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Endocrinol."],"abstract":"<jats:sec><jats:title>Context<\/jats:title><jats:p>Although a monoallelic mutation in the calcium-sensing receptor (<jats:italic>CASR<\/jats:italic>) gene causes familial hypocalciuric hypercalcemia (FHH), the functional characterization of the identified <jats:italic>CASR<\/jats:italic> mutation linked to the clinical response to calcimimetics therapy is still limited.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>A 45-year-old male presenting with moderate hypercalcemia, hypocalciuria, and inappropriately high parathyroid hormone (PTH) had a good response to cinacalcet (total serum calcium (Ca<jats:sup>2+<\/jats:sup>) from 12.5 to 10.1 mg\/dl). We identified the genetic mutation and characterized the functional and pathophysiological mechanisms, and then linked the mutation to calcimimetics treatment <jats:italic>in vitro<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Design<\/jats:title><jats:p>Sanger sequencing of the <jats:italic>CASR<\/jats:italic>, <jats:italic>GNA11<\/jats:italic>, and <jats:italic>AP2S1<\/jats:italic> genes was performed in his family. The simulation model was used to predict the function of the identified mutant. <jats:italic>In vitro<\/jats:italic> studies, including immunoblotting, immunofluorescence, a cycloheximide chase study, Calbryte\u2122 520 Ca<jats:sup>2+<\/jats:sup> detection, and half-maximal effective concentration (EC<jats:sub>50<\/jats:sub>), were examined.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>This proband was found to carry a <jats:italic>de novo<\/jats:italic> heterozygous missense <jats:italic>I554N<\/jats:italic> in the cysteine-rich domain of <jats:italic>CASR<\/jats:italic>, which was pathogenic based on the different software prediction models and ACGME criteria. The simulation model showed that <jats:italic>CASR I554N<\/jats:italic> mutation decreased its binding energy with Ca<jats:sup>2+<\/jats:sup>. Human <jats:italic>CASR I554N<\/jats:italic> mutation attenuated the stability of CASR protein, reduced the expression of p-ERK 1\/2, and blunted the intracellular Ca<jats:sup>2+<\/jats:sup> response to gradient extracellular Ca<jats:sup>2+<\/jats:sup> (eCa<jats:sup>2+<\/jats:sup>) concentration. The EC<jats:sub>50<\/jats:sub> study also demonstrated the correctable effect of calcimimetics on the function of the <jats:italic>CASR I554N<\/jats:italic> mutation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This novel <jats:italic>CASR I554N<\/jats:italic> mutation causing FHH attenuates CASR stability, its binding affinity with Ca<jats:sup>2+<\/jats:sup>, and the response to eCa<jats:sup>2+<\/jats:sup> corrected by therapeutic calcimimetics.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2024.1291160","type":"journal-article","created":{"date-parts":[[2024,2,29]],"date-time":"2024-02-29T05:07:57Z","timestamp":1709183277000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Identification and characterization of a novel CASR mutation causing familial hypocalciuric 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