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Combined blockade of mTOR and p21-activated kinases pathways prevents tumour growth in KRAS-mutated colorectal cancer

British Journal of Cancer volume 129, pages 1071–1082 (2023)Cite this article

Subjects

Abstract

Background

The identification of novel therapeutic strategies for metastatic colorectal cancer (mCRC) patients harbouring KRAS mutations represents an unmet clinical need. In this study, we aimed to clarify the role of p21-activated kinases (Paks) as therapeutic target for KRAS-mutated CRC.

Methods

Paks expression and activation levels were evaluated in a cohort of KRAS-WT or -mutated CRC patients by immunohistochemistry. The effects of Paks inhibition on tumour cell proliferation and signal transduction were assayed by RNAi and by the use of three pan-Paks inhibitors (PF-3758309, FRAX1036, GNE-2861), evaluating CRC cells, spheroids and tumour xenografts’ growth.

Results

Paks activation positively correlated with KRAS mutational status in both patients and cell lines. Moreover, genetic modulation or pharmacological inhibition of Paks led to a robust impairment of KRAS-mut CRC cell proliferation. However, Paks prolonged blockade induced a rapid tumour adaptation through the hyper-activation of the mTOR/p70S6K pathway. The addition of everolimus (mTOR inhibitor) prevented the growth of KRAS-mut CRC tumours in vitro and in vivo, reverting the adaptive tumour resistance to Paks targeting.

Conclusions

In conclusion, our results suggest the simultaneous blockade of mTOR and Pak pathways as a promising alternative therapeutic strategy for patients affected by KRAS-mut colorectal cancer.

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Fig. 1: Expression and activation of p21-activated kinases in CRC patients and cell lines.
Fig. 2: Effect of PAKs downregulation and pharmacological inhibition on cell proliferation and signalling activation in CRC cell lines.
Fig. 3: Prolonged exposure of HCT116 and SW480 CRC cells to Paks inhibitors led to a rapid tumor adaptation.
Fig. 4: Dissecting the effects of PAK1, 2 and 4 or KRAS knockdown on the mTOR pathway activation and cell proliferation.
Fig. 5: Overcoming CRC tumour adaptation to PF309 through mTOR inhibition.
Fig. 6: The combination of mTOR and Paks’ pathway blockade prevents HCT116 xenografts growth.

Data availability

All relevant data are included in the manuscript and its Supplementary materials file or available from the corresponding author upon reasonable request.

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Funding

This work was supported by AIRC IG 21339 grant (RB); MFAG 21505 – 2018 grant (LF) and Lilly (LF); MFAG 27826 – 2022 grant (AS). DE was supported by AIRC fellowship for Italy – 26795.

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Author notes

  1. These authors contributed equally: Stefania Belli, Ada Pesapane.

Authors and Affiliations

  1. Department of Clinical Medicine and Surgery, University of Naples “Federico II”, 80131, Naples, Italy

    Stefania Belli, Ada Pesapane, Alberto Servetto, Daniela Esposito, Fabiana Napolitano, Claudia Maria Ascione, Alessandra Allotta, Luigi Formisano & Roberto Bianco

  2. CEINGE Biotecnologie Avanzate S.C.aR.L., Via G. Salvatore 486, 80145, Naples, Italy

    Nicola Zambrano

  3. Pathology Unit, University of Campania “L. Vanvitelli”, Naples, Italy

    Federica Zito Marino & Renato Franco

  4. Department of Precision Medicine, University of Campania “L. Vanvitelli”, Naples, Italy

    Teresa Troiani

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Contributions

Conceptualisation: SB, AP, LF and RB. Methodology: SB, AP, LF and RB. Writing—original draft preparation: SB, AP, AS LF. Investigation: SB, AP, DE, AA, FZM. Writing—review and editing: SB, AP, AS, DE, PC, CMA, AA, NZ, FZM, RF, TT, LF and RB. Supervision: LF and RB. Funding acquisition: AS, LF and RB. All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Luigi Formisano or Roberto Bianco.

Ethics declarations

Competing interests

AS reports honoraria from Eli Lilly, MSD, and Janssen and travel support from Bristol-Myers Squibb and AstraZeneca. LF declares the following competing interests: consultant and advisory board for Seagen, Amgen, BMS, MSD, Jansen and Pierre Fabre Pharma. RB declares the following competing interests: consultant and advisory board for BMS, MSD, Pfizer, AstraZeneca, Lilly and Novartis. The remaining authors declare no competing interests.

Ethics approval and consent to participate

CRC samples were retrospectively analysed at the Pathology Unit, University of Campania “L. Vanvitelli”. All patients agreed to participate in the study based on informed consent. Research Ethics Committee of the University of Campania “L. Vanvitelli” – AORN “Ospedale dei Colli” approved the study (reference n.0019530/2016). The study was performed in accordance with the Declaration of Helsinki.

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Belli, S., Pesapane, A., Servetto, A. et al. Combined blockade of mTOR and p21-activated kinases pathways prevents tumour growth in KRAS-mutated colorectal cancer. Br J Cancer 129, 1071–1082 (2023). https://doi.org/10.1038/s41416-023-02390-z

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