{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T06:54:24Z","timestamp":1768805664150,"version":"3.49.0"},"reference-count":38,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2016,6,1]],"date-time":"2016-06-01T00:00:00Z","timestamp":1464739200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2016,6,1]],"date-time":"2016-06-01T00:00:00Z","timestamp":1464739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Ossification of the posterior longitudinal ligament of the spine (OPLL), which is characterized by ectopic bone formation in the spinal ligaments, can cause spinal-cord compression. To date, at least 11 susceptibility genes have been genetically linked to OPLL. In order to identify potential deleterious alleles in these OPLL-associated genes, we designed a capture array encompassing all coding regions of the target genes for next-generation sequencing (NGS) in a cohort of 55 unrelated patients with OPLL. By bioinformatics analyses, we successfully identified three novel and five extremely rare variants (MAF\u2009&lt;\u20090.005). These variants were predicted to be deleterious by commonly used various algorithms, thereby resulting in missense mutations in four OPLL-associated genes (i.e., <jats:italic>COL6A1<\/jats:italic>, <jats:italic>COL11A2<\/jats:italic>, <jats:italic>FGFR1<\/jats:italic> and <jats:italic>BMP2<\/jats:italic>). Furthermore, potential effects of the patient with p.Q89E of <jats:italic>BMP2<\/jats:italic> were confirmed by a markedly increased BMP2 level in peripheral blood samples. Notably, seven of the variants were found to be associated with the patients with continuous subtype changes by cervical spinal radiological analyses. Taken together, our findings revealed for the first time that deleterious coding variants of the four OPLL-associated genes are potentially pathogenic in the patients with OPLL.<\/jats:p>","DOI":"10.1038\/srep26962","type":"journal-article","created":{"date-parts":[[2016,6,1]],"date-time":"2016-06-01T09:54:32Z","timestamp":1464774872000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Targeted next-generation sequencing reveals multiple deleterious variants in OPLL-associated genes"],"prefix":"10.1038","volume":"6","author":[{"given":"Xin","family":"Chen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Guo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Cai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fengshan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shengfa","family":"Pan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaobo","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Feifei","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yinze","family":"Diao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanbin","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhen","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoguang","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhongqiang","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhongjun","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Sun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jie","family":"Du","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2016,6,1]]},"reference":[{"key":"BFsrep26962_CR1","doi-asserted-by":"crossref","unstructured":"Tsuyama, N. Ossification of the posterior longitudinal ligament of the spine. Clin Orthop Relat Res 71\u201384 (1984).","DOI":"10.1097\/00003086-198404000-00010"},{"key":"BFsrep26962_CR2","doi-asserted-by":"publisher","first-page":"405","DOI":"10.2183\/pjab.90.405","volume":"90","author":"S Ikegawa","year":"2014","unstructured":"Ikegawa, S. Genomic study of ossification of the posterior longitudinal ligament of the spine. Proc Jpn Acad Ser B Phys Biol Sci 90, 405\u2013412 (2014).","journal-title":"Proc Jpn Acad Ser B Phys Biol Sci"},{"key":"BFsrep26962_CR3","first-page":"285","volume":"9","author":"Zhongshi Li.","year":"1999","unstructured":"Zhongshi Li. et al. A morbidity research about cervical spondylotic ossification of posterior longitudinal ligament in patients with cervibrachialgia in the North China. Chinese Journal of Spine and Spinal Cord 9, 285\u2013286 (1999).","journal-title":"Chinese Journal of Spine and Spinal Cord"},{"key":"BFsrep26962_CR4","doi-asserted-by":"publisher","first-page":"1184","DOI":"10.1097\/00007632-198911000-00009","volume":"14","author":"K Terayama","year":"1989","unstructured":"Terayama, K. Genetic studies on ossification of the posterior longitudinal ligament of the spine. Spine (Phila Pa 1976) 14, 1184\u20131191 (1989).","journal-title":"Spine (Phila Pa 1976)"},{"key":"BFsrep26962_CR5","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1038\/956","volume":"19","author":"A Okawa","year":"1998","unstructured":"Okawa, A. et al. Mutation in Npps in a mouse model of ossification of the posterior longitudinal ligament of the spine. Nat Genet 19, 271\u2013273 (1998).","journal-title":"Nat Genet"},{"key":"BFsrep26962_CR6","doi-asserted-by":"publisher","first-page":"1460","DOI":"10.1086\/301868","volume":"62","author":"H Koga","year":"1998","unstructured":"Koga, H. et al. Genetic mapping of ossification of the posterior longitudinal ligament of the spine. Am J Hum Genet 62, 1460\u20131467 (1998).","journal-title":"Am J Hum Genet"},{"key":"BFsrep26962_CR7","doi-asserted-by":"publisher","first-page":"812","DOI":"10.1086\/378593","volume":"73","author":"T Tanaka","year":"2003","unstructured":"Tanaka, T. et al. Genomewide linkage and linkage disequilibrium analyses identify COL6A1, on chromosome 21, as the locus for ossification of the posterior longitudinal ligament of the spine. Am J Hum Genet 73, 812\u2013822 (2003).","journal-title":"Am J Hum Genet"},{"key":"BFsrep26962_CR8","doi-asserted-by":"publisher","first-page":"956","DOI":"10.1007\/s00586-008-0651-8","volume":"17","author":"H Wang","year":"2008","unstructured":"Wang, H. et al. Association of bone morphogenetic protein-2 gene polymorphisms with susceptibility to ossification of the posterior longitudinal ligament of the spine and its severity in Chinese patients. Eur Spine J 17, 956\u2013964 (2008).","journal-title":"Eur Spine J"},{"key":"BFsrep26962_CR9","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1359\/jbmr.2002.17.1.128","volume":"17","author":"K Furushima","year":"2002","unstructured":"Furushima, K. et al. Large-scale screening for candidate genes of ossification of the posterior longitudinal ligament of the spine. J Bone Miner Res 17, 128\u2013137 (2002).","journal-title":"J Bone Miner Res"},{"key":"BFsrep26962_CR10","doi-asserted-by":"publisher","first-page":"e40587","DOI":"10.1371\/journal.pone.0040587","volume":"7","author":"Y Ren","year":"2012","unstructured":"Ren, Y. et al. Association of a BMP9 haplotype with ossification of the posterior longitudinal ligament (OPLL) in a Chinese population. PLoS One 7, e40587 (2012).","journal-title":"PLoS One"},{"key":"BFsrep26962_CR11","doi-asserted-by":"publisher","first-page":"1264","DOI":"10.1097\/00007632-200106010-00017","volume":"26","author":"M Kamiya","year":"2001","unstructured":"Kamiya, M., Harada, A., Mizuno, M., Iwata, H. & Yamada, Y. Association between a polymorphism of the transforming growth factor-beta1 gene and genetic susceptibility to ossification of the posterior longitudinal ligament in Japanese patients. Spine (Phila Pa 1976) 26, 1264\u20131266, discussion 1266\u20131267 (2001).","journal-title":"Spine (Phila Pa 1976)"},{"key":"BFsrep26962_CR12","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1007\/s00439-006-0170-9","volume":"119","author":"T Horikoshi","year":"2006","unstructured":"Horikoshi, T. et al. A large-scale genetic association study of ossification of the posterior longitudinal ligament of the spine. Hum Genet 119, 611\u2013616 (2006).","journal-title":"Hum Genet"},{"key":"BFsrep26962_CR13","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1016\/j.jocn.2012.05.031","volume":"20","author":"DW Jekarl","year":"2013","unstructured":"Jekarl, D. W. et al. TGFBR2 gene polymorphism is associated with ossification of the posterior longitudinal ligament. J Clin Neurosci 20, 453\u2013456 (2013).","journal-title":"J Clin Neurosci"},{"key":"BFsrep26962_CR14","doi-asserted-by":"publisher","first-page":"7","DOI":"10.3340\/jkns.2012.52.1.7","volume":"52","author":"JK Jun","year":"2012","unstructured":"Jun, J. K. & Kim, S. M. Association study of fibroblast growth factor 2 and fibroblast growth factor receptors gene polymorphism in korean ossification of the posterior longitudinal ligament patients. J Korean Neurosurg Soc 52, 7\u201313 (2012).","journal-title":"J Korean Neurosurg Soc"},{"key":"BFsrep26962_CR15","doi-asserted-by":"publisher","first-page":"1765","DOI":"10.1097\/00007632-200208150-00015","volume":"27","author":"N Ogata","year":"2002","unstructured":"Ogata, N. et al. Association of bone metabolism regulatory factor gene polymorphisms with susceptibility to ossification of the posterior longitudinal ligament of the spine and its severity. Spine (Phila Pa 1976) 27, 1765\u20131771 (2002).","journal-title":"Spine (Phila Pa 1976)"},{"key":"BFsrep26962_CR16","doi-asserted-by":"publisher","first-page":"32628","DOI":"10.1074\/jbc.M002594200","volume":"275","author":"Y Zhang","year":"2000","unstructured":"Zhang, Y. & Chen, Q. Changes of matrilin forms during endochondral ossification. Molecular basis of oligomeric assembly. J Biol Chem 275, 32628\u201332634 (2000).","journal-title":"J Biol Chem"},{"key":"BFsrep26962_CR17","doi-asserted-by":"publisher","first-page":"1998","DOI":"10.1074\/jbc.M203250200","volume":"278","author":"LC Gowen","year":"2003","unstructured":"Gowen, L. C. et al. Targeted disruption of the osteoblast\/osteocyte factor 45 gene (OF45) results in increased bone formation and bone mass. J Biol Chem 278, 1998\u20132007 (2003).","journal-title":"J Biol Chem"},{"key":"BFsrep26962_CR18","doi-asserted-by":"publisher","first-page":"9484","DOI":"10.1073\/pnas.85.24.9484","volume":"85","author":"EA Wang","year":"1988","unstructured":"Wang, E. A. et al. Purification and characterization of other distinct bone-inducing factors. Proc Natl Acad Sci USA 85, 9484\u20139488 (1988).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep26962_CR19","doi-asserted-by":"publisher","first-page":"1012","DOI":"10.1038\/ng.3045","volume":"46","author":"M Nakajima","year":"2014","unstructured":"Nakajima, M. et al. A genome-wide association study identifies susceptibility loci for ossification of the posterior longitudinal ligament of the spine. Nat Genet 46, 1012\u20131016 (2014).","journal-title":"Nat Genet"},{"key":"BFsrep26962_CR20","doi-asserted-by":"publisher","first-page":"127","DOI":"10.11005\/jbm.2014.21.2.127","volume":"21","author":"S Ikegawa","year":"2014","unstructured":"Ikegawa, S. Genetics of ossification of the posterior longitudinal ligament of the spine: a mini review. J Bone Metab 21, 127\u2013132 (2014).","journal-title":"J Bone Metab"},{"key":"BFsrep26962_CR21","doi-asserted-by":"publisher","first-page":"825","DOI":"10.1038\/ng.2646","volume":"45","author":"GL Carvill","year":"2013","unstructured":"Carvill, G. L. et al. Targeted resequencing in epileptic encephalopathies identifies de novo mutations in CHD2 and SYNGAP1. Nat Genet 45, 825\u2013830 (2013).","journal-title":"Nat Genet"},{"key":"BFsrep26962_CR22","doi-asserted-by":"publisher","first-page":"1035","DOI":"10.1002\/humu.22332","volume":"34","author":"B Sikkema-Raddatz","year":"2013","unstructured":"Sikkema-Raddatz, B. et al. Targeted next-generation sequencing can replace Sanger sequencing in clinical diagnostics. Hum Mutat 34, 1035\u20131042 (2013).","journal-title":"Hum Mutat"},{"key":"BFsrep26962_CR23","doi-asserted-by":"publisher","first-page":"13115","DOI":"10.1038\/srep13115","volume":"5","author":"J Guo","year":"2015","unstructured":"Guo, J. et al. Wide mutation spectrum and frequent variant Ala27Thr of FBN1 identified in a large cohort of Chinese patients with sporadic TAAD. Sci Rep 5, 13115 (2015).","journal-title":"Sci Rep"},{"key":"BFsrep26962_CR24","doi-asserted-by":"publisher","first-page":"E4","DOI":"10.3171\/2010.12.FOCUS10277","volume":"30","author":"MY Wang","year":"2011","unstructured":"Wang, M. Y. & Thambuswamy, M. Ossification of the posterior longitudinal ligament in non-Asians: demographic, clinical and radiographic findings in 43 patients. Neurosurg Focus 30, E4 (2011).","journal-title":"Neurosurg Focus"},{"key":"BFsrep26962_CR25","doi-asserted-by":"publisher","first-page":"354","DOI":"10.1097\/00007632-198107000-00005","volume":"6","author":"K Hirabayashi","year":"1981","unstructured":"Hirabayashi, K., Miyakawa, J., Satomi, K., Maruyama, T. & Wakano, K. Operative results and postoperative progression of ossification among patients with ossification of cervical posterior longitudinal ligament. Spine (Phila Pa 1976) 6, 354\u2013364 (1981).","journal-title":"Spine (Phila Pa 1976)"},{"key":"BFsrep26962_CR26","doi-asserted-by":"publisher","first-page":"948","DOI":"10.1359\/jbmr.2001.16.5.948","volume":"16","author":"S Maeda","year":"2001","unstructured":"Maeda, S. et al. Functional impact of human collagen alpha2(XI) gene polymorphism in pathogenesis of ossification of the posterior longitudinal ligament of the spine. J Bone Miner Res 16, 948\u2013957 (2001).","journal-title":"J Bone Miner Res"},{"key":"BFsrep26962_CR27","doi-asserted-by":"publisher","first-page":"S33","DOI":"10.1097\/00007632-199203001-00007","volume":"17","author":"H Kawaguchi","year":"1992","unstructured":"Kawaguchi, H. et al. Immunohistochemical demonstration of bone morphogenetic protein-2 and transforming growth factor-beta in the ossification of the posterior longitudinal ligament of the cervical spine. Spine (Phila Pa 1976) 17, S33\u201336 (1992).","journal-title":"Spine (Phila Pa 1976)"},{"key":"BFsrep26962_CR28","doi-asserted-by":"publisher","first-page":"e106598","DOI":"10.1371\/journal.pone.0106598","volume":"9","author":"JM Li","year":"2014","unstructured":"Li, J. M. et al. Uniaxial cyclic stretch promotes osteogenic differentiation and synthesis of BMP2 in the C3H10T1\/2 cells with BMP2 gene variant of rs2273073 (T\/G). PLoS One 9, e106598 (2014).","journal-title":"PLoS One"},{"key":"BFsrep26962_CR29","doi-asserted-by":"publisher","first-page":"178","DOI":"10.18632\/oncotarget.235","volume":"2","author":"J He","year":"2011","unstructured":"He, J. et al. IgH gene rearrangements as plasma biomarkers in Non- Hodgkin\u2019s lymphoma patients. Oncotarget 2, 178\u2013185 (2011).","journal-title":"Oncotarget"},{"key":"BFsrep26962_CR30","doi-asserted-by":"crossref","unstructured":"Wu, J. et al. Recurrent GNAS mutations define an unexpected pathway for pancreatic cyst development. Sci Transl Med 3, 92ra66 (2011).","DOI":"10.1126\/scitranslmed.3002543"},{"key":"BFsrep26962_CR31","doi-asserted-by":"publisher","first-page":"1754","DOI":"10.1093\/bioinformatics\/btp324","volume":"25","author":"H Li","year":"2009","unstructured":"Li, H. & Durbin, R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 25, 1754\u20131760 (2009).","journal-title":"Bioinformatics"},{"key":"BFsrep26962_CR32","doi-asserted-by":"publisher","first-page":"617","DOI":"10.1016\/j.ajhg.2009.04.007","volume":"84","author":"DC Guo","year":"2009","unstructured":"Guo, D. C. et al. Mutations in smooth muscle alpha-actin (ACTA2) cause coronary artery disease, stroke and Moyamoya disease, along with thoracic aortic disease. Am J Hum Genet 84, 617\u2013627 (2009).","journal-title":"Am J Hum Genet"},{"key":"BFsrep26962_CR33","doi-asserted-by":"publisher","first-page":"2078","DOI":"10.1093\/bioinformatics\/btp352","volume":"25","author":"H Li","year":"2009","unstructured":"Li, H. et al. The Sequence Alignment\/Map format and SAMtools. Bioinformatics 25, 2078\u20132079 (2009).","journal-title":"Bioinformatics"},{"key":"BFsrep26962_CR34","doi-asserted-by":"publisher","first-page":"e164","DOI":"10.1093\/nar\/gkq603","volume":"38","author":"K Wang","year":"2010","unstructured":"Wang, K., Li, M. & Hakonarson, H. ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res 38, e164 (2010).","journal-title":"Nucleic Acids Res"},{"key":"BFsrep26962_CR35","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1038\/nmeth0410-248","volume":"7","author":"IA Adzhubei","year":"2010","unstructured":"Adzhubei, I. A. et al. A method and server for predicting damaging missense mutations. Nat Methods 7, 248\u2013249 (2010).","journal-title":"Nat Methods"},{"key":"BFsrep26962_CR36","doi-asserted-by":"publisher","first-page":"W452","DOI":"10.1093\/nar\/gks539","volume":"40","author":"NL Sim","year":"2012","unstructured":"Sim, N. L. et al. SIFT web server: predicting effects of amino acid substitutions on proteins. Nucleic Acids Res 40, W452\u2013457 (2012).","journal-title":"Nucleic Acids Res"},{"key":"BFsrep26962_CR37","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1038\/nmeth.2890","volume":"11","author":"JM Schwarz","year":"2014","unstructured":"Schwarz, J. M., Cooper, D. N., Schuelke, M. & Seelow, D. MutationTaster2: mutation prediction for the deep-sequencing age. Nat Methods 11, 361\u2013362 (2014).","journal-title":"Nat Methods"},{"key":"BFsrep26962_CR38","doi-asserted-by":"publisher","first-page":"e1001025","DOI":"10.1371\/journal.pcbi.1001025","volume":"6","author":"EV Davydov","year":"2010","unstructured":"Davydov, E. V. et al. Identifying a high fraction of the human genome to be under selective constraint using GERP++. PLoS Comput Biol 6, e1001025 (2010).","journal-title":"PLoS Comput Biol"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep26962","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep26962.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep26962.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,4]],"date-time":"2023-01-04T16:11:50Z","timestamp":1672848710000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep26962"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,6,1]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2016,7,18]]}},"alternative-id":["BFsrep26962"],"URL":"https:\/\/doi.org\/10.1038\/srep26962","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,6,1]]},"assertion":[{"value":"24 December 2015","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 May 2016","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 June 2016","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"26962"}}