{"doi":"10.1101/2024.05.23.595624","title":"Targeting both GD2 and B7-H3 using bispecific antibody improves tumor selectivity for GD2-positive tumors","abstract":"Abstract Objectives Disialoganglioside 2 (GD2), overexpressed by cancers such as melanoma and neuroblastoma, is a tumor antigen for targeted therapy. The delivery of conventional IgG antibody technologies targeting GD2 is limited clinically by its co-expression on nerves that contributes to toxicity presenting as severe neuropathic pain. To improve the tumor selectivity of current GD2-targeting approaches, a next-generation bispecific antibody targeting GD2 and B7-H3 (CD276) was generated. Methods Differential expression of human B7-H3 (hB7-H3) was transduced into GD2 + B78 murine melanoma cells and confirmed by flow cytometry. We assessed the avidity and selectivity of our GD2-B7-H3 targeting bispecific antibodies (INV34-6, INV33-2, and INV36-6) towards GD2 + /hB7-H3 - B78 cells relative to GD2 + /hB7-H3 + B78 cells using flow cytometry and competition binding assays, comparing results an anti-GD2 antibody (dinutuximab, DINU). The bispecific antibodies, DINU, and a non-targeted bispecific control (bsAb CTRL) were conjugated with deferoxamine for radiolabeling with Zr-89 (t 1/2 = 78.4 h). Using positron emission tomography (PET) studies, we evaluated the in vivo avidity and selectivity of the GD2-B7-H3 targeting bispecific compared to bsAb CTRL and DINU using GD2 + /hB7-H3 + and GD2 + /hB7-H3 - B78 tumor models. Results Flow cytometry and competition binding assays showed that INV34-6 bound with high avidity to GD2 + /hB7-H3 + B78 cells with high avidity but not GD2 + /hB7-H3 + B78 cells. In comparison, no selectivity between cell types was observed for DINU. PET in mice bearing the GD2 + /hB7-H3 - and GD2 + /hB7-H3 + B78 murine tumor showed similar biodistribution in normal tissues for [ 89 Zr]Zr-Df-INV34-6, [ 89 Zr]Zr-Df-bsAb CTRL, and [ 89 Zr]Zr-Df-DINU. Importantly, [ 89 Zr]Zr-Df-INV34-6 tumor uptake was selective to GD2 + /hB7-H3 + B78 over GD2 + /hB7-H3 - B78 tumors, and substantially higher to GD2 + /hB7-H3 + B78 than the non-targeted [ 89 Zr]Zr-Df-bsAb CTRL control. [ 89 Zr]Zr-Df-DINU displayed similar uptake in both GD2 + tumor models, with uptake comparable to [ 89 Zr]Zr-Df-INV34-6 in the GD2 + /hB7-H3 + B78 model. Conclusion The GD2-B7-H3 targeting bispecific antibodies successfully improved selectivity to cells expressing both antigens. This approach should address the severe toxicities associated with GD2-targeting therapies by reducing off-tumor GD2 binding in nerves. Continued improvements in bispecific antibody technologies will continue to transform the therapeutic biologics landscape. Graphical Abstract","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":486170,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9577,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":339889,"name":"Amy K. Erbe","orcid":"0000-0003-1175-4062","position":1,"is_corresponding":false},{"id":1329961,"name":"Nathan B Clemons","orcid":null,"position":2,"is_corresponding":false},{"id":705266,"name":"Arika S. Feils","orcid":"0000-0002-9349-4534","position":3,"is_corresponding":false},{"id":1251729,"name":"Yadira Medina-Guevara","orcid":"0000-0002-1700-5436","position":4,"is_corresponding":false},{"id":718756,"name":"Justin J. Jeffery","orcid":null,"position":5,"is_corresponding":false},{"id":244586,"name":"Todd E. Barnhart","orcid":"0000-0002-9981-2150","position":6,"is_corresponding":false},{"id":244590,"name":"Jonathan W. Engle","orcid":"0000-0002-3399-7228","position":7,"is_corresponding":false},{"id":339504,"name":"Paul M. Sondel","orcid":"0000-0002-0981-8875","position":8,"is_corresponding":false},{"id":621689,"name":"Reinier Hernandez","orcid":"0000-0002-0729-2179","position":9,"is_corresponding":false},{"id":244583,"name":"Zachary T. Rosenkrans","orcid":"0000-0002-7958-3645","position":0,"is_corresponding":true}],"reference_count":35,"raw_metadata":null,"created_at":"2026-07-19T02:07:57.049469Z","pmid":"38853889","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}