{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T14:01:44Z","timestamp":1793109704105,"version":"4.5.0"},"reference-count":70,"publisher":"BMJ","issue":"3","license":[{"start":{"date-parts":[[2025,3,26]],"date-time":"2025-03-26T00:00:00Z","timestamp":1742947200000},"content-version":"unspecified","delay-in-days":25,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["104-2320-B-400-017-MY3"],"award-info":[{"award-number":["104-2320-B-400-017-MY3"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["107-2628-B-400-002, 108-2628-B-400 -002"],"award-info":[{"award-number":["107-2628-B-400-002, 108-2628-B-400 -002"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["109-2326-B-400-007, 110-2326-B-400-004"],"award-info":[{"award-number":["109-2326-B-400-007, 110-2326-B-400-004"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["111-2326-B-400-004, 112-2326-B-400 -002"],"award-info":[{"award-number":["111-2326-B-400-004, 112-2326-B-400 -002"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["113-2326-B-400-007"],"award-info":[{"award-number":["113-2326-B-400-007"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005564","name":"Gilead Sciences","doi-asserted-by":"publisher","award":["2018 Gilead Sciences RSP in liver disease-Asia\/N\/A"],"award-info":[{"award-number":["2018 Gilead Sciences RSP in liver disease-Asia\/N\/A"]}],"id":[{"id":"10.13039\/100005564","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2025,3,10]]},"published-print":{"date-parts":[[2025,3]]},"abstract":"<jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Impairment of Akt signaling has been observed in antigen-specific cytotoxic T lymphocytes (CTLs) during chronic viral infections or tumor progression. Despite numerous studies emphasizing Akt\u2019s role in driving CTL effector functions, there is limited exploration of using Akt molecules in T-cell engineering to enhance their antiviral or antitumor capabilities for therapeutic purposes. Some studies even conclude that inhibiting Akt activation during the in vitro expansion process can prevent T-cell exhaustion and boost the antitumor effector functions of chimeric antigen receptor-T cells in vivo. Given the unique expression patterns and functions of the three Akt isoforms in immune cells, we proposed that Akt isoforms in CTLs may regulate effector functions and T-cell exhaustion distinctly.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>In this study, we genetically modified tumor\/virus-antigen-specific T-cell receptor tg CTLs to ectopically express Akt isoforms via retroviral transduction. We subsequently conducted western blotting, flow cytometry, and RNA sequencing analysis to assess their Akt expression, expression of immune checkpoints, antitumor\/antivirus functionalities, and transcriptome. Additionally, we employed a persistent Hepatitis B Virus mouse model and a syngeneic hepatocellular carcinoma mouse model for further evaluation of their antivirus\/antitumor efficacies.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>We found that both Akt1 and Akt2 overexpression enhanced the cytotoxic capabilities of mouse CTLs, although with different dynamics. Specifically, Akt2 signaling in CTLs accelerated effector functions, leading to a rapid attack on tumor cells. Conversely, Akt1 signaling triggered calcium influx and subsequent nuclear factor of activated T cells (NFAT) activation, while Akt2 signaling suppressed calcium influx, preventing excessive NFAT expression and nuclear translocation. This repression of NFAT transcriptional activity by Akt2 signaling during prolonged antigen stimulation subsequently led to reduced expression of transcription factors associated with T-cell exhaustion, such as Egr2, Nr4a, Tox, and immune checkpoints. Consequently, Akt2-overexpressed CTLs displayed reduced T-cell exhaustion within the tumor microenvironment and efficiently eradicated tumors.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>These findings highlight the essential role of Akt signaling in enabling tumor-specific CTLs to eliminate cancer cells in the solid TME, with Akt isoforms differentially regulating the calcium\u2013calcineurin\u2013NFAT signaling pathway. This discovery suggests the potential of AKT2 in T-cell engineering technology to enhance the survival and effector functions of adoptively transferred T cells for treating liver malignancies or chronic viral infections.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1136\/jitc-2024-009827","type":"journal-article","created":{"date-parts":[[2025,3,27]],"date-time":"2025-03-27T02:57:40Z","timestamp":1743044260000},"page":"e009827","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":7,"title":["Differential regulation of calcium-NFAT signaling pathway by Akt isoforms: unraveling effector dynamics and exhaustion of cytotoxic T lymphocytes in tumor microenvironment"],"prefix":"10.1136","volume":"13","author":[{"given":"Wen-Ling","family":"Chen","sequence":"first","affiliation":[{"name":"National Health Research Institutes, Institute of Molecular and Genomic Medicine, Miaoli, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6647-7440","authenticated-orcid":false,"given":"Yong-Lin","family":"Chang","sequence":"additional","affiliation":[{"name":"National Health Research Institutes, Institute of Molecular and Genomic Medicine, Miaoli, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8706-2439","authenticated-orcid":false,"given":"Su-Fang","family":"Lin","sequence":"additional","affiliation":[{"name":"National Health Research Institutes, National Institute of Cancer Research, Miaoli, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrike","family":"Protzer","sequence":"additional","affiliation":[{"name":"Institute of Virology, Technical University of Munich\/ Helmholtz Munich, School of Medicine and Health, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masanori","family":"Isogawa","sequence":"additional","affiliation":[{"name":"Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hung-Chih","family":"Yang","sequence":"additional","affiliation":[{"name":"National Taiwan University Graduate Institute of Microbiology, Taipei City, Taiwan"},{"name":"National Taiwan University Graduate Institute of Clinical Medicine, Taipei City, Taiwan"},{"name":"Division of Gastroenterology and Hepatology, Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2847-4932","authenticated-orcid":false,"given":"Li-Rung","family":"Huang","sequence":"additional","affiliation":[{"name":"National Health Research Institutes, Institute of Molecular and Genomic Medicine, Miaoli, Taiwan"},{"name":"National Taiwan University Graduate Institute of Clinical Medicine, Taipei City, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"239","published-online":{"date-parts":[[2025,3,26]]},"reference":[{"key":"2025100302134926000_13.3.e009827.1","doi-asserted-by":"crossref","DOI":"10.20900\/immunometab20210007","article-title":"Diverse Roles of Akt in T cells","volume":"3","author":"Abdullah","year":"2021","journal-title":"Immunometabolism"},{"key":"2025100302134926000_13.3.e009827.2","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(98)70274-X"},{"key":"2025100302134926000_13.3.e009827.3","doi-asserted-by":"publisher","DOI":"10.1126\/science.1106148"},{"key":"2025100302134926000_13.3.e009827.4","doi-asserted-by":"publisher","DOI":"10.1038\/nri3278"},{"key":"2025100302134926000_13.3.e009827.5","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1103568"},{"key":"2025100302134926000_13.3.e009827.6","doi-asserted-by":"crossref","DOI":"10.3389\/fimmu.2013.00020","article-title":"Role of PI3K\/Akt signaling in memory CD8 T cell differentiation","volume":"4","author":"Kim","year":"2013","journal-title":"Front Immunol"},{"key":"2025100302134926000_13.3.e009827.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.10.013"},{"key":"2025100302134926000_13.3.e009827.8","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1611299114"},{"key":"2025100302134926000_13.3.e009827.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2007.09.006"},{"key":"2025100302134926000_13.3.e009827.10","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2035"},{"key":"2025100302134926000_13.3.e009827.11","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02844-06"},{"key":"2025100302134926000_13.3.e009827.12","doi-asserted-by":"publisher","DOI":"10.1002\/hep.23936"},{"key":"2025100302134926000_13.3.e009827.13","doi-asserted-by":"publisher","DOI":"10.1038\/nri3862"},{"key":"2025100302134926000_13.3.e009827.14","doi-asserted-by":"crossref","DOI":"10.1038\/ncomms7692","article-title":"PD-1 alters T-cell metabolic reprogramming by inhibiting glycolysis and promoting lipolysis and fatty acid oxidation","volume":"6","author":"Patsoukis","year":"2015","journal-title":"Nat Commun"},{"key":"2025100302134926000_13.3.e009827.15","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-17-0381"},{"key":"2025100302134926000_13.3.e009827.16","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.25.21.9543-9553.2005"},{"key":"2025100302134926000_13.3.e009827.17","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-065X.2009.00767.x"},{"key":"2025100302134926000_13.3.e009827.18","doi-asserted-by":"crossref","DOI":"10.3389\/fimmu.2022.990874","article-title":"Akt isoforms in the immune system","volume":"13","author":"Guerau-de-Arellano","year":"2022","journal-title":"Front Immunol"},{"key":"2025100302134926000_13.3.e009827.19","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1119038109"},{"key":"2025100302134926000_13.3.e009827.20","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1262"},{"key":"2025100302134926000_13.3.e009827.21","doi-asserted-by":"publisher","DOI":"10.15252\/embr.201541905"},{"key":"2025100302134926000_13.3.e009827.22","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1701204"},{"key":"2025100302134926000_13.3.e009827.23","doi-asserted-by":"crossref","DOI":"10.1080\/2162402X.2015.1005448","article-title":"Akt1 and -2 inhibition diminishes terminal differentiation and enhances central memory CD8+ T-cell proliferation and survival","volume":"4","author":"Abu Eid","year":"2015","journal-title":"Oncoimmunology"},{"key":"2025100302134926000_13.3.e009827.24","doi-asserted-by":"publisher","DOI":"10.1016\/j.cellsig.2012.01.001"},{"key":"2025100302134926000_13.3.e009827.25","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1003490","article-title":"CD40 activation rescues antiviral CD8+ T cells from PD-1-mediated exhaustion","volume":"9","author":"Isogawa","year":"2013","journal-title":"PLoS Pathog"},{"key":"2025100302134926000_13.3.e009827.26","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2012.03.006"},{"key":"2025100302134926000_13.3.e009827.27","doi-asserted-by":"crossref","DOI":"10.1186\/s40425-018-0462-3","article-title":"A novel spontaneous hepatocellular carcinoma mouse model for studying T-cell exhaustion in the tumor microenvironment","volume":"6","author":"Liu","year":"2018","journal-title":"J Immunother Cancer"},{"key":"2025100302134926000_13.3.e009827.28","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0506580102"},{"key":"2025100302134926000_13.3.e009827.29","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btr260"},{"key":"2025100302134926000_13.3.e009827.30","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M302847200"},{"key":"2025100302134926000_13.3.e009827.31","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.259549"},{"key":"2025100302134926000_13.3.e009827.32","doi-asserted-by":"publisher","DOI":"10.1160\/TH13-06-0484"},{"key":"2025100302134926000_13.3.e009827.33","unstructured":"Huang L , Lin S . Data from: mRNA-seq analysis of mouse cytotoxic T lymphocytes (CTLs) over-expressing Akt isoforms. Gene Expression Omnibus 2020. Available: https:\/\/www.ncbi.nlm.nih.gov\/geo\/query\/acc.cgi?acc=GSE151131"},{"key":"2025100302134926000_13.3.e009827.34","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0511137103"},{"key":"2025100302134926000_13.3.e009827.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2012.08.021"},{"key":"2025100302134926000_13.3.e009827.36","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20071641"},{"key":"2025100302134926000_13.3.e009827.37","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1201305"},{"key":"2025100302134926000_13.3.e009827.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2015.01.006"},{"key":"2025100302134926000_13.3.e009827.39","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1038\/s41586-019-0985-x","article-title":"NR4A transcription factors limit CAR T cell function in solid tumours","volume":"567","author":"Chen","year":"2019","journal-title":"Nature New Biol"},{"key":"2025100302134926000_13.3.e009827.40","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1905675116"},{"key":"2025100302134926000_13.3.e009827.41","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20160485"},{"key":"2025100302134926000_13.3.e009827.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.01.012"},{"key":"2025100302134926000_13.3.e009827.43","doi-asserted-by":"publisher","DOI":"10.1038\/ni.1689"},{"key":"2025100302134926000_13.3.e009827.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2013.07.013"},{"key":"2025100302134926000_13.3.e009827.45","doi-asserted-by":"publisher","DOI":"10.1126\/science.275.5308.1930"},{"key":"2025100302134926000_13.3.e009827.46","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-00612-6"},{"key":"2025100302134926000_13.3.e009827.47","doi-asserted-by":"crossref","DOI":"10.3389\/fimmu.2018.02747","article-title":"Revisiting the Concept of Targeting NFAT to Control T Cell Immunity and Autoimmune Diseases","volume":"9","author":"Lee","year":"2018","journal-title":"Front Immunol"},{"key":"2025100302134926000_13.3.e009827.48","doi-asserted-by":"crossref","DOI":"10.1084\/jem.20201599","article-title":"NFAT-dependent and -independent exhaustion circuits program maternal CD8 T cell hypofunction in pregnancy","volume":"219","author":"Lewis","year":"2022","journal-title":"J Exp Med"},{"key":"2025100302134926000_13.3.e009827.49","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-023-01614-x"},{"key":"2025100302134926000_13.3.e009827.50","first-page":"387","article-title":"IL-6 enhances the nuclear translocation of DNA cytosine-5-methyltransferase 1 (DNMT1) via phosphorylation of the nuclear localization sequence by the AKT kinase","volume":"4","author":"Hodge","year":"2007","journal-title":"Cancer Genomics Proteomics"},{"key":"2025100302134926000_13.3.e009827.51","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.22684"},{"key":"2025100302134926000_13.3.e009827.52","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.7842"},{"key":"2025100302134926000_13.3.e009827.53","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcancer.2019.04.001"},{"key":"2025100302134926000_13.3.e009827.54","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2014.06.004"},{"key":"2025100302134926000_13.3.e009827.55","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.25.15.6592-6602.2005"},{"key":"2025100302134926000_13.3.e009827.56","doi-asserted-by":"publisher","DOI":"10.1038\/nri2152"},{"key":"2025100302134926000_13.3.e009827.57","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2007.04.011"},{"key":"2025100302134926000_13.3.e009827.58","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(01)00233-3"},{"key":"2025100302134926000_13.3.e009827.59","doi-asserted-by":"crossref","DOI":"10.1002\/advs.202103508","article-title":"Inhibition of Calcium Signaling Prevents Exhaustion and Enhances Anti\u2010Leukemia Efficacy of CAR\u2010T Cells via SOCE\u2010Calcineurin\u2010NFAT and Glycolysis Pathways","volume":"9","author":"Shao","year":"2022","journal-title":"Adv Sci (Weinh)"},{"key":"2025100302134926000_13.3.e009827.60","doi-asserted-by":"crossref","first-page":"1113882","DOI":"10.3389\/fimmu.2023.1113882","article-title":"The current landscape of CAR T-cell therapy for solid tumors: Mechanisms, research progress, challenges, and counterstrategies","volume":"14","author":"Daei Sorkhabi","year":"2023","journal-title":"Front Immunol"},{"key":"2025100302134926000_13.3.e009827.61","doi-asserted-by":"publisher","DOI":"10.1038\/s41571-023-00832-4"},{"key":"2025100302134926000_13.3.e009827.62","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.copbio.2020.01.009","article-title":"All systems go: converging synthetic biology and combinatorial treatment for CAR-T cell therapy","volume":"65","author":"Lanitis","year":"2020","journal-title":"Curr Opin Biotechnol"},{"key":"2025100302134926000_13.3.e009827.63","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1038\/s41586-019-1805-z","article-title":"c-Jun overexpression in CAR T cells induces exhaustion resistance","volume":"576","author":"Lynn","year":"2019","journal-title":"Nature New Biol"},{"key":"2025100302134926000_13.3.e009827.64","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-021-00964-8"},{"key":"2025100302134926000_13.3.e009827.65","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2022.09.013"},{"key":"2025100302134926000_13.3.e009827.66","doi-asserted-by":"publisher","DOI":"10.1186\/s13045-023-01504-7"},{"key":"2025100302134926000_13.3.e009827.67","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.95103","article-title":"Inhibition of AKT signaling uncouples T cell differentiation from expansion for receptor-engineered adoptive immunotherapy","volume":"2","author":"Klebanoff","year":"2017","journal-title":"JCI Insight"},{"key":"2025100302134926000_13.3.e009827.68","first-page":"2379","article-title":"Akt inhibition at the initial stage of CAR-T preparation enhances the CAR-positive expression rate, memory phenotype and in vivo efficacy","volume":"9","author":"Zhang","year":"2019","journal-title":"Am J Cancer Res"},{"key":"2025100302134926000_13.3.e009827.69","doi-asserted-by":"publisher","DOI":"10.1126\/science.292.5522.1728"},{"key":"2025100302134926000_13.3.e009827.70","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2004.11.003"}],"container-title":["Journal for ImmunoTherapy of Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jitc-2024-009827","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T09:14:01Z","timestamp":1759482841000},"score":1,"resource":{"primary":{"URL":"https:\/\/jitc.bmj.com\/lookup\/doi\/10.1136\/jitc-2024-009827"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3]]},"references-count":70,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,3,26]]},"published-print":{"date-parts":[[2025,3]]}},"alternative-id":["10.1136\/jitc-2024-009827"],"URL":"https:\/\/doi.org\/10.1136\/jitc-2024-009827","relation":{},"ISSN":["2051-1426"],"issn-type":[{"value":"2051-1426","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3]]}}}