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Cellular and Molecular Biology

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

British Journal of Cancer volume 130, pages 201–212 (2024)Cite this article

Subjects

Abstract

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.

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Fig. 1: Inhibition of ac4C modification promotes myeloid cell and B cell infiltrations and accelerates HCC development.
Fig. 2: NAT10 expression in myeloid cells is required for effective immunotherapy.
Fig. 3: Inhibition of ac4C modification promotes MDSC infiltration in the TME of HCC.
Fig. 4: NAT10 induces M1 macrophages and reduces MDSCs.
Fig. 5: NAT10 inhibits PDL1 expression in myeloid cells and enhances CTL-mediated tumour cell killing.
Fig. 6: Ac4C modification promotes PDL1 mRNA degradation in myeloid cells.
Fig. 7: Ac4C+CD11b+ infiltration relates to better prognosis in HCC patients.

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Data availability

The study data are available from the GEO database and are publicly available as of the date of publication (RNA-sequence (GSE220196), single-cell RNA-sequence (GSE244613) and ac4C-RIP-sequence (GSE227526)).

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Funding

The National Natural Science Foundation of China (Nos. 92159202 and 82273177). The Key Research and Development Plan of the Zhejiang Province (Nos. 2019C03050 and 2021C03118).

Author information

Author notes

  1. These authors contributed equally: Nan Xu, Jianyong Zhuo, Yiyuan Chen, Renyi Su, Huan Chen.

Authors and Affiliations

  1. Zhejiang University School of Medicine, Hangzhou, 310058, China

    Nan Xu, Jianyong Zhuo, Renyi Su, Zhensheng Zhang, Zhengxing Lian, Di Lu, Xuyong Wei, Xiao Xu, Shuai Wang & Qiang Wei

  2. The Fourth Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, 310006, China

    Yiyuan Chen, Huan Chen, Zhengxing Lian, Di Lu, Xuyong Wei & Qiang Wei

  3. NHC Key Laboratory of Combined Multi-organ Transplantation, Hangzhou, 310003, China

    Di Lu, Xuyong Wei, Shusen Zheng, Xiao Xu & Qiang Wei

  4. Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, China

    Shusen Zheng

  5. Shulan (Hangzhou) Hospital, Zhejiang Shuren University School of Medicine, Hangzhou, 310022, China

    Shusen Zheng

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Contributions

QW, SW, and XX conceived and supervised the study. NX, JYZ, YYC, HC, ZSZ, ZXL, and XYW performed all the experiments. SW, RYS, DL, and NX analysed the data. NX and SW wrote the manuscript. SSZ and XX reviewed the manuscript.

Corresponding authors

Correspondence to Xiao Xu, Shuai Wang or Qiang Wei.

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Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

All the individuals signed informed consent forms for the use of their clinical specimens in the present study according to the principles expressed in the Declaration of Helsinki and approved by the Institutional Research Ethics Committees of the Affiliated Hangzhou First People’s Hospital and the First Affiliated Hospital, Zhejiang University School of Medicine.

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Xu, N., Zhuo, J., Chen, Y. et al. Downregulation of N4-acetylcytidine modification in myeloid cells attenuates immunotherapy and exacerbates hepatocellular carcinoma progression. Br J Cancer 130, 201–212 (2024). https://doi.org/10.1038/s41416-023-02510-9

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