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Chimeric antigen receptors with human scFvs preferentially induce T cell anti-tumor activity against tumors with high B7H6 expression

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Abstract

B7H6 is emerging as a promising tumor antigen that is known to be expressed on a wide array of tumors and is reported to stimulate anti-tumor responses from the immune system. As such, B7H6 presents a good target for tumor-specific immunotherapies. B7H6-specific chimeric antigen receptors (CAR) based on a murine antibody showed successful targeting and elimination of tumors expressing B7H6. However, mouse single chain variable fragments (scFvs) have the potential to induce host anti-CAR responses that may limit efficacy, so human scFvs specific for B7H6 were selected by yeast surface display. In this study, we validate the functionality of these human scFvs when formatted into chimeric antigen receptors. The data indicate that T cells expressing these B7H6-specific human scFvs as CARs induced potent anti-tumor activity in vitro and in vivo against tumors expressing high amounts of B7H6. Importantly, these human scFv-based CARs are sensitive to changes in B7H6 expression which may potentially spare non-tumor cells that express B7H6 and provides the foundation for future clinical development.

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Abbreviations

B6:

C57BL/6

CAR:

Chimeric antigen receptor

CYP:

Cytoplasmic

H:

Hinge

MFI:

Mean fluorescent intensity

NK:

Natural killer

scFv:

Single-chain fragment variable

TM:

Transmembrane

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Acknowledgements

The authors thank the staff of the Center for Comparative Medicine Research for assistance with animal care and the Shared Molecular Biology Resource at the Norris Cotton Cancer Center for sequencing. The authors also thank the National Cancer Institute Biological Resource Branch for recombinant human IL-2.

Funding

This work was supported by funds from The Elmer R. Pfefferkorn and Allan U. Munck Education and Research Fund, the Center of Biomedical Research Excellence (8P30GM103415), and the Center for Synthetic Immunity.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, The Geisel School of Medicine at Dartmouth, One Medical Center Drive, Lebanon, NH, 03756, USA

    Albert T. Gacerez, Casey K. Hua, Margaret E. Ackerman & Charles L. Sentman

  2. Center for Synthetic Immunity, The Geisel School of Medicine at Dartmouth, Lebanon, NH, 03756, USA

    Albert T. Gacerez & Charles L. Sentman

  3. Thayer School of Engineering, Dartmouth College, Hanover, NH, 03755, USA

    Casey K. Hua & Margaret E. Ackerman

Authors
  1. Albert T. Gacerez
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  4. Charles L. Sentman
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Contributions

All authors have contributed to the design and interpretation of experiments and the writing and editing of the manuscript. Albert T. Gacerez was responsible for acquisition of data.

Corresponding author

Correspondence to Charles L. Sentman.

Ethics declarations

Conflict of interest

All authors have patent applications in antibody-based therapies and cell engineering. These conflicts are managed under the policies of Dartmouth College.

Ethical Approval and ethical standards

All procedures involving mice were in accordance with the ethical standards set forth by the Dartmouth College’s Institution Animal Care and Use Committee.

Animal source

C57BL/6 (B6) mice from Charles River.

Cell line authentication

ATCC authenticated K562 on August 25, 2016 and SH-SY5Y on October 27, 2016. Murine cell lines were obtained from 2001 to 2006. RMA cell lines were obtained from Michael Bennett (UT Southwestern Medical Center) and B16F10 cell lines were obtained from Richard Barth (Geisel School of Medicine at Dartmouth).

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Gacerez, A.T., Hua, C.K., Ackerman, M.E. et al. Chimeric antigen receptors with human scFvs preferentially induce T cell anti-tumor activity against tumors with high B7H6 expression. Cancer Immunol Immunother 67, 749–759 (2018). https://doi.org/10.1007/s00262-018-2124-1

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  • DOI: https://doi.org/10.1007/s00262-018-2124-1

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