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Antifungal agent Terbinafine restrains tumor growth in preclinical models of hepatocellular carcinoma via AMPK-mTOR axis

Oncogene volume 40, pages 5302–5313 (2021)Cite this article

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Abstract

The prognosis of hepatocellular carcinoma (HCC) remains unsatisfactory due to limited effective treatment options. In this work, we investigated the therapeutic efficacy of Terbinafine for HCC and the underlying mechanism. The influence of Terbinafine on cell growth, 3D spheroid formation, clonogenic survival, and protein synthesis was investigated in human HCC cell lines. Co-immunoprecipitation, immunofluorescence, and other techniques were employed to explore how Terbinafine exerts its anticancer effect. Subcutaneous tumorigenicity assay, orthotopic and patient-derived xenograft (PDX) HCC models were used to evaluate the anticancer effect of Terbinafine monotherapy and the combinatorial treatment with Terbinafine and sorafenib against HCC. The anticancer activity of Terbinafine was Squalene epoxidase (SQLE)-independent. Instead, Terbinafine robustly suppressed the proliferation of HCC cells by inhibiting mTORC1 signaling via activation of AMPK. Terbinafine alone or in combination with sorafenib delayed tumor progression and markedly prolonged the survival of tumor-bearing mice. The synergy between Terbinafine and sorafenib was due to concomitant inhibition of mTORC1 and induction of severe persistent DNA double-strand breaks (DSBs), which led to the delayed proliferation and accelerated cell death. Terbinafine showed promising anticancer efficacy in preclinical models of HCC and may serve as a potential therapeutic strategy for HCC.

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Fig. 1: Terbinafine suppresses mTORC1 signaling.
Fig. 2: AMPK mediates mTORC1 inhibition by Terbinafine.
Fig. 3: Synthetic lethality with a combination of Terbinafine and sorafenib in HCC cells.
Fig. 4: Terbinafine synergizes with sorafenib in suppressing mTORC1 signaling.
Fig. 5: Combinatorial treatment with Terbinafine and sorafenib elicits severe DSBs and induces cell death.
Fig. 6: Combined Terbinafine and sorafenib treatment in vivo impairs tumor growth and prolongs survival.
Fig. 7: Terbinafine plus sorafenib combinatorial treatment causes regression of patient-derived xenograft (PDX) in vivo.

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Acknowledgements

We thank Professor Yu Li of our institute (SINH, CAS) for the gift of an anti-puromycin monoclonal antibody (#EQ0001, Kefafast). We also appreciate the New World Group for their Charitable Foundation to establish the Institute for Nutritional Sciences, SIBS, CAS-New World Joint laboratory, which has given full support to this study. In addition, the Author Gratefully Acknowledges the Support of the SA-SIBS Scholarship Program.

Funding

This work was supported by the National Key R&D Program of China (2018YFC1603002 and 2018YFC1604404), National Natural Science Foundation of China (81730083 and 82030084) to Dong Xie; and National Natural Science Foundation of China (31771538 and 81972757), Youth Innovation Promotion Association of Chinese Academy of Sciences fund (2017324) to Jing-Jing Li.

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Author notes

  1. These authors contributed equally: Er-Bin Zhang, Xiuping Zhang.

Authors and Affiliations

  1. CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 200031, Shanghai, China

    Er-Bin Zhang, Fengkun Zhang, Tian-Wei Chen, Ning Ma, Qian-Zhi Ni, Yi-Kang Wang, Qian-Wen Zheng, Hui-Jun Cao, Ji Xia, Bing Zhu, Sheng Xu, Xufen Ding, Dong Xie & Jing-Jing Li

  2. Department of Hepatic Surgery VI, Eastern Hepatobiliary Surgery Hospital, Naval Medical University, 2000438, Shanghai, China

    Xiuping Zhang, Kang Wang & Shuqun Cheng

  3. Faculty of Hepato-Pancreato-Biliary Surgery, Chinese People Liberation Army (PLA) General Hospital; Institute of Hepatobiliary Surgery of Chinese PLA, 100853, Beijing, China

    Xiuping Zhang

  4. School of Life Science and Technology, ShanghaiTech University, 201210, Shanghai, China

    Qian-Wen Zheng & Dong Xie

  5. First People’s Hospital of Huzhou, First Affiliated Hospital of Huzhou University, 313000, Huzhou, Zhejiang, China

    Xiang Wang

  6. Department of Thoracic Surgery, Section of Esophageal Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University, 200030, Shanghai, China

    Zhigang Li

  7. NHC Key Laboratory of Food Safety Risk Assessment, China National Center for Food Safety Risk Assessment, 100022, Beijing, China

    Dong Xie

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Contributions

E.Z. contributed to conceptualization, investigation, formal analysis, validation, project administration, and writing—original draft; X.Z. contributed to the investigation, formal analysis, validation, and project administration; K.W., F.Z., T.C., N.M., Q.N., Y.W., Q.Z., H.C., J.X., B.Z., S.X., X.D., X.W., and Z.L. contributed to the formal analysis, methodology, and resources; S.C., D.X., J.L. contributed to the funding acquisition, supervision, and writing—review & editing. All authors read and approved the final paper.

Corresponding authors

Correspondence to Shuqun Cheng, Dong Xie or Jing-Jing Li.

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Mice were manipulated according to the guidelines for the care and use of laboratory animals and were approved by the Institutional Biomedical Research Ethics Committee of the Shanghai Institutes for Biological Sciences (#ER-SIBS-251902). For Patient-Derived Xenograft (PDX) model studies, written informed consent was obtained from each patient.

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Zhang, EB., Zhang, X., Wang, K. et al. Antifungal agent Terbinafine restrains tumor growth in preclinical models of hepatocellular carcinoma via AMPK-mTOR axis. Oncogene 40, 5302–5313 (2021). https://doi.org/10.1038/s41388-021-01934-y

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