title: Retargeting T cells to GD2 pentasaccharide on human tumors using Bispecific humanized antibody.
abstract: Anti-disialoganglioside GD2 IgG antibodies have shown clinical efficacy in solid tumors that lack human leukocyte antigens (e.g., neuroblastoma) by relying on Fc-dependent cytotoxicity. However, there are pain side effects secondary to complement activation. T-cell retargeting bispecific antibodies (BsAb) also have clinical potential, but it is thus far only effective against liquid tumors. In this study, a fully humanized hu3F8-BsAb was developed, in which the anti-CD3 huOKT3 single-chain Fv fragment (ScFv) was linked to the carboxyl end of the anti-GD2 hu3F8 IgG1 light chain, and was aglycosylated at N297 of Fc to prevent complement activation and cytokine storm. In vitro, hu3F8-BsAb activated T cells through classic immunologic synapses, inducing GD2-specific tumor cytotoxicity at femtomolar EC50 with >10⁵-fold selectivity over normal tissues, releasing Th1 cytokines (TNFα, IFNγ, and IL2) when GD2⁺ tumors were present. In separate murine neuroblastoma and melanoma xenograft models, intravenous hu3F8-BsAb activated T cells in situ and recruited intravenous T cells for tumor ablation, significantly prolonging survival from local recurrence or from metastatic disease. Hu3F8-BsAb, but not control BsAb, drove T cells and monocytes to infiltrate tumor stroma. These monocytes were necessary for sustained T-cell proliferation and/or survival and contributed significantly to the antitumor effect. The in vitro and in vivo antitumor properties of hu3F8-BsAb and its safety profile support its further clinical development as a cancer therapeutic, and provide the rationale for exploring aglycosylated IgG-scFv as a structural platform for retargeting human T cells.
authors.affiliation: Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, New York.
authors.forename: Hong
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, New York.
authors.forename: Ming
authors.initials: M
authors.lastname: Cheng
authors.affiliation: Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, New York.
authors.forename: Hongfen
authors.initials: H
authors.lastname: Guo
authors.affiliation: Department of Immunology, Memorial Sloan Kettering Cancer Center, New York, New York.
authors.forename: Yuedan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Immunology, Memorial Sloan Kettering Cancer Center, New York, New York.
authors.forename: Morgan
authors.initials: M
authors.lastname: Huse
authors.affiliation: Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, New York. cheungn@mskcc.org.
authors.forename: Nai-Kong V
authors.initials: NK
authors.lastname: Cheung
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000502:physiology; D005732:Gangliosides / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008545:Melanoma / Q000188:drug therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D009000:Monocytes / Q000276:immunology; D009447:Neuroblastoma / Q000188:drug therapy*; D057127:Single-Chain Antibodies / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D018033:Antibodies, Bispecific; D017252:CD3 Complex; D005732:Gangliosides; D007074:Immunoglobulin G; D057127:Single-Chain Antibodies; C019403:ganglioside, GD2