{"doi":"10.1136/jitc-2025-011807","title":"Comparison of axicabtagene ciloleucel and tisagenlecleucel patient CAR-T cell products by single-cell RNA sequencing","abstract":"BACKGROUND: Autologous CD19 chimeric antigen receptor (CAR) T-cell therapy leads to durable responses and improved survival in patients with relapsed or refractory large B-cell lymphoma (R/R LBCL). Among approved CAR T-cell products, axicabtagene ciloleucel (axi-cel; CD19/CD28) has greater real-world efficacy and cytokine-associated toxicity than tisagenlecleucel (tisa-cel; CD19/4-1BB), for reasons that are poorly understood. METHODS: Here we report single-cell RNA sequencing (scRNA-seq) of 57 pre-infusion CAR T-cell products from axi-cel (n=39) and tisa-cel (n=18) patients treated as standard-of-care for R/R LBCL, and their biological associations with clinical outcomes. In vitro CAR manufacturing conditions mimicking those known for axi-cel and tisa-cel were performed using CD19/CD28z or CD19/4-1BBz constructs. RESULTS: ScRNA-seq revealed that axi-cel and tisa-cel are markedly different products. Axi-cel is comprised of more CD4 central memory, CD8 central memory, and CD8 effectors, whereas tisa-cel is comprised of more proliferative CD4 and CD8 cells. Across multiple T-cell subsets, axi-cel had greater expression of immune response pathways and protein synthesis and trafficking pathways versus tisa-cel. On comparison of infusion product CAR transgene-positive (CAR+) cells to CAR transgene-negative (CAR-) T cells, axi-cel CAR+ cells had vastly different gene expression than axi-cel CAR- cells. Unexpectedly, tisa-cel CAR+ cells were highly similar to tisa-cel CAR- cells. Under recapitulated CAR-T manufacturing conditions known to be used for axi-cel and tisa-cel, we found that CAR+ cells differed from CAR- cells early after manufacturing yet became more similar to CAR- cells after prolonged expansion. Prolonged time in expansion culture, as used during tisa-cel manufacturing, greatly decreased naïve and central memory T-cell subsets. CONCLUSIONS: Following manufacture, axi-cel is less differentiated and has greater immune activation compared with tisa-cel, potentially accounting for its greater efficacy and toxicity in patients. Our data support the conclusion that tisa-cel is adversely affected by its manufacturing rather than by the CAR construct.","journal":"Journal for ImmunoTherapy of Cancer","year":2025,"id":524002,"datarank":0.24141568686511508,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.0,"self_endowment_contribution":0.24141568686511508,"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":0.9488,"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":108395,"name":"Michael D. Jain","orcid":"0000-0002-7789-1257","position":1,"is_corresponding":false},{"id":851800,"name":"Meghan Menges","orcid":"0009-0007-7446-9492","position":2,"is_corresponding":false},{"id":1396865,"name":"Ling Cen","orcid":"0000-0001-5961-024X","position":3,"is_corresponding":false},{"id":1396866,"name":"Jerald Noble","orcid":"0000-0003-2571-9428","position":4,"is_corresponding":false},{"id":743068,"name":"Reginald Atkins","orcid":"0009-0007-0104-7344","position":5,"is_corresponding":false},{"id":1377695,"name":"Turab Mohammad","orcid":null,"position":6,"is_corresponding":false},{"id":432856,"name":"Christina A. Bachmeier","orcid":"0000-0002-9832-3670","position":7,"is_corresponding":false},{"id":1110643,"name":"Samira Naderinezhad","orcid":"0000-0002-0703-2334","position":8,"is_corresponding":false},{"id":432855,"name":"Kayla Reid","orcid":"0000-0002-6585-0781","position":9,"is_corresponding":false},{"id":1179368,"name":"Salvatore Corallo","orcid":"0009-0001-2450-4022","position":10,"is_corresponding":false},{"id":785380,"name":"Sean J. Yoder","orcid":null,"position":11,"is_corresponding":false},{"id":627233,"name":"Chaomei Zhang","orcid":null,"position":12,"is_corresponding":false},{"id":1397501,"name":"Lanmin Zhang","orcid":null,"position":13,"is_corresponding":false},{"id":635260,"name":"Julieta Abraham-Miranda","orcid":"0000-0001-6982-564X","position":14,"is_corresponding":false},{"id":432857,"name":"Bijal Shah","orcid":"0000-0003-4328-6021","position":15,"is_corresponding":false},{"id":11139,"name":"Julio C. Chávez","orcid":"0000-0002-2045-6238","position":16,"is_corresponding":false},{"id":362028,"name":"Rebecca S. Hesterberg","orcid":"0000-0003-0472-7517","position":17,"is_corresponding":false},{"id":1397502,"name":"Luis Cuadrado Delgado","orcid":null,"position":18,"is_corresponding":false},{"id":792135,"name":"Constanza Savid-Frontera","orcid":"0000-0001-8614-3528","position":19,"is_corresponding":false},{"id":242942,"name":"Paulo C. Rodrı́guez","orcid":"0000-0001-7480-6566","position":20,"is_corresponding":false},{"id":226779,"name":"John L. Cleveland","orcid":"0000-0002-7561-9065","position":21,"is_corresponding":false},{"id":377318,"name":"Xuefeng Wang","orcid":"0000-0001-5775-408X","position":22,"is_corresponding":false},{"id":432861,"name":"Marco L. Davila","orcid":"0000-0002-6270-3065","position":23,"is_corresponding":false},{"id":11097,"name":"Frederick L. Locke","orcid":"0000-0001-9063-6691","position":24,"is_corresponding":false},{"id":1351526,"name":"Xiaoqing Yu","orcid":"0000-0002-3255-9285","position":0,"is_corresponding":true}],"reference_count":31,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:50:07.396155Z","pmid":"40730421","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":[]}