的细胞间受体-配体相互作用。miniPDX/PDO模型证实了TLS驱动的MSS型CRC对PD-1阻断的敏感性。
CRC特异性的TLS特征可识别出预后良好且对化疗和PD-1阻断反应增强的患者，包括对免疫检查点阻断耐药的MSS型CRC。TLS丰度反映了免疫原性TME，为个性化治疗策略提供了一种生物标志物。
