{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T19:07:08Z","timestamp":1773860828740,"version":"3.50.1"},"reference-count":51,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2019,8,24]],"date-time":"2019-08-24T00:00:00Z","timestamp":1566604800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National MPS Society Research Grant","award":["NA"],"award-info":[{"award-number":["NA"]}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30GM114736"],"award-info":[{"award-number":["P30GM114736"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mucopolysaccharidosis IVA (MPS IVA) is caused by a deficiency of the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS). Conventional enzyme replacement therapy (ERT) is approved for MPS IVA. However, the fact that the infused enzyme cannot penetrate avascular lesions in cartilage leads to minimal impact on the bone lesion. Moreover, short half-life, high cost, instability, and narrow optimal pH range remain unmet challenges in ERT. Thermostable keratanase, endo-\u03b2-N-acetylglucosaminidase, has a unique character of a wide optimal pH range of pH 5.0\u20137.0. We hypothesized that this endoglycosidase degrades keratan sulfate (KS) polymer in circulating blood and, therefore, ameliorates the accumulation of KS in multiple tissues. We propose a novel approach, Substrate Degradation Enzyme Therapy (SDET), to treat bone lesion of MPS IVA. We assessed the effect of thermostable keratanase on blood KS level and bone pathology using Galns knock-out MPS IVA mice. After a single administration of 2 U\/kg (= 0.2 mg\/kg) of the enzyme at 8 weeks of age via intravenous injection, the level of serum KS was significantly decreased to normal range level, and this suppression was maintained for at least 4 weeks. We administered 2 U\/kg of the enzyme to MPS IVA mice every fourth week for 12 weeks (total of 3 times) at newborns or 8 weeks of age. After a third injection, serum mono-sulfated KS levels were kept low for 4 weeks, similar to that in control mice, and at 12 weeks, bone pathology was markedly improved when SDET started at newborns, compared with untreated MPS IVA mice. Overall, thermostable keratanase reduces the level of KS in blood and provides a positive impact on cartilage lesions, demonstrating that SDET is a novel therapeutic approach to MPS IVA.<\/jats:p>","DOI":"10.3390\/ijms20174139","type":"journal-article","created":{"date-parts":[[2019,8,26]],"date-time":"2019-08-26T04:38:23Z","timestamp":1566794303000},"page":"4139","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Development of Substrate Degradation Enzyme Therapy for Mucopolysaccharidosis IVA Murine Model"],"prefix":"10.3390","volume":"20","author":[{"given":"Kazuki","family":"Sawamoto","sequence":"first","affiliation":[{"name":"Nemours\/Alfred I. duPont Hospital for Children, 1600 Rockland Rd., Wilmington, DE 19803, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0673-2160","authenticated-orcid":false,"given":"Shunji","family":"Tomatsu","sequence":"additional","affiliation":[{"name":"Nemours\/Alfred I. duPont Hospital for Children, 1600 Rockland Rd., Wilmington, DE 19803, USA"},{"name":"Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501-1193, Japan"},{"name":"Department of Pediatrics, Thomas Jefferson University, Philadelphia, PA 19107, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,8,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/S0002-9610(29)90496-7","article-title":"Chondro-osteo-dystrophy, roentgenopgraphic & clinical features of a child with dislocation of vertebrae","volume":"7","author":"Brailsford","year":"1929","journal-title":"Am. J. Surg."},{"key":"ref_2","first-page":"129","article-title":"Sur une forme de dystrophie osseuse familial","volume":"32","author":"Morquio","year":"1929","journal-title":"Arch. M\u00e9d. Enfants Paris."},{"key":"ref_3","unstructured":"Scriver, C.R., Beaudet, A.L., Sly, W.S., and Valle, D. (2001). The mucopolysaccharidoses. The Metabolic and Molecular Bases of Inherited Disease, McGraw-Hill. [8th ed.]."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1016\/j.ymgme.2016.11.007","article-title":"Mucopolysaccharidosis IVA and glycosaminoglycans","volume":"120","author":"Khan","year":"2017","journal-title":"Mol. Genet. Metab."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.ymgme.2018.05.004","article-title":"Diagnosis and prognosis of Mucopolysaccharidosis IVA","volume":"125","author":"Peracha","year":"2018","journal-title":"Mol. Genet. Metab."},{"key":"ref_6","unstructured":"Tomatsu, S. (2018). Mucopolysaccharidosis type IVA: Clinical features, biochemistry, diagnosis, genetics, and treatment. Mucopolysaccharidoses Update, Nova Science Publishers."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/j.ymgme.2015.09.007","article-title":"Obstructive airway in Morquio A syndrome, the past, the present and the future","volume":"117","author":"Tomatsu","year":"2016","journal-title":"Mol. Genet. Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ymgme.2018.03.004","article-title":"Growth impairment in mucopolysaccharidoses","volume":"124","author":"Melbouci","year":"2018","journal-title":"Mol. Genet. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1007\/s10545-007-0529-7","article-title":"International Morquio A Registry: Clinical manifestation and natural course of Morquio A disease","volume":"30","author":"Tomatsu","year":"2007","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_10","first-page":"65","article-title":"Current and emerging treatments and surgical interventions for Morquio A syndrome: A review","volume":"2012","author":"Tomatsu","year":"2012","journal-title":"Res. Rep. Endocr. Disord."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e329","DOI":"10.1016\/j.athoracsur.2016.02.113","article-title":"Surgical Reconstruction for Severe Tracheal Obstruction in Morquio A Syndrome","volume":"102","author":"Pizarro","year":"2016","journal-title":"Ann. Thorac. Surg."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1517\/21678707.2015.1086640","article-title":"Enzyme replacement therapy for treating mucopolysaccharidosis type IVA (Morquio A syndrome): Effect and limitations","volume":"3","author":"Tomatsu","year":"2015","journal-title":"Expert Opin. Orphan Drugs"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1080\/21678707.2016.1214572","article-title":"Current therapies for Morquio A syndrome and their clinical outcomes","volume":"4","author":"Sawamoto","year":"2016","journal-title":"Expert Opin. Orphan Drugs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1007\/s40262-014-0173-y","article-title":"Pharmacokinetic and pharmacodynamic evaluation of elosulfase alfa, an enzyme replacement therapy in patients with Morquio A syndrome","volume":"53","author":"Qi","year":"2014","journal-title":"Clin. Pharmacokinet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1937","DOI":"10.2147\/DDDT.S68562","article-title":"Impact of enzyme replacement therapy and hematopoietic stem cell transplantation in patients with Morquio A syndrome","volume":"9","author":"Tomatsu","year":"2015","journal-title":"Drug Des. Devel. Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.ymgme.2014.12.001","article-title":"Therapies for the bone in mucopolysaccharidoses","volume":"114","author":"Tomatsu","year":"2015","journal-title":"Mol. Genet. Metab."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1038\/s10038-019-0604-6","article-title":"Effect of enzyme replacement therapy on the growth of patients with Morquio A","volume":"64","author":"Doherty","year":"2019","journal-title":"J. Hum. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1093\/hmg\/ddm353","article-title":"Enzyme replacement therapy in a murine model of Morquio A syndrome","volume":"17","author":"Tomatsu","year":"2008","journal-title":"Hum. Mol. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1094","DOI":"10.1038\/mt.2010.32","article-title":"Enhancement of drug delivery: Enzyme-replacement therapy for murine Morquio A syndrome","volume":"18","author":"Tomatsu","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_20","first-page":"42","article-title":"30 months follow-up of an early enzyme replacement therapy in a severe Morquio A patient: About one case","volume":"9","author":"Wiedemann","year":"2016","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.ymgme.2015.09.011","article-title":"Hematopoietic stem cell transplantation for Morquio A syndrome","volume":"117","author":"Yabe","year":"2016","journal-title":"Mol. Genet. Metab."},{"key":"ref_22","first-page":"31","article-title":"Long-term therapeutic efficacy of allogenic bone marrow transplantation in a patient with mucopolysaccharidosis IVA","volume":"1","author":"Chinen","year":"2014","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2104","DOI":"10.1016\/j.bbmt.2016.08.015","article-title":"Allogeneic Hematopoietic Stem Cell Transplantation in Thirty-Four Pediatric Cases of Mucopolysaccharidosis-A Ten-Year Report from the China Children Transplant Group","volume":"22","author":"Wang","year":"2016","journal-title":"Biol. Blood Marrow Transplant."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e226","DOI":"10.1016\/j.bbmt.2019.02.012","article-title":"Hematopoietic Stem Cell Transplantation for Mucopolysaccharidoses: Past, Present, and Future","volume":"25","author":"Taylor","year":"2019","journal-title":"Biol. Blood Marrow Transplant."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1016\/j.ymgme.2009.03.008","article-title":"Deficiency in N-acetylgalactosamine-6-sulfate sulfatase results in collagen perturbations in cartilage of Morquio syndrome A patients","volume":"97","author":"Bank","year":"2009","journal-title":"Mol. Genet. Metab."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1073\/pnas.0912937107","article-title":"Involvement of the Toll-like receptor 4 pathway and use of TNF-alpha antagonists for treatment of the mucopolysaccharidoses","volume":"107","author":"Simonaro","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1016\/j.mehy.2010.08.006","article-title":"Morquio disease: The role of cartilage canals in the pathogenesis of chondrogenic dwarfism","volume":"75","author":"Zustin","year":"2010","journal-title":"Med. Hypotheses"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Frohbergh, M., Ge, Y., Meng, F., Karabul, N., Solyom, A., Lai, A., Iatridis, J., Schuchman, E.H., and Simonaro, C.M. (2014). Dose responsive effects of subcutaneous pentosan polysulfate injection in mucopolysaccharidosis type VI rats and comparison to oral treatment. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100882"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"25766","DOI":"10.1074\/jbc.M212183200","article-title":"Purification, characterization, and molecular cloning of a novel keratan sulfate hydrolase, endo-beta-N-acetylglucosaminidase, from Bacillus circulans","volume":"278","author":"Yamagishi","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"17091","DOI":"10.1523\/JNEUROSCI.5120-10.2011","article-title":"Keratan Sulfate Restricts Neural Plasticity after Spinal Cord Injury","volume":"31","author":"Imagama","year":"2011","journal-title":"J. Neurosci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.ymgme.2014.05.013","article-title":"Enzyme replacement therapy in newborn mucopolysaccharidosis IVA mice: Early treatment rescues bone lesions?","volume":"114","author":"Tomatsu","year":"2015","journal-title":"Mol. Genet. Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5233","DOI":"10.4049\/jimmunol.168.10.5233","article-title":"Receptor-mediated monitoring of tissue well-being via detection of soluble heparan sulfate by Toll-like receptor 4","volume":"168","author":"Johnson","year":"2002","journal-title":"J. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"18265","DOI":"10.1074\/jbc.M606352200","article-title":"Recognition of hyaluronan released in sterile injury involves a unique receptor complex dependent on Toll-like receptor 4, CD44, and MD-2","volume":"282","author":"Taylor","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1016\/j.joca.2007.04.008","article-title":"A molecular and histological characterization of cartilage from patients with Morquio syndrome","volume":"15","author":"Roseti","year":"2007","journal-title":"Osteoarthr. Cartil."},{"key":"ref_35","first-page":"1","article-title":"Di-sulfated Keratan Sulfate as a Novel Biomarker for Mucopolysaccharidosis II, IVA, and IVB","volume":"21","author":"Shimada","year":"2015","journal-title":"JIMD. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1203\/01.PDR.0000113767.60140.E9","article-title":"Development and testing of new screening method for keratan sulfate in mucopolysaccharidosis IVA","volume":"55","author":"Tomatsu","year":"2004","journal-title":"Pediatr. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"S35","DOI":"10.1007\/s10545-009-9013-x","article-title":"Validation of keratan sulfate level in mucopolysaccharidosis type IVA by liquid chromatography-tandem mass spectrometry","volume":"33","author":"Tomatsu","year":"2010","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"69","DOI":"10.4137\/BMI.S7451","article-title":"Comparison of liquid chromatography-tandem mass spectrometry and sandwich ELISA for determination of keratan sulfate in plasma and urine","volume":"6","author":"Hintze","year":"2011","journal-title":"Biomark. Insights"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.ymgme.2018.04.011","article-title":"Glycosaminoglycans analysis in blood and urine of patients with mucopolysaccharidosis","volume":"125","author":"Khan","year":"2018","journal-title":"Mol. Genet. Metab."},{"key":"ref_40","first-page":"100455","article-title":"Biomarkers in patients with mucopolysaccharidosis type II and IV","volume":"19","author":"Fujitsuka","year":"2019","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/0006-2944(75)90064-2","article-title":"Lack of relationship between blood and urine levels of glycosaminoglycans and lysomal enzymes","volume":"12","author":"Erickson","year":"1975","journal-title":"Biochem. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.ymgme.2017.12.007","article-title":"Glycosaminoglycan fragments as a measure of disease burden in the mucopolysaccharidosis type I mouse","volume":"123","author":"Saville","year":"2018","journal-title":"Mol. Genet. Metab."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.ebiom.2018.10.038","article-title":"Association of immune response with efficacy and safety outcomes in adults with phenylketonuria administered pegvaliase in phase 3 clinical trials","volume":"37","author":"Gupta","year":"2018","journal-title":"EBioMedicine"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/S0021-9258(18)56635-7","article-title":"A submicrodetermination of glucose","volume":"181","author":"Park","year":"1949","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3349","DOI":"10.1093\/hmg\/ddg366","article-title":"Mouse model of N-acetylgalactosamine-6-sulfate sulfatase deficiency (Galns\u2212\/\u2212) produced by targeted disruption of the gene defective in Morquio A disease","volume":"12","author":"Tomatsu","year":"2003","journal-title":"Hum. Mol. Genet."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"31237","DOI":"10.1074\/jbc.M801586200","article-title":"Tetrasulfated disaccharide unit in heparan sulfate: Enzymatic formation and tissue distribution","volume":"283","author":"Mochizuki","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1002\/bmc.760","article-title":"Analytical method for determination of disaccharides derived from keratan sulfates in human serum and plasma by high-performance liquid chromatography\/turbo-ionspray ionization tandem mass spectrometry","volume":"21","author":"Oguma","year":"2007","journal-title":"Biomed. Chromatogr."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.ab.2007.05.016","article-title":"Analytical method for the determination of disaccharides derived from keratan, heparan, and dermatan sulfates in human serum and plasma by high-performance liquid chromatography\/turbo ionspray ionization tandem mass spectrometry","volume":"368","author":"Oguma","year":"2007","journal-title":"Anal. Biochem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1007\/8904_2014_311","article-title":"Chondroitin 6-Sulfate as a Novel Biomarker for Mucopolysaccharidosis IVA and VII","volume":"16","author":"Shimada","year":"2014","journal-title":"JIMD. Rep."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/s10545-016-9981-6","article-title":"Newborn screening for mucopolysaccharidoses: A pilot study of measurement of glycosaminoglycans by tandem mass spectrometry","volume":"40","author":"Kubaski","year":"2017","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"3321","DOI":"10.1093\/hmg\/ddi364","article-title":"Development of MPS IVA mouse (Galnstm (hC79S.mC76S) slu) tolerant to human N-acetylgalactosamine-6-sulfate sulfatase","volume":"14","author":"Tomatsu","year":"2005","journal-title":"Hum. Mol. Genet."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/17\/4139\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:13:47Z","timestamp":1760188427000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/17\/4139"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,8,24]]},"references-count":51,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2019,9]]}},"alternative-id":["ijms20174139"],"URL":"https:\/\/doi.org\/10.3390\/ijms20174139","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,8,24]]}}}