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Prior evidence highlighted a significant contribution of the non\u2010canonical NF\u2010\u03baB signalling pathway in initiation and aggressiveness of different tumour types. Lymphotoxin\u2010\u03b2 (LT\u03b2) stimulates the NF\u2010\u03baB\u2010inducing kinase (NIK), resulting in the activation of the transcription factor RelB. However, the functional contribution of the non\u2010canonical NF\u2010\u03baB signalling pathway via the LT\u03b2\/NIK\/RelB axis in CCA carcinogenesis and progression has not been established.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Human CCA\u2010derived cell lines and organoids were examined to determine the expression of NF\u2010\u03baB pathway components upon activation or inhibition. Proliferation and cell death were analysed using real\u2010time impedance measurement and flow cytometry. Immunoblot, qRT\u2010PCR, RNA sequencing and in\u00a0situ hybridization were employed to analyse gene and protein expression. Four in\u00a0vivo models of iCCA were used to probe the activation and regulation of the non\u2010canonical NF\u2010\u03baB pathway.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Exposure to LT\u03b11\/\u03b22 activates the LT\u03b2\/NIK\/RelB axis and promotes proliferation in CCA. Inhibition of NIK with the small molecule inhibitor B022 efficiently suppresses RelB expression in patient\u2010derived CCA organoids and nuclear co\u2010translocation of RelB and p52 stimulated by LT\u03b11\/\u03b22 in CCA cell lines. In murine CCA, RelB expression is significantly increased and LT\u03b2 is the predominant ligand of the non\u2010canonical NF\u2010\u03baB signalling pathway.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our study confirms that the non\u2010canonical NF\u2010\u03baB axis LT\u03b2\/NIK\/RelB drives cholangiocarcinogenesis and represents a candidate therapeutic target.<\/jats:p><\/jats:sec>","DOI":"10.1111\/liv.16069","type":"journal-article","created":{"date-parts":[[2024,8,22]],"date-time":"2024-08-22T02:20:33Z","timestamp":1724293233000},"page":"2950-2963","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Lymphotoxin beta\u2010activated <scp>LTBR\/NIK\/RELB<\/scp> axis drives proliferation in cholangiocarcinoma"],"prefix":"10.1111","volume":"44","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5595-7787","authenticated-orcid":false,"given":"Kaiyu","family":"Xu","sequence":"first","affiliation":[{"name":"Department of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg Heidelberg University Hospital  Heidelberg Germany"},{"name":"Liver Cancer Center Heidelberg Heidelberg University Hospital  Heidelberg Germany"},{"name":"Medical Faculty Heidelberg Heidelberg University  Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annika","family":"Kessler","sequence":"additional","affiliation":[{"name":"Department of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg Heidelberg University Hospital  Heidelberg Germany"},{"name":"Liver Cancer Center Heidelberg Heidelberg University Hospital  Heidelberg Germany"},{"name":"Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology University 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