{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T13:38:32Z","timestamp":1762609112248,"version":"3.37.3"},"reference-count":24,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2023,1,3]],"date-time":"2023-01-03T00:00:00Z","timestamp":1672704000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2023,1,3]],"date-time":"2023-01-03T00:00:00Z","timestamp":1672704000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["16-CE14-0010-01"],"award-info":[{"award-number":["16-CE14-0010-01"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Biallelic pathogenic variants in the <jats:italic>SLC34A3<\/jats:italic> gene, encoding for the NPT2c cotransporter, cause Hereditary Hypophosphatemic Rickets with Hypercalciuria (HHRH). However, the associated phenotype is highly variable. In addition, mice deleted for <jats:italic>Slc34a3<\/jats:italic> exhibit a different phenotype compared to humans, without urinary phosphate leakage. The mechanisms by which <jats:italic>SLC34A3<\/jats:italic> variants disrupt phosphate\/calcium metabolism are un-completely understood. In this study we explored these mechanisms in vitro using <jats:italic>SLC34A3<\/jats:italic> variants identified in patients with urinary phosphate leakage. We analyzed the consequences of these variants on NPT2c function and the link with the phenotype of the patients. We studied 20 patients with recurrent nephrolithiasis and low serum phosphate concentration harboring variants in the <jats:italic>SLC34A3<\/jats:italic> gene. Half of the patients carried homozygous or composite heterozygous variants. Three patients had in addition variants in <jats:italic>SLC34A1<\/jats:italic> and <jats:italic>SLC9A3R1<\/jats:italic> genes. All these patients benefited from a precise analysis of their phenotype. We generated 13 of these mutants by site-directed mutagenesis. Then we carried out transient transfections of these mutants in HEK cells and measured their phosphate uptake capacity under different conditions. Among the 20 patients included, 3 had not only mutations in NPT2c but also in NPT2a or NHERF1 genes. Phosphate uptake was decreased in 8 NPT2c mutants studied and normal for 5. Four variants were initially categorized as variants of uncertain significance. Expression of the corresponding mutants showed that one did not modify phosphate transport, two reduced it moderately and one abolished it. Co-transfection of the NPT2c mutants with the wild-type plasmid of NPT2c or NPT2a did not reveal dominant negative effect of the mutants on NPT2c-mediated phosphate transport. A detailed analysis of patient phenotypes did not find a link between the severity of the disorder and the level of phosphate transport impairment. NPT2c mutations classified as ACMG3 identified in patients with renal phosphate leak should be characterized by in vitro study to check if they alter NPT2c-mediated phosphate transport since phosphate uptake capacity may not be affected. In addition, research for mutations in NHERF1 and NPT2a genes should always be associated to NPT2c sequencing.<\/jats:p>","DOI":"10.1038\/s41598-022-25995-5","type":"journal-article","created":{"date-parts":[[2023,1,3]],"date-time":"2023-01-03T11:04:45Z","timestamp":1672743885000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Relationship between clinical phenotype and in vitro analysis of 13 NPT2c\/SCL34A3 mutants"],"prefix":"10.1038","volume":"13","author":[{"given":"Fran\u00e7ois","family":"Brazier","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie","family":"Courbebaisse","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amandine","family":"David","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Bergerat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christine","family":"Leroy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta","family":"Lindner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G\u00e9rard","family":"Maruani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Camille","family":"Saint Jacques","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emmanuel","family":"Letavernier","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marguerite","family":"Hureaux","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosa","family":"Vargas-Poussou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dominique","family":"Pri\u00e9","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2023,1,3]]},"reference":[{"key":"25995_CR1","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1056\/NEJM198503073121003","volume":"312","author":"M Tieder","year":"1985","unstructured":"Tieder, M. et al. Hereditary hypophosphatemic rickets with hypercalciuria. N. Engl. J. Med. 312, 611\u2013617 (1985).","journal-title":"N. Engl. J. Med."},{"key":"25995_CR2","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1056\/NEJM198701153160302","volume":"316","author":"M Tieder","year":"1987","unstructured":"Tieder, M. et al. \u201cIdiopathic\u201d hypercalciuria and hereditary hypophosphatemic rickets. Two phenotypical expressions of a common genetic defect. N. Engl. J. Med. 316, 125\u2013129 (1987).","journal-title":"N. Engl. J. Med."},{"key":"25995_CR3","doi-asserted-by":"publisher","first-page":"5372","DOI":"10.1073\/pnas.95.9.5372","volume":"95","author":"L Beck","year":"1998","unstructured":"Beck, L. et al. Targeted inactivation of Npt2 in mice leads to severe renal phosphate wasting, hypercalciuria, and skeletal abnormalities. Proc. Natl. Acad. Sci. 95, 5372\u20135377 (1998).","journal-title":"Proc. Natl. Acad. Sci."},{"key":"25995_CR4","doi-asserted-by":"publisher","first-page":"2366","DOI":"10.1681\/ASN.2013101085","volume":"25","author":"D Dasgupta","year":"2014","unstructured":"Dasgupta, D. et al. Mutations in SLC34A3\/NPT2c are associated with kidney stones and nephrocalcinosis. J. Am. Soc. Nephrol. 25, 2366\u20132375 (2014).","journal-title":"J. Am. Soc. Nephrol."},{"key":"25995_CR5","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1086\/499409","volume":"78","author":"C Bergwitz","year":"2006","unstructured":"Bergwitz, C. et al. SLC34A3 mutations in patients with hereditary hypophosphatemic rickets with hypercalciuria predict a key role for the sodium-phosphate cotransporter NaPi-IIc in maintaining phosphate homeostasis. Am. J. Hum. Genet. 78, 179\u2013192 (2006).","journal-title":"Am. J. Hum. Genet."},{"key":"25995_CR6","doi-asserted-by":"publisher","first-page":"4433","DOI":"10.1210\/jc.2009-1535","volume":"94","author":"AL Tencza","year":"2009","unstructured":"Tencza, A. L. et al. Hypophosphatemic rickets with hypercalciuria due to mutation in SLC34A3\/type IIc sodium-phosphate cotransporter: Presentation as hypercalciuria and nephrolithiasis. J. Clin. Endocrinol. Metab. 94, 4433\u20134438 (2009).","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"25995_CR7","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1681\/ASN.2008020177","volume":"20","author":"H Segawa","year":"2009","unstructured":"Segawa, H. et al. Type IIc sodium-dependent phosphate transporter regulates calcium metabolism. J. Am. Soc. Nephrol. 20, 104\u2013113 (2009).","journal-title":"J. Am. Soc. Nephrol."},{"key":"25995_CR8","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1182\/blood-2012-05-427302","volume":"121","author":"A Forand","year":"2013","unstructured":"Forand, A. et al. EKLF-driven PIT1 expression is critical for mouse erythroid maturation in vivo and in vitro. Blood 121, 666\u2013678 (2013).","journal-title":"Blood"},{"key":"25995_CR9","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1016\/j.cca.2015.09.021","volume":"451","author":"J-C Souberbielle","year":"2015","unstructured":"Souberbielle, J.-C. et al. Serum calcitriol concentrations measured with a new direct automated assay in a large population of adult healthy subjects and in various clinical situations. Clin. Chim. Acta Int. J. Clin. Chem. 451, 149\u2013153 (2015).","journal-title":"Clin. Chim. Acta Int. J. Clin. Chem."},{"key":"25995_CR10","doi-asserted-by":"publisher","first-page":"1128","DOI":"10.1056\/NEJMoa0802836","volume":"359","author":"Z Karim","year":"2008","unstructured":"Karim, Z. et al. NHERF1 mutations and responsiveness of renal parathyroid hormone. N. Engl. J. Med. 359, 1128\u20131135 (2008).","journal-title":"N. Engl. J. Med."},{"key":"25995_CR11","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1086\/499410","volume":"78","author":"B Lorenz-Depiereux","year":"2006","unstructured":"Lorenz-Depiereux, B. et al. Hereditary hypophosphatemic rickets with hypercalciuria is caused by mutations in the sodium-phosphate cotransporter gene SLC34A3. Am. J. Hum. Genet. 78, 193\u2013201 (2006).","journal-title":"Am. J. Hum. Genet."},{"key":"25995_CR12","doi-asserted-by":"publisher","first-page":"4022","DOI":"10.1210\/jc.2005-2840","volume":"91","author":"S Ichikawa","year":"2006","unstructured":"Ichikawa, S. et al. Intronic deletions in the SLC34A3 gene cause hereditary hypophosphatemic rickets with hypercalciuria. J. Clin. Endocrinol. Metab. 91, 4022\u20134027 (2006).","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"25995_CR13","doi-asserted-by":"publisher","first-page":"1100","DOI":"10.1016\/j.bone.2012.02.015","volume":"50","author":"Y Yu","year":"2012","unstructured":"Yu, Y. et al. Novel NaPi-IIc mutations causing HHRH and idiopathic hypercalciuria in several unrelated families: Long-term follow-up in one kindred. Bone 50, 1100\u20131106 (2012).","journal-title":"Bone"},{"key":"25995_CR14","doi-asserted-by":"publisher","first-page":"886","DOI":"10.1053\/j.ajkd.2018.12.026","volume":"73","author":"A Bertholet-Thomas","year":"2019","unstructured":"Bertholet-Thomas, A. et al. Fluconazole as a new therapeutic tool to manage patients with NPTIIc (SLC34A3) mutation: A case report. Am. J. Kidney Dis. 73, 886\u2013889 (2019).","journal-title":"Am. J. Kidney Dis."},{"key":"25995_CR15","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1007\/s00240-019-01116-2","volume":"47","author":"R Sch\u00f6nauer","year":"2019","unstructured":"Sch\u00f6nauer, R. et al. Evaluating pathogenicity of SLC34A3-Ser192Leu, a frequent European missense variant in disorders of renal phosphate wasting. Urolithiasis 47, 511\u2013519 (2019).","journal-title":"Urolithiasis"},{"key":"25995_CR16","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1681\/ASN.2014040388","volume":"26","author":"J Halbritter","year":"2015","unstructured":"Halbritter, J. et al. Fourteen monogenic genes account for 15% of nephrolithiasis\/nephrocalcinosis. J. Am. Soc. Nephrol. 26, 543\u2013551 (2015).","journal-title":"J. Am. Soc. Nephrol."},{"key":"25995_CR17","doi-asserted-by":"publisher","first-page":"604","DOI":"10.1681\/ASN.2014101025","volume":"27","author":"KP Schlingmann","year":"2016","unstructured":"Schlingmann, K. P. et al. Autosomal-recessive mutations in SLC34A1 encoding sodium-phosphate cotransporter 2A cause idiopathic infantile hypercalcemia. J. Am. Soc. Nephrol. 27, 604\u2013614 (2016).","journal-title":"J. Am. Soc. Nephrol."},{"key":"25995_CR18","doi-asserted-by":"publisher","first-page":"2289","DOI":"10.1007\/s00467-016-3443-0","volume":"31","author":"D Dinour","year":"2016","unstructured":"Dinour, D. et al. Loss of function of NaPiIIa causes nephrocalcinosis and possibly kidney insufficiency. Pediatr. Nephrol. (Berlin, Germany) 31, 2289\u20132297 (2016).","journal-title":"Pediatr. Nephrol. (Berlin, Germany)"},{"key":"25995_CR19","doi-asserted-by":"publisher","DOI":"10.14814\/phy2.13715","volume":"6","author":"A Fearn","year":"2018","unstructured":"Fearn, A. et al. Clinical, biochemical, and pathophysiological analysis of SLC34A1 mutations. Physiol. Rep. 6, e13715 (2018).","journal-title":"Physiol. Rep."},{"key":"25995_CR20","doi-asserted-by":"publisher","first-page":"1650","DOI":"10.1515\/cclm-2018-1208","volume":"57","author":"E de Paolis","year":"2019","unstructured":"de Paolis, E., Scaglione, G. L., de Bonis, M., Minucci, A. & Capoluongo, E. CYP24A1 and SLC34A1 genetic defects associated with idiopathic infantile hypercalcemia: From genotype to phenotype. Clin. Chem. Lab. Med. 57, 1650\u20131667 (2019).","journal-title":"Clin. Chem. Lab. Med."},{"unstructured":"The Genome Aggregation Database (gnomAD). https:\/\/gnomad.broadinstitute.org\/ (2019).","key":"25995_CR21"},{"unstructured":"Genomnis. Human Splicing Finder. (2020).","key":"25995_CR22"},{"key":"25995_CR23","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1038\/gim.2015.30","volume":"17","author":"S Richards","year":"2015","unstructured":"Richards, S. et al. Standards and guidelines for the interpretation of sequence variants: A joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet. Med. 17, 405\u2013424 (2015).","journal-title":"Genet. Med."},{"key":"25995_CR24","doi-asserted-by":"publisher","first-page":"383","DOI":"10.1038\/jid.2008.270","volume":"129","author":"GM Boyle","year":"2009","unstructured":"Boyle, G. M. et al. Macrophage inhibitory cytokine-1 is overexpressed in malignant melanoma and is associated with tumorigenicity. J. Investig. Dermatol. 129, 383\u2013391 (2009).","journal-title":"J. Investig. Dermatol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-022-25995-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-022-25995-5","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-022-25995-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,6]],"date-time":"2023-01-06T22:04:13Z","timestamp":1673042653000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-022-25995-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,3]]},"references-count":24,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["25995"],"URL":"https:\/\/doi.org\/10.1038\/s41598-022-25995-5","relation":{},"ISSN":["2045-2322"],"issn-type":[{"type":"electronic","value":"2045-2322"}],"subject":[],"published":{"date-parts":[[2023,1,3]]},"assertion":[{"value":"6 May 2022","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 December 2022","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 January 2023","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"85"}}