{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T09:12:51Z","timestamp":1789981971053,"version":"4.0.1"},"reference-count":33,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,10,19]],"date-time":"2017-10-19T00:00:00Z","timestamp":1508371200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,10,19]],"date-time":"2017-10-19T00:00:00Z","timestamp":1508371200000},"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>Keratoconus (KC) is a common degenerative corneal disease, and heredity plays a key role in its development. Although few genes are known to cause KC, a large proportion of disease-causing genes remain to be revealed. Here, we report the identification of <jats:italic>TUBA3D<\/jats:italic> as a novel gene linked to KC. Using whole-exome sequencing of a twins pedigree, a novel de novo mutation (c.31\u2009C\u2009&gt;\u2009T, p.Gln11stop) in <jats:italic>TUBA3D<\/jats:italic> gene was identified. A screening performed in 200 additional unrelated patients with KC revealed another two mutations (c.201insTT, p.Val68Leufs*2; c.*2\u2009G\u2009&gt;\u2009A) in two patients. <jats:italic>TUBA3D<\/jats:italic> was expressed highly in the cornea, and the twins had lower <jats:italic>TUBA3D<\/jats:italic> expression and higher UPA and MMP1 expressions than the normal parents. Through function prediction and <jats:italic>in vitro<\/jats:italic> cell experiment, we further demonstrated that the mutant proteins of <jats:italic>TUBA3D<\/jats:italic> were unstable and could lead to human corneal fibroblast cells performing higher MMPs expression and oxidative stress. These changes thus reduce the amount of extracellular matrices within corneas and undoubtedly play a major role in stromal thinning, which is characteristic of KC corneas. Our study showed that <jats:italic>TUBA3D<\/jats:italic> is a new gene that causes KC, thus supporting the evidence that this protein has an additional function into the human cornea.<\/jats:p>","DOI":"10.1038\/s41598-017-13162-0","type":"journal-article","created":{"date-parts":[[2017,10,13]],"date-time":"2017-10-13T11:21:04Z","timestamp":1507893664000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["De novo mutations of TUBA3D are associated with keratoconus"],"prefix":"10.1038","volume":"7","author":[{"given":"Xiao-dan","family":"Hao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang-yang","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Su-xia","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei-yun","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,10,19]]},"reference":[{"key":"13162_CR1","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/0039-6257(84)90094-8","volume":"28","author":"JH Krachmer","year":"1984","unstructured":"Krachmer, J. H., Feder, R. S. & Belin, M. W. Keratoconus and related noninflammatory corneal thinning disorders. Surv. Ophthalmol. \n                           28, 293\u2013322 (1984).","journal-title":"Surv. Ophthalmol."},{"key":"13162_CR2","doi-asserted-by":"publisher","first-page":"581","DOI":"10.1097\/ICO.0b013e31820cd61d","volume":"31","author":"YO Kok","year":"2012","unstructured":"Kok, Y. O., Tan, G. F. & Loon, S. C. Review: keratoconus in Asia. Cornea \n                           31, 581\u2013593 (2012).","journal-title":"Cornea"},{"key":"13162_CR3","doi-asserted-by":"publisher","first-page":"1288","DOI":"10.1038\/eye.2008.305","volume":"23","author":"I Rahman","year":"2009","unstructured":"Rahman, I., Carley, F., Hillarby, C., Brahma, A. & Tullo, A. B. Penetrating keratoplasty: indications, outcomes, and complications. Eye (Lond) \n                           23, 1288\u20131294 (2009).","journal-title":"Eye (Lond)"},{"key":"13162_CR4","doi-asserted-by":"publisher","first-page":"1291","DOI":"10.1136\/bjo.2008.148981","volume":"93","author":"L Xie","year":"2009","unstructured":"Xie, L. et al. Major shifts in corneal transplantation procedures in north China: 5316 eyes over 12 years. Br. J. Ophthalmol. \n                           93, 1291\u20131295 (2009).","journal-title":"Br. J. Ophthalmol."},{"key":"13162_CR5","doi-asserted-by":"publisher","first-page":"106","DOI":"10.1111\/j.1755-3768.2012.02400.x","volume":"91","author":"K Nielsen","year":"2013","unstructured":"Nielsen, K., Hjortdal, J., Pihlmann, M. & Corydon, T. J. Update on the keratoconus genetics. Acta Ophthalmol \n                           91, 106\u2013113 (2013).","journal-title":"Acta Ophthalmol"},{"key":"13162_CR6","doi-asserted-by":"publisher","first-page":"146","DOI":"10.1111\/cxo.12024","volume":"96","author":"KP Burdon","year":"2013","unstructured":"Burdon, K. P. & Vincent, A. L. Insights into keratoconus from a genetic perspective. Clin. Exp. Optom. \n                           96, 146\u2013154 (2013).","journal-title":"Clin. Exp. Optom."},{"key":"13162_CR7","unstructured":"Willoughby, C. E. & Lechner, J. In Corneal Disease: Recent Developments in \n                           Diagnosis and Therapy (eds Thomas Reinhard & Frank Larkin) Ch. 3, (Springer Heidelberg New York Dordrecht London, 2013)."},{"key":"13162_CR8","first-page":"1","volume":"178","author":"A Ihalainen","year":"1986","unstructured":"Ihalainen, A. Clinical and epidemiological features of keratoconus genetic and external factors in the pathogenesis of the disease. Acta Ophthalmol Suppl \n                           178, 1\u201364 (1986).","journal-title":"Acta Ophthalmol Suppl"},{"key":"13162_CR9","first-page":"3160","volume":"43","author":"H Tyynismaa","year":"2002","unstructured":"Tyynismaa, H. et al. A locus for autosomal dominant keratoconus: linkage to 16q22.3-q23.1 in Finnish families. Invest. Ophthalmol. Vis. Sci. \n                           43, 3160\u20133164 (2002).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"13162_CR10","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1136\/jmg.2003.012872","volume":"41","author":"F Brancati","year":"2004","unstructured":"Brancati, F. et al. A locus for autosomal dominant keratoconus maps to human chromosome 3p14-q13. J Med Genet \n                           41, 188\u2013192 (2004).","journal-title":"J Med Genet"},{"key":"13162_CR11","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1097\/01.GIM.0000170772.41860.54","volume":"7","author":"YG Tang","year":"2005","unstructured":"Tang, Y. G. et al. Genomewide linkage scan in a multigeneration Caucasian pedigree identifies a novel locus for keratoconus on chromosome 5q14.3-q21.1. Genet Med \n                           7, 397\u2013405 (2005).","journal-title":"Genet Med"},{"key":"13162_CR12","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1016\/j.gene.2012.04.061","volume":"503","author":"T Guan","year":"2012","unstructured":"Guan, T., Liu, C., Ma, Z. & Ding, S. The point mutation and polymorphism in keratoconus candidate gene TGFBI in Chinese population. Gene \n                           503, 137\u2013139 (2012).","journal-title":"Gene"},{"key":"13162_CR13","first-page":"37","volume":"37","author":"JA Karolak","year":"2016","unstructured":"Karolak, J. A., Polakowski, P., Szaflik, J., Szaflik, J. P. & Gajecka, M. Molecular Screening of Keratoconus Susceptibility Sequence Variants in VSX1, TGFBI, DOCK9, STK24, and IPO5 Genes in Polish Patients and Novel TGFBI Variant Identification. Ophthalmic. Genet. \n                           37, 37\u201343 (2016).","journal-title":"Ophthalmic. Genet."},{"key":"13162_CR14","doi-asserted-by":"publisher","first-page":"628","DOI":"10.1016\/j.ajhg.2011.09.014","volume":"89","author":"AE Hughes","year":"2011","unstructured":"Hughes, A. E. et al. Mutation altering the miR-184 seed region causes familial keratoconus with cataract. Am. J. Hum. Genet. \n                           89, 628\u2013633 (2011).","journal-title":"Am. J. Hum. Genet."},{"key":"13162_CR15","doi-asserted-by":"publisher","first-page":"5266","DOI":"10.1167\/iovs.13-12035","volume":"54","author":"J Lechner","year":"2013","unstructured":"Lechner, J. et al. Mutational analysis of MIR184 in sporadic keratoconus and myopia. Invest. Ophthalmol. Vis. Sci. \n                           54, 5266\u20135272 (2013).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"13162_CR16","doi-asserted-by":"publisher","first-page":"3812","DOI":"10.1093\/nar\/gkg509","volume":"31","author":"PC Ng","year":"2003","unstructured":"Ng, P. C. & Henikoff, S. SIFT: Predicting amino acid changes that affect protein function. Nucleic Acids Res \n                           31, 3812\u20133814 (2003).","journal-title":"Nucleic Acids Res"},{"key":"13162_CR17","doi-asserted-by":"publisher","first-page":"575","DOI":"10.1038\/nmeth0810-575","volume":"7","author":"JM Schwarz","year":"2010","unstructured":"Schwarz, J. M., Rodelsperger, C., Schuelke, M. & Seelow, D. MutationTaster evaluates disease-causing potential of sequence alterations. Nat Methods \n                           7, 575\u2013576 (2010).","journal-title":"Nat Methods"},{"key":"13162_CR18","doi-asserted-by":"publisher","first-page":"2714","DOI":"10.1002\/pmic.201000799","volume":"11","author":"S de Mateo","year":"2011","unstructured":"de Mateo, S., Castillo, J., Estanyol, J. M., Ballesca, J. L. & Oliva, R. Proteomic characterization of the human sperm nucleus. Proteomics \n                           11, 2714\u20132726 (2011).","journal-title":"Proteomics"},{"key":"13162_CR19","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1097\/01.ico.0000208820.32614.00","volume":"25","author":"Z Mackiewicz","year":"2006","unstructured":"Mackiewicz, Z. et al. Collagenolytic proteinases in keratoconus. Cornea \n                           25, 603\u2013610 (2006).","journal-title":"Cornea"},{"key":"13162_CR20","doi-asserted-by":"crossref","unstructured":"Du, G. et al. Induction of matrix metalloproteinase-1 by tumor necrosis factor-alpha is mediated by interleukin-6 in cultured fibroblasts of keratoconus. Exp Biol Med (Maywood) (2016).","DOI":"10.1177\/1535370216650940"},{"key":"13162_CR21","doi-asserted-by":"publisher","first-page":"185","DOI":"10.3109\/02713680903477824","volume":"35","author":"SA Balasubramanian","year":"2010","unstructured":"Balasubramanian, S. A., Pye, D. C. & Willcox, M. D. Are proteinases the reason for keratoconus? Curr. Eye. Res. \n                           35, 185\u2013191 (2010).","journal-title":"Curr. Eye. Res."},{"key":"13162_CR22","first-page":"2293","volume":"42","author":"A Behndig","year":"2001","unstructured":"Behndig, A., Karlsson, K., Johansson, B. O., Brannstrom, T. & Marklund, S. L. Superoxide dismutase isoenzymes in the normal and diseased human cornea. Invest. Ophthalmol. Vis. Sci. \n                           42, 2293\u20132296 (2001).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"13162_CR23","doi-asserted-by":"publisher","first-page":"1902","DOI":"10.1167\/iovs.05-0828","volume":"47","author":"M Chwa","year":"2006","unstructured":"Chwa, M. et al. Increased stress-induced generation of reactive oxygen species and apoptosis in human keratoconus fibroblasts. Invest. Ophthalmol. Vis. Sci. \n                           47, 1902\u20131910 (2006).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"13162_CR24","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 \n                           25, 1754\u20131760 (2009).","journal-title":"Bioinformatics"},{"key":"13162_CR25","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 \n                           25, 2078\u20132079 (2009).","journal-title":"Bioinformatics"},{"key":"13162_CR26","doi-asserted-by":"publisher","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 \n                           38, e164 (2010).","journal-title":"Nucleic Acids Res"},{"key":"13162_CR27","doi-asserted-by":"crossref","unstructured":"Adzhubei, I., Jordan, D. M. & Sunyaev, S. R. Predicting functional effect of human missense mutations using PolyPhen-2. Curr Protoc Hum Genet Chapter 7, Unit720 (2013).","DOI":"10.1002\/0471142905.hg0720s76"},{"key":"13162_CR28","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1038\/ng.2892","volume":"46","author":"M Kircher","year":"2014","unstructured":"Kircher, M. et al. A general framework for estimating the relative pathogenicity of human genetic variants. Nat. Genet. \n                           46, 310\u2013315 (2014).","journal-title":"Nat. Genet."},{"key":"13162_CR29","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1038\/nature12929","volume":"506","author":"M Fromer","year":"2014","unstructured":"Fromer, M. et al. De novo mutations in schizophrenia implicate synaptic networks. Nature \n                           506, 179\u2013184 (2014).","journal-title":"Nature"},{"key":"13162_CR30","doi-asserted-by":"publisher","first-page":"2731","DOI":"10.1093\/molbev\/msr121","volume":"28","author":"K Tamura","year":"2011","unstructured":"Tamura, K. et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. \n                           28, 2731\u20132739 (2011).","journal-title":"Mol. Biol. Evol."},{"key":"13162_CR31","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 \n                           11, 361\u2013362 (2014).","journal-title":"Nat Methods"},{"key":"13162_CR32","doi-asserted-by":"publisher","first-page":"W252","DOI":"10.1093\/nar\/gku340","volume":"42","author":"M Biasini","year":"2014","unstructured":"Biasini, M. et al. SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res \n                           42, W252\u2013258 (2014).","journal-title":"Nucleic Acids Res"},{"key":"13162_CR33","doi-asserted-by":"publisher","first-page":"8031","DOI":"10.1167\/iovs.14-14098","volume":"55","author":"P Chen","year":"2014","unstructured":"Chen, P. et al. Mutations in the ABCA3 gene are associated with cataract-microcornea syndrome. Invest. Ophthalmol. Vis. Sci. \n                           55, 8031\u20138043 (2014).","journal-title":"Invest. Ophthalmol. Vis. Sci."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-13162-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-13162-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-13162-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T12:15:58Z","timestamp":1671797758000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-13162-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,10,19]]},"references-count":33,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["13162"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-13162-0","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,10,19]]},"assertion":[{"value":"21 April 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 September 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 October 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"13570"}}