{"doi":"10.1101/2024.01.31.575774","title":"Metabolic priming of GD2 <i>TRAC</i> -CAR T cells during manufacturing promotes memory phenotypes while enhancing persistence","abstract":"ABSTRACT Manufacturing Chimeric Antigen Receptor (CAR) T cell therapies is complex, with limited understanding of how media composition impact T-cell phenotypes. CRISPR/Cas9 ribonucleoproteins can precisely insert a CAR sequence while disrupting the endogenous T cell receptor alpha constant ( TRAC ) gene resulting in TRAC -CAR T cells with an enriched stem cell memory T-cell population, a process that could be further optimized through modifications to the media composition. In this study we generated anti-GD2 TRAC -CAR T cells using “metabolic priming” (MP), where the cells were activated in glucose/glutamine low media and then expanded in glucose/glutamine high media. T cell products were evaluated using spectral flow cytometry, metabolic assays, cytokine production, cytotoxicity assays in vitro and potency against human GD2+ xenograft neuroblastoma models in vivo . Compared to standard TRAC -CAR T cells, MP TRAC -CAR T cells showed less glycolysis, higher CCR7/CD62L expression, more bound NAD(P)H activity and reduced IFN-γ, IL-2, IP-10, IL-1β, IL-17, and TGFβ production at the end of manufacturing ex vivo , with increased central memory CAR T cells and better persistence observed in vivo . Metabolic priming with media during CAR T cell biomanufacturing can minimize glycolysis and enrich memory phenotypes ex vivo , which could lead to better responses against solid tumors in vivo .","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":485415,"datarank":0.26876392038420827,"base_score":1.791759469228055,"endowment":1.791759469228055,"self_citation_contribution":0.26876392038420827,"citation_network_contribution":0.0,"self_endowment_contribution":0.26876392038420827,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9588,"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":264988,"name":"Dan L. Pham","orcid":null,"position":1,"is_corresponding":false},{"id":277445,"name":"Matthew H. Forsberg","orcid":"0000-0002-0514-6354","position":2,"is_corresponding":false},{"id":1214109,"name":"Madison Bugel","orcid":null,"position":3,"is_corresponding":false},{"id":1214110,"name":"Anna Tommasi","orcid":null,"position":4,"is_corresponding":false},{"id":1214111,"name":"Anthony Lauer","orcid":null,"position":5,"is_corresponding":false},{"id":1214112,"name":"Jolanta Vidugirienė","orcid":null,"position":6,"is_corresponding":false},{"id":1214113,"name":"Brookelyn Hrdlicka","orcid":null,"position":7,"is_corresponding":false},{"id":1214114,"name":"Alexandria McHale","orcid":null,"position":8,"is_corresponding":false},{"id":853022,"name":"Quaovi H. Sodji","orcid":"0000-0002-9469-1388","position":9,"is_corresponding":false},{"id":108082,"name":"Melissa C. Skala","orcid":"0000-0002-6320-7637","position":10,"is_corresponding":false},{"id":277448,"name":"Christian M. Capitini","orcid":"0000-0002-2276-6731","position":11,"is_corresponding":false},{"id":253458,"name":"Krishanu Saha","orcid":"0000-0003-2837-0858","position":12,"is_corresponding":false},{"id":858830,"name":"Dan Cappabianca","orcid":"0009-0005-2354-4363","position":0,"is_corresponding":true}],"reference_count":63,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:07:52.536246Z","pmid":"38562720","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":[]}