{"doi":"10.1101/2024.09.03.611042","title":"Efficient multiplex non-viral engineering and expansion of polyclonal γδ CAR-T cells for immunotherapy","abstract":"ABSTRACT Gamma delta (γδ) T cells are defined by their unique ability to recognize a limited repertoire of non-peptide, non-MHC-associated antigens on transformed and pathogen-infected cells. In addition to their lack of alloreactivity, γδ T cells exhibit properties distinct from other lymphocyte subsets, prompting significant interest in their development as an off-the-shelf cellular immunotherapeutic. However, their low abundance in circulation, heterogeneity, limited methods for ex vivo expansion, and under-developed methodologies for genetic modification have hindered basic study and clinical application of γδ T cells. Here, we implement a feeder-free, scalable approach for ex vivo manufacture of polyclonal, non-virally modified, gene edited chimeric antigen receptor (CAR)-γδ T cells in support of therapeutic application. Engineered CAR-γδ T cells demonstrate high function in vitro and and in vivo. Longitudinal in vivo pharmacokinetic profiling of adoptively transferred polyclonal CAR-γδ T cells uncover subset-specific responses to IL-15 cytokine armoring and multiplex base editing. Our results present a robust platform for genetic modification of polyclonal CAR-γδ T cells and present unique opportunities to further define synergy and the contribution of discrete, engineered CAR-γδ T cell subsets to therapeutic efficacy in vivo .","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":484733,"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":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9512,"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":6709,"name":"Matthew J. Johnson","orcid":"0000-0002-1313-1742","position":1,"is_corresponding":false},{"id":1326893,"name":"Jihyun Kim","orcid":"0009-0000-5769-016X","position":2,"is_corresponding":false},{"id":821950,"name":"Sophia Wenthe","orcid":null,"position":3,"is_corresponding":false},{"id":1215855,"name":"Joshua Krueger","orcid":"0009-0002-7658-149X","position":4,"is_corresponding":false},{"id":1215854,"name":"Bryce Wick","orcid":"0000-0002-2844-0898","position":5,"is_corresponding":false},{"id":633216,"name":"Mitchell G. Kluesner","orcid":"0000-0001-5828-9204","position":6,"is_corresponding":false},{"id":500951,"name":"Andrew Crane","orcid":"0000-0003-3452-2766","position":7,"is_corresponding":false},{"id":331508,"name":"Jason Bell","orcid":"0000-0003-4869-6476","position":8,"is_corresponding":false},{"id":273414,"name":"Joseph G. Skeate","orcid":"0000-0002-1765-5949","position":9,"is_corresponding":false},{"id":254329,"name":"Branden S. Moriarity","orcid":"0000-0002-3031-3767","position":10,"is_corresponding":false},{"id":254328,"name":"Beau R. Webber","orcid":"0000-0002-3950-8747","position":11,"is_corresponding":false},{"id":1216478,"name":"Jacob E. Bridge","orcid":null,"position":0,"is_corresponding":true}],"reference_count":85,"raw_metadata":null,"created_at":"2026-07-19T02:07:42.971417Z","pmid":"39464114","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":[]}