靶向 CD73 可抑制肝内胆管癌的肿瘤进展，并增强抗 PD-1 治疗的抗肿瘤活性。

Targeting CD73 limits tumor progression and enhances anti-tumor activity of anti-PD-1 therapy in intrahepatic cholangiocarcinoma.

机构信息

Department of Liver Surgery and Transplantation, Zhongshan Hospital, Fudan University, Shanghai, 200032, China.

Department of Hepatic Oncology, Xiamen Clinical Research Center for Cancer Therapy, Zhongshan Hospital, Fudan University (Xiamen Branch), Xiamen, 361015, China.

出版信息

J Cancer Res Clin Oncol. 2024 Jul 13;150(7):348. doi: 10.1007/s00432-024-05869-1.

DOI:10.1007/s00432-024-05869-1

PMID:39002018

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11246275/

Abstract

BACKGROUND & AIMS: Patients with intrahepatic cholangiocarcinoma (iCCA) respond poorly to immune checkpoint blockades (ICBs). In this study, we aimed to dissect the potential mechanisms underlying poor response to ICBs and explore a rational ICB-based combination therapy in iCCA.

METHODS

scRNA-seq dataset GSE151530 was analyzed to investigate the differentially expressed genes in malignant cells following ICBs therapy. RNA-seq analysis and western blot assays were performed to examine the upstream and downstream signaling pathways of CD73. Subcutaneous tumor xenograft models were utilized to investigate the impact of CD73 on iCCA growth. Plasmid AKT/NICD-induced spontaneous murine iCCAs were used to explore the therapeutic efficacy of CD73 enzymatic inhibitor AB680 combined with PD-1 blockade. Time-of-flight mass cytometry (CyTOF) was conducted to identify the tumor-infiltrating immune cell populations and their functional changes in murine iCCAs treated with AB680 in combination with PD-1 antibody.

RESULTS

scRNA-seq analysis identified elevated CD73 expression in malignant cells in response to ICBs therapy. Mechanistically, ICBs therapy upregulated CD73 expression in malignant cells via TNF-α/NF-κB signaling pathway. In vivo studies revealed that CD73 inhibition suppressed the growth of subcutaneous tumors, and achieved synergistic depression effects with gemcitabine and cisplatin (GC). Adenosine produced by CD73 activates AKT/GSK3β/β-catenin signaling axis in iCCA cells. CD73 inhibitor AB680 potentiates anti-tumor efficacy of PD-1 antibody in murine iCCAs. CyTOF analysis showed that AB680 combined with anti-PD-1 therapy promoted the infiltration of CD8 T, CD4 T cells, and NK cells in murine iCCAs, while simultaneously decreased the proportions of macrophages and neutrophils. Moreover, AB680 combined with anti-PD-1 significantly upregulated the expression of Granzyme B, Tbet and co-stimulatory molecule ICOS in infiltrating CD8 T cells.

CONCLUSIONS

CD73 inhibitor AB680 limits tumor progression and potentiates therapeutic efficacy of GC chemotherapy or anti-PD-1 treatment in iCCA. AB680 combined with anti-PD-1 therapy effectively elicits anti-tumor immune response.

摘要

背景与目的

肝内胆管癌（iCCA）患者对免疫检查点阻断（ICB）的反应较差。在这项研究中，我们旨在剖析导致对 ICB 反应较差的潜在机制，并探索 iCCA 中合理的基于 ICB 的联合治疗方法。

方法

分析 scRNA-seq 数据集 GSE151530，以研究 ICB 治疗后恶性细胞中差异表达的基因。进行 RNA-seq 分析和 Western blot 检测，以研究 CD73 的上游和下游信号通路。利用皮下肿瘤异种移植模型研究 CD73 对 iCCA 生长的影响。利用质粒 AKT/NICD 诱导自发的小鼠 iCCAs 来探索 CD73 酶抑制剂 AB680 与 PD-1 阻断联合治疗的疗效。利用飞行时间质谱流式细胞术（CyTOF）鉴定 AB680 联合 PD-1 抗体治疗后小鼠 iCCAs 中肿瘤浸润免疫细胞群及其功能变化。

结果

scRNA-seq 分析鉴定出 ICB 治疗后恶性细胞中 CD73 表达升高。在机制上，ICB 治疗通过 TNF-α/NF-κB 信号通路上调恶性细胞中 CD73 的表达。体内研究表明，CD73 抑制抑制了皮下肿瘤的生长，并与吉西他滨和顺铂（GC）联合产生协同抑制作用。CD73 产生的腺苷激活 iCCA 细胞中的 AKT/GSK3β/β-catenin 信号轴。CD73 抑制剂 AB680 增强了 PD-1 抗体在小鼠 iCCAs 中的抗肿瘤疗效。CyTOF 分析表明，AB680 联合抗 PD-1 治疗促进了小鼠 iCCAs 中 CD8 T、CD4 T 细胞和 NK 细胞的浸润，同时降低了巨噬细胞和中性粒细胞的比例。此外，AB680 联合抗 PD-1 显著上调浸润 CD8 T 细胞中 Granzyme B、Tbet 和共刺激分子 ICOS 的表达。