{"doi":"10.1101/2025.10.23.684052","title":"Chimeric Antigen Receptors Transmit Piconewton Forces that are Coupled with T Cell Function","abstract":"Chimeric antigen receptor (CAR) T cells are engineered to display a receptor that binds antigens expressed on the surface of cancerous cells, which leads to cancer cytotoxicity. Recently, the T cell field has come to recognize the role of small, piconewton level forces in establishing specificity and cytotoxicity in naïve T cells with the αβ TCR, raising the possibility that these forces could be present in CAR T cells. Using DNA-based tension probes, we reveal 8-19 pN mechanical forces with ∼1 sec timescales transmitted by CAR T cells to their target antigens. CAR-antigen force magnitude is independent of CAR expression level and shows heterogeneity across different T cell donors, suggesting utility as a biomarker of T cell fitness. Using an established exhaustion model, we show strong correlation between CAR exhaustion, cytotoxic capacity, and CAR-antigen force, suggesting that CAR mechanics provide a biomarker of CAR potency complementary to functional assays. Pharmacological inhibition studies demonstrate that CAR forces are driven by actin, Zap70 and Src family proximal kinases. Titration of dasatinib, a clinically used tyrosine kinase inhibitor also dampens both CAR-antigen tension and CAR function in a dose-dependent manner. Structural engineering of the CAR confirms that force levels are modulated by the scFv receptor and co-stimulatory domains, but force transmission requires CD3ζ ITAMs. Taken together, this work shows that CAR T cells transmit pN force to their cognate antigens which holds potential significance in the design and screening of CAR therapeutic candidates and for predicting treatment outcomes in a personalized fashion.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":579033,"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":0.9467,"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":1489289,"name":"Areeba A. Hashmi","orcid":null,"position":1,"is_corresponding":false},{"id":554914,"name":"Anna M. Davis","orcid":null,"position":2,"is_corresponding":false},{"id":1330942,"name":"Alexander Foote","orcid":null,"position":3,"is_corresponding":false},{"id":1068743,"name":"Gianna M. Branella","orcid":"0000-0003-3025-7977","position":4,"is_corresponding":false},{"id":1167509,"name":"Jhordan Rogers","orcid":"0000-0002-0329-859X","position":5,"is_corresponding":false},{"id":1009640,"name":"Emily C Sullivan","orcid":"0000-0002-1684-3382","position":6,"is_corresponding":false},{"id":1440012,"name":"Mehdi Rahman","orcid":null,"position":7,"is_corresponding":false},{"id":492435,"name":"Sunil S. Raikar","orcid":"0000-0003-2903-9542","position":8,"is_corresponding":false},{"id":313824,"name":"Khalid Salaita","orcid":"0000-0003-4138-3477","position":9,"is_corresponding":false},{"id":1217992,"name":"Rachel Fitzgerald","orcid":"0000-0003-2905-6895","position":0,"is_corresponding":true}],"reference_count":48,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:58:24.957414Z","pmid":"41279584","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":[]}