Heparan sulfate as a regulator of endochondral ossification and osteochondroma development

Matrix Biology

Volumn 34, Issue , 2014, Pages 55-63

  Jochmann, Katja ^a   Bachvarova, Velina ^a   Vortkamp, Andrea ^a  

^a UNIVERSITY OF DUISBURG ESSEN   (Germany)

Author keywords

Chondrocyte differentiation; Endochondral ossification; Exostoses; Heparan sulfate; Ihh; Indian hedgehog; Osteochondroma; Signaling range

Indexed keywords

EXOSTOSIN-1; EXOSTOSIN-2; HEPARAN SULFATE; N ACETYLGLUCOSAMINYLTRANSFERASE;

BONE DEVELOPMENT; CARTILAGE CELL; CELL DIFFERENTIATION; CELL PROLIFERATION; CHONDROGENESIS; GENETICS; HUMAN; METABOLISM; OSTEOCHONDROMA; SIGNAL TRANSDUCTION;

CELL DIFFERENTIATION; CELL PROLIFERATION; CHONDROCYTES; CHONDROGENESIS; HEPARITIN SULFATE; HUMANS; N-ACETYLGLUCOSAMINYLTRANSFERASES; OSTEOCHONDROMA; OSTEOGENESIS; SIGNAL TRANSDUCTION;

EID: 84894428448     PISSN: 0945053X     EISSN: 15691802     Source Type: Journal    
DOI: 10.1016/j.matbio.2013.11.003     Document Type: Review

References (86)

1. Ahn J., Ludecke H.J., Lindow S., Horton W.A., Lee B., Wagner M.J., Horsthemke B., Wells D.E. Cloning of the putative tumour suppressor gene for hereditary multiple exostoses (EXT1). Nat. Genet. 1995, 11(2):137-143.
2. Ai X., Do A.T., Kusche-Gullberg M., Lindahl U., Lu K., Emerson C.P. Substrate specificity and domain functions of extracellular heparan sulfate 6-O-endosulfatases, QSulf1 and QSulf2. J. Biol. Chem. 2006, 281(8):4969-4976.
3. Benoist-Lasselin C., de Margerie E., Gibbs L., Cormier S., Silve C., Nicolas G., LeMerrer M., Mallet J.F., Munnich A., Bonaventure J., et al. Defective chondrocyte proliferation and differentiation in osteochondromas of MHE patients. Bone 2006, 39(1):17-26.
4. Bernfield M., Gotte M., Park P.W., Reizes O., Fitzgerald M.L., Lincecum J., Zako M. Functions of cell surface heparan sulfate proteoglycans. Annu. Rev. Biochem. 1999, 68:729-777.
5. Bishop J.R., Schuksz M., Esko J.D. Heparan sulphate proteoglycans fine-tune mammalian physiology. Nature 2007, 446(7139):1030-1037.
6. Bovee J.V., Cleton-Jansen A.M., Wuyts W., Caethoven G., Taminiau A.H., Bakker E., Van Hul W., Cornelisse C.J., Hogendoorn P.C. EXT-mutation analysis and loss of heterozygosity in sporadic and hereditary osteochondromas and secondary chondrosarcomas. Am. J. Hum. Genet. 1999, 65(3):689-698.
7. Bovee J.V., van den Broek L.J., Cleton-Jansen A.M., Hogendoorn P.C. Up-regulation of PTHrP and Bcl-2 expression characterizes the progression of osteochondroma towards peripheral chondrosarcoma and is a late event in central chondrosarcoma. Lab. Invest. 2000, 80(12):1925-1934.
8. Bullock S.L., Fletcher J.M., Beddington R.S., Wilson V.A. Renal agenesis in mice homozygous for a gene trap mutation in the gene encoding heparan sulfate 2-sulfotransferase. Genes Dev. 1998, 12(12):1894-1906.
9. Cardin A.D., Weintraub H.J. Molecular modeling of protein-glycosaminoglycan interactions. Arteriosclerosis 1989, 9(1):21-32.
10. Chang S.C., Mulloy B., Magee A.I., Couchman J.R. Two distinct sites in sonic Hedgehog combine for heparan sulfate interactions and cell signaling functions. J. Biol. Chem. 2011, 286(52):44391-44402.
11. Clement A., Wiweger M., von der Hardt S., Rusch M.A., Selleck S.B., Chien C.B., Roehl H.H. Regulation of zebrafish skeletogenesis by ext2/dackel and papst1/pinscher. PLoS Genet. 2008, 4(7):e1000136.
12. Cortes M., Baria A.T., Schwartz N.B. Sulfation of chondroitin sulfate proteoglycans is necessary for proper Indian hedgehog signaling in the developing growth plate. Development 2009, 136(10):1697-1706.
13. Cuellar A., Reddi A.H. Cell biology of osteochondromas: bone morphogenic protein signalling and heparan sulphates. Int. Orthop. 2013, 37(8):1591-1596.
14. Day T.F., Guo X., Garrett-Beal L., Yang Y. Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev. Cell 2005, 8(5):739-750.
15. Echtermeyer F., Bertrand J., Dreier R., Meinecke I., Neugebauer K., Fuerst M., Lee Y.J., Song Y.W., Herzog C., Theilmeier G., et al. Syndecan-4 regulates ADAMTS-5 activation and cartilage breakdown in osteoarthritis. Nat. Med. 2009, 15(9):1072-1076.
16. Esko J.D., Lindahl U. Molecular diversity of heparan sulfate. J. Clin. Invest. 2001, 108(2):169-173.
17. Esko J.D., Selleck S.B. Order out of chaos: assembly of ligand binding sites in heparan sulfate. Annu. Rev. Biochem. 2002, 71:435-471.
18. Farshi P., Ohlig S., Pickhinke U., Hoing S., Jochmann K., Lawrence R., Dreier R., Dierker T., Grobe K. Dual roles of the Cardin-Weintraub motif in multimeric Sonic hedgehog. J. Biol. Chem. 2011, 286(26):23608-23619.
19. Francannet C., Cohen-Tanugi A., Le Merrer M., Munnich A., Bonaventure J., Legeai-Mallet L. Genotype-phenotype correlation in hereditary multiple exostoses. J. Med. Genet. 2001, 38(7):430-434.
20. Franz-Odendaal T.A. Induction and patterning of intramembranous bone. Front. Biosci. (Landmark Ed.) 2011, 16:2734-2746.
21. Gao B., Guo J., She C., Shu A., Yang M., Tan Z., Yang X., Guo S., Feng G., He L. Mutations in IHH, encoding Indian hedgehog, cause brachydactyly type A-1. Nat. Genet. 2001, 28(4):386-388.
22. Gao B., Hu J., Stricker S., Cheung M., Ma G., Law K.F., Witte F., Briscoe J., Mundlos S., He L., et al. A mutation in Ihh that causes digit abnormalities alters its signalling capacity and range. Nature 2009, 458(7242):1196-1200.
23. Grobe K., Inatani M., Pallerla S.R., Castagnola J., Yamaguchi Y., Esko J.D. Cerebral hypoplasia and craniofacial defects in mice lacking heparan sulfate Ndst1 gene function. Development 2005, 132(16):3777-3786.
24. Habuchi H., Nagai N., Sugaya N., Atsumi F., Stevens R.L., Kimata K. Mice deficient in heparan sulfate 6-O-sulfotransferase-1 exhibit defective heparan sulfate biosynthesis, abnormal placentation, and late embryonic lethality. J. Biol. Chem. 2007, 282(21):15578-15588.
25. Hall C.R., Cole W.G., Haynes R., Hecht J.T. Reevaluation of a genetic model for the development of exostosis in hereditary multiple exostosis. Am. J. Med. Genet. 2002, 112(1):1-5.
26. Hameetman L., Rozeman L.B., Lombaerts M., Oosting J., Taminiau A.H., Cleton-Jansen A.M., Bovee J.V., Hogendoorn P.C. Peripheral chondrosarcoma progression is accompanied by decreased Indian Hedgehog signalling. J. Pathol. 2006, 209(4):501-511.
27. Hill T.P., Spater D., Taketo M.M., Birchmeier W., Hartmann C. Canonical Wnt/beta-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev. Cell 2005, 8(5):727-738.
28. Holmborn K., Habicher J., Kasza Z., Eriksson A.S., Filipek-Gorniok B., Gopal S., Couchman J.R., Ahlberg P.E., Wiweger M., Spillmann D., et al. On the roles and regulation of chondroitin sulfate and heparan sulfate in zebrafish pharyngeal cartilage morphogenesis. J. Biol. Chem. 2012, 287(40):33905-33916.
29. Holst C.R., Bou-Reslan H., Gore B.B., Wong K., Grant D., Chalasani S., Carano R.A., Frantz G.D., Tessier-Lavigne M., Bolon B., et al. Secreted sulfatases Sulf1 and Sulf2 have overlapping yet essential roles in mouse neonatal survival. PLoS One 2007, 2(6):e575.
30. Hu Z., Yu M., Hu G. NDST-1 modulates BMPR and PTHrP signaling during endochondral bone formation in a gene knockout model. Bone 2007, 40(6):1462-1474.
31. Huegel J., Mundy C., Sgariglia F., Nygren P., Billings P.C., Yamaguchi Y., Koyama E., Pacifici M. Perichondrium phenotype and border function are regulated by Ext1 and heparan sulfate in developing long bones: a mechanism likely deranged in Hereditary Multiple Exostoses. Dev. Biol. 2013, 377(1):100-112.
32. Jager M., Westhoff B., Portier S., Leube B., Hardt K., Royer-Pokora B., Gossheger G., Krauspe R. Clinical outcome and genotype in patients with hereditary multiple exostoses. J. Orthop. Res. 2007, 25(12):1541-1551.
33. Jasuja R., Allen B.L., Pappano W.N., Rapraeger A.C., Greenspan D.S. Cell-surface heparan sulfate proteoglycans potentiate chordin antagonism of bone morphogenetic protein signaling and are necessary for cellular uptake of chordin. J. Biol. Chem. 2004, 279(49):51289-51297.
34. Jennes I., Pedrini E., Zuntini M., Mordenti M., Balkassmi S., Asteggiano C.G., Casey B., Bakker B., Sangiorgi L., Wuyts W. Multiple osteochondromas: mutation update and description of the multiple osteochondromas mutation database (MOdb). Hum. Mutat. 2009, 30(12):1620-1627.
35. Jones K.B., Piombo V., Searby C., Kurriger G., Yang B., Grabellus F., Roughley P.J., Morcuende J.A., Buckwalter J.A., Capecchi M.R., et al. A mouse model of osteochondromagenesis from clonal inactivation of Ext1 in chondrocytes. Proc. Natl. Acad. Sci. U. S. A. 2010, 107(5):2054-2059.
36. Kaneko H., Ishijima M., Futami I., Tomikawa-Ichikawa N., Kosaki K., Sadatsuki R., Yamada Y., Kurosawa H., Kaneko K., Arikawa-Hirasawa E. Synovial perlecan is required for osteophyte formation in knee osteoarthritis. Matrix Biol. 2013, 32(3-4):178-187.
37. Koziel L., Kunath M., Kelly O.G., Vortkamp A. Ext1-dependent heparan sulfate regulates the range of Ihh signaling during endochondral ossification. Dev. Cell 2004, 6(6):801-813.
38. Kronenberg H.M. Developmental regulation of the growth plate. Nature 2003, 423(6937):332-336.
39. Lanske B., Karaplis A.C., Lee K., Luz A., Vortkamp A., Pirro A., Karperien M., Defize L.H., Ho C., Mulligan R.C., et al. PTH/PTHrP receptor in early development and Indian hedgehog-regulated bone growth. Science 1996, 273(5275):663-666.
40. Ledin J., Staatz W., Li J.P., Gotte M., Selleck S., Kjellen L., Spillmann D. Heparan sulfate structure in mice with genetically modified heparan sulfate production. J. Biol. Chem. 2004, 279(41):42732-42741.
41. Legeai-Mallet L., Rossi A., Benoist-Lasselin C., Piazza R., Mallet J.F., Delezoide A.L., Munnich A., Bonaventure J., Zylberberg L. EXT 1 gene mutation induces chondrocyte cytoskeletal abnormalities and defective collagen expression in the exostoses. J. Bone Miner. Res. 2000, 15(8):1489-1500.
42. Li J.P., Gong F., Hagner-McWhirter A., Forsberg E., Abrink M., Kisilevsky R., Zhang X., Lindahl U. Targeted disruption of a murine glucuronyl C5-epimerase gene results in heparan sulfate lacking L-iduronic acid and in neonatal lethality. J. Biol. Chem. 2003, 278(31):28363-28366.
43. Lind T., Tufaro F., McCormick C., Lindahl U., Lidholt K. The putative tumor suppressors EXT1 and EXT2 are glycosyltransferases required for the biosynthesis of heparan sulfate. J. Biol. Chem. 1998, 273(41):26265-26268.
44. Lortat-Jacob H., Burhan I., Scarpellini A., Thomas A., Imberty A., Vives R.R., Johnson T., Gutierrez A., Verderio E.A. Transglutaminase-2 interaction with heparin: identification of a heparin binding site that regulates cell adhesion to fibronectin-transglutaminase-2 matrix. J. Biol. Chem. 2012, 287(22):18005-18017.
45. Ma G., Yu J., Xiao Y., Chan D., Gao B., Hu J., He Y., Guo S., Zhou J., Zhang L., et al. Indian hedgehog mutations causing brachydactyly type A1 impair Hedgehog signal transduction at multiple levels. Cell Res. 2011, 21(9):1343-1357.
46. Maes C., Kobayashi T., Selig M.K., Torrekens S., Roth S.I., Mackem S., Carmeliet G., Kronenberg H.M. Osteoblast precursors, but not mature osteoblasts, move into developing and fractured bones along with invading blood vessels. Dev. Cell 2010, 19(2):329-344.
47. Matsumoto K., Irie F., Mackem S., Yamaguchi Y. A mouse model of chondrocyte-specific somatic mutation reveals a role for Ext1 loss of heterozygosity in multiple hereditary exostoses. Proc. Natl. Acad. Sci. U. S. A. 2010, 107(24):10932-10937.
48. Matsumoto Y., Matsumoto K., Irie F., Fukushi J., Stallcup W.B., Yamaguchi Y. Conditional ablation of the heparan sulfate-synthesizing enzyme Ext1 leads to dysregulation of bone morphogenic protein signaling and severe skeletal defects. J. Biol. Chem. 2010, 285(25):19227-19234.
49. Minina E., Kreschel C., Naski M.C., Ornitz D.M., Vortkamp A. Interaction of FGF, Ihh/Pthlh, and BMP signaling integrates chondrocyte proliferation and hypertrophic differentiation. Dev. Cell 2002, 3(3):439-449.
50. Mitchell K.J., Pinson K.I., Kelly O.G., Brennan J., Zupicich J., Scherz P., Leighton P.A., Goodrich L.V., Lu X., Avery B.J., et al. Functional analysis of secreted and transmembrane proteins critical to mouse development. Nat. Genet. 2001, 28(3):241-249.
51. Mundy C., Yasuda T., Kinumatsu T., Yamaguchi Y., Iwamoto M., Enomoto-Iwamoto M., Koyama E., Pacifici M. Synovial joint formation requires local Ext1 expression and heparan sulfate production in developing mouse embryo limbs and spine. Dev. Biol. 2011, 351(1):70-81.
52. Ohlig S., Farshi P., Pickhinke U., van den Boom J., Hoing S., Jakuschev S., Hoffmann D., Dreier R., Scholer H.R., Dierker T., et al. Sonic hedgehog shedding results in functional activation of the solubilized protein. Dev. Cell 2011, 20(6):764-774.
53. Ohlig S., Pickhinke U., Sirko S., Bandari S., Hoffmann D., Dreier R., Farshi P., Gotz M., Grobe K. An emerging role of Sonic hedgehog shedding as a modulator of heparan sulfate interactions. J. Biol. Chem. 2012, 287(52):43708-43719.
54. Otsuki S., Taniguchi N., Grogan S.P., D'Lima D., Kinoshita M., Lotz M. Expression of novel extracellular sulfatases Sulf-1 and Sulf-2 in normal and osteoarthritic articular cartilage. Arthritis Res. Ther. 2008, 10(3):R61.
55. Otsuki S., Hanson S.R., Miyaki S., Grogan S.P., Kinoshita M., Asahara H., Wong C.H., Lotz M.K. Extracellular sulfatases support cartilage homeostasis by regulating BMP and FGF signaling pathways. Proc. Natl. Acad. Sci. U. S. A. 2010, 107(22):10202-10207.
56. Paine-Saunders S., Viviano B.L., Economides A.N., Saunders S. Heparan sulfate proteoglycans retain Noggin at the cell surface: a potential mechanism for shaping bone morphogenetic protein gradients. J. Biol. Chem. 2002, 277(3):2089-2096.
57. Pallerla S.R., Pan Y., Zhang X., Esko J.D., Grobe K. Heparan sulfate Ndst1 gene function variably regulates multiple signaling pathways during mouse development. Dev. Dyn. 2007, 236(2):556-563.
58. Pedrini E., Jennes I., Tremosini M., Milanesi A., Mordenti M., Parra A., Sgariglia F., Zuntini M., Campanacci L., Fabbri N., et al. Genotype-phenotype correlation study in 529 patients with multiple hereditary exostoses: identification of "protective" and "risk" factors. J. Bone Joint Surg. Am. 2011, 93(24):2294-2302.
59. Porter D.E., Lonie L., Fraser M., Dobson-Stone C., Porter J.R., Monaco A.P., Simpson A.H. Severity of disease and risk of malignant change in hereditary multiple exostoses. A genotype-phenotype study. J. Bone Joint Surg. Br. 2004, 86(7):1041-1046.
60. Ratzka A., Kalus I., Moser M., Dierks T., Mundlos S., Vortkamp A. Redundant function of the heparan sulfate 6-O-endosulfatases Sulf1 and Sulf2 during skeletal development. Dev. Dyn. 2008, 237(2):339-353.
61. Ratzka A., Mundlos S., Vortkamp A. Expression patterns of sulfatase genes in the developing mouse embryo. Dev. Dyn. 2010, 239(6):1779-1788.
62. Reijnders C.M., Waaijer C.J., Hamilton A., Buddingh E.P., Dijkstra S.P., Ham J., Bakker E., Szuhai K., Karperien M., Hogendoorn P.C., et al. No haploinsufficiency but loss of heterozygosity for EXT in multiple osteochondromas. Am. J. Pathol. 2010, 177(4):1946-1957.
63. Rubin J.B., Choi Y., Segal R.A. Cerebellar proteoglycans regulate sonic hedgehog responses during development. Development 2002, 129(9):2223-2232.
64. Sarrazin S., Lamanna W.C., Esko J.D. Heparan sulfate proteoglycans. Cold Spring Harb. Perspect. Biol. 2011, 3(7).
65. Schmale G.A., Conrad E.U., Raskind W.H. The natural history of hereditary multiple exostoses. J. Bone Joint Surg. Am. 1994, 76(7):986-992.
66. Sgariglia F., Candela M.E., Huegel J., Jacenko O., Koyama E., Yamaguchi Y., Pacifici M., Enomoto-Iwamoto M. Epiphyseal abnormalities, trabecular bone loss and articular chondrocyte hypertrophy develop in the long bones of postnatal Ext1-deficient mice. Bone 2013, 57(1):220-231.
67. Stattin E.L., Linden B., Lonnerholm T., Schuster J., Dahl N. Brachydactyly type A1 associated with unusual radiological findings and a novel Arg158Cys mutation in the Indian hedgehog (IHH) gene. Eur. J. Med. Genet. 2009, 52(5):297-302.
68. Stickens D., Clines G., Burbee D., Ramos P., Thomas S., Hogue D., Hecht J.T., Lovett M., Evans G.A. The EXT2 multiple exostoses gene defines a family of putative tumour suppressor genes. Nat. Genet. 1996, 14(1):25-32.
69. Stickens D., Zak B.M., Rougier N., Esko J.D., Werb Z. Mice deficient in Ext2 lack heparan sulfate and develop exostoses. Development 2005, 132(22):5055-5068.
70. Sugahara K., Schwartz N.B. Defect in 3'-phosphoadenosine 5'-phosphosulfate formation in brachymorphic mice. Proc. Natl. Acad. Sci. U. S. A. 1979, 76(12):6615-6618.
71. Szuhai K., Jennes I., de Jong D., Bovee J.V., Wiweger M., Wuyts W., Hogendoorn P.C. Tiling resolution array-CGH shows that somatic mosaic deletion of the EXT gene is causative in EXT gene mutation negative multiple osteochondromas patients. Hum. Mutat. 2011, 32(2):E2036-E2049.
72. Tiet T.D., Hopyan S., Nadesan P., Gokgoz N., Poon R., Lin A.C., Yan T., Andrulis I.L., Alman B.A., Wunder J.S. Constitutive hedgehog signaling in chondrosarcoma up-regulates tumor cell proliferation. Am. J. Pathol. 2006, 168(1):321-330.
73. Vanhoenacker F.M., Van Hul W., Wuyts W., Willems P.J., De Schepper A.M. Hereditary multiple exostoses: from genetics to clinical syndrome and complications. Eur. J. Radiol. 2001, 40(3):208-217.
74. Vortkamp A., Lee K., Lanske B., Segre G.V., Kronenberg H.M., Tabin C.J. Regulation of rate of cartilage differentiation by Indian hedgehog and PTH-related protein [see comments]. Science 1996, 273(5275):613-622.
75. Wang W., Zhong B., Sun J., Cao J., Tian J., Zhong N., Zhao W., Tian L., Xu P., Guo D., et al. Down-regulated HS6ST2 in osteoarthritis and Kashin-Beck disease inhibits cell viability and influences expression of the genes relevant to aggrecan metabolism of human chondrocytes. Rheumatology (Oxford) 2011, 50(12):2176-2186.
76. Warda M., Toida T., Zhang F., Sun P., Munoz E., Xie J., Linhardt R.J. Isolation and characterization of heparan sulfate from various murine tissues. Glycoconj. J. 2006, 23(7-8):555-563.
77. Whalen D.M., Malinauskas T., Gilbert R.J., Siebold C. Structural insights into proteoglycan-shaped Hedgehog signaling. Proc. Natl. Acad. Sci. U. S. A. 2013, 110(41):16420-16425.
78. Witt R.M., Hecht M.L., Pazyra-Murphy M.F., Cohen S.M., Noti C., van Kuppevelt T.H., Fuller M., Chan J.A., Hopwood J.J., Seeberger P.H., et al. Heparan sulfate proteoglycans containing a glypican 5 core and 2-O-sulfo-iduronic acid function as Sonic Hedgehog co-receptors to promote proliferation. J. Biol. Chem. 2013, 288(36):26275-26288.
79. Wuelling M., Vortkamp A. Chondrocyte proliferation and differentiation. Endocr. Dev. 2011, 21:1-11.
80. Wuyts W., Van Hul W., De Boulle K., Hendrickx J., Bakker E., Vanhoenacker F., Mollica F., Ludecke H.J., Sayli B.S., Pazzaglia U.E., et al. Mutations in the EXT1 and EXT2 genes in hereditary multiple exostoses. Am. J. Hum. Genet. 1998, 62(2):346-354.
81. Yamada S., Busse M., Ueno M., Kelly O.G., Skarnes W.C., Sugahara K., Kusche-Gullberg M. Embryonic fibroblasts with a gene trap mutation in Ext1 produce short heparan sulfate chains. J. Biol. Chem. 2004, 279(31):32134-32141.
82. Yang W., Wang J., Moore D.C., Liang H., Dooner M., Wu Q., Terek R., Chen Q., Ehrlich M.G., Quesenberry P.J., et al. Ptpn11 deletion in a novel progenitor causes metachondromatosis by inducing hedgehog signalling. Nature 2013, 499(7459):491-495.
83. Yasuda T., Mundy C., Kinumatsu T., Shibukawa Y., Shibutani T., Grobe K., Minugh-Purvis N., Pacifici M., Koyama E. Sulfotransferase Ndst1 is needed for mandibular and TMJ development. J. Dent. Res. 2010, 89(10):1111-1116.
84. Yoon B.S., Ovchinnikov D.A., Yoshii I., Mishina Y., Behringer R.R., Lyons K.M. Bmpr1a and Bmpr1b have overlapping functions and are essential for chondrogenesis in vivo. Proc. Natl. Acad. Sci. U. S. A. 2005, 102(14):5062-5067.
85. Yoon B.S., Pogue R., Ovchinnikov D.A., Yoshii I., Mishina Y., Behringer R.R., Lyons K.M. BMPs regulate multiple aspects of growth-plate chondrogenesis through opposing actions on FGF pathways. Development 2006, 133(23):4667-4678.
86. Zak B.M., Schuksz M., Koyama E., Mundy C., Wells D.E., Yamaguchi Y., Pacifici M., Esko J.D. Compound heterozygous loss of Ext1 and Ext2 is sufficient for formation of multiple exostoses in mouse ribs and long bones. Bone 2011, 48(5):979-987.