Mikkelsen, KasperHarwood, Seandean LykkeCompte, MartaMerino, NekaneMølgaard, KasperLykkemark, SimonÁlvarez Méndez, Ana MaríaBlanco, Francisco J.Álvarez Vallina, Luis2024-10-022024-10-022019-07-31Mikkelsen K, Harwood SL, Compte M, Merino N, Mølgaard K, Lykkemark S, Alvarez-Mendez A, Blanco FJ and Álvarez-Vallina L (2019) Carcinoembryonic Antigen (CEA)-Specific 4-1BB-Costimulation Induced by CEA-Targeted 4-1BB-Agonistic Trimerbodies. Front. Immunol. 10:1791. doi: 10.3389/fimmu.2019.0179110.3389/fimmu.2019.01791https://hdl.handle.net/20.500.14352/1085404-1BB (CD137) is an inducible costimulatory receptor that promotes expansion and survival of activated T cells; and IgG-based 4-1BB-agonistic monoclonal antibodies exhibited potent antitumor activity in clinical trials. However, the clinical development of those antibodies is restricted by major off-tumor toxicities associated with FcgR interactions. We have recently generated an EGFR-targeted 4-1BB-agonistic trimerbody that demonstrated strong antitumor activity and did not induce systemic inflammatory cytokine secretion and hepatotoxicity associated with first-generation 4-1BB agonists. Here, we generate a bispecific 4-1BB-agonistic trimerbody targeting the carcinoembryonic antigen (CEA) that is highly expressed in cancers of diverse origins. The CEA-targeted anti-4-1BB-agonistic trimerbody consists of three 4-1BB-specific single-chain fragment variable antibodies and three anti-CEA single-domain antibodies positioned around a murine collagen XVIII-derived homotrimerization domain. The trimerbody was produced as a homogenous, non-aggregating, soluble protein purifiable by standard affinity chromatographic methods. The purified trimerbody was found to be trimeric in solution, very efficient at recognizing 4-1BB and CEA, and potently costimulating T cells in vitro in the presence of CEA. Therefore, trimerbody-based tumor-targeted 4-1BB costimulation is a broadly applicable and clinically feasible approach to enhance the costimulatory environment of disseminated tumor lesions.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/journal articlehttps://doi.org/10.3389/fimmu.2019.01791https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2019.01791/fullopen access61cancer immunotherapyimmunostimulatory antibodiescostimulation4-1BB4-1BB agoniststrimerbodiesCEACEA-targeted 4-1BB agonistsCiencias Biomédicas32 Ciencias Médicas