Downregulation of N4-acetylcytidine modification in myeloid cells attenuates immunotherapy and exacerbates hepatocellular carcinoma progression

• Published in: British journal of cancer 2024-02, Vol.130 (2), p.201-212
• Main Authors: Xu, Nan, Zhuo, Jianyong, Chen, Yiyuan, Su, Renyi, Chen, Huan, Zhang, Zhensheng, Lian, Zhengxing, Lu, Di, Wei, Xuyong, Zheng, Shusen, Xu, Xiao, Wang, Shuai, Wei, Qiang
• Format: Article
• Language: English
• Subjects: 631/67/1059/2325; 692/308/575; Animals; Biomedical and Life Sciences; Biomedicine; Cancer Research; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - therapy; CD11b antigen; Cytidine - analogs & derivatives; Cytotoxicity; Down-Regulation; Drug Resistance; Epidemiology; Flow cytometry; Gene expression; Hepatocellular carcinoma; Humans; Immunotherapy; Infiltration; Liver cancer; Liver Neoplasms - drug therapy; Liver Neoplasms - therapy; Lymphocytes T; Macrophages; Metastases; Mice; Molecular Medicine; Molecular modelling; mRNA stability; Myeloid cells; Myeloid-Derived Suppressor Cells - metabolism; Oncology; Phenotypes; RNA modification; RNA, Messenger - genetics; Transplantation
• ISSN: 0007-0920; 1532-1827; 1532-1827
• DOI: 10.1038/s41416-023-02510-9

Background N4-acetylcytidine (ac4C) is a conserved and abundant mRNA modification that controls protein expression by affecting translation efficiency and mRNA stability. Whether the ac4C modification of mRNA regulates hepatocellular carcinoma (HCC) development or affects the immunotherapy of HCC is unknown. Methods By constructing an orthotopic transplantation mouse HCC model and isolating tumour-infiltrated immunocytes, we evaluated the ac4C modification intensity using flow cytometry. Remodelin hydrobromide (REM), an ac4C modification inhibitor, was systematically used to understand the extensive role of ac4C modification in immunocyte phenotypes. Single-cell RNA-seq was performed to comprehensively evaluate the changes in the tumour-infiltrating immunocytes and identify targeted cell clusters. RNA-seq and RIP-seq analyses were performed to elucidate the underlying molecular mechanisms. Tyramide Signal Amplification (TSA) analysis on the HCC tissue microarray was performed to explore the clinical relatedness of our findings. Results Ac4C modification promoted M1 macrophage infiltration and reduced myeloid-derived suppressor cell MDSCs infiltration in HCC. The inhibition of ac4C modification induces PDL1 expression by stabilising mRNA in the myeloid cells, thereby attenuating the CTL-mediated tumour cell-killing ability. High infiltration of ac4C+CD11b+ cells is positively related to a better prognosis in patients with HCC. Conclusions Ac4C modification of myeloid cells enhanced the HCC immunotherapy by suppressing PDL1 expression.