{"doi":"10.1101/2024.04.16.589780","title":"Directed evolution-based discovery of ligands for in vivo restimulation of CAR-T cells","abstract":"Chimeric antigen receptor (CAR) T cell therapy targeting CD19 elicits remarkable clinical efficacy in B-cell malignancies, but many patients relapse due to failed expansion and/or progressive loss of CAR-T cells. We recently reported a strategy to potently restimulate CAR-T cells in vivo, enhancing their functionality by administration of a vaccine-like stimulus comprised of surrogate peptide ligands for a CAR linked to a lymph node-targeting amphiphilic PEG-lipid (termed CAR-T-vax). Here, we demonstrate a general strategy to generate and optimize peptide mimotopes enabling CAR-T-vax generation for any CAR. Using the clinical CD19 CAR FMC63 as a test case, we employed yeast surface display to identify peptide binders to soluble IgG versions of FMC63, which were subsequently affinity matured by directed evolution. CAR-T vaccines using these optimized mimotopes triggered marked expansion of both murine CD19 CAR-T cells in a syngeneic model and human CAR-T cells in a humanized mouse model of B cell acute lymphoblastic leukemia (B-ALL), and enhanced control of leukemia progression. This approach thus enables vaccine boosting to be applied to any clinically-relevant CAR-T cell product.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":488343,"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":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9523,"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":1017131,"name":"Alexandra Neeser","orcid":null,"position":1,"is_corresponding":false},{"id":913347,"name":"Ranjani Ramasubramanian","orcid":"0000-0002-9620-4934","position":2,"is_corresponding":false},{"id":816178,"name":"Anna Romanov","orcid":"0000-0002-1239-4946","position":3,"is_corresponding":false},{"id":1333046,"name":"Ryan Tannir","orcid":null,"position":4,"is_corresponding":false},{"id":315904,"name":"Naveen K. Mehta","orcid":null,"position":5,"is_corresponding":false},{"id":309813,"name":"Benjamin Cossette","orcid":null,"position":6,"is_corresponding":false},{"id":633121,"name":"Duncan M. Morgan","orcid":"0000-0002-3253-1362","position":7,"is_corresponding":false},{"id":1332735,"name":"Beatriz Gonçalves","orcid":"0000-0002-5220-7383","position":8,"is_corresponding":false},{"id":1333047,"name":"Ina Sukaj","orcid":null,"position":9,"is_corresponding":false},{"id":1030566,"name":"Elisa Bergaggio","orcid":"0000-0002-9960-3674","position":10,"is_corresponding":false},{"id":1332736,"name":"Stephan Kadauke","orcid":"0000-0003-2996-8034","position":11,"is_corresponding":false},{"id":623304,"name":"Regina M. Myers","orcid":"0000-0002-2542-2061","position":12,"is_corresponding":false},{"id":1032169,"name":"Luca Paruzzo","orcid":"0000-0002-6505-0194","position":13,"is_corresponding":false},{"id":1027063,"name":"Guido Ghilardi","orcid":"0000-0002-0095-3579","position":14,"is_corresponding":false},{"id":975286,"name":"Austin Cozzone","orcid":null,"position":15,"is_corresponding":false},{"id":273372,"name":"Stephen J. Schuster","orcid":"0000-0002-3376-8978","position":16,"is_corresponding":false},{"id":1333048,"name":"Noelle Frey","orcid":null,"position":17,"is_corresponding":false},{"id":689275,"name":"Libin Zhang","orcid":"0000-0002-4611-1852","position":18,"is_corresponding":false},{"id":528344,"name":"Parisa Yousefpour","orcid":"0000-0003-0431-8131","position":19,"is_corresponding":false},{"id":239103,"name":"Wuhbet Abraham","orcid":null,"position":20,"is_corresponding":false},{"id":811690,"name":"Heikyung Suh","orcid":"0000-0003-0787-298X","position":21,"is_corresponding":false},{"id":256650,"name":"Marco Ruella","orcid":"0000-0003-4301-5811","position":22,"is_corresponding":false},{"id":496143,"name":"Stephan A. Grupp","orcid":"0000-0001-8030-7595","position":23,"is_corresponding":false},{"id":42115,"name":"Roberto Chiarle","orcid":"0000-0003-1564-8531","position":24,"is_corresponding":false},{"id":314110,"name":"K. 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