Department of Oncology, Johns Hopkins University School of Medicine, 1650 Orleans Street, CRB1 Room 351, Baltimore, MD, 21231, USA.
The Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
J Hematol Oncol. 2022 Mar 28;15(1):37. doi: 10.1186/s13045-022-01253-z.
Immunotherapy has demonstrated a limited clinical efficacy in approximately 5% of cholangiocarcinoma. The main challenges for an effective immunotherapy response in cholangiocarcinoma arise from the tumor microenvironment, which is poorly understood.
For a comprehensive analysis of the tumor microenvironment in cholangiocarcinoma, we performed multiplex immunohistochemistry with two 15-marker immune panels and Nanostring assays for a comprehensive analysis of 104 surgically resected cholangiocarcinomas including intrahepatic, hilar, and distal cholangiocarcinoma. We also validated some key findings with a batch integration analysis of published single cell RNA sequencing data.
This study found that natural killer cells occupy the largest immune cell compartment in cholangiocarcinoma. Granzyme-BCD8 effector T cells are significantly associated with better overall survival in both intrahepatic and distal cholangiocarcinoma. Above 85% of intrahepatic cholangiocarcinomas with higher density of PD-1EOMESCD8 effector T cells are associated with long-term survival. However, only the density of PD-1EOMESCD8 T cells in the tumor areas, but not in the peripheries of the tumors, is prognostic. In all three cholangiocarcinoma subtypes, T regulator cells are significantly associated with a poor prognosis; however, M1 and M2 tumor-associated macrophages or PD-L1 tumor-associated macrophage demonstrate different prognostic values. Combining PD-L1 M1 or M2, PD-L1 M1 or M2 tumor-associated macrophages, and T regulator cells to subgroup intrahepatic and distal cholangiocarcinoma, the prognosis is significantly better distinguished. Moreover, PD-L1 M2 tumor-associated macrophages is associated with a good prognosis in intrahepatic and distal cholangiocarcinoma, suggesting this subtype of M2 tumor-associated macrophages may be antitumoral. Interestingly, lower densities of various types of immunosuppressive cells are associated with decreased infiltration of effector T cells in distal and hilar cholangiocarcinoma, but not in intrahepatic cholangiocarcinoma. In intrahepatic cholangiocarcinoma, PD-L1 tumor-associated macrophages exert their immunosuppressive function likely through promoting T cell exhaustion.
This study suggests that the densities of Granzyme-BCD8 effector T cells and non-exhausted PD-1EOMESCD8 T cells and the PD-L1 status in the tumor-associated macrophages are prognostic makers in cholangiocarcinomas. The study also supports targeting PD-L1 tumor-associated macrophages as the immunotherapy for cholangiocarcinoma.
免疫疗法在大约 5%的胆管癌患者中显示出有限的临床疗效。胆管癌中有效的免疫治疗反应的主要挑战来自肿瘤微环境，但该环境尚未得到充分了解。
为了全面分析胆管癌的肿瘤微环境，我们对 104 例手术切除的胆管癌进行了双重 15 标志物免疫组化分析，并使用 Nanostring 检测进行了全面分析，包括肝内、肝门和远端胆管癌。我们还使用已发表的单细胞 RNA 测序数据的批量整合分析验证了一些关键发现。
本研究发现自然杀伤细胞在胆管癌中占据最大的免疫细胞区室。颗粒酶-B+CD8 效应 T 细胞与肝内和远端胆管癌的总生存期显著相关。超过 85%的 PD-1+EOMES+CD8 效应 T 细胞密度较高的肝内胆管癌与长期生存相关。然而，只有肿瘤区域而不是肿瘤周围的 PD-1+EOMES+CD8 T 细胞密度具有预后意义。在所有三种胆管癌亚型中，T 调节细胞与不良预后显著相关；然而，M1 和 M2 肿瘤相关巨噬细胞或 PD-L1 肿瘤相关巨噬细胞具有不同的预后价值。将 PD-L1+M1 或 M2、PD-L1+M1 或 M2 肿瘤相关巨噬细胞和 T 调节细胞组合成亚组，可显著更好地区分肝内和远端胆管癌的预后。此外，PD-L1+M2 肿瘤相关巨噬细胞与肝内和远端胆管癌的良好预后相关，表明这种 M2 肿瘤相关巨噬细胞亚型可能具有抗肿瘤作用。有趣的是，在远端和肝门胆管癌中，各种类型的免疫抑制细胞密度较低与效应 T 细胞浸润减少相关，但在肝内胆管癌中则不然。在肝内胆管癌中，PD-L1 肿瘤相关巨噬细胞通过促进 T 细胞耗竭发挥其免疫抑制功能。
本研究表明，Granzyme-B+CD8 效应 T 细胞和非耗竭 PD-1+EOMES+CD8 T 细胞的密度以及肿瘤相关巨噬细胞中的 PD-L1 状态是胆管癌的预后标志物。该研究还支持将 PD-L1 肿瘤相关巨噬细胞作为胆管癌的免疫治疗靶点。
