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Early response in phosphorylation of ribosomal protein S6 is associated with sensitivity to trametinib in colorectal cancer cells

Abstract

Mutations in RAS or BRAF are associated with poor prognosis and resistance to epidermal growth factor receptor (EGFR)-targeted therapy in colorectal cancer (CRC). Despite their common ability to activate downstream genes such as MEK and ERK, the therapeutic benefit of MEK inhibitors for patients with RAS/BRAF mutant CRC is limited, highlighting the need for biomarkers to predict the efficacy of MEK inhibition. Previously, we reported that a change in phosphorylation of ribosomal protein S6 (pS6) after MEK inhibition was significantly associated with sensitivity to MEK inhibition in gastric cancer cells. Here, we investigated the value of the response in pS6 for predicting the efficacy of trametinib, a MEK inhibitor, in patients with RAS/BRAF mutant CRC using patient-derived CRC organoids. We found that a subset of CRC cell lines and organoids were sensitive to trametinib. The change in phosphorylated ERK, a downstream molecule of the RAS/RAF/MEK pathway, was not significantly associated with trametinib sensitivity. On the other hand, only those with sensitivity showed a reduction of pS6 levels in response to trametinib. The change in pS6 after trametinib treatment was detectable by Western blotting, immunohistochemistry or immunocytochemistry. We also demonstrated an impact of MEK inhibition on pS6 in vivo using a xenograft model. Our data suggest that, in combination with patient-derived organoids, immunostaining-based detection of pS6 could be useful for prediction of trametinib sensitivity.

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Fig. 1: Suppression of pS6 after MEK inhibition is related to sensitivity to MEK inhibition in a subset of RAS/BRAF mutant CRC cell lines.
Fig. 2: Correlation between efficacy of MEK inhibition and pS6 response to treatment in CRC cells with acquired trametinib resistance.
Fig. 3: Establishment of patient-derived CRC organoids.
Fig. 4: Response of the pS6 level after MEK inhibition is also related to trametinib sensitivity in CRC organoids.

Data availability

The data that support the findings of this study are available from the corresponding authors, upon reasonable request.

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Acknowledgements

We would like to thank Ms Mami Kimoto for their excellent assistance with experiments. We are also grateful to Hans Clevers, Tomohiro Mizutani, Else Driehuis and Stieneke van den Brink for critical advice on organoid experiments.

Funding

This work was supported at the Discretion of the President of Oita University (2019) and partly by JSPS KAKENHI Grant Number 18K15283.

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Conceptualization: YH, YT; Formal analysis and investigation: YH, YK; Writing—original draft preparation: YH, YT; Writing—review and editing: DK, SK, NH, CN, TU, TH, KM, KH, TO, MK; Funding acquisition: YH; Resources: TH, TA, YU, HS, TE; Supervision: MI, MM, KM. All authors have approved the final version of the paper.

Corresponding authors

Correspondence to Yuka Hirashita or Yoshiyuki Tsukamoto.

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The authors declare no competing interests.

Ethical approval

This study was approved by the ethics committee of Oita University Faculty of Medicine (approval number: 1541).

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Hirashita, Y., Tsukamoto, Y., Kudo, Y. et al. Early response in phosphorylation of ribosomal protein S6 is associated with sensitivity to trametinib in colorectal cancer cells. Lab Invest (2021). https://doi.org/10.1038/s41374-021-00590-w

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