{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T06:32:15Z","timestamp":1781850735426,"version":"3.54.5"},"reference-count":44,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T00:00:00Z","timestamp":1749600000000},"content-version":"vor","delay-in-days":163,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"AZV","award":["NU22-05-00374"],"award-info":[{"award-number":["NU22-05-00374"]}]},{"name":"OPVVV","award":["CZ.02.1.01\/0.0\/0.0\/16_025\/0007428"],"award-info":[{"award-number":["CZ.02.1.01\/0.0\/0.0\/16_025\/0007428"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,12,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Cancer immunotherapy with transgenic T-cell receptor-engineered T cells (TCR-T) enables the targeting of intracellular tumor-specific antigens; in contrast, chimeric antigen receptor-modified T cells (CAR-T) mediate tumor cell killing via the recognition of surface antigens. In the case of acute myeloid leukemia, the lack of leukemia-specific surface antigens limits the efficacy of CAR-T cells; therefore, TCR-T cells may represent a more targeted immunotherapy approach. However, the tumor immunosuppressive environment eliminates the best-functioning, high-avidity TCR-T cells, thus creating a need for novel, enhanced TCR-T cells.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>The piggyBac transposon vector used for gene modification of T cells expresses a T-cell receptor specific to the WT1 tumour antigen, an NFAT promoter-regulated CAR specific to GM-CSF receptor, a CD3xCD33 bispecific T-cell engager, and a truncated EGFR suicide gene system. The transgenic T cells were generated by electroporation using a single expression vector, and the efficiency of these engineered TCR-T cells was evaluated using models that utilized AML cell lines and primary AML cells.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>The NFAT-driven GM-CSF CAR significantly enhances the antileukemic activity of WT1-specific TCR-T cells, which importantly maintain specificity for their HLA\/peptide antigenic complex. Next, by inserting the CD3xCD33 bispecific T-cell engager into the transposon vector, both TCR-T cells and recruited non-transfected bystander T cells can efficiently target the CD33 antigen, providing more robust antileukemic effects.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>The presented strategy, utilizing a single piggyBac transposon vector, enables the complex redirection of T-cell specificity against acute myeloid leukemia by inserting TCR, CAR, BiTE constructs, along with a tEGFR gene suicide system.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/immadv\/ltaf022","type":"journal-article","created":{"date-parts":[[2025,6,5]],"date-time":"2025-06-05T07:51:12Z","timestamp":1749109872000},"source":"Crossref","is-referenced-by-count":2,"title":["Targeting of acute myeloid leukemia by five-gene engineered T cells expressing transgenic T-cell receptor specific to WT1, chimeric antigenic receptor specific to GM-CSF receptor, bispecific T-cell engager specific to CD33, and tEGFR suicide gene system"],"prefix":"10.1093","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4855-4550","authenticated-orcid":false,"given":"Krist\u00fdna","family":"\u0160milauerov\u00e1","sequence":"first","affiliation":[{"name":"Institute of Hematology and Blood Transfusion, Department of Immunotherapy , Prague ,","place":["Czechia"]},{"name":"Faculty of Natural Sciences, Charles University , Prague ,","place":["Czechia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"\u0160tach","sequence":"additional","affiliation":[{"name":"Institute of Hematology and Blood Transfusion, Department of Immunotherapy , Prague ,","place":["Czechia"]},{"name":"Faculty of Natural Sciences, Charles University , Prague 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