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Translational Therapeutics

SIRT1 is an actionable target to restore p53 function in HPV-associated cancer therapy

British Journal of Cancer volume 129, pages 1863–1874 (2023)Cite this article

Subjects

Abstract

Background

Our aim was to evaluate the efficacy and anti-cancer action of a precision medicine approach involving a novel SIRT1-dependent pathway that, when disrupted, leads to the restoration of a functional p53 in human papillomavirus (HPV)-transformed cells.

Methods

The anticancer potential of inhibiting SIRT1 was evaluated by examining the effects of the specific SIRT1 inhibitor EX527 (also known as Selisistat) or genetic silencing, either individually or in conjunction with standard chemotherapeutic agents, on a range of HPV+ cancer cells and a preclinical mouse model of HPV16-induced cancer.

Results

We show that SIRT1 inhibition restores a transcriptionally active K382-acetylated p53 in HPV+ but not HPV- cell lines, which in turn promotes G0/G1 cell cycle arrest and inhibits clonogenicity specifically in HPV+ cells. Additionally, EX527 treatment increases the sensitivity of HPV+ cells to sublethal doses of standard genotoxic agents. The enhanced sensitivity to cisplatin as well as p53 restoration were also observed in an in vivo tumorigenicity assay using syngeneic C3.43 cells harbouring an integrated HPV16 genome, injected subcutaneously into C57BL/6J mice.

Conclusions

Our findings uncover an essential role of SIRT1 in HPV-driven oncogenesis, which may have direct translational implications for the treatment of this type of cancer.

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Fig. 1: SIRT1 gene expression and status in HPV+ vs HPV- head and neck squamous cell carcinoma (HNSCC).
Fig. 2: Pharmacological and genetic inhibition of SIRT1 activity restores Ac-p53 (K382) protein expression.
Fig. 3: Restoration of a functional Ac-p53 (K382) inhibits the growth of HPV+ cells.
Fig. 4: SIRT1 inhibition enhances the sensitivity of HPV+ cells to genotoxic agents.
Fig. 5: EX527 enhances cisplatin cytotoxic activity in an in vivo syngeneic mouse model of HPV16-induced cancer.
Fig. 6: Schematic model depicting the molecular circuitry governing p53 functions via SIRT1-mediated acetylation in HPV+ cells.

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Data availability

All data generated or analysed during this study are included either in this article or in additional files, and the links to public datasets are reported in the manuscript.

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Acknowledgements

We thank Marcello Arsura for critically reviewing the manuscript. NOKE6/E7 cells were kindly provided by Martina Niebler and Frank Rösl, German Cancer Research Center (DKFZ)-Heidelberg and C3.43 cells by Martin Kast, University of Southern California in Los Angeles. We are also grateful to Michela Salvo from the Histology Research Core Facility at University of Piemonte Orientale (Novara, Italy) for technical support in tissue processing and histological analysis, Nausicaa Clemente from the Department of Health Sciences at University of Piemonte Orientale (Novara, Italy) for technical support to perform in vivo experiments, and Valeria Caneparo at CAAD, Center for Translational Research and Autoimmune and Allergic Disease, for assistance in SA-β-gal analysis.

Funding

This work was supported by the Italian Ministry for University and Research-MIUR (20178ALPCM-PRIN 2017 to MG), the AGING Project–Department of Excellence–DIMET, University of Piemonte Orientale, Associazione Italiana per la Ricerca sul Cancro-AIRC (25767-IG 2021 to MG), Next Generation EU—PNRR M6C2—Investimento 2.1 Valorizzazione e potenziamento della ricerca biomedica del SSN (PNRR-MAD-2022-12376570 to MG), and by the Italian Ministry of Foreign Affairs and International Cooperation (PGRBR22GR03 to AV). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

Authors and Affiliations

  1. Virology Unit, Department of Translational Medicine, Eastern Piedmont University, Novara, Italy

    Irene Lo Cigno, Federica Calati, Carlo Girone, Cinzia Borgogna & Marisa Gariglio

  2. HPV Unit, UOSD Tumor Immunology and Immunotherapy, IRCCS Regina Elena National Cancer Institute, Rome, Italy

    Aldo Venuti

  3. Pathology Unit, Department of Health Sciences, Eastern Piedmont University, Novara, Italy

    Renzo Boldorini

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  1. Irene Lo Cigno
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Contributions

ILC: Conceptualisation, data curation, investigation, methodology, project administration, software, validation, writing–original draft. FC: Conceptualisation, data curation, investigation, methodology, writing–original draft. CG: Data curation, investigation, methodology. CB: Formal analysis, project administration, supervision, writing–review and editing. AV: Resources, validation, writing–review and editing. RB: Investigation, validation, writing–review and editing. MG: Conceptualisation, supervision, funding acquisition, writing–original draft, writing–review and editing.

Corresponding author

Correspondence to Marisa Gariglio.

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Competing interests

ILC, FC and MG report the Italian Patent No. 102020000023281 issued on October 2nd, 2020 (“SIRT1 INHIBITOR OR ANTAGONIST FOR USE IN PREVENTING AND/OR TREATING AN HPV-INDUCED TUMOUR”). This patent is pending for the international patent application approval (PCT/IB2021/059055). All the other authors declare no competing financial interests.

Ethics approval

The mice used for the in vivo tumorigenicity assays were housed in our animal facilities in accordance with “The Guide for the Care and Use of Laboratory Animals”, and the experimentation was approved by the Italian Ministry of Health (Agreement No. 219/2020-PR).

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Lo Cigno, I., Calati, F., Girone, C. et al. SIRT1 is an actionable target to restore p53 function in HPV-associated cancer therapy. Br J Cancer 129, 1863–1874 (2023). https://doi.org/10.1038/s41416-023-02465-x

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