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Activation of CEACAM1 with an agonistic monoclonal antibody results in inhibition of melanoma cells

Cancer Gene Therapy volume 29, pages 1676–1685 (2022)Cite this article

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A Correction to this article was published on 03 February 2023

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Abstract

Inhibitory receptors (IRs), such as the carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), are cell surface molecules expressed on both normal epithelial, endothelial, and hematopoietic cells and on neoplastic cells. IRs are usually used by cancer cells to inhibit immune cell functions. Thus, CEACAM1 positive tumor cells can interact homophilically with CEACAM1 expressed on T and NK cells to inhibit their antibody-dependent cell-mediated cytotoxicity (ADCC). In this study, we investigated the effect of agonistic/activating anti-CEACAM1 monoclonal antibody (mAb) on melanoma cell lines in vitro and in vivo, following our hypothesis that activation of CEACAM1 on melanoma cells by distinct mAbs may induce inhibition of cancer cell proliferation and/or their death. To address this, we established an activating anti-CEACAM1 mAb (CCM5.01) and characterized its binding to the CEACAM1 receptor. Using this mAb, we assessed the expression of CEACAM1 on four different human melanoma cell lines by western blot and flow cytometry and determined its effect on cell viability in vitro by MTT assay. Furthermore, we evaluated the mAb mechanism of action and found that binding of CEACAM1 with CCM5.01 induced SHP1 phosphorylation and p53 activation resulting in melanoma cell apoptosis. For in vivo studies, a xenograft model of melanoma was performed by injection of Mel-14 cells subcutaneously (s.c.) in SCID/Beige mice followed by intraperitoneal (i.p.) injection of CCM5.01 or of IgG1 isotype control every other day. CCM5.01 treated mice showed a slight but not significant decrease in tumor weight in comparison to the control group. Based on the obtained data, we suggest that activating CEACAM1 on melanoma cells might be a promising novel approach to fight cancers expressing this IR.

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Fig. 1: Binding characteristics of CCM5.01 and CCM5.19 mAbs.
Fig. 2: CEACAM1 activation reduces melanoma cell viability and induces apoptosis.
Fig. 3: CCM5.01 mAb binding of CEACAM1 on Mel-14 cell line induces p53 and SHP1 phosphorylation thus mediating a decrease in cell survival.
Fig. 4: Evaluation of anti-tumor efficacy of CCM5.01 mAb in Mel-14 xenografted SCID/Beige mice.

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Data availability

The datasets used and/or analyzed during this study are available from the corresponding author on request.

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Acknowledgements

The authors thank Ofra Moshel, PhD (The Core Research Facility, The Institute of Drug Research, The Hebrew University of Jerusalem) who provided advice and technical help for FC and MST experiments and analyses.

Funding

This work was partially funded by grants from Aimwell Charitable Trust (UK), INTEGRA holdings, the Israel Cancer Association (No. 20210024) to FL-S, and the Israeli Scholarship Education Foundation (ISEF) to IZ.

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Author notes

  1. These authors contributed equally: Bernhard B. Singer, Francesca Levi-Schaffer.

Authors and Affiliations

  1. Pharmacology & Experimental Therapeutics Unit, Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel

    Ilan Zaffran, Nadine Landolina, Pratibha Gaur & Francesca Levi-Schaffer

  2. Center for Proteomics, Faculty of Medicine, University of Rijeka, Rijeka, Croatia

    Tihana Lenac Rovis & Stipan Jonjic

  3. The Lautenberg Center for General and Tumor Immunology, The Hebrew University Hadassah Medical School, IMRIC, Jerusalem, Israel

    Ofer Mandelboim

  4. Institute of Anatomy, Medical Faculty, University Duisburg-Essen, Essen, Germany

    Bernhard B. Singer

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Contributions

Designing research studies: FL-S. Conducting experiments: IZ, NL, PG, TLR. Acquiring data: IZ, NL, TLR, SJ, BBS. Analyzing data: IZ, NL, PG, TLR, SJ, BBS, FL-S. Providing reagents: OM, SJ, BBS, FL-S. Writing the manuscript: IZ, FL-S, BBS. Revising the manuscript: IZ, BBS, FL-S.

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Correspondence to Francesca Levi-Schaffer.

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Zaffran, I., Landolina, N., Gaur, P. et al. Activation of CEACAM1 with an agonistic monoclonal antibody results in inhibition of melanoma cells. Cancer Gene Ther 29, 1676–1685 (2022). https://doi.org/10.1038/s41417-022-00486-x

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