{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T06:03:06Z","timestamp":1785391386361,"version":"3.55.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Glioblastoma (GBM), the most prevalent and aggressive malignant tumor of the nervous system, presents a formidable clinical challenge due to its rapid progression and extremely poor prognosis. T cell receptor fusion constructs (TRuC) -T cells represent a highly promising engineered T-cell therapy beyond CAR-T technology. Early prediction of TRuC-T cell cytotoxicity against GBM is crucial for clinical application.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We developed B7-H3-targeting TRuC-T cells and incorporated NF-\u03baB- or NFAT-responsive transcriptional elements into the TRuC construct. Using flow cytometry, luciferase assays, and\n                      <jats:italic>in vitro<\/jats:italic>\n                      \/\n                      <jats:italic>vivo<\/jats:italic>\n                      functional studies, we evaluated whether TRuC-T cells exert cytotoxicity via NF-\u03baB and NFAT signaling pathways.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      B7-H3 TRuC-T cells effectively suppressed GBM growth both\n                      <jats:italic>in vitro<\/jats:italic>\n                      and\n                      <jats:italic>in vivo<\/jats:italic>\n                      . Co-culture of TRuC-T cells carrying NF-\u03baB- or NFAT-reporters with target cells induced GFP expression, indicating pathway activation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>Upon engagement with B7-H3, both NFAT and NF-\u03baB signaling pathways are activated and participate in the regulation of B7-H3-targeting TRuC-T cells. Our data demonstrate that NF-\u03baB\/NFAT transcriptional reporters integrated into these TRuC constructs can be utilized to monitor target recognition and may serve as early indicators of cytotoxic efficacy. Overall, this system suggests a potential avenue for GBM therapy and offers an initial framework for the functional assessment of TRuC-T cell therapeutic potency.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1876463","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T05:41:14Z","timestamp":1785390074000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["NF-\u03baB\/NFAT signaling contributes to B7-H3 TRuC-T cell cytotoxicity and supports a reporter platform for glioblastoma immunotherapy"],"prefix":"10.3389","volume":"17","author":[{"given":"Junyue","family":"Lin","sequence":"first","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peiyuan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanhong","family":"Lu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qianjin","family":"Yu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiahao","family":"Zheng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Yang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuanhao","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng","family":"Wei","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]},{"name":"Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, The First Affiliated Hospital of Wenzhou Medical University","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jimin","family":"Gao","sequence":"additional","affiliation":[{"name":"Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University","place":["Wenzhou, China"]},{"name":"Zhejiang Qixin Biotech","place":["Wenzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ai","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Geriatric, Affiliated Hangzhou First People\u2019s Hospital, Westlake University School of Medicine","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,7,30]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"665","DOI":"10.1016\/j.ccell.2025.03.019","article-title":"CAR-T therapy in solid tumors","volume":"43","author":"Du","year":"2025","journal-title":"Cancer Cell"},{"key":"B2","doi-asserted-by":"publisher","first-page":"449","DOI":"10.1186\/s12967-023-04292-3","article-title":"Harnessing the potential of CAR-T cell therapy: progress, challenges, and future directions in hematological and solid tumor treatments","volume":"21","author":"Dagar","year":"2023","journal-title":"J Transl Med"},{"key":"B3","doi-asserted-by":"publisher","first-page":"1005","DOI":"10.1016\/j.trecan.2025.08.014","article-title":"CAR T cell persistence in cancer","volume":"11","author":"Mueller","year":"2025","journal-title":"Trends Cancer"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2025.1564998","article-title":"Challenges in the preclinical design and assessment of CAR-T cells","volume":"16","author":"Tomai","year":"2025","journal-title":"Front Immunol"},{"key":"B5","article-title":"Chimeric-antigen-receptor (CAR) T cells and the factors influencing their therapeutic efficacy","volume":"2","author":"Kravets","year":"2017","journal-title":"J Immunol Res Ther"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1186\/s40364-022-00434-9","article-title":"Strategies to enhance CAR-T persistence","volume":"10","author":"Liu","year":"2022","journal-title":"biomark Res"},{"key":"B7","doi-asserted-by":"publisher","first-page":"2087","DOI":"10.1038\/s41467-019-10097-0","article-title":"Synthetic TRuC receptors engaging the complete T cell receptor for potent anti-tumor response","volume":"10","author":"Baeuerle","year":"2019","journal-title":"Nat Commun"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.omto.2022.04.003","article-title":"Chimeric anti-GPC3 sFv-CD3\u03f5 receptor-modified T cells with IL7 co-expression for the treatment of solid tumors","volume":"25","author":"Sun","year":"2022","journal-title":"Mol Ther Oncolytics"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1080\/2162402X.2023.2182058","article-title":"Mesothelin-targeting T cells bearing a novel T cell receptor fusion construct (TRuC) exhibit potent antitumor efficacy against solid tumors","volume":"12","author":"Ding","year":"2023","journal-title":"Oncoimmunology"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-023-02452-y","article-title":"Mesothelin-targeting T cell receptor fusion construct cell therapy in refractory solid tumors: phase 1\/2 trial interim results","volume":"29","author":"Hassan","year":"2023","journal-title":"Nat Med"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/s1097-2765(03)00107-2","article-title":"Death and NF-kappaB in T cell activation: life at the edge","volume":"11","author":"Green","year":"2003","journal-title":"Mol Cell"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-065X.2009.00817.x","article-title":"Regulation of T-cell tolerance by calcium\/NFAT signaling","volume":"231","author":"Baine","year":"2009","journal-title":"Immunol Rev"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2013.02.002","article-title":"A new look at T cell receptor signaling to nuclear factor-\u03baB","volume":"34","author":"Paul","year":"2013","journal-title":"Trends Immunol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3390\/cells10061335","article-title":"Challenges with methods for detecting and studying the transcription factor nuclear factor kappa B (NF-\u03baB) in the central nervous system","volume":"10","author":"Mostafizar","year":"2021","journal-title":"Cells"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1054-1","article-title":"CAR T cell trogocytosis and cooperative killing regulate tumour antigen escape","volume":"568","author":"Hamieh","year":"2019","journal-title":"Nature"},{"key":"B16","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1038\/s41392-023-01708-w","article-title":"Trogocytosis of CAR molecule regulates CAR-T cell dysfunction and tumor antigen escape","volume":"8","author":"Zhai","year":"2023","journal-title":"Signal Transduct Target Ther"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2024-010702","article-title":"Tailoring CAR surface density and dynamics to improve CAR-T cell therapy","volume":"13","author":"Hinckley-Boned","year":"2025","journal-title":"J Immunother Cancer"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/j.critrevonc.2008.02.005","article-title":"Glioblastoma in adults","volume":"67","author":"Brandes","year":"2008","journal-title":"Crit Rev Oncol Hematol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3390\/cells14070494","article-title":"Therapeutic targets in glioblastoma: molecular pathways, emerging strategies, and future directions","volume":"14","author":"Tang","year":"2025","journal-title":"Cells"},{"key":"B20","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1007\/s44272-025-00044-0","article-title":"Management of cytokine release syndrome (CRS) following CAR T-cell therapy: a comprehensive review","volume":"4","author":"Zhang","year":"2025","journal-title":"Clin Cancer Bull"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3390\/cancers14235983","article-title":"Efficacy, safety, and challenges of CAR T-cells in the treatment of solid tumors","volume":"14","author":"Chen","year":"2022","journal-title":"Cancers"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1186\/s12943-023-01751-9","article-title":"New frontiers in immune checkpoint B7-H3 (CD276) research and drug development","volume":"22","author":"Getu","year":"2023","journal-title":"Mol Cancer"},{"key":"B23","doi-asserted-by":"publisher","first-page":"1209","DOI":"10.3390\/cells14151209","article-title":"B7-H3 in Cancer Immunotherapy\u2014Prospects and Challenges: A Review of the Literature","volume":"14","author":"Mielcarska","year":"2025","journal-title":"Cells"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1111\/cas.12915","article-title":"Development of DS-5573a: a novel afucosylated mA b directed at B7-H3 with potent antitumor activity","volume":"107","author":"Nagase-Zembutsu","year":"2016","journal-title":"Cancer Sci"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-022-01765-8","article-title":"Engineered cellular immunotherapies in cancer and beyond","volume":"28","author":"Finck","year":"2022","journal-title":"Nat Med"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adf3700","article-title":"TCR-engineered T cell therapy in solid tumors: state of the art and perspectives","volume":"9","author":"Baulu","year":"2023","journal-title":"Sci Adv"},{"key":"B27","doi-asserted-by":"publisher","first-page":"256","DOI":"10.1007\/s00262-024-03808-0","article-title":"Antitumor effects of intracranial injection of B7-H3-targeted Car-T and Car-Nk cells in a patient-derived glioblastoma xenograft model","volume":"73","author":"Tachi","year":"2024","journal-title":"Cancer Immunol Immunother"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-024-03451-3","article-title":"Intracerebroventricular B7-H3-targeting CAR T cells for diffuse intrinsic pontine glioma: a phase 1 trial","volume":"31","author":"Vitanza","year":"2025","journal-title":"Nat Med"},{"key":"B29","doi-asserted-by":"publisher","first-page":"7196","DOI":"10.1038\/s41467-025-61427-4","article-title":"Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function","volume":"16","author":"Barisa","year":"2025","journal-title":"Nat Commun"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2024-009485","article-title":"Preclinical development of three novel CARs targeting CD79b for the treatment of non-Hodgkin's lymphoma and characterization of the loss of the target antigen","volume":"12","author":"Esquinas","year":"2024","journal-title":"J Immunother Cancer"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2024.114949","article-title":"TCR-mimicking STAR conveys superior sensitivity over CAR in targeting tumors with low-density neoantigens","volume":"43","author":"Huang","year":"2024","journal-title":"Cell Rep"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.aay8248","article-title":"4-1BB costimulation promotes CAR T cell survival through noncanonical NF-\u03baB signaling","volume":"13","author":"Philipson","year":"2020","journal-title":"Sci Signal"},{"key":"B33","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1186\/s12943-025-02446-z","article-title":"Enhancement of CAR-T cell efficacy and persistence via CD30 costimulation and NF-\u03baB signaling","volume":"24","author":"Su","year":"2025","journal-title":"Mol Cancer"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1136\/jitc-2025-011971","article-title":"Increased NFAT activity with dual CAR stimulation in CD19xCD22 CAR T-cells is associated with decreased exhaustion and improved survival","volume":"13","author":"Rankin","year":"2025","journal-title":"J Immunother Cancer"},{"key":"B35","doi-asserted-by":"publisher","first-page":"456","DOI":"10.1016\/j.immuni.2020.07.011","article-title":"Chimeric antigen receptor designed to prevent ubiquitination and downregulation showed durable antitumor efficacy","volume":"53","author":"Li","year":"2020","journal-title":"Immunity"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.7150\/thno.68966","article-title":"A multimodal imaging workflow for monitoring CAR T cell therapy against solid tumor from whole-body to single-cell level","volume":"12","author":"Pfeifer","year":"2022","journal-title":"Theranostics"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1007\/s12026-011-8216-z","article-title":"Regulation of NF-\u03baB induction by TCR\/CD28","volume":"50","author":"Cheng","year":"2011","journal-title":"Immunol Res"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1038\/87704","article-title":"Constitutive pre-TCR signaling promotes differentiation through Ca2+ mobilization and activation of NF-kappaB and NFAT","volume":"2","author":"Aifantis","year":"2001","journal-title":"Nat Immunol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceca.2017.01.014","article-title":"Calcium-NFAT transcriptional signalling in T cell activation and T cell exhaustion","volume":"63","author":"Hogan","year":"2017","journal-title":"Cell Calcium"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.3390\/biom16060757","article-title":"Two faces of NFAT transcription factors in lymphocytes\u2014a personal account","volume":"16","author":"Serfling","year":"2026","journal-title":"Biomolecules"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.cir-21-0626","article-title":"Regulatory programs of B-cell activation and germinal center reaction allow B-ALL escape from CD19 CAR T-cell therapy","volume":"10","author":"Im","year":"2022","journal-title":"Cancer Immunol Res"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1876463\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T05:41:16Z","timestamp":1785390076000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1876463\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,30]]},"references-count":41,"alternative-id":["10.3389\/fimmu.2026.1876463"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2026.1876463","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,30]]},"article-number":"1876463"}}