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Genome sequencing of human GBC has demonstrated that loss\u2010of\u2010function mutations of E74\u2010like ETS transcription factor 3 (<jats:italic>ELF3<\/jats:italic>) are frequently observed, with ELF3 considered to be a tumour suppressor in GBC. To clarify the underlying molecular mechanisms by which ELF3 suppresses GBC development, we performed <jats:italic>in vivo<\/jats:italic> analysis using a combination of autochthonous and allograft mouse models. We first evaluated the clinical significance of ELF3 expression in human GBC tissues and found that low ELF3 expression was associated with advanced clinical stage and deep tumour invasion. For <jats:italic>in vivo<\/jats:italic> analysis, we generated <jats:italic>Pdx1\u2010Cre<\/jats:italic>; <jats:italic>Kras<\/jats:italic><jats:sup>G12D<\/jats:sup>; <jats:italic>Trp53<\/jats:italic><jats:sup>R172H<\/jats:sup>; <jats:italic>Elf3<\/jats:italic><jats:sup>f\/f<\/jats:sup> (KPCE) mice and <jats:italic>Pdx1\u2010Cre<\/jats:italic>; <jats:italic>Kras<\/jats:italic><jats:sup>G12D<\/jats:sup>; <jats:italic>Trp53<\/jats:italic><jats:sup>R172H<\/jats:sup>; <jats:italic>Elf3<\/jats:italic><jats:sup>wt\/wt<\/jats:sup> (KPC) mice as a control and analysed their gallbladders histologically. KPCE mice developed larger papillary lesions in the gallbladder than those developed by KPC mice. Organoids established from the gallbladders of KPCE and KPC mice were analysed <jats:italic>in vitro<\/jats:italic>. RNA sequencing showed upregulated expression of epiregulin (<jats:italic>Ereg<\/jats:italic>) in KPCE organoids, and western blotting revealed that EGFR\/mechanical targets of rapamycin complex 1 (mTORC1) were upregulated in KPCE organoids. In addition, ChIP assays on <jats:italic>Elf3<\/jats:italic>\u2010overexpressing KPCE organoids showed that ELF3 directly regulated <jats:italic>Ereg<\/jats:italic>. <jats:italic>Ereg<\/jats:italic> deletion in KPCE organoids (using CRISPR\/Cas9) induced EGFR\/mTORC1 downregulation, indicating that ELF3 controlled EGFR\/mTORC1 activity through regulation of <jats:italic>Ereg<\/jats:italic> expression. We also generated allograft mouse models using KPCE and KPC organoids and found that KPCE organoid allograft tumours exhibited poorly differentiated structures with mTORC1 upregulation and mesenchymal phenotype, which were suppressed by <jats:italic>Ereg<\/jats:italic> deletion. Furthermore, EGFR\/mTORC1 inhibition suppressed cell proliferation and epithelial\u2013mesenchymal transition in KPCE organoids. Our results suggest that ELF3 suppresses GBC development via downregulation of EREG\/EGFR\/mTORC1 signalling. EGFR\/mTORC1 inhibition is a potential therapeutic option for GBC with <jats:italic>ELF3<\/jats:italic> mutation. \u00a9 2023 The Authors. <jats:italic>The Journal of Pathology<\/jats:italic> published by John Wiley &amp; Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.<\/jats:p>","DOI":"10.1002\/path.6144","type":"journal-article","created":{"date-parts":[[2023,6,24]],"date-time":"2023-06-24T12:48:46Z","timestamp":1687610926000},"page":"28-42","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["<scp>ELF3<\/scp> suppresses gallbladder cancer development through downregulation of the <scp>EREG<\/scp>\/<scp>EGFR<\/scp>\/<scp>mTOR<\/scp> complex 1 signalling pathway"],"prefix":"10.1002","volume":"261","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1794-3091","authenticated-orcid":false,"given":"Takeharu","family":"Nakamura","sequence":"first","affiliation":[{"name":"Department of 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