{"doi":"10.1101/2025.10.05.680562","title":"Base-editing a single missense mutation in A20 enhances CAR-T cell efficacy","abstract":"Abstract T cell exhaustion limits the efficacy of cancer immunotherapies. Here, we performed genome-wide loss-of-function screening in repetitively stimulated human T cells and identified the mulitfunctional ubiquitin-modifying protein A20/TNFAIP3 as a major negative regulator of exhausted T cell persistence. Protein large language modeling, deep base-editing mutagenesis, and studies in immunocompetent mice with domain-specific inactivating mutations revealed A20’s non-enzymatic M1 ubiquitin-binding zinc finger 7 (A20 ZF7 ) motif as critical to suppression of anti-tumor immunity. A20 ZF7 -deficient CD8 + tumor-infiltrating lymphocytes (TILs) resisted terminal exhaustion and circumvented an unappreciated mechanism restraining perforin degranulation in terminally exhausted cells. Human chimeric antigen receptor (CAR)-T cells engineered via base-editing to inactivate A20 ZF7 via a single missense mutation also resisted exhaustion, secreted more perforin and robustly suppressed cancer in vivo . These studies pinpoint A20 ZF7 as a novel T cell checkpoint and reveal precision base-editing of missense mutations as an effective approach to enhance CAR-T cell therapy.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":577166,"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":0.959,"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":843110,"name":"Stefanie Bachl","orcid":null,"position":1,"is_corresponding":false},{"id":1486710,"name":"Luis R Sandoval","orcid":null,"position":2,"is_corresponding":false},{"id":1486711,"name":"Carter Ching","orcid":null,"position":3,"is_corresponding":false},{"id":1400355,"name":"Christopher Bowman","orcid":"0000-0002-4107-4116","position":4,"is_corresponding":false},{"id":636378,"name":"Nupura Kale","orcid":"0000-0002-3185-8646","position":5,"is_corresponding":false},{"id":1486712,"name":"Manu Prabandham","orcid":null,"position":6,"is_corresponding":false},{"id":842014,"name":"Morgan E. 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