{"doi":"10.1101/2025.09.21.677627","title":"Bicistronic CAR T-cells Against CD70 & Active Integrin β2 Overcome Antigen Heterogeneity and Preserve Safety in Acute Myeloid Leukemia","abstract":"The surface antigen landscape of acute myeloid leukemia (AML) displays significant heterogeneity and overlap with healthy hematopoietic cells. This imparts a substantial hurdle to the development of AML-targeting chimeric antigen receptor (CAR) T-cells that can avoid on-target, off-tumor toxicity. Here, we develop a dual-antigen targeting CAR-T against CD70 and the active conformation of integrin β2 (aITGB2), each previously reported as promising AML targets due to minimal off-tumor expression. We show an OR-gated approach for these antigens significantly increases the proportion of AML blasts that can be targeted, in part using a novel ex vivo co-culture method to restore surface protein homeostasis following a freeze-thaw cycle. We test dual-targeting CAR-T constructs with different combinations of costimulatory domains, identifying constructs with superior anti-tumor cytotoxicity in vitro against AML cell line and patient derived xenograft models. We further show significantly improved in vivo tumor clearance and survival for a dual targeting CAR in murine models of AML tumor heterogeneity. Finally, we show that this dual-targeting CAR does not increase off-tumor toxicity, especially against hematopoietic stem and progenitor cells. Together, these findings demonstrate a promising clinically-translatable approach for the treatment of AML without the notable toxicity liabilities associated with other leading CAR-T targets for this disease.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":575532,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9487,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1303519,"name":"H. A. Johnson","orcid":"0009-0002-1527-7910","position":1,"is_corresponding":false},{"id":1483791,"name":"Nicole Lei","orcid":null,"position":2,"is_corresponding":false},{"id":1483792,"name":"Jeremiah Wong","orcid":null,"position":3,"is_corresponding":false},{"id":1483793,"name":"Nabeel Razi","orcid":null,"position":4,"is_corresponding":false},{"id":1037061,"name":"Adila Izgutdina","orcid":"0000-0002-3296-3261","position":5,"is_corresponding":false},{"id":53177,"name":"Corynn Kasap","orcid":"0000-0002-3667-8097","position":6,"is_corresponding":false},{"id":1326003,"name":"Nikhil Chilakapati","orcid":"0009-0004-3456-8144","position":7,"is_corresponding":false},{"id":351615,"name":"Jose Rivera","orcid":"0000-0002-6588-4914","position":8,"is_corresponding":false},{"id":357627,"name":"Paul Phojanakong","orcid":null,"position":9,"is_corresponding":false},{"id":841673,"name":"Juan A. Cámara","orcid":"0000-0001-7494-1127","position":10,"is_corresponding":false},{"id":842934,"name":"Fernando Salangsang","orcid":null,"position":11,"is_corresponding":false},{"id":323012,"name":"Veronica Steri","orcid":"0000-0003-2499-6618","position":12,"is_corresponding":false},{"id":953122,"name":"Aaron C. Logan","orcid":"0000-0002-1179-5879","position":13,"is_corresponding":false},{"id":805865,"name":"Justin Eyquem","orcid":"0000-0001-8262-1190","position":14,"is_corresponding":false},{"id":337815,"name":"Benjamin J. Huang","orcid":"0000-0001-6996-0833","position":15,"is_corresponding":false},{"id":28927,"name":"Arun P. Wiita","orcid":"0000-0002-7465-6964","position":16,"is_corresponding":false},{"id":1272961,"name":"Amrik S. Kang","orcid":"0000-0003-0445-5654","position":0,"is_corresponding":true}],"reference_count":0,"raw_metadata":null,"created_at":"2026-07-19T02:57:48.486077Z","pmid":"41001002","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":[]}