{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T17:52:45Z","timestamp":1790790765890,"version":"4.1.0"},"reference-count":48,"publisher":"BMJ","issue":"7","license":[{"start":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T00:00:00Z","timestamp":1784073600000},"content-version":"unspecified","delay-in-days":14,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"Regenerative Medicine Minnesota Grant","award":["RMM 2025 02"],"award-info":[{"award-number":["RMM 2025 02"]}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30CA77598"],"award-info":[{"award-number":["P30CA77598"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R37CA266344"],"award-info":[{"award-number":["R37CA266344"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"American Cancer Society Research Professor Award","award":["123939"],"award-info":[{"award-number":["123939"]}]},{"DOI":"10.13039\/100007249","name":"University of Minnesota","doi-asserted-by":"publisher","award":["Academic Investment Research Program"],"award-info":[{"award-number":["Academic Investment Research Program"]}],"id":[{"id":"10.13039\/100007249","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Minnesota Partnership for Biotechnology and Medical Genomics Award","award":["CON000000113821"],"award-info":[{"award-number":["CON000000113821"]}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2026,6,28]]},"published-print":{"date-parts":[[2026,7]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Genes that enhance T-cell function represent promising targets for improving engineered T-cell therapies for cancer. While extensive CRISPR knockout screens have identified key genes enhancing T-cell persistence, employing\n                      <jats:italic>Sleeping Beauty<\/jats:italic>\n                      (\n                      <jats:italic>SB<\/jats:italic>\n                      ) insertional mutagenesis, which induces both gain-of-function (GOF) and loss-of-function (LOF) mutations via the generation of fusion transcripts with endogenous genes, may uncover additional critical factors that previous approaches have overlooked.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We developed transgenic mice carrying\n                      <jats:underline>D<\/jats:underline>\n                      oxycycline (Dox)-\n                      <jats:underline>i<\/jats:underline>\n                      nducible\n                      <jats:italic>\n                        <jats:underline>SB<\/jats:underline>\n                      <\/jats:italic>\n                      mutag\n                      <jats:underline>e<\/jats:underline>\n                      nesis s\n                      <jats:underline>y<\/jats:underline>\n                      stem (DiSBey) in primary T cells. Using DiSBey, we conducted screens for genetic alterations enhancing T-cell persistence under chronic antigen exposure. Specifically, CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells from Dox-fed DiSBey mice were subjected to repeated anti-CD3 stimulation over 18 days to mimic chronic antigenic stimulation. We then identified\n                      <jats:italic>SB<\/jats:italic>\n                      transposon genomic insertion sites and corresponding fusion transcripts from the persistent DiSBey CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells using enhanced-specificity tagmentation sequencing and RNA sequencing, respectively.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Under chronic stimulation,\n                      <jats:italic>SB<\/jats:italic>\n                      -mutagenized CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells exhibited improved persistence and reduced terminal exhaustion phenotype. Across six independent screens, we identified 38 genes that were recurrently targeted by the\n                      <jats:italic>SB<\/jats:italic>\n                      transposon T2\/Onc2 and differentially expressed under chronic anti-CD3 stimulation. T2\/Onc2 insertions into\n                      <jats:italic>Bach2<\/jats:italic>\n                      and\n                      <jats:italic>Elmo1<\/jats:italic>\n                      were recurrently identified at the genomic level and were associated with altered nascent transcript expression.\n                      <jats:italic>Bach2<\/jats:italic>\n                      , known as a key regulator of T-cell memory formation and resistance to chronic viral infection-induced exhaustion but less characterized in engineered T cells for cancer therapy, was found to counteract exhaustion in vitro and enhance in vivo tumor persistence in the B16-Ova tumor model. Further, we showed that ectopic\n                      <jats:italic>Bach2<\/jats:italic>\n                      expression levels influence engineered T-cell differentiation lineage, as low\n                      <jats:italic>Bach2<\/jats:italic>\n                      overexpression retained more functional progenitor exhausted T cells and exhibited improved therapeutic efficacy. Finally, in human CART19-28\u03b6 cells,\n                      <jats:italic>BACH2<\/jats:italic>\n                      overexpression enhanced cytotoxicity and tumor control following chronic cancer stimulation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      Controllable\n                      <jats:italic>SB<\/jats:italic>\n                      mutagenesis using DiSBey mice provides a novel platform for functional screening of genes that improve T-cell phenotypes important for their use as therapies. Our findings highlight a dose-dependent role of BACH2 in enhancing the function of engineered T cells under conditions of chronic antigenic stimulation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2025-013536","type":"journal-article","created":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T12:10:44Z","timestamp":1784117444000},"page":"e013536","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":1,"title":["<i>Sleeping Beauty<\/i>\n                    mutagenesis identifies\n                    <i>BACH2<\/i>\n                    and other regulators of CD8\n                    <sup>+<\/sup>\n                    T-cell exhaustion, persistence in vivo, and CAR-T cell function under tumor-associated chronic antigen stimulation"],"prefix":"10.1136","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-9841-1874","authenticated-orcid":false,"given":"Chang-Jung","family":"Lee","sequence":"first","affiliation":[{"name":"University of Minnesota Twin Cities Department of Genetics Cell Biology and Development, Minneapolis, Minnesota, USA"},{"name":"University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alex T","family":"Larsson","sequence":"additional","affiliation":[{"name":"University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tyler A","family":"Jubenville","sequence":"additional","affiliation":[{"name":"University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wendy 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