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Translational Therapeutics

APR-246 induces apoptosis and enhances chemo-sensitivity via activation of ROS and TAp73-Noxa signal in oesophageal squamous cell cancer with TP53 missense mutation

British Journal of Cancer volume 125, pages 1523–1532 (2021)Cite this article

Subjects

Abstract

Background

Mutations in p53, identified in 90% of oesophageal squamous cell carcinoma (ESCC), are associated with unfavourable prognosis and chemo-resistance. APR-246 induces apoptosis by restoring transcriptional ability of mutant p53, and may be a promising therapeutic agent to overcome chemo-resistance in ESCC.

Methods

In ESCC cell lines differing in p53 status, we performed in vitro cell viability and apoptosis assays, evaluated reactive oxygen species (ROS) generation, and assessed signal changes by western blot after APR-246 administration with/without chemo-agent. Antitumour effects and signal changes were evaluated in in vivo experiments using xenograft and patient-derived xenograft (PDX) mouse models.

Results

APR-246 administration induced significant apoptosis by upregulating p73 and Noxa via ROS induction in ESCC cell lines harbouring p53 missense mutations. Moreover, APR-246 plus chemotherapy exerted combined antitumour effects in ESCC with p53 missense mutations. This effect was also mediated through enhanced ROS activity, leading to massive apoptosis via upregulation of p73 and Noxa. These findings were confirmed by xenograft and PDX models with p53 mutant ESCC.

Conclusion

APR-246 strongly induced apoptosis by inducing ROS activity and p73-Noxa signalling, specifically in ESCC with p53 missense mutation. This antitumour effect was further enhanced by combination with 5-FU, which we first confirmed in ESCC preclinical model.

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Fig. 1: APR-246 alone induces massive apoptosis via ROS induction leading to upregulation of p73-Noxa in ESCC with p53 missense mutation.
Fig. 2: APR-246-induced apoptosis was further enhanced by its combination with 5-FU in ESCC with p53 missense mutation.
Fig. 3: p73-Noxa upregulation as an essential pathway in combination therapy with APR-246 and 5-FU in ESCC with p53 missense mutation.
Fig. 4: Combination treatment with APR-246 and 5-FU yields synergic anticancer effect in a xenograft mouse model of ESCC with p53 missense mutation.
Fig. 5: PDX mouse model confirmed ROS-TAp73-Noxa-mediated apoptosis by treatment with APR-246 plus 5-FU without any adverse events.

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Data availability

No data sets were generated or analysed during the current study.

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Funding

This work was supported by a grants-in-aid of the SGH foundation (Kyoto, Japan) and by the Public Trust Surgery Research Fund (Tokyo, Japan) and Japan Society for the Promotion of Science (JSPS).

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Authors and Affiliations

  1. Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan

    Teruyuki Kobayashi, Tomoki Makino, Kotaro Yamashita, Takuro Saito, Koji Tanaka, Tsuyoshi Takahashi, Yukinori Kurokawa, Makoto Yamasaki, Kiyokazu Nakajima, Hidetoshi Eguchi & Yuichiro Doki

  2. Department of Pathology, Graduate School of Medicine, Osaka University, Osaka, Japan

    Eiichi Morii

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Contributions

TK and TM contributed to the design and analysis of the results and the writing of the manuscript. KY, TS, KT, TT, YK, MY, and KN contributed to the implementation of the research and analysis of the results. EM, HE, and YD contributed to the design and implementation of the research.

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Correspondence to Tomoki Makino.

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The authors declare no competing interests.

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The Osaka University Animal Experiments Committee had given ethical approval for the animal studies (ethical approval number: 27-061-002).

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Kobayashi, T., Makino, T., Yamashita, K. et al. APR-246 induces apoptosis and enhances chemo-sensitivity via activation of ROS and TAp73-Noxa signal in oesophageal squamous cell cancer with TP53 missense mutation. Br J Cancer 125, 1523–1532 (2021). https://doi.org/10.1038/s41416-021-01561-0

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