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Dual targeting ovarian cancer by Muc16 CAR T cells secreting a bispecific T cell engager antibody for an intracellular tumor antigen WT1

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Abstract

Epithelial ovarian cancer is the most lethal of gynecological cancers. The therapeutic efficacy of chimeric antigen receptor (CAR) T cell directed against single antigens is limited by the heterogeneous target antigen expression in epithelial ovarian tumors. To overcome this limitation, we describe an engineered cell with both dual targeting and orthogonal cytotoxic modalities directed against two tumor antigens that are highly expressed on ovarian cancer cells: cell surface Muc16 and intracellular WT1. Muc16-specific CAR T cells (4H11) were engineered to secrete a bispecific T cell engager (BiTE) constructed from a TCR mimic antibody (ESK1) reactive with the WT1-derived epitope RMFPNAPYL (RMF) presented by HLA-A2 molecules. The secreted ESK1 BiTE recruited and redirected other T cells to WT1 on the tumor cells. We show that ESK1 BiTE-secreting 4H11 CAR T cells exhibited enhanced anticancer activity against cancer cells with low Muc16 expression, compared to 4H11 CAR T cells alone, both in vitro and in mouse tumor models. Dual orthogonal cytotoxic modalities with different specificities targeting both surface and intracellular tumor-associated antigens present a promising strategy to overcome resistance to CAR T cell therapy in epithelial ovarian cancer and other cancers.

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Funding

This work was supported by NIH RO1 CA55349, PO1 CA23766, P30 CA008748 and R35 CA241894 to DAS, and NCI 1R50CA265328-01A1 to TD. REO was funded in part by the National Institutes of Health/National Cancer Institute Cancer Center Support Grant (P30 CA008748) and was funded in part by P30 CA008748.

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Authors and Affiliations

  1. Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, 10065, USA

    Sung Soo Mun, Jeremy Meyerberg, Leila Peraro, Tatyana Korontsvit, Thomas Gardner, Manish Malviya, Tao Dao & David A. Scheinberg

  2. Gynecologic Medical Oncology Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    Chrisann Kyi & Roisin E. O’Cearbhaill

  3. Weill Cornell Medical College, New York, NY, USA

    Chrisann Kyi, Roisin E. O’Cearbhaill & David A. Scheinberg

  4. Eureka Therapeutics, Inc., Emeryville, CA, USA

    Cheng Liu

Authors
  1. Sung Soo Mun
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  2. Jeremy Meyerberg
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  3. Leila Peraro
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  4. Tatyana Korontsvit
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  9. Cheng Liu
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  10. Tao Dao
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  11. David A. Scheinberg
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Contributions

TD and DAS conceived the idea and wrote the manuscript. SSM, TG, LP, and TD co-designed the study, analyzed data, and participated in the manuscript preparation. SSM, JPM, LP, TK, and MM performed experiments. CL designed and provided the ESK1-BITE and the related materials. REO and CK provided clinical samples, expertise in ovarian cancer study, and edited the manuscript.

Corresponding author

Correspondence to David A. Scheinberg.

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Conflict of interest

DAS is on a board of, has equity in, receives research funding from, has licensed technology with, or income from Lantheus, Sellas, Iovance, Arvinas, Clade, Pfizer, Actinium, Oncopep, Repertoire, Sapience, Atengen, CoImmune, and Eureka Therapeutics. TD is a consultant of Eureka Therapeutics. REO reports personal fees from Tesaro/GSK, Regeneron, Seattle Genetics, Fresenius Kabi, Gynecologic Oncology Foundation, Bayer, Curio/Onclive, R-Pharm, and Immunogen, other from Hitech Health, outside the submitted work. She is a non-compensated steering committee member for the PRIMA, Moonstone (Tesaro/GSK) and DUO-O (AstraZeneca) studies and non-compensated advisor for Carina Biotech. Her institute receives funding for clinical research from Bayer/Celgene/Juno, Tesaro/GSK, Merck, Ludwig Cancer Institute, Abbvie/StemCentrx, Regeneron, TCR2 Therapeutics, Atara Biotherapeutics, MarkerTherapeutics, Syndax Pharmaceuticals, Genmab/Seagen Therapeutics, Sellas Therapeutics, Genentech, Kite Pharma, and Gynecologic Oncology Foundation. LP is now an employee of Beam. TG is now an employee of Arsenal. All other authors declare no conflict or competing interests.

Ethical standards

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Memorial Sloan Kettering Cancer Center Institutional Review Board (IRB Human Subject Assurance Number FW00004998) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Mun, S.S., Meyerberg, J., Peraro, L. et al. Dual targeting ovarian cancer by Muc16 CAR T cells secreting a bispecific T cell engager antibody for an intracellular tumor antigen WT1. Cancer Immunol Immunother 72, 3773–3786 (2023). https://doi.org/10.1007/s00262-023-03529-w

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