Chung Shin-Yi, Yeh Yi-Chen, Huang Chien-Jung, Chiang Nai-Jung, Hsu Dennis Shin-Shian, Chan Ming-Hsien, Lu Meng-Lun, Hsu Tzu-Sheng, Hung Yi-Ping, Yeh Chun-Nan, Hsiao Michael, Chang Yu-Chan, Wang Yu-Chao, Chen Ming-Huang
Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan.
Institute of Biomedical Informatics, National Yang Ming Chiao Tung University, Taipei, Taiwan.
J Immunother Cancer. 2025 Jan 27;13(1):e010173. doi: 10.1136/jitc-2024-010173.
Cholangiocarcinoma is a challenging malignancy with limited responses to conventional therapies, particularly immune checkpoint inhibitor therapy. Tumor-infiltrating lymphocytes (TILs) and tertiary lymphoid structures (TLSs) are key components of the tumor microenvironment (TME) and have been implicated in the immune response to cancer. However, the role and difference of TLSs and TILs in patients with cholangiocarcinoma remains unclear. This study elucidates their contributions to the TME.
We examined 16 tumor samples from a single-arm, phase II trial of nivolumab plus modified gemcitabine and S-1 and various datasets. Immunohistochemistry and RNA sequencing were employed to assess TLSs and TILs presence and activity. Differential gene expression and signature of immune cell composition were examined by GeoMx Digital Spatial Profiler and Cancer Transcriptome Altas analysis.
TLS-positive (N=7) patients demonstrated significantly better immunotherapy outcomes compared with TLS-negative (N=9) patients, including higher objective response rates (71% vs 0%) and disease control rates (100% vs 67%). The presence of TLSs correlated with improved progression-free and overall survival (p=0.03). TLSs were associated with "inflamed" tumors characterized by substantial immune infiltration, particularly involving T and B cells. Gene expression analyses identified significant upregulation of B cell-related genes in TLSs. Additionally, TLSs exhibited higher properties of memory B cells and myeloid dendritic cells but lower levels of innate immune cells compared with TILs. T cells within TLSs showed elevated expression of precursor-exhausted-related genes and lower cytotoxicity signature. Furthermore, TILs in TLS-positive tumors had higher levels of exhaustion signatures compared with TILs in TLS-negative tumors. Clinical data corroborated these findings, with higher PD-L1 and LAG-3 expression in TLS-positive tumors.
Our findings revealed that TILs in TLS-positive tumors have more exhausted T cell signature and PD-1 and LAG-3 protein expression in CCA which support our clinical finding. TLSs can predict favorable immunotherapy responses in patients with cholangiocarcinoma, highlighting their potential as a biomarker and therapeutic target to enhance treatment efficacy.
胆管癌是一种具有挑战性的恶性肿瘤，对传统疗法，尤其是免疫检查点抑制剂疗法的反应有限。肿瘤浸润淋巴细胞（TILs）和三级淋巴结构（TLSs）是肿瘤微环境（TME）的关键组成部分，并与癌症免疫反应有关。然而，TLSs和TILs在胆管癌患者中的作用和差异仍不清楚。本研究阐明了它们对TME的贡献。
我们检查了来自纳武单抗加改良吉西他滨和S-1的单臂II期试验的16个肿瘤样本以及各种数据集。采用免疫组织化学和RNA测序来评估TLSs和TILs的存在和活性。通过GeoMx数字空间分析器和癌症转录组图谱分析来检查差异基因表达和免疫细胞组成特征。
与TLS阴性（N=9）患者相比，TLS阳性（N=7）患者表现出明显更好的免疫治疗结果，包括更高的客观缓解率（71%对0%）和疾病控制率（100%对67%）。TLSs的存在与无进展生存期和总生存期的改善相关（p=0.03）。TLSs与以大量免疫浸润为特征的“炎症性”肿瘤相关，特别是涉及T细胞和B细胞。基因表达分析确定了TLSs中B细胞相关基因的显著上调。此外，与TILs相比，TLSs表现出更高的记忆B细胞和髓样树突状细胞特性，但先天免疫细胞水平较低。TLSs内的T细胞显示出前体耗竭相关基因的表达升高和较低的细胞毒性特征。此外，与TLS阴性肿瘤中的TILs相比，TLS阳性肿瘤中的TILs具有更高水平的耗竭特征。临床数据证实了这些发现，TLS阳性肿瘤中PD-L1和LAG-3表达更高。
我们的研究结果表明，TLS阳性肿瘤中的TILs在CCA中具有更多的耗竭T细胞特征以及PD-1和LAG-3蛋白表达，这支持了我们的临床发现。TLSs可以预测胆管癌患者的良好免疫治疗反应，突出了它们作为生物标志物和治疗靶点以提高治疗效果的潜力。
