{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T04:00:05Z","timestamp":1792555205123,"version":"4.5.0"},"reference-count":40,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,6,4]],"date-time":"2025-06-04T00:00:00Z","timestamp":1748995200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Ectonucleotide Pyrophosphatase\/Phosphodiesterase 1 (<jats:italic>ENPP1<\/jats:italic>) plays a key role in mineralization processes, and mutations in this gene are associated with various severe diseases. Clinical case reports have implicated the <jats:italic>ENPP1<\/jats:italic> Y451C mutation in diffuse idiopathic skeletal hyperostosis patients, but its precise impact on bone mineralization and ectopic calcification remains unclear.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We used bioinformatics tools and <jats:italic>in vitro<\/jats:italic> functional assays to assess the impact of the <jats:italic>ENPP1<\/jats:italic> Y451C mutation on protein structure and enzymatic activity. Furthermore, we generated a knock-in mouse model (<jats:italic>Enpp1<jats:sup>Y433C<\/jats:sup><\/jats:italic>) to evaluate microarchitecture or signs of ectopic calcification by Micro-CT.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Bioinformatics analysis and <jats:italic>in vitro<\/jats:italic> assays showed that the Y451C mutation affects the <jats:italic>ENPP1<\/jats:italic> protein\u2019s structure, reducing enzymatic activity by approximately 50%. We successfully generated the <jats:italic>Enpp1<jats:sup>Y433C<\/jats:sup><\/jats:italic> knock-in mouse model. However, no significant differences were observed in body phenotype or biochemical markers in <jats:italic>Enpp1<jats:sup>Y433C<\/jats:sup><\/jats:italic> mice at 3, 5, and 10 months, compared to wild-type controls. Similarly, no significant changes were observed in bone microarchitecture or signs of ectopic calcification.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The <jats:italic>ENPP1<\/jats:italic> Y451C mutation significantly reduces enzymatic activity <jats:italic>in vitro<\/jats:italic>, yet the <jats:italic>Enpp1<jats:sup>Y433C<\/jats:sup><\/jats:italic> knock-in mouse model shows no significant abnormalities in mineralization, providing additional evidence for the pathogenicity assessment of <jats:italic>ENPP1<\/jats:italic> Y451C variant. Given that these results are from mouse models, further studies are required to clarify its pathogenicity in humans.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2025.1566392","type":"journal-article","created":{"date-parts":[[2025,6,4]],"date-time":"2025-06-04T04:14:08Z","timestamp":1749010448000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Impact of the ENPP1 mutation on bone mineralization and ectopic calcification: evidence from in vitro and in vivo 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