Afatinib combined with anti-PD1 enhances immunotherapy of hepatocellular carcinoma via ERBB2/STAT3/PD-L1 signaling

Chao Yu; Xinyi Zhang; Min Wang; Gaoxin Xu; Siqi Zhao; Yongheng Feng; Chao Pan; Weijun Yang; Jin Zhou; Longcheng Shang; Yong Ma

doi:10.3389/fonc.2023.1198118

Afatinib combined with anti-PD1 enhances immunotherapy of hepatocellular carcinoma via ERBB2/STAT3/PD-L1 signaling

Front Oncol. 2023 May 30:13:1198118. doi: 10.3389/fonc.2023.1198118. eCollection 2023.

Authors

Chao Yu¹, Xinyi Zhang¹, Min Wang¹, Gaoxin Xu¹, Siqi Zhao¹, Yongheng Feng¹, Chao Pan¹, Weijun Yang¹, Jin Zhou¹, Longcheng Shang¹, Yong Ma¹

Affiliation

¹ Department of General Surgery, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.

Abstract

Background: Afatinib is mainly used to treat advanced non-small cell lung cancer, but its therapeutic effect on hepatocellular carcinoma is still unclear.

Methods: Over 800 drugs were screened by CCK8 technology and afatinib was found to have a significant inhibitory effect on liver cancer cells. The expression of PDL1 in tumor cells treated with drugs were detected by qRT-PCR and Weston Blot experiments. The effects of afatinib on the growth, migration and invasion of HCC cells were evaluated using wound healing, Transwell, and cell cloning assays. The in vivo effects of afatinib in combination with anti-PD1 were evaluated in C57/BL6J mice with subcutaneous tumorigenesis. Bioinformatics analysis was performed to explore the specific mechanism of afatinib's inhibition of ERBB2 in improving the expression level of PD-L1, which was subsequently verified through experiments.

Results: Afatinib was found to have a significant inhibitory effect on liver cancer cells, as confirmed by in vitro experiments, which demonstrated that it could significantly suppress the growth, invasion and migration of HCC cells. qRT PCR and Weston Blot experiments also showed that Afatinib can enhance the expression of PD-L1 in tumor cells. In addition, in vitro experiments confirmed that afatinib can significantly enhance the immunotherapeutic effect of hepatocellular carcinoma. Afatinib's ability to increase PD-L1 expression is mediated by STAT3 activation following its action on HCC cells.

Conclusion: Afatinib enhances PD-L1 expression in tumor cells through the STAT3/PD-L1 pathway. The combination of afatinib and anti-PD1 treatment significantly increases the immunotherapeutic effect of HCC.

Keywords: ERBB2; PD-L1; afatinib; hepatocellular carcinoma (HCC); immunotherapy.

Grants and funding

This study was supported by the National Natural Science Foundation (No. 82203762), Chen Xiaoping Foundation for the Development of Science and Technology of Hubei Province (No. CXPJJH12000009-08), and Xinghuo Talent Program of Nanjing First Hospital.