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Ginkgetin targets GRP78 to induce dual pathways of ER stress and immune activation in osteosarcoma

Oncogene volume 45, pages 1042–1056 (2026)Cite this article

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Abstract

Osteosarcoma is an aggressive malignancy characterized by rapid proliferation and a propensity for metastasis. The endoplasmic reticulum (ER) chaperone GRP78, a critical regulator of osteosarcoma progression, represents a promising therapeutic target. In this study, we identified the natural compound ginkgetin (Gink) as a novel GRP78 inhibitor. Cellular thermal shift assays, surface plasmon resonance, and mutagenesis studies revealed that Gink directly binds to GRP78, with K296 serving as a key interaction site. In vitro, Gink suppressed osteosarcoma cell proliferation, migration, and invasion while inducing apoptosis and autophagy by activating the PERK-eIF2α-ATF4 pathway. Co-immunoprecipitation assays showed that Gink competitively disrupted GRP78-PERK interaction. In orthotopic and patient-derived xenograft models, Gink treatment markedly attenuated tumor growth and metastasis. Gink also reprogrammed the tumor immune microenvironment by reducing M2 macrophage polarization and synergizing with anti-PD1 therapy to enhance CD8+ T-cell activity. Accordingly, Gink could be developed as a GRP78-targeting agent that triggers ER stress and immune activation, offering a dual-pronged therapeutic strategy against osteosarcoma.

Ginkgetin (Gink) directly binds to GRP78 in a competitive manner, disrupting the interaction between GRP78 and PERK. This leads to PERK activation and phosphorylation, which in turn phosphorylates eIF2α to trigger ATF4 transcription. Ultimately, this cascade induces apoptosis and autophagy, inhibiting cancer progression. Additionally, Gink suppresses M2 macrophage polarization and enhances CD8+ T cell cytotoxicity, both of which contribute to the prevention of cancer development.

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Fig. 1: Screening Gink as a GRP78-targeting drug from a natural compound library.
Fig. 2: Gink binds to the K296 residue of GRP78 and suppresses its ATPase activity.
Fig. 3: Gink targets GRP78-induced early ER stress.
Fig. 4: Gink activates the PERK-eIF2α-ATF4 pathway to induce autophagy and apoptosis.
Fig. 5: Gink inhibits osteosarcoma cell proliferation and migration.
Fig. 6: Gink inhibits osteosarcoma proliferation and metastasis in vivo.
Fig. 7: Gink influences M2 macrophage polarization and acts synergistically with anti-PD1 immunotherapy.

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All data and materials are available upon request by contacting the corresponding author.

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Acknowledgements

We thank all members of Shanghai Bone Tumor Institute for their assistance. This work was supported by grants from the National Key Research and Development Program of China (NO. 2021YFC2400600/2021YFC2400605) and The Science and Technology Plan Project of Inner Mongolia Autonomous Region (NO. 2023YFSH0001).

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  1. These authors contributed equally: Wenyuan Xu, Tongtong Liu.

Authors and Affiliations

  1. Department of Orthopedics, Shanghai Bone Tumor Institution, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Wenyuan Xu, Tongtong Liu, Xinglong Ma, He Dong, Yinghao Cao, Zhuoying Wang, Yingqi Hua, Zhengdong Cai, Mengxiong Sun & Tao Zhang

  2. School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China

    Guanyi Li

  3. Precision Research Center for Refractory Diseases, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Jingjie Li

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Contributions

WYX was responsible for methodology, validation, formal analysis, and writing-original draft. TTL was responsible for methodology, formal analysis, writing-original draft. XLM contributed to formal analysis and data curation. HD and YHC created visualizations. ZYW and GYL conducted investigation. YQH and ZDC provided resources. MXS and JJL were responsible for conceptualization, project administration, writing-review & editing. TZ contributed to conceptualization, supervision, project administration, writing-review & editing.

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Correspondence to Mengxiong Sun, Jingjie Li or Tao Zhang.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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All methods were performed in accordance with the relevant guidelines and regulations. Tumor specimens were obtained with written informed consent from the patients and with approval from the Ethics Committee of Shanghai General Hospital. Animal experiments were approved by the Animal Ethical and Welfare Committee of the Shanghai General Hospital and were performed following institutional guidelines (Approval number: 2024AWS306).

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Xu, W., Liu, T., Ma, X. et al. Ginkgetin targets GRP78 to induce dual pathways of ER stress and immune activation in osteosarcoma. Oncogene 45, 1042–1056 (2026). https://doi.org/10.1038/s41388-026-03704-0

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