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However, resistance, either primary or acquired, remains a major clinical challenge. To better understand the underlying resistance mechanisms to BTK inhibitors, we established an ibrutinib-resistant model from a patient-derived splenic marginal zone lymphoma (MZL) cell line (VL51) through prolonged drug exposure. Resistant cells exhibited a 15-fold increase in ibrutinib\u2019s IC50, along with distinct morphological changes, mitochondrial activation, and cross-resistance to covalent, non-covalent BTK inhibitors and BTK degraders. Integrated transcriptomic, epigenomic, and proteomic analyses identified overexpression and secretion of IL-16 as a key feature of resistance, driven by chromatin remodeling and activation of the FLI1 transcription factor. IL-16 conferred ibrutinib resistance via CD9-mediated activation of the NF-\u03baB and AKT signaling pathways and was found to be elevated in the serum of ibrutinib-refractory CLL patients. Functional studies showed that targeting the IL-16\/CD9 axis using neutralizing antibodies or CD9-binding peptides restored sensitivity to BTK inhibitors and R-CHOP chemotherapy in MZL, mantle cell lymphoma, and diffuse large B-cell lymphoma models. These findings reveal a novel, targetable resistance mechanism with potential therapeutic implications for overcoming BTK inhibitor resistance in B-cell lymphomas.<\/jats:p>","DOI":"10.1101\/2025.05.07.652612","type":"posted-content","created":{"date-parts":[[2025,5,10]],"date-time":"2025-05-10T12:05:16Z","timestamp":1746878716000},"source":"Crossref","is-referenced-by-count":5,"title":["IL-16 production is a mechanism of resistance to BTK inhibitors and R-CHOP in lymphomas"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3123-6203","authenticated-orcid":false,"given":"Alberto J.","family":"Arribas","sequence":"first","affiliation":[{"name":"Institute of Oncology Research, Faculty of Biomedical Sciences, USI, Bellinzona, Switzerland"},{"name":"SIB Swiss Institute of Bioinformatics, Lausanne, 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