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IgA Fc-folate conjugate activates and recruits neutrophils to directly target triple-negative breast cancer cells

  • Preclinical study
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Abstract

Purpose

According to the American Cancer Society, 1 in 8 women in the U.S. will develop breast cancer, with triple-negative breast cancer (TNBC) comprising 15–20% of all breast cancer cases. TNBC is an aggressive subtype due to its high metastatic potential and lack of targeted therapy. Recently, folate receptor alpha (FRA) is found to be expressed on 80% of TNBC with high expression correlating with poor prognosis. In this study, we examined whether binding IgA Fc-folate molecules to FRA receptors on TNBC cells can elicit and induce neutrophils (PMNs), by binding their FcαR1 receptors, to destroy TNBC cells.

Methods

FRA was analyzed on TNBC cells and binding assays were performed using 3H-folate. Fc-folate was synthesized by linking Fc fragments of IgA via amine groups to folate. Binding specificity and antibody-dependent cellular cytotoxicity (ADCC) potential of Fc-folate to FcαR1 were confirmed by measuring PMN adhesion and myeloperoxidase (MPO) release in a cell-based ELISA. Fc-folate binding to FRA-expressing TNBC cells inducing PMNs to destroy these cells was determined using 51Cr-release and calcein-labeling assays.

Results

Our results demonstrate expression of FRA on TNBC cells at levels consistent with folate binding. Fc-folate binds with high affinity to FRA compared to whole IgA-folate and induces MPO release from PMN when bound to FcαR1. Fc-folate inhibited binding of 3H-folate to TNBC cells and induced significant cell lysis of TNBC cells when incubated in the presence of PMNs.

Conclusion

These findings support the hypothesis that an IgA Fc-folate conjugate can destroy TNBC cells by eliciting PMN-mediated ADCC.

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Acknowledgements

We are grateful to Dr. Stephen Ethier for kindly providing us with his SUM cell lines and Dr. Larry Matherly for kindly providing us with the iGROV1 cell line. We are also thankful to Dr. Larry Lum for all his help, insight, and advice throughout this study.

Funding

A portion of this work was supported by an award from the Barbara Ann Karmanos Cancer Institute Tumor Microenvironment Program and by an NIH STTR award (R41CA213548) funded by the National Cancer Institute.

Author information

Authors and Affiliations

  1. College of Osteopathic Medicine, Michigan State University, 4707 St. Antoine, Box 402, Detroit, MI, 48201, USA

    Eric D. Frontera & Rafa M. Khansa

  2. Department of Hematology/Oncology, University of Virginia Cancer Center, 1300 Jefferson Park Ave., I MSB 7191 West Complex I, Charlottesville, VA, 22903, USA

    Dana L. Schalk

  3. Wayne State University, 42 West Warren Avenue, Detroit, MI, 48202, USA

    Lauren E. Leakan

  4. Michael and Marian Ilitch Department of Surgery, Wayne State University School of Medicine, 4100 John R St., Mailcode HW08AO, Detroit, MI, 48201, USA

    Tracey J. Guerin-Edbauer, David H. Gorski & Cecilia L. Speyer

  5. Department of Oncology, Wayne State University School of Medicine, 4100 John R St., Detroit, MI, 48201, USA

    Manohar Ratnam

  6. Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, 4100 John R St., Detroit, MI, 48201, USA

    Manohar Ratnam & David H. Gorski

  7. Tumor Microenvironment Program, Barbara Ann Karmanos Cancer Institute, 4100 John R St., Detroit, MI, 48201, USA

    Cecilia L. Speyer

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  1. Eric D. Frontera
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  3. Dana L. Schalk
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  8. Cecilia L. Speyer
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Correspondence to Cecilia L. Speyer.

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All procedures performed in studies involving human participants were performed in accordance with the Declaration of Helsinki and have been approved following Expedited Review (IRB #123016MP4E) by the Chairperson for the Wayne State University Institutional Review Board (MP4).

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Frontera, E.D., Khansa, R.M., Schalk, D.L. et al. IgA Fc-folate conjugate activates and recruits neutrophils to directly target triple-negative breast cancer cells. Breast Cancer Res Treat 172, 551–560 (2018). https://doi.org/10.1007/s10549-018-4941-5

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  • DOI: https://doi.org/10.1007/s10549-018-4941-5

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