170476 20260422085545.0 doi 10.1016/j.biopha.2026.119371 sideral 148982 ART-2026-148982 eng Conde-Giménez, María (orcid)0000-0003-4358-9110 Variant-dependent pharmacological rescue of phenylalanine hydroxylase supports a precision therapeutic strategy for phenylketonuria 2026 Phenylketonuria (PKU) is an inherited metabolic disorder caused by pathogenic variants in phenylalanine hydroxylase (PAH), leading to toxic phenylalanine accumulation and severe neurological complications if untreated. Current pharmacological treatment relies on tetrahydrobiopterin (BH4), which benefits only a subset of patients, highlighting a major unmet need for alternative therapies. Here, we combined high-throughput screening, computational modelling, and drug repurposing to identify pharmacological chaperones capable of rescuing PAH function. We evaluated 26 structurally diverse small molecules in HEK293T cells expressing wild-type PAH or one of eight PKU-associated variants spanning phenotypes from mild to classical disease. Chaperoning efficacy was strongly variant-dependent, and for every variant tested at least one compound produced a greater activity increase than BH4 under identical assay conditions. Notably, belinostat, a clinically approved histone deacetylase inhibitor, emerged as the most effective compound for several clinically severe variants. Mechanistically, functional rescue consistently correlated with an increased population of tetrameric, catalytically competent PAH, as quantified by mass photometry. The crystal structure of the PAH–belinostat complex (PDB ID: 9T1O), together with structural models for all compounds, provide a framework for rational optimization. These results establish a preclinical proof-of-concept for genotype-guided pharmacological chaperone therapy in PKU and support the feasibility of personalized, variant-specific treatment strategies. Access copy available to the general public Unrestricted info:eu-repo/grantAgreement/ES/DGA/E45-20R info:eu-repo/grantAgreement/ES/DGA/E45-23R info:eu-repo/grantAgreement/EC/H2020/101004806/EU/MOlecular-Scale Biophysics Research Infrastructure/MOSBRI This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 101004806-MOSBRI info:eu-repo/grantAgreement/ES/MICINN/PID2019-107293GB-I00 info:eu-repo/grantAgreement/ES/MICINN/PID2022-141068NB-I00 info:eu-repo/semantics/openAccess by-nc-nd https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Salillas, Sandra (orcid)0000-0003-0195-5434 Galiana-Cameo, María (orcid)0000-0002-2043-4864 Martínez-Oliván, Juan E. (orcid)0000-0002-8784-7735 Mahía, Alejandro (orcid)0000-0002-8839-6796 Ledesma, Manuel Galano-Frutos, Juan José (orcid)0000-0002-1896-7805 Maity, Ritwik (orcid)0000-0003-4058-2888 Velázquez-Campoy, Adrián Universidad de Zaragoza (orcid)0000-0001-5702-4538 Díaz-de-Villegas, María D. Hurtado-Guerrero, Ramón (orcid)0000-0002-3122-9401 Sancho, Javier 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 199 (2026), 119371 [15 pp.] Biomed. pharmacother. BIOMEDICINE & PHARMACOTHERAPY 0753-3322 8365144 http://zaguan.unizar.es/record/170476/files/texto_completo.pdf Versión publicada 2409289 http://zaguan.unizar.es/record/170476/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:170476 articulos driver 2026-04-22-08:33:14 ARTICLE