title: T lymphocytes expressing a CD16 signaling receptor exert antibody-dependent cancer cell killing.
abstract: To expand applications for T-cell-based immunotherapy in cancer, we designed a receptor that binds the Fc portion of human immunoglobulins and delivers activation signals. The construct included the high-affinity CD16 (FCGR3A) V158 variant, CD8α hinge, and transmembrane domains, along with signaling domains from CD3ζ and 4-1BB (TNFRSF9), forming a chimeric receptor termed CD16V-BB-ζ. After retrovirus-mediated expression in human T cells, CD16V-BB-ζ bound humanized antibodies with higher affinity than a control receptor containing the more common F158 variant. Engagement of CD16V-BB-ζ provoked T-cell activation, exocytosis of lytic granules, and sustained proliferation, with a mean cell recovery after 4-week coculture with Daudi lymphoma cells and rituximab of nearly 70-fold relative to input cells. In contrast, unbound antibody alone produced no effect. CD16V-BB-ζ T cells specifically killed lymphoma cells and primary chronic lymphocytic leukemia cells in combination with rituximab at a low effector:target ratio, even when assayed on mesenchymal cells. Trastuzumab triggered CD16V-BB-ζ-mediated killing of HER2 (ERBB2)(+) breast and gastric cancer cells; similar results were obtained with an anti-GD2 antibody in neuroblastoma and osteosarcoma cells. Furthermore, coadministration of CD16V-BB-ζ T cells with immunotherapeutic antibodies exerted considerable antitumor activity in vivo. Signaling mediated by 4-1BB-CD3ζ induced higher T-cell activation, proliferation, and cytotoxicity than CD3ζ or FcεRIγ, and the receptor was expressed effectively after mRNA electroporation without viral vectors, facilitating clinical translation. Our results offer preclinical proof of concept for CD16V-BB-ζ as a universal, next-generation chimeric receptor with the potential to augment the efficacy of antibody therapies for cancer.
authors.affiliation: Authors' Affiliations: Departments of Pediatrics and Surgery, and Cancer Science Institute, National University of Singapore, Singapore; Department of Pediatrics, Niigata University, Niigata, Japan; and Department of Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee.
authors.forename: Ko
authors.initials: K
authors.lastname: Kudo
authors.forename: Chihaya
authors.initials: C
authors.lastname: Imai
authors.forename: Paolo
authors.initials: P
authors.lastname: Lorenzini
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kamiya
authors.forename: Koji
authors.initials: K
authors.lastname: Kono
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Davidoff
authors.forename: Wee Joo
authors.initials: WJ
authors.lastname: Chng
authors.forename: Dario
authors.initials: D
authors.lastname: Campana
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009369:Neoplasms / Q000276:immunology* / Q000628:therapy*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis / Q000276:immunology; D017452:Receptors, IgG / Q000096:biosynthesis / Q000276:immunology; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology*; D014183:Transplantation, Heterologous
chemical_list: D000911:Antibodies, Monoclonal; D016693:Receptors, Antigen, T-Cell, alpha-beta; D017452:Receptors, IgG