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Impaired autophagy by cepharanthine induces immunogenic cell death and enhances anti-PD-1 response in MSS-type colorectal cancer

Oncogene volume 44, pages 3171–3182 (2025)Cite this article

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Abstract

Blockade of the PD-1/PD-L1 axis has demonstrated remarkable success in treating colorectal cancer (CRC) with high microsatellite instability (MSI-H). However, the metastatic CRC with microsatellite stability (MSS) does not respond to this approach. A recent study revealed that rare neoantigens in MSS-type CRC cells can be recognized by antigen-specific CD8+ T cells but not by dendritic cells (DCs). Thus, increasing neoantigen availability by DCs may improve the efficacy of PD-1/PD-L1 inhibitors in MSS-type CRC. Here, we conducted a drug library screening for ‘eat me’ signal, represented by cell surface calreticulin (CRT) exposure, in MSS-type CT26 cells. Cells treated with identified cepharanthine (CEP) presented hallmarks of immunogenic cell death (ICD), characterized by increased cell surface CRT exposure, the release of HMGB1 and ATP, increased susceptibility to phagocytosis, and the ability of vaccines to elicit immunogenic potential in vivo. Mechanistically, CEP blocked autophagic flux by inhibiting autophagsome-lysosome fusion, leading to endoplasmic reticulum (ER) stress and ICD activation. Moreover, CEP upregulated PD-L1 expression on tumour cells, impeding the antitumour immune response in vivo. The combination of CEP and anti-PD-1 therapy provided therapeutic benefit to MSS-type CRC tumours, with an increased proportion of activated DCs and IFNγ+ CD8+ T cells and a decreased proportion of regulatory T cells within the tumour. Based on the above observation, subsequent clinical trials can be conducted to achieve the clinical goal of increasing the survival benefit of MSS-CRC patients.

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Fig. 1: Identification of CEP as a potent ICD inducer in MSS-type CT26 cells.
Fig. 2: CEP is a bona fide ICD inducer in MSS-type CRC cells.
Fig. 3: CEP-treated MSS-type CRC cells activate DCs.
Fig. 4: CEP inhibits autophagic flux in MSS-type CRC cells.
Fig. 5: CEP triggers ER stress in MSS-type CRC cells.
Fig. 6: CEP induces immune escape in vivo.
Fig. 7: CEP enhances anti-PD-1 treatment efficacy in MSS-type CRC tumours.
Fig. 8: Illustration of the mechanism of CEP-induced ICD and the subsequent activation of the antitumour immune response.

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On reasonable request to the corresponding author.

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Acknowledgements

We thank the members of the C.Xu laboratories for their helpful advice and discussion.

Funding

This work was supported by the special key projects for Natural Science Foundation of Chongqing (2024NSCQ-KJFZMSX0220), the Scientific Research Grant of Chengdu Municipal Health Commission (2022099), the Medical Research Project of Sichuan Province (Q22011) granted to MZ, and the National Key R&D Programme of China (Grant No. 2023YFC3402100), National Natural Science Foundation of China (Grant No. 92259102), Sichuan Natural Science Foundation of China (Grant No. 2024NSFSC0057), and Chongqing Science and Technology Bureau (Grant No. CSTB2024TIAD-KPX0029) to CX.

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Authors and Affiliations

  1. Yu-Yue Pathology Scientific Research Center, Chongqing, PR China

    Ming Zhao & Chuan Xu

  2. Jinfeng Laboratory, Chongqing, PR China

    Ming Zhao & Chuan Xu

  3. Department of Gynecology, Clinical Medical College and the First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan, PR China

    Juan Nie

  4. Department of Gastroenterology, Clinical Medical College and the First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan, PR China

    Ao Ye, Xuemei Li, Ziyi Yang & Jun Li

  5. Collaborative Innovation Centre of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry, Guangxi Medical University, Nanning, Guangxi, PR China

    Chang Liu

  6. Clinical Medical College, Chengdu Medical College, Chengdu, Sichuan, PR China

    Runyu Zhou

  7. Department of Oncology & Cancer Institute, Sichuan Academy of Medical Sciences, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, PR China

    Chuan Xu

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Contributions

MZ and CX conceptualised and designed the experiment. MZ, JN, ZYY, XML, AY, and CL carried out the experiment. RYZ and XML participated in the data collection. MZ, RYZ, and JL helped data analysis. CX revised the manuscript.

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Correspondence to Chuan Xu.

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All methods were performed in accordance with the relevant guidelines and regulations. The mouse experiment was approved by the Animal Welfare Ethics Committee at Chengdu Medical College (IACUC-24-034).

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Zhao, M., Nie, J., Ye, A. et al. Impaired autophagy by cepharanthine induces immunogenic cell death and enhances anti-PD-1 response in MSS-type colorectal cancer. Oncogene 44, 3171–3182 (2025). https://doi.org/10.1038/s41388-025-03488-9

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