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Here, we describe small molecule inhibitors of USP25 and USP28, leading to cancer cell cycle arrest and death. However, genetic deletion of USP25\/28 does not replicate this effect. An integrated \u2013omics approach revealed off-target effects for thienopyridine carboxamide compounds upon the translation apparatus. Chemoproteomics and biochemical analyses suggested binding of the compound to a region near the ribosome complex polypeptide exit tunnel. Structural analysis of a USP28-inhibitor complex enabled the design of modified USP25\/28 inhibitor molecules which minimized translation-related off-target effects. In distinction to earlier compounds, the optimized inhibitors were non-toxic to breast cancer cells yet retained potent anti-proliferative activity in squamous lung carcinoma cells, where USP28 is associated with disease progression. Together, our results demonstrate that refined USP25\/28 inhibitors can selectively suppress tumor growth by targeting the TP63\u2013FBW7\u2013c-MYC signaling axis, offering a more precise therapeutic strategy for treating squamous lung cancers whilst minimizing undesired cytotoxicity.<\/jats:p>","DOI":"10.64898\/2026.03.17.712179","type":"posted-content","created":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T14:45:14Z","timestamp":1774017914000},"source":"Crossref","is-referenced-by-count":0,"title":["Refined USP25\/28 inhibitors with improved selectivity towards c-Myc driven squamous lung cancer cells"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1693-9664","authenticated-orcid":false,"given":"Ad\u00e1n","family":"Pinto-Fern\u00e1ndez","sequence":"first","affiliation":[{"name":"Chinese Academy of Medical Sciences Oxford Institute, Nuffield Department of Medicine, Oxford, UK"},{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1927-9577","authenticated-orcid":false,"given":"Claire","family":"Heride","sequence":"additional","affiliation":[{"name":"Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Crown St., Liverpool L69 3BX, UK"},{"name":"Cancer Research Horizons, 2 Redman Place, London E20 1JQ, UK"}]},{"given":"Andrew P","family":"Turnbull","sequence":"additional","affiliation":[{"name":"Cancer Research Horizons, 2 Redman Place, London E20 1JQ, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5735-5094","authenticated-orcid":false,"given":"Dora","family":"Pedroso","sequence":"additional","affiliation":[{"name":"Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Crown St., Liverpool L69 3BX, UK"},{"name":"GIMM - Gulbenkian Institute for Molecular Medicine, Edif\u00edcio Egas Moniz Avenida Professor Egas Moniz, 1649-028 Lisbon, Portugal"}]},{"given":"Victoria","family":"Smith","sequence":"additional","affiliation":[{"name":"Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Crown St., Liverpool L69 3BX, UK"}]},{"given":"Lisa","family":"Mullee","sequence":"additional","affiliation":[{"name":"Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Crown St., Liverpool L69 3BX, UK"}]},{"given":"Wojciech W","family":"Krajewski","sequence":"additional","affiliation":[{"name":"Cancer Research Horizons, 2 Redman Place, London E20 1JQ, UK"}]},{"given":"Cameron","family":"Bell","sequence":"additional","affiliation":[{"name":"Cancer Research Horizons, 2 Redman Place, London E20 1JQ, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6308-2491","authenticated-orcid":false,"given":"Anthony","family":"Varca","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, MA, USA"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4267-1876","authenticated-orcid":false,"given":"Thomas","family":"Charlton","sequence":"additional","affiliation":[{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]},{"given":"Dylan","family":"Jones","sequence":"additional","affiliation":[{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6906-4510","authenticated-orcid":false,"given":"Tom","family":"McAllister","sequence":"additional","affiliation":[{"name":"Chemistry Research Laboratory, University of Oxford, Oxford, UK; Chemistry, School of Natural and Environmental Sciences, Newcastle University, Newcastle Upon Tyne, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9715-5951","authenticated-orcid":false,"given":"Roman","family":"Fischer","sequence":"additional","affiliation":[{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8076-1480","authenticated-orcid":false,"given":"Elena","family":"Navarro-Guerrero","sequence":"additional","affiliation":[{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6495-7026","authenticated-orcid":false,"given":"Daniel","family":"Ebner","sequence":"additional","affiliation":[{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1169-5082","authenticated-orcid":false,"given":"Akane","family":"Kawamura","sequence":"additional","affiliation":[{"name":"Chemistry Research Laboratory, University of Oxford, Oxford, UK; Chemistry, School of Natural and Environmental Sciences, Newcastle University, Newcastle Upon Tyne, UK"}]},{"given":"Sun","family":"Kim","sequence":"additional","affiliation":[{"name":"FORMA Therapeutics, Watertown, Boston, MA, USA"},{"name":"Incyte, Philadelphia, USA"}]},{"given":"Dave","family":"Guerin","sequence":"additional","affiliation":[{"name":"FORMA Therapeutics, Watertown, Boston, MA, USA"},{"name":"Molecular Design Consulting LLC, Boston, USA"}]},{"given":"Tim R","family":"Hammonds","sequence":"additional","affiliation":[{"name":"Cancer Research Horizons, 2 Redman Place, London E20 1JQ, UK"},{"name":"University of Leicester, UK"}]},{"given":"Jeffrey","family":"Kearns","sequence":"additional","affiliation":[{"name":"FORMA Therapeutics, Watertown, Boston, MA, USA"},{"name":"Novartis Institutes for BioMedical Research (NIBR), Cambridge MA, USA"}]},{"given":"Neil","family":"Jones","sequence":"additional","affiliation":[{"name":"Cancer Research Horizons, 2 Redman Place, London E20 1JQ, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4562-1823","authenticated-orcid":false,"given":"Sara J","family":"Buhrlage","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, MA, USA"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8092-4320","authenticated-orcid":false,"given":"David","family":"Komander","sequence":"additional","affiliation":[{"name":"Ubiquitin Signalling Division, The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, AU"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4735-9814","authenticated-orcid":false,"given":"Sylvie","family":"Urb\u00e9","sequence":"additional","affiliation":[{"name":"Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Crown St., Liverpool L69 3BX, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3355-9479","authenticated-orcid":false,"given":"Michael J","family":"Clague","sequence":"additional","affiliation":[{"name":"Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Crown St., Liverpool L69 3BX, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8160-2446","authenticated-orcid":false,"given":"Benedikt M","family":"Kessler","sequence":"additional","affiliation":[{"name":"Chinese Academy of Medical Sciences Oxford Institute, Nuffield Department of Medicine, Oxford, UK"},{"name":"Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, Oxford, UK"}]}],"member":"54368","reference":[{"key":"2026032712453241000_2026.03.17.712179v2.1","doi-asserted-by":"crossref","unstructured":"Wu, X. et al. 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