{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T03:17:52Z","timestamp":1792552672898,"version":"4.5.0"},"reference-count":37,"publisher":"Oxford University Press (OUP)","issue":"11","license":[{"start":{"date-parts":[[2014,5,13]],"date-time":"2014-05-13T00:00:00Z","timestamp":1399939200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,11,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:sec>\n                  <jats:title>\u2003<\/jats:title>\n                  <jats:p>PHOSPHO1 and tissue-nonspecific alkaline phosphatase (TNAP) have nonredundant functions during skeletal mineralization. Although TNAP deficiency (Alpl\u2212\/\u2212 mice) leads to hypophosphatasia, caused by accumulation of the mineralization inhibitor inorganic pyrophosphate (PPi), comparably elevated levels of PPi in Phospho1\u2212\/\u2212 mice do not explain their stunted growth, spontaneous fractures, bowed long bones, osteomalacia, and scoliosis. We have previously shown that elevated PPi in Alpl\u2212\/\u2212 mice is accompanied by elevated osteopontin (OPN), another potent mineralization inhibitor, and that the amount of OPN correlates with the severity of hypophosphatasia in mice. Here we demonstrate that plasma OPN is elevated and OPN expression is upregulated in the skeleton, particularly in the vertebrae, of Phospho1\u2212\/\u2212 mice. Liquid chromatography\/tandem mass spectrometry showed an increased proportion of phosphorylated OPN (p-OPN) peptides in Phospho1\u2212\/\u2212 mice, suggesting that accumulation of p-OPN causes the skeletal abnormalities in Phospho1\u2212\/\u2212 mice. We also show that ablation of the OPN gene, Spp1, leads to improvements in the skeletal phenotype in Phospho1\u2212\/\u2212 as they age. In particular, their scoliosis is ameliorated at 1 month of age and is completely rescued at 3 months of age. There is also improvement in the long bone defects characteristic of Phospho1\u2212\/\u2212 mice at 3 months of age. Mineralization assays comparing [Phospho1\u2212\/\u2212; Spp1\u2212\/\u2212], Phospho1\u2212\/\u2212, and Spp1\u2212\/\u2212 chondrocytes display corrected mineralization by the double knockout cells. Expression of chondrocyte differentiation markers was also normalized in the [Phospho1\u2212\/\u2212; Spp1\u2212\/\u2212] mice. Thus, although Alpl and Phospho1 deficiencies lead to similar skeletal phenotypes and comparable changes in the expression levels of PPi and OPN, there is a clear dissociation in the hierarchical roles of these potent inhibitors of mineralization, with elevated PPi and elevated p-OPN levels causing the respective skeletal phenotypes in Alpl\u2212\/\u2212 and Phospho1\u2212\/\u2212 mice. \u00a9 2014 American Society for Bone and Mineral Research.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1002\/jbmr.2281","type":"journal-article","created":{"date-parts":[[2014,5,13]],"date-time":"2014-05-13T22:17:44Z","timestamp":1400019464000},"page":"2369-2381","source":"Crossref","is-referenced-by-count":53,"title":["Ablation of Osteopontin Improves the Skeletal Phenotype of <i>Phospho1<\/i>\n                  <i>\u2212\/\u2212<\/i> Mice"],"prefix":"10.1093","volume":"29","author":[{"given":"Manisha C","family":"Yadav","sequence":"first","affiliation":[{"name":"Sanford Children's Health Research CenterSanford-Burnham Medical Research InstituteLa JollaCAUSA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmen","family":"Huesa","sequence":"additional","affiliation":[{"name":"The Roslin Institute and R(D)SVSUniversity of Edinburgh, Easter BushMidlothianScotlandUK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonoko","family":"Narisawa","sequence":"additional","affiliation":[{"name":"Sanford Children's Health Research CenterSanford-Burnham Medical Research InstituteLa JollaCAUSA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc F","family":"Hoylaerts","sequence":"additional","affiliation":[{"name":"Department of Cardiovascular SciencesCenter for Molecular and Vascular BiologyUniversity of LeuvenLeuvenBelgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Moreau","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Faculty of Dentistry, Department of BiochemistryFaculty of Medicine, Universit\u00e9 de Montr\u00e9al, CHU Sainte-Justine Research CenterMontrealCanada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin","family":"Farquharson","sequence":"additional","affiliation":[{"name":"The Roslin Institute and R(D)SVSUniversity of Edinburgh, Easter BushMidlothianScotlandUK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Luis","family":"Mill\u00e1n","sequence":"additional","affiliation":[{"name":"Sanford Children's Health Research CenterSanford-Burnham Medical Research InstituteLa JollaCAUSA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2014,5,13]]},"reference":[{"key":"2024020318472369000_jbmr2281-bib-0001","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1016\/j.bone.2006.05.014","article-title":"The presence of PHOSPHO1 in matrix vesicles and its developmental expression prior to skeletal mineralization","volume":"39","author":"Stewart","year":"2006","journal-title":"Bone."},{"key":"2024020318472369000_jbmr2281-bib-0002","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1359\/jbmr.070108","article-title":"Functional involvement of PHOSPHO1 in matrix vesicle-mediated skeletal mineralization","volume":"22","author":"Roberts","year":"2007","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0003","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1002\/jbmr.195","article-title":"Loss of skeletal mineralization by the simultaneous ablation of PHOSPHO1 and alkaline phosphatase function: a unified model of the mechanisms of initiation of skeletal calcification","volume":"26","author":"Yadav","year":"2011","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0004","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1016\/j.bone.2011.01.010","article-title":"PHOSPHO1 is essential for mechanically competent mineralization and the avoidance of spontaneous fractures","volume":"48","author":"Huesa","year":"2011","journal-title":"Bone."},{"key":"2024020318472369000_jbmr2281-bib-0005","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1177\/0022034513490958","article-title":"Compounded PHOSPHO1\/ALPL deficiencies reduce dentin mineralization","volume":"92","author":"McKee","year":"2013","journal-title":"J Dent Res."},{"key":"2024020318472369000_jbmr2281-bib-0006","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1007\/s00223-012-9672-8","article-title":"The role of phosphatases in the initiation of skeletal mineralization","volume":"93","author":"Mill\u00e1n","year":"2013","journal-title":"Calcif Tissue Int."},{"key":"2024020318472369000_jbmr2281-bib-0007","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1101\/gad.1276205","article-title":"Unique coexpression in osteoblasts of broadly expressed genes accounts for the spatial restriction of ECM mineralization to bone","volume":"19","author":"Murshed","year":"2005","journal-title":"Genes Dev."},{"key":"2024020318472369000_jbmr2281-bib-0008","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1359\/jbmr.071213","article-title":"Enzyme replacement therapy for murine hypophosphatasia","volume":"23","author":"Mill\u00e1n","year":"2008","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0009","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/S0169-6009(08)80225-5","article-title":"Osteopontin hydroxyapatite interactions in vitro. Inhibition of hydroxyapatite formation and growth in a gelatin gel","volume":"22","author":"Boskey","year":"1993","journal-title":"Bone Miner."},{"key":"2024020318472369000_jbmr2281-bib-0010","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1042\/bj3000723","article-title":"Modulation of crystal formation by bone phosphoproteins; structural specificity of the osteopontin-mediated inhibition of hydroxyapatite formation","volume":"300","author":"Hunter","year":"1994","journal-title":"Biochem J."},{"key":"2024020318472369000_jbmr2281-bib-0011","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1177\/10454411000110030101","volume":"11","author":"Sodek","year":"2000","journal-title":"Osteopontin. Crit Rev Oral Biol Med."},{"key":"2024020318472369000_jbmr2281-bib-0012","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1007\/s00223-013-9698-6","article-title":"Role of osteopontin in modulation of hydroxyapatite formation","volume":"93","author":"Hunter","year":"2013","journal-title":"Calcif Tissue Int."},{"key":"2024020318472369000_jbmr2281-bib-0013","doi-asserted-by":"crossref","first-page":"8819","DOI":"10.1073\/pnas.83.23.8819","article-title":"Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence","volume":"83","author":"Oldberg","year":"1986","journal-title":"Proc Natl Acad Sci USA."},{"key":"2024020318472369000_jbmr2281-bib-0014","doi-asserted-by":"crossref","first-page":"2040","DOI":"10.1002\/pro.5560041009","article-title":"Posttranslational modifications of bovine osteopontin: identification of twenty-eight phosphorylation and three O-glycosylation sites","volume":"4","author":"Sorensen","year":"1995","journal-title":"Protein Sci."},{"key":"2024020318472369000_jbmr2281-bib-0015","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1359\/jbmr.090832","article-title":"Phosphorylation-dependent inhibition of mineralization by osteopontin ASARM peptides is regulated by PHEX cleavage","volume":"25","author":"Addison","year":"2010","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0016","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s00223-001-1121-z","article-title":"Osteopontin deficiency increases mineral content and mineral crystallinity in mouse bone","volume":"71","author":"Boskey","year":"2002","journal-title":"Calcif Tissue Int."},{"key":"2024020318472369000_jbmr2281-bib-0017","doi-asserted-by":"crossref","first-page":"1377","DOI":"10.1359\/jbmr.060619","article-title":"Elevated skeletal osteopontin levels contribute to the hypophosphatasia phenotype in Akp2\u2212\/\u2212 mice","volume":"21","author":"Harmey","year":"2006","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0018","doi-asserted-by":"crossref","first-page":"994","DOI":"10.1359\/jbmr.2003.18.6.994","article-title":"Linked deficiencies in extracellular PPi and osteopontin mediate pathologic calcification associated with defective PC-1 and ANK expression","volume":"18","author":"Johnson","year":"2003","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0019","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1016\/S0002-9440(10)63208-7","article-title":"Concerted regulation of inorganic pyrophosphate and osteopontin by Akp2, Enpp1, and Ank\u2014an integrated model of the pathogenesis of mineralization disorders","volume":"164","author":"Harmey","year":"2004","journal-title":"Am J Pathol."},{"key":"2024020318472369000_jbmr2281-bib-0020","doi-asserted-by":"crossref","first-page":"15872","DOI":"10.1074\/jbc.M701116200","article-title":"Pyrophosphate inhibits mineralization of osteoblast cultures by binding to mineral, up-regulating osteopontin and inhibiting alkaline phosphatase activity","volume":"282","author":"Addison","year":"2007","journal-title":"J Biol Chem."},{"key":"2024020318472369000_jbmr2281-bib-0021","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1359\/jbmr.1998.13.7.1101","article-title":"Mice lacking osteopontin show normal development and bone structure but display altered osteoclast formation in vitro","volume":"13","author":"Rittling","year":"1998","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0022","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1002\/1096-9896(2000)9999:9999<::AID-PATH722>3.0.CO;2-Y","article-title":"Abnormal vitamin B6 metabolism in alkaline phosphatase knock-out mice causes multiple abnormalities, but not the impaired bone mineralization","volume":"193","author":"Narisawa","year":"2001","journal-title":"J Pathol."},{"key":"2024020318472369000_jbmr2281-bib-0023","doi-asserted-by":"crossref","first-page":"9445","DOI":"10.1073\/pnas.142063399","article-title":"Tissue-nonspecific alkaline phosphatase and plasma cell membrane glycoprotein-1 are central antagonistic regulators of bone mineralization","volume":"99","author":"Hessle","year":"2002","journal-title":"Proc Natl Acad Sci USA."},{"key":"2024020318472369000_jbmr2281-bib-0024","doi-asserted-by":"crossref","first-page":"2900","DOI":"10.1016\/S0021-9258(18)61592-3","article-title":"Isolation, characterization, and biosynthesis of phosphorylated glycoprotein from rat bone","volume":"262","author":"Prince","year":"1987","journal-title":"J Biol Chem."},{"key":"2024020318472369000_jbmr2281-bib-0025","doi-asserted-by":"crossref","first-page":"1587","DOI":"10.1002\/jbmr.1901","article-title":"In vivo overexpression of tissue-nonspecific alkaline phosphatase increases skeletal mineralization and affects the phosphorylation status of osteopontin","volume":"28","author":"Narisawa","year":"2013","journal-title":"J Bone Miner Res."},{"key":"2024020318472369000_jbmr2281-bib-0026","author":"Moreau"},{"key":"2024020318472369000_jbmr2281-bib-0027","doi-asserted-by":"crossref","first-page":"2035","DOI":"10.1016\/S0002-9440(10)64482-3","article-title":"Osteopontin inhibits mineral deposition and promotes regression of ectopic calcification","volume":"161","author":"Steitz","year":"2002","journal-title":"Am J Pathol."},{"key":"2024020318472369000_jbmr2281-bib-0028","doi-asserted-by":"crossref","first-page":"F1935","DOI":"10.1152\/ajprenal.00383.2007","article-title":"Renal calcinosis and stone formation in mice lacking osteopontin, Tamm-Horsfall protein, or both","volume":"293","author":"Mo","year":"2007","journal-title":"Am J Physiol Renal Physiol."},{"key":"2024020318472369000_jbmr2281-bib-0029","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1084\/jem.20020911","article-title":"Inactivation of the osteopontin gene enhances vascular calcification of matrix Gla protein\u2013deficient mice: evidence for osteopontin as an inducible inhibitor of vascular calcification in vivo","volume":"196","author":"Speer","year":"2002","journal-title":"J Exp Med."},{"key":"2024020318472369000_jbmr2281-bib-0030","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1097\/01.ASN.0000040593.93815.9D","article-title":"Osteopontin is a critical inhibitor of calcium oxalate crystal formation and retention in renal tubules","volume":"14","author":"Wesson","year":"2003","journal-title":"J Am Soc Nephrol."},{"key":"2024020318472369000_jbmr2281-bib-0031","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1007\/s00223-002-1090-x","article-title":"Osteopontin facilitates bone resorption, decreasing bone mineral crystallinity and content during calcium deficiency","volume":"73","author":"Shapses","year":"2003","journal-title":"Calc Tissue Int."},{"key":"2024020318472369000_jbmr2281-bib-0032","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1159\/000020591","article-title":"Osteopontin function in pathology: lessons from osteopontin deficient mice","volume":"7","author":"Rittling","year":"1999","journal-title":"Exp Nephrol."},{"key":"2024020318472369000_jbmr2281-bib-0033","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/S0014-5793(97)01498-1","article-title":"Osteopontin involvement in integrin-mediated cell signaling and regulation of expression of alkaline phosphatase during early differentiation of UMR cells","volume":"420","author":"Liu","year":"1997","journal-title":"FEBS Lett."},{"key":"2024020318472369000_jbmr2281-bib-0034","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.bone.2009.08.055","article-title":"Inorganic pyrophosphatase induces type I collagen in osteoblasts","volume":"46","author":"Polewski","year":"2010","journal-title":"Bone."},{"key":"2024020318472369000_jbmr2281-bib-0035","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1016\/S0022-3476(82)80061-9","article-title":"Infantile hypophosphatasia: enzyme replacement therapy by intravenous infusion of alkaline phosphatase-rich plasma from patients with Paget bone disease","volume":"101","author":"Whyte","year":"1982","journal-title":"J Pediatr."},{"key":"2024020318472369000_jbmr2281-bib-0036","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1016\/S0022-3476(84)80079-7","article-title":"Enzyme replacement therapy for infantile hypophosphatasia attempted by intravenous infusions of alkaline phosphatase-rich Paget plasma: results in three additional patients","volume":"105","author":"Whyte","year":"1984","journal-title":"J Pediatr."},{"issue":"10","key":"2024020318472369000_jbmr2281-bib-0037","doi-asserted-by":"crossref","first-page":"904","DOI":"10.1056\/NEJMoa1106173","article-title":"Enzyme-replacement therapy in life-threatening hypophosphatasia","volume":"366","author":"Whyte","year":"2012","journal-title":"N Engl J Med."}],"container-title":["Journal of Bone and Mineral Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjbmr.2281","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/academic.oup.com\/jbmr\/article-pdf\/29\/11\/2369\/56565331\/jbmr2281.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jbmr\/article-pdf\/29\/11\/2369\/56565331\/jbmr2281.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,13]],"date-time":"2024-03-13T14:52:23Z","timestamp":1710341543000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jbmr\/article\/29\/11\/2369-2381\/7598888"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,5,13]]},"references-count":37,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2014,5,13]]},"published-print":{"date-parts":[[2014,11,1]]}},"URL":"https:\/\/doi.org\/10.1002\/jbmr.2281","relation":{},"ISSN":["0884-0431","1523-4681"],"issn-type":[{"value":"0884-0431","type":"print"},{"value":"1523-4681","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2014,11,1]]},"published":{"date-parts":[[2014,5,13]]}}}