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The EGFR-specific antibody cetuximab combined with chemotherapy triggers immunogenic cell death

Nature Medicine volume 22, pages 624–631 (2016)Cite this article

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Abstract

Cetuximab is a monoclonal antibody that is effective in the treatment of metastatic colorectal cancer (mCRC). Cetuximab blocks epidermal growth factor receptor (EGFR)-ligand interaction and inhibits downstream RAS–ERK activation. However, only some activating mutations in RAS affect cetuximab efficacy, and it is not clear what else mediates treatment success. Here we hypothesized that cetuximab induces immunogenic cell death (ICD) that activates a potent antitumor response. We found that cetuximab, in combination with chemotherapy, fostered ICD in CRC cells, which we measured via the endoplasmic reticulum (ER) stress response and an increase in phagocytosis by dendritic cells. ICD induction depended on the mutational status of the EGFR signaling pathway and on the inhibition of the splicing of X-box binding protein 1 (XBP1), an unfolded protein response (UPR) mediator. We confirmed the enhanced immunogenicity elicited by cetuximab in a mouse model of human EGFR-expressing CRC. Overall, we demonstrate a new, immune-related mechanism of action of cetuximab that may help to tailor personalized medicine.

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Figure 1: Cetuximab triggers ER stress response and the translocation of CRT and ERp57 to the cell surface.
Figure 2: Cetuximab-treated tumor cells are readily phagocytosed by DCs.
Figure 3: Cetuximab-treated hEGFR-CT26 cells induce a potent anti-tumor response in vivo.
Figure 4: Treatment with F + C, in combination with PLX4032 and trametinib, induces phagocytosis of BRAFV600E Lim1215 cells.
Figure 5: Cetuximab does not block the splicing of XBP1 and cell adaptation in Lim1215 BRAFV600E cells.

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Acknowledgements

This work has been funded by grants of the Italian Association for Cancer Research (AIRC) (IG11863 to M.R.) and the Italian Ministry of Health (Ricerca finalizzata) (RF-2011-02348920 to M.R.). We thank S. Chiocca for editing the manuscript.

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Authors and Affiliations

  1. Department of Experimental Oncology, European Institute of Oncology, Milan, Italy

    Chiara Pozzi, Alessandro Cuomo, Ilaria Spadoni, Alessio Silvola, Tiziana Bonaldi, Pier Paolo Di Fiore, Giuseppe Penna & Maria Rescigno

  2. Division of Medical Oncology, Unit of Gastrointestinal and Neuroendocrine Tumors, European Institute of Oncology, Milan, Italy

    Elena Magni, Paola Simona Ravenda & Maria Giulia Zampino

  3. IFOM, Fondazione Istituto FIRC (Fondazione Italiana per la Ricerca sul Cancro) di Oncologia Molecolare, Milan, Italy

    Alexia Conte, Sara Sigismund, Pier Paolo Di Fiore & Alberto Bardelli

  4. Candiolo Cancer Institute–Fondazione Piemontese per l′Oncologia (FPO), Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Candiolo, Italy

    Carlotta Cancelliere

  5. Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy

    Pier Paolo Di Fiore & Maria Rescigno

  6. Department of Oncology, University of Torino, Torino, Italy

    Alberto Bardelli

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  1. Chiara Pozzi
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Contributions

C.P. did most of the experiments. C.P. and G.P. analyzed the data. M.R. designed the study, analyzed the data and wrote the paper, with editorial input from M.G.Z. and P.S.R. E.M., A.B. and P.P.D.F. participated in the study design. I.S. performed confocal analyses. T.B. designed the SILAC experiments. A.S. and A. Cuomo performed and analyzed the data from SILAC analysis. S.S. and A. Conte performed some of the biochemical studies. C.C. generated isogenic Lim1215 cell lines.

Corresponding author

Correspondence to Maria Rescigno.

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The authors declare no competing financial interests.

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Supplementary Text and Figures

Supplementary Figures 1–8 and Supplementary Tables 3–4 (PDF 1210 kb)

Supplementary Table 1

SILAC analysis (XLSX 315 kb)

Supplementary Table 2

SILAC analysis (XLSX 224 kb)

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Pozzi, C., Cuomo, A., Spadoni, I. et al. The EGFR-specific antibody cetuximab combined with chemotherapy triggers immunogenic cell death. Nat Med 22, 624–631 (2016). https://doi.org/10.1038/nm.4078

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