Huang Lingli, Zhong Qian, Huang Silan, Yang Kejia, Cai Yuchen, Guo Guifang
VIP Department, Sun Yat-sen University Cancer Center, Guangzhou, P.R. China.
State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Guangzhou, P.R. China.
Hepatol Commun. 2025 Mar 13;9(4). doi: 10.1097/HC9.0000000000000674. eCollection 2025 Apr 1.
The absence of representative Epstein-Barr virus-associated intrahepatic cholangiocarcinoma (EBVaICC) cell lines has limited our understanding of the molecular and immunological characteristics of this cancer subtype.
We reviewed patients with metastatic cholangiocarcinoma at Sun Yat-sen University Cancer Center from January 2015 to August 2023. Among them, 22 patients with EBVaICC and 66 patients with non-EBVaICC who received anti-PD1 treatment were included. Additionally, 2 EBV-positive ICC cell lines, RBE-EBV and HuH28-EBV, were developed through cell-to-cell infection. Stable EBV infection and responsiveness to viral reactivation were confirmed. Transcriptomic and bioinformatics analyses were performed, and in vitro experiments examined the immune effects of EBV-positive ICC. Key immune-related genes and cytokines were validated by reverse transcription quantitative polymerase chain reaction and ELISA in cell lines and patient plasma samples.
In this study, we found that patients with EBVaICC showed enhanced immune responses and improved overall and progression-free survival compared to patients with non-EBVaICC. We first successfully established and validated 2 EBV-positive ICC cell lines (RBE-EBV and HuH28-EBV). These cell lines were confirmed for stable EBV infection and displayed responsiveness to viral reactivation, making them suitable for future studies. Transcriptomic analyses and in vitro studies revealed that EBV activated the cGAS-STING pathway, resulting in MHC-I upregulation and CXCL10 secretion in ICC cells, which collectively enhanced CD8+ T cell chemotaxis and cytotoxicity. Furthermore, ELISA analysis showed higher plasma levels of CXCL10 and IFN-γ in patients with EBVaICC, suggesting a potential role for EBV in enhancing immunotherapy sensitivity in this subtype.
The established EBV-positive ICC cell lines revealed enhanced immunogenicity driven by cGAS-STING pathway activation, providing valuable models for future research and insights into the mechanisms of improved immunotherapy sensitivity in EBVaICC.
缺乏具有代表性的爱泼斯坦-巴尔病毒相关肝内胆管癌（EBVaICC）细胞系限制了我们对该癌症亚型分子和免疫特征的理解。
我们回顾了2015年1月至2023年8月在中山大学肿瘤防治中心接受治疗的转移性胆管癌患者。其中，纳入了22例接受抗PD1治疗的EBVaICC患者和66例非EBVaICC患者。此外，通过细胞间感染建立了2株EBV阳性的ICC细胞系，即RBE-EBV和HuH28-EBV。证实了稳定的EBV感染及对病毒再激活的反应性。进行了转录组学和生物信息学分析，并通过体外实验研究了EBV阳性ICC的免疫效应。通过逆转录定量聚合酶链反应和酶联免疫吸附测定法在细胞系和患者血浆样本中验证了关键免疫相关基因和细胞因子。
在本研究中，我们发现与非EBVaICC患者相比，EBVaICC患者表现出增强的免疫反应以及更好的总生存期和无进展生存期。我们首次成功建立并验证了2株EBV阳性的ICC细胞系（RBE-EBV和HuH28-EBV）。这些细胞系被证实存在稳定的EBV感染，并对病毒再激活有反应，适合未来的研究。转录组学分析和体外研究表明，EBV激活了cGAS-STING通路，导致ICC细胞中MHC-I上调和CXCL10分泌，共同增强了CD8+T细胞的趋化性和细胞毒性。此外，酶联免疫吸附测定分析显示，EBVaICC患者血浆中CXCL10和IFN-γ水平较高，提示EBV在增强该亚型免疫治疗敏感性方面可能发挥作用。
所建立的EBV阳性ICC细胞系揭示了由cGAS-STING通路激活驱动的增强免疫原性，为未来研究以及深入了解EBVaICC免疫治疗敏感性改善机制提供了有价值的模型。
