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Sensitizing solid tumors to CAR-mediated cytotoxicity using synthetic antigens

Gamboa, L.; Zamat, A. H.; Vanover, D.; Thiveaud, C. A.; Peck, H. E.; Phuengkham, H.; Sivakumar, A.; Harris, A. M.; Dahotre, S. N.; Su, F.-Y.; Santangelo, P. J.; Kwong, G. A.

2021-12-12 bioengineering
10.1101/2021.12.11.472238 bioRxiv
Show abstract

CAR T cell immunotherapy relies on CAR targeting of tumor-associated antigens, yet heterogenous antigen expression, interpatient variation, and off-tumor expression by healthy cells remain barriers. Here, we develop synthetic antigens to sensitize solid tumors for recognition and elimination by CAR T cells. Unlike tumor-associated antigens, we design synthetic antigens that are orthogonal to endogenous proteins to eliminate off-tumor targeting and that have a small genetic footprint to facilitate efficient tumor delivery to tumors by viral vectors. Using the RSV-F camelid single-domain antibody (VHH) as a synthetic antigen, we show that adoptive transfer of VHH CAR T cells to mice bearing VHH expressing tumors reduced tumor burden in multiple syngeneic mouse models of cancer, improved survival, induced epitope spread, and protected against tumor rechallenge. Our work supports in situ delivery of synthetic antigens to treat antigen low or negative tumors with CAR T cells.

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