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Therapeutic bispecific T-cell engager antibody targeting the intracellular oncoprotein WT1

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.

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Figure 1: ESK1-BiTE induces T-cell activation and cytotoxicity in WT1+/HLA A*02:01+ tumor cells.
Figure 2: ESK1-BiTE effectively treats SET-2 AML in NSG mice.
Figure 3: Therapy of primary ALL and mesothelioma with ESK1-BiTE.
Figure 4: ESK1-BiTE induces secondary T-cell responses to epitopes other than WT1-RMF in the context of HLA-A*02:01 molecules.
Figure 5: Generation of long-lived, cytotoxic effector cells.
Figure 6: ESK1-BiTE induces secondary T-cell responses to other than WT1-RMF epitopes in the context of HLA A*02:01 molecules.

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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.

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Authors

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

Correspondence to David A Scheinberg.

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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.

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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

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