{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T02:48:27Z","timestamp":1793155707258,"version":"4.5.0"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T00:00:00Z","timestamp":1728518400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["21-65-00004"],"award-info":[{"award-number":["21-65-00004"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Adoptive cell therapy using TCR-engineered T-cells is one of the most effective strategies against tumor cells. The TCR T-cell approach has been well tested against a variety of blood neoplasms but is yet to be deeply tested against solid tumors. Among solid tumors, cancer-testis antigens are the most prominent targets for tumor-specific therapy, as they are usually found on cells that lie behind blood-tissue barriers.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We have employed a novel efficient protocol for MAGE-A3-specific T-cell clonal expansion, performed single-cell multi-omic analysis of the expanded T-cells via BD Rhapsody, engineered a selected T-cell receptor into a lentiviral construct, and tested it in an <jats:italic>in vitro<\/jats:italic> LDH-cytotoxicity test.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results and discussion<\/jats:title><jats:p>We have observed a 191-fold increase in the MAGE-A3-specific T-cell abundance, obtained a dominant T-cell receptor via single-cell multi-omic BD Rhapsody data analysis in the TCRscape bioinformatics tool, and observed potent cytotoxicity of the dominant-clonotype transduced TCR T-cells against a MAGE-A3-positive tumor. We have demonstrated the efficiency of our T-cell enrichment protocol in obtaining potent anti-tumor T-cells and their T-cell receptors, especially when paired with the modern single-cell analysis methods.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2024.1470130","type":"journal-article","created":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T04:31:40Z","timestamp":1728534700000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["Modified Dendritic cell-based T-cell expansion protocol and single-cell multi-omics allow for the selection of the most expanded and in vitro-effective clonotype via profiling of thousands of MAGE-A3-specific T-cells"],"prefix":"10.3389","volume":"15","author":[{"given":"Sergey","family":"Sennikov","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Volynets","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saleh","family":"Alrhmoun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roman","family":"Perik-Zavodskii","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga","family":"Perik-Zavodskaia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Fisher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Lopatnikova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Shevchenko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kirill","family":"Nazarov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Philippova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alaa","family":"Alsalloum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vasily","family":"Kurilin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Silkov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,10,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3390\/cancers16020246","article-title":"Melanoma antigen family A (MAGE A) as promising biomarkers and therapeutic targets in bladder cancer","volume":"16","author":"Verma","year":"2024","journal-title":"Cancers (Basel)"},{"key":"B2","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1034\/J.1600-065X.2002.18803.X","article-title":"Cancer\/testis antigens: An expanding family of targets for cancer immunotherapy","volume":"188","author":"Scanlan","year":"2002","journal-title":"Immunol Rev"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2010.08.029","article-title":"MAGE-RING protein complexes comprise a family of E3 ubiquitin ligases","volume":"39","author":"Doyle","year":"2010","journal-title":"Mol Cell"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2017.03.005","article-title":"A comprehensive guide to the MAGE family of ubiquitin ligases","volume":"429","author":"Lee","year":"2017","journal-title":"J Mol Biol"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1007\/s40291-020-00476-5","article-title":"Prognostic value of melanoma-associated antigen-A (MAGE-A) gene expression in various human cancers: A systematic review and meta-analysis of 7428 patients and 44 studies","volume":"24","author":"Poojary","year":"2020","journal-title":"Mol Diagn Ther"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ceb.2015.08.002","article-title":"The MAGE protein family And cancer","volume":"37","author":"Weon","year":"2015","journal-title":"Curr Opin Cell Biol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-09-0737","article-title":"The prioritization of cancer antigens: A national cancer institute pilot project for the acceleration of translational research","volume":"15","author":"Cheever","year":"2009","journal-title":"Clin Cancer Res"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1097\/CJI.0b013e3182829903","article-title":"Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy","volume":"36","author":"Morgan","year":"2013","journal-title":"J Immunother"},{"key":"B9","unstructured":"HomeClinicalTrials.gov"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2013-03-490565","article-title":"Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma","volume":"122","author":"Linette","year":"2013","journal-title":"Blood"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1098\/rsos.150016","article-title":"The growth threshold conjecture: a theoretical framework for understanding T-cell tolerance","volume":"2","author":"Arias","year":"2015","journal-title":"R Soc Open Sci"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1038\/nri3307","article-title":"Transcriptional control of effector and memory CD8+ T cell differentiation","volume":"12","author":"Kaech","year":"2012","journal-title":"Nat Rev Immunol"},{"key":"B13","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1186\/s12896-017-0379-9","article-title":"Overview of methodologies for T-cell receptor repertoire analysis","volume":"17","author":"Rosati","year":"2017","journal-title":"BMC Biotechnol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/FIMMU.2024.1431303\/BIBTEX","article-title":"Single-cell multi-omics reveal stage of differentiation and trajectory-dependent immunity-related gene expression patterns in human erythroid cells","volume":"15","author":"Perik-Zavodskii","year":"2024","journal-title":"Front Immunol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-021-01201-8","article-title":"High-throughput and single-cell T cell receptor sequencing technologies","volume":"18","author":"Pai","year":"2021","journal-title":"Nat Methods"},{"key":"B16","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1007\/978-1-0716-2756-3_2","article-title":"BD Rhapsody\u2122 single-cell analysis system workflow: from sample to multimodal single-cell sequencing data","volume":"2584","author":"Ulbrich","year":"2023","journal-title":"Methods Mol Biol"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2022-SITC2022.0030","article-title":"30 Single cell multiomic profiling of the antigen-specific immune response using antigen specific dCODE Dextramer\u00ae (RiO) reagents and BD\u00ae AbSeq Reagents on the BD Rhapsody\u2122 single-cell analysis system","volume":"10","author":"Jacobsen","year":"2022","journal-title":"J Immunother Cancer"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines12030699","article-title":"Development of cell technologies based on dendritic cells for immunotherapy of oncological diseases","volume":"12","author":"Kurilin","year":"2024","journal-title":"Biomedicines"},{"key":"B19","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1186\/s12865-017-0219-7","article-title":"Generation of populations of antigen-specific cytotoxic T cells using DCs transfected with DNA construct encoding HER2\/neu tumor antigen epitopes","volume":"18","author":"Kuznetsova","year":"2017","journal-title":"BMC Immunol"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3892\/or.2019.7435","article-title":"Autologous dendritic cells and activated cytotoxic T\u2212cells as combination therapy for breast cancer","volume":"43","author":"Shevchenko","year":"2019","journal-title":"Oncol Rep"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3892\/OL.2017.7403","article-title":"Use of antigen-primed dendritic cells for inducing antitumor immune responses in vitro in patients with non-small cell lung cancer","volume":"15","author":"Obleukhova","year":"2018","journal-title":"Oncol Lett"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3389\/FGENE.2023.866407","article-title":"An HLA map of the world: A comparison of HLA frequencies in 200 worldwide populations reveals diverse patterns for class I and class II","volume":"14","author":"Arrieta-Bola\u00f1os","year":"2023","journal-title":"Front Genet"},{"key":"B23","unstructured":"GitHub - IdoSpringer\/ERGO-II: ERGO-II, an updated version of ERGO including more features for TCR-peptide binding prediction"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1093\/BIOINFORMATICS\/BTX286","article-title":"McPAS-TCR: a manually curated catalogue of pathology-associated T cell receptor sequences","volume":"33","author":"Tickotsky","year":"2017","journal-title":"Bioinformatics"},{"key":"B25","unstructured":"GitHub - Perik-Zavodskii\/TCRscape: TCRscape - a tool for simultaneous multimodal gene expression and clonotype analysis of single T-cells profiled via the BD Rhapsody system"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1016\/0166-0934(89)90132-8","article-title":"Protamine sulfate as an effective alternative to polybrene in retroviral-mediated gene-transfer: implications for human gene therapy","volume":"23","author":"Cornetta","year":"1989","journal-title":"J Virol Methods"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2567.2011.03547.x","article-title":"T-cell receptor affinity in thymic development","volume":"135","author":"Moran","year":"2012","journal-title":"Immunology"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1002\/eji.200535098","article-title":"The effects of thymic selection on the range of T cell cross-reactivity","volume":"35","author":"Chao","year":"2005","journal-title":"Eur J Immunol"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1038\/nri2469","article-title":"Affinity threshold for thymic selection through a T-cell receptor\u2013co-receptor zipper","volume":"9","author":"Palmer","year":"2009","journal-title":"Nat Rev Immunol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1126\/science.abl5282","article-title":"Tuning T cell receptor sensitivity through catch bond engineering","volume":"376","author":"Zhao","year":"2022","journal-title":"Science (80-)"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.crmeth.2023.100459","article-title":"TCR sequencing and cloning methods for repertoire analysis and isolation of tumor-reactive TCRs","volume":"3","author":"Genolet","year":"2023","journal-title":"Cell Rep Methods"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0141561","article-title":"Multiplex identification of antigen-specific T cell receptors using a combination of immune assays and immune receptor sequencing","volume":"10","author":"Klinger","year":"2015","journal-title":"PloS One"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1016\/j.kint.2023.01.023","article-title":"Identification of proteinase 3 autoreactive CD4+T cells and their T-cell receptor repertoires in antineutrophil cytoplasmic antibody\u2013associated vasculitis","volume":"103","author":"Sharma","year":"2023","journal-title":"Kidney Int"},{"key":"B34","doi-asserted-by":"publisher","first-page":"24512","DOI":"10.1038\/s41598-021-04291-8","article-title":"Biased TCR gene usage in citrullinated Tenascin C specific T-cells in rheumatoid arthritis","volume":"11","author":"Sharma","year":"2021","journal-title":"Sci Rep"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1111\/ajt.12431","article-title":"TCR repertoire analysis by next generation sequencing allows complex differential diagnosis of T cell\u2013related pathology","volume":"13","author":"Dziubianau","year":"2013","journal-title":"Am J Transplant"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2024.1470130\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T04:31:43Z","timestamp":1728534703000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2024.1470130\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,10]]},"references-count":35,"alternative-id":["10.3389\/fimmu.2024.1470130"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2024.1470130","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,10]]},"article-number":"1470130"}}