Abstract
Intracellular tumor antigens presented on the cell surface in the context of human leukocyte antigen (HLA) molecules have been targeted by T cell–based therapies, but there has been little progress in developing small-molecule drugs or antibodies directed to these antigens. Here we describe a bispecific T-cell engager (BiTE) antibody derived from a T-cell receptor (TCR)-mimic monoclonal antibody (mAb) ESK1, which binds a peptide derived from the intracellular oncoprotein WT1 presented on HLA-A*02:01. Despite the very low density of the complexes at the cell surface, ESK1-BiTE selectively activated and induced proliferation of cytolytic human T cells that killed cells from multiple leukemias and solid tumors in vitro and in mice. We also discovered that in an autologous in vitro setting, ESK1-BiTE induced a robust secondary CD8 T-cell response specific for tumor-associated antigens other than WT1. Our study provides an approach that targets tumor-specific intracellular antigens without using cell therapy and suggests that epitope spreading could contribute to the therapeutic efficacy of this BiTE.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Coulie, P.G., Van den Eynde, B.J., van der Bruggen, P. & Boon, T. Tumour antigens recognized by T lymphocytes: at the core of cancer immunotherapy. Nat. Rev. Cancer 14, 135–146 (2014).
Morris, E. et al. Generation of tumor-specific T-cell therapies. Blood Rev. 20, 61–69 (2006).
Dao, T. et al. Targeting the intracellular WT1 oncogene product with a therapeutic human antibody. Sci. Transl. Med. 5, 176ra33 (2013).
Veomett, N. et al. Therapeutic efficacy of an Fc-enhanced TCR-like antibody to the intracellular WT1 oncoprotein. Clin. Cancer Res. 20, 4036–4046 (2014).
Dubrovsky, L. et al. A TCR-mimic antibody to WT1 bypasses tyrosine kinase inhibitor resistance in human BCR-ABL+ leukemias. Blood 123, 3296–3304 (2014).
Curran, K.J., Pegram, H.J. & Brentjens, R.J. Chimeric antigen receptors for T cell immunotherapy: current understanding and future directions. J. Gene Med. 14, 405–415 (2012).
Sadelain, M., Brentjens, R. & Rivière, I. The basic principles of chimeric antigen receptor design. Cancer Discov. 3, 388–398 (2013).
Frankel, S.R. & Baeuerle, P.A. Targeting T cells to tumor cells using bispecific antibodies. Curr. Opin. Chem. Biol. 17, 385–392 (2013).
Nagorsen, D. & Baeuerle, P.A. Immunomodulatory therapy of cancer with T cell-engaging BiTE antibody blinatumomab. Exp. Cell Res. 317, 1255–1260 (2011).
Brischwein, K. et al. Strictly target cell-dependent activation of T cells by bispecific single-chain antibody constructs of the BiTE class. J. Immunother. 30, 798–807 (2007).
Brentjens, R.J. et al. CD19-targeted T cells rapidly induce molecular remissions in adults with chemotherapy-refractory acute lymphoblastic leukemia. Sci. Transl. Med. 5, 177ra38 (2013).
Hoffmann, P. et al. Serial killing of tumor cells by cytotoxic T cells redirected with a CD19-/CD3-bispecific single-chain antibody construct. Int. J. Cancer 115, 98–104 (2005).
Mittendorf, E.A., Holmes, J.P., Ponniah, S. & Peoples, G.E. The E75 HER2/neu peptide vaccine. Cancer Immunol. Immunother. 57, 1511–1521 (2008).
Doubrovina, E. et al. Mapping of novel peptides of WT-1 and presenting HLA alleles that induce epitope-specific HLA-restricted T cells with cytotoxic activity against WT-1(+) leukemias. Blood 120, 1633–1646 (2012).
Pedersen, A.E. et al. Wild type p53-specific antibody and T-cell responses in cancer patients. J. Immunother. 34, 629–640 (2011).
Kessler, J.H. et al. Efficient identification of novel HLA-A(*)0201-presented cytotoxic T lymphocyte epitopes in the widely expressed tumor antigen PRAME by proteasome-mediated digestion analysis. J. Exp. Med. 193, 73–88 (2001).
Quintarelli, C. et al. High-avidity cytotoxic T lymphocytes specific for a new PRAME-derived peptide can target leukemic and leukemic-precursor cells. Blood 117, 3353–3362 (2011).
Dao, T. et al. Identification of a human cyclin D1-derived peptide that induces human cytotoxic CD4 T cells. PLoS One 4, e6730 (2009).
Peralbo, E., Alonso, C. & Solana, R. Invariant NKT and NK-like lymphocytes: Two different T cell subtypes that are differentially affected by aging. Exp. Gerontol. 42, 703–708 (2007).
Oka, Y. et al. WT1 peptide cancer vaccine for patients with hematopoietic malignancies and solid cancers. Scientific World Journal 7, 649–665 (2007).
Cheever, M.A. et al. The prioritization of cancer antigens: a national cancer institute pilot project for the acceleration of translational research. Clin. Cancer Res. 15, 5323–5337 (2009).
Chapuis, A.G. et al. Transferred WT1-reactive CD8+ T cells can mediate antileukemic activity and persist in post-transplant patients. Sci. Transl. Med. 5, 174ra27 (2013).
Sergeeva, A. et al. An anti-PR1/HLA-A2 T-cell receptor-like antibody mediates complement-dependent cytotoxicity against acute myeloid leukemia progenitor cells. Blood 117, 4262–4272 (2011).
Corbière, V. et al. Antigen spreading contributes to MAGE vaccination-induced regression of melanoma metastases. Cancer Res. 71, 1253–1262 (2011).
Hunder, N.N. et al. Treatment of metastatic melanoma with autologous CD4+ T cells against NY-ESO-1. N. Engl. J. Med. 358, 2698–2703 (2008).
Abès, R., Gélizé, E., Fridman, W.H. & Teillaud, J.L. Long-lasting antitumor protection by anti-CD20 antibody through cellular immune response. Blood 116, 926–934 (2010).
Selenko, N. et al. CD20 antibody (C2B8)-induced apoptosis of lymphoma cells promotes phagocytosis by dendritic cells and cross-priming of CD8+ cytotoxic T cells. Leukemia 15, 1619–1626 (2001).
Hilchey, S.P. et al. Rituximab immunotherapy results in the induction of a lymphoma idiotype-specific T-cell response in patients with follicular lymphoma: support for a “vaccinal effect” of rituximab. Blood 113, 3809–3812 (2009).
Ott, P.A., Hodi, F.S. & Robert, C. CTLA-4 and PD-1/PD-L1 blockade: new immunotherapeutic modalities with durable clinical benefit in melanoma patients. Clin. Cancer Res. 19, 5300–5309 (2013).
Tumeh, P.C. et al. PD-1 blockade induces responses by inhibiting adaptive immune resistance. Nature 515, 568–571 (2014).
Gubin, M.M. et al. Checkpoint blockade cancer immunotherapy targets tumour-specific mutant antigens. Nature 515, 577–581 (2014).
Tran, E. et al. Cancer immunotherapy based on mutation-specific CD4+ T cells in a patient with epithelial cancer. Science 344, 641–645 (2014).
Inoue, K. et al. Aberrant overexpression of the Wilms tumor gene (WT1) in human leukemia. Blood 89, 1405–1412 (1997).
Oka, Y. et al. WT1 peptide cancer vaccine for patients with hematopoietic malignancies and solid cancers. Scientific World Journal 7, 649–665 (2007).
Brischwein, K. et al. MT110: a novel bispecific single-chain antibody construct with high efficacy in eradicating established tumors. Mol. Immunol. 43, 1129–1143 (2006).
Fan, L. et al. Improving the efficiency of CHO cell line generation using glutamine synthetase gene knockout cells. Biotechnol. Bioeng. 109, 1007–1015 (2012).
Doubrovina, E. et al. Adoptive immunotherapy with unselected or EBV-specific T cells for biopsy-proven EBV+ lymphomas after allogeneic hematopoietic cell transplantation. Blood 119, 2644–2656 (2012).
Acknowledgements
The study was supported by US National Institutes of Health grant R01 CA 55349, P01 CA23766, MARF, P30 CA008748, Memorial Sloan Kettering Cancer Center's (MSKCC's) Experimental Therapeutics Center and the Lymphoma Foundation and Tudor and Glades funds. We thank D Levine, F. Dao and M. Mattar for their efforts and help in collecting clinical samples. We also thank the MSKCC Small-Animal Imaging Core Facility, R. Gejman for statistical analyses, T.-Y. Kuo for helpful discussions for Renilla transduction, A. Selvakumar and A. Yeh for their expert HLA typing.
Author information
Authors and Affiliations
Contributions
T.D., D.A.S. and R.J.O'R. designed the experiments, interpreted the data and wrote the manuscript. D.P., E.D. and M.D.d.M.G. participated in the design of some experiments. T.D., D.P., A.S., T.K., V.Z., N.V., L.D., M.C., V.P. and M.D.d.M.G. performed the experiments. Y.X., J.X., S.Y. and C.L. engineered T-BiTEs. D.A.S. is the principal investigator.
Corresponding author
Ethics declarations
Competing interests
T.D., L.D. and D.A.S. are inventors of technology described in this paper and licensed by Memorial Sloan Kettering Cancer Center to Novartis.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–9 (PDF 1945 kb)
Rights and permissions
About this article
Cite this article
Dao, T., Pankov, D., Scott, A. et al. Therapeutic bispecific T-cell engager antibody targeting the intracellular oncoprotein WT1. Nat Biotechnol 33, 1079–1086 (2015). https://doi.org/10.1038/nbt.3349
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nbt.3349
This article is cited by
-
T cell receptor therapeutics: immunological targeting of the intracellular cancer proteome
Nature Reviews Drug Discovery (2023)
-
Facile repurposing of peptide–MHC-restricted antibodies for cancer immunotherapy
Nature Biotechnology (2023)
-
Immunotherapy in hematologic malignancies: achievements, challenges and future prospects
Signal Transduction and Targeted Therapy (2023)
-
Dual targeting ovarian cancer by Muc16 CAR T cells secreting a bispecific T cell engager antibody for an intracellular tumor antigen WT1
Cancer Immunology, Immunotherapy (2023)
-
CD19-targeted BiTE expression by an oncolytic vaccinia virus significantly augments therapeutic efficacy against B-cell lymphoma
Blood Cancer Journal (2022)