{"database":"GEO","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE302nnn/GSE302682/"]},"type":"primary"},"statusCodeValue":200,"statusCode":"OK"}],"scores":null,"additional":{"omics_type":["Transcriptomics"],"species":["Mus musculus"],"gds_type":["Expression profiling by high throughput sequencing"],"full_dataset_link":["https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE302682"],"repository":["GEO"],"entry_type":["GSE"],"additional_accession":[]},"is_claimable":false,"name":"Sleeping Beauty mutagenesis identifies BACH2 and other regulators promoting CD8+ T cell persistence and effector function under tumor-associated chronic antigen stimulation - RNAseq","description":"● Background: Genes that enhance T cell function could represent promising targets for improving engineered T cell therapies for cancer. While extensive CRISPR knockout screens have identified key genes that enhance T cell persistence, employing Sleeping Beauty (SB) insertional mutagenesis, which induces both gain- (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. ● Methods: We developed transgenic mouse models carrying Doxycycline (Dox)-inducible SB engineered system (DiSBey) in primary T cells. Using DiSBey, we conducted screens to identify genes that enhance T cell persistence under chronic antigen exposure. Specifically, CD8⁺ 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 SB transposon genomic insertion sites and corresponding fusion transcripts from the persistent DiSBey CD8⁺ T cells using enhanced-specificity tagmentation sequencing (esTag-seq) and RNA-seq, respectively. ● Results: Under chronic stimulation, SB-mutagenized CD8⁺ T cells exhibited improved persistence and reduced terminal exhaustion phenotype. Across six independent screens, we identified 38 genes that were recurrently targeted by the SB transposon T2/Onc2 and differentially expressed under chronic anti-CD3 stimulation stress. Among these, T2/Onc2 insertions into Bach2 and Elmo1 were repeatedly found at the genomic level and were associated with altered nascent transcript expression. Bach2, previously recognized as a key regulator of T cell memory formation and resistance to chronic viral infection but less characterized in engineered T cells for cancer therapy, was found to enhance in vivo persistence in the B16-Ova tumor model. We further demonstrated that ectopic Bach2 expression levels influence engineered T cell differentiation lineage. A Bach2low signature allowed differentiation into both KLRG1⁺ and CD62L⁺ phenotypes, whereas Bach2high restricted differentiation predominantly to the CD62L⁺ subset. Finally, in human CART19-28ζ cells, BACH2 overexpression enhanced cytotoxicity and improved tumor control following chronic cancer stimulation in vivo. ● Conclusions: Controllable SB mutagenesis using DiSBey mice provides a novel platform for functional screening of genes that improve T cell therapeutic phenotypes. Our findings highlight a dose-dependent role of BACH2 in enhancing the function of engineered T cells under conditions of chronic antigenic stimulation.","dates":{"publication":"2026/07/22"},"accession":"GSE302682","cross_references":{"GSM":["GSM9108790","GSM9108791","GSM9108792","GSM9108757","GSM9108758","GSM9108759","GSM9108793","GSM9108794","GSM9108795","GSM9108796","GSM9108752","GSM9108797","GSM9108753","GSM9108754","GSM9108798","GSM9108799","GSM9108755","GSM9108756","GSM9108780","GSM9108781","GSM9108782","GSM9108783","GSM9108784","GSM9108785","GSM9108786","GSM9108787","GSM9108788","GSM9108789","GSM9108770","GSM9108809","GSM9108779","GSM9108812","GSM9108813","GSM9108814","GSM9108815","GSM9108816","GSM9108817","GSM9108771","GSM9108772","GSM9108773","GSM9108774","GSM9108775","GSM9108776","GSM9108777","GSM9108810","GSM9108811","GSM9108778","GSM9108768","GSM9108801","GSM9108802","GSM9108769","GSM9108803","GSM9108804","GSM9108805","GSM9108806","GSM9108807","GSM9108808","GSM9108760","GSM9108761","GSM9108762","GSM9108763","GSM9108764","GSM9108765","GSM9108766","GSM9108800","GSM9108767"],"GPL":["30172","23479"],"GSE":["302682"],"taxon":["Mus musculus"],"PMID":["[42457339]"]}}