Distinct PD-L1/PD1 Profiles and Clinical Implications in Intrahepatic Cholangiocarcinoma Patients with Different Risk Factors

Jia-Cheng Lu, Hai-Ying Zeng, Qi-Man Sun, Qing-Nan Meng, Xiao-Yong Huang, Peng-Fei Zhang, Xuan Yang, Rui Peng, Chao Gao, Chuan-Yuan Wei, Ying-Hao Shen, Jia-Bing Cai, Rui-Zhao Dong, Ying-Hong Shi, Hui-Chuan Sun, Yujiang G Shi, Jian Zhou, Jia Fan, Ai-Wu Ke, Liu-Xiao Yang, Guo-Ming Shi, Jia-Cheng Lu, Hai-Ying Zeng, Qi-Man Sun, Qing-Nan Meng, Xiao-Yong Huang, Peng-Fei Zhang, Xuan Yang, Rui Peng, Chao Gao, Chuan-Yuan Wei, Ying-Hao Shen, Jia-Bing Cai, Rui-Zhao Dong, Ying-Hong Shi, Hui-Chuan Sun, Yujiang G Shi, Jian Zhou, Jia Fan, Ai-Wu Ke, Liu-Xiao Yang, Guo-Ming Shi

Abstract

Rationale: PD1/PD-L1 immune checkpoint inhibitors have shown promising results for several malignancies. However, PD1/PD-L1 signaling and its therapeutic significance remains largely unknown in intrahepatic cholangiocarcinoma (ICC) cases with complex etiology. Methods: We investigated the expression and clinical significance of CD3 and PD1/PD-L1 in 320 ICC patients with different risk factors. In addition, we retrospectively analyzed 7 advanced ICC patients who were treated with PD1 inhibitor. Results: The cohort comprised 233 patients with HBV infection, 18 patients with hepatolithiasis, and 76 patients with undetermined risk factors. PD-L1 was mainly expressed in tumor cells, while CD3 and PD1 were expressed in infiltrating lymphocytes of tumor tissues. PD1/PD-L1 signals were activated in tumor tissues, and expression was positively correlated with HBV infection and lymph node invasion. More PD1+ T cells and higher PD-L1 expression were observed in tumor tissues of ICC patients with HBV infection compared to patients with hepatolithiasis or undetermined risk factors. More PD1+ T cells and/or high PD-L1 expression negatively impacted the prognosis of patients with HBV infection but not those with hepatolithiasis. Multivariate analysis showed PD1/PD-L1 expression was an independent indicator of ICC patient prognosis. Advanced ICC patients with HBV infection and less PD1+ T cells tended to have good response to anti-PD1 therapy. Conclusion: Hyperactivated PD1/PD-L1 signals in tumor tissues are a negative prognostic marker for ICCs after resection. HBV infection- and hepatolithiasis-related ICCs have distinct PD1/PD-L1 profiles. Further, PD1+ T cells could be used as a biomarker to predict prognosis and assay the efficiency of anti-PD1 immunotherapy in ICC patients with HBV infection.

Keywords: hepatitis B virus; hepatolithiasis; immune checkpoint blockage; intrahepatic cholangiocarcinoma; programmed cell death protein 1; programmed cell death protein ligand 1.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Figure 1
Figure 1
PD1/PD-L1 expression in tumor tissues of 320 ICCs. (A) and (B) Two representative ICC tissues showing different levels of PD-L1 expression (200×). (C) Semi-quantitative analysis of PD-L1 staining in tumor tissues (T) and peritumor tissues (PT) (**p < 0.01, Wilcoxon signed-rank test). (D) and (E) Two representative ICC tissues with different numbers of PD1- and CD3-positive cells (200×). (F) Statistical analysis of PD1+ T cells in tumor tissues (T) and peritumor tissues (PT) (**p < 0.01, paired Student's t-test).
Figure 2
Figure 2
Prognostic implication of PD1/PD-L1 axis in ICC patients. (A) Kaplan-Meier estimate of overall survival in the whole cohort with different PD-L1 levels (log-rank test). (B) Kaplan-Meier estimate of cumulative recurrence in the whole cohort with different PD-L1 levels (log-rank test). (C) Kaplan-Meier estimate of overall survival in the whole cohort with different PD1 levels (log-rank test). (D) Kaplan-Meier estimate of cumulative recurrence in the whole cohort with different PD1 levels (log-rank test). (E) Kaplan-Meier estimate of overall survival in the whole cohort with combined PD1/PD-L1 levels (double-positive refers to PD1high/PD-L1high; single positive refers to PD1high/PD-L1low or PD1low/PD-L1high; double negative refers to PD1low/PD-L1low) (log-rank test). (F) Kaplan-Meier estimate of cumulative recurrence in the whole cohort with combined PD1/PD-L1 levels (double positive refers to PD1high/PD-L1high; single positive refers to PD1high/PD-L1low or PD1low/PD-L1high; double negative refers to PD1low/PD-L1low) (log-rank test).
Figure 3
Figure 3
Implication of PD1/PD-L1 expression in ICC patients with different risk factors. (A) Kaplan-Meier estimate of overall survival in the whole cohort with different risk factors (*p < 0.05, log-rank test). (B) Kaplan-Meier estimate of cumulative recurrence in the whole cohort with different risk factors (*p < 0.05, log-rank test). (C) Kaplan-Meier estimate of overall survival of ICC patients with only HBV infection, grouped by PD1 level (log-rank test). (D) Kaplan-Meier estimate of cumulative recurrence in ICC patients with only HBV infection, grouped by PD1 level (log-rank test). (E) Kaplan-Meier estimate of overall survival of ICC patients with only HBV infection, grouped by PD-L1 level (log-rank test). (F) Kaplan-Meier estimate of cumulative recurrence in ICC patients with only HBV infection, grouped by PD-L1 level (log-rank test). (G) Kaplan-Meier estimate of overall survival of ICC patients with only hepatolithiasis, grouped by PD1 level (log-rank test). (H) Kaplan-Meier estimate of cumulative recurrence in ICC patients with only hepatolithiasis, grouped by PD1 level (log-rank test).
Figure 4
Figure 4
CD3, PD1, and PD-L1 expression and tumor responses to anti-PD1 immunotherapy in 7 advanced ICC patients. (A) Representative ICC tissues stained for CD3, PD1, and PD-L1 (200×). (B) Percentage change in tumor burden after anti-PD1 immunotherapy.

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