Runx2 is essential for the transdifferentiation of chondrocytes into osteoblasts

English version

Autoři: Xin Qin ^aff001; Qing Jiang ^aff001; Kenichi Nagano ^aff002; Takeshi Moriishi ^aff003; Toshihiro Miyazaki ^aff003; Hisato Komori ^aff001; Kosei Ito ^aff004; Klaus von der Mark ^aff005; Chiharu Sakane ^aff006; Hitomi Kaneko ^aff001; Toshihisa Komori ^aff001
Působiště autorů: Basic and Translational Research Center for Hard Tissue Disease, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ^aff001; Department of Oral Pathology and Bone Metabolism, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ^aff002; Department of Cell Biology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ^aff003; Department of Molecular Bone Biology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ^aff004; Department of Experimental Medicine I, Nikolaus-Fiebiger Center of Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany ^aff005; Division of Comparative Medicine, Life Science Support Center, Nagasaki University, Nagasaki, Japan ^aff006
Vyšlo v časopise: Runx2 is essential for the transdifferentiation of chondrocytes into osteoblasts. PLoS Genet 16(11): e1009169. doi:10.1371/journal.pgen.1009169
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1009169

Souhrn

Chondrocytes proliferate and mature into hypertrophic chondrocytes. Vascular invasion into the cartilage occurs in the terminal hypertrophic chondrocyte layer, and terminal hypertrophic chondrocytes die by apoptosis or transdifferentiate into osteoblasts. Runx2 is essential for osteoblast differentiation and chondrocyte maturation. Runx2-deficient mice are composed of cartilaginous skeletons and lack the vascular invasion into the cartilage. However, the requirement of Runx2 in the vascular invasion into the cartilage, mechanism of chondrocyte transdifferentiation to osteoblasts, and its significance in bone development remain to be elucidated. To investigate these points, we generated Runx2^fl/flCre mice, in which Runx2 was deleted in hypertrophic chondrocytes using Col10a1 Cre. Vascular invasion into the cartilage was similarly observed in Runx2^fl/fl and Runx2^fl/flCre mice. Vegfa expression was reduced in the terminal hypertrophic chondrocytes in Runx2^fl/flCre mice, but Vegfa was strongly expressed in osteoblasts in the bone collar, suggesting that Vegfa expression in bone collar osteoblasts is sufficient for vascular invasion into the cartilage. The apoptosis of terminal hypertrophic chondrocytes was increased and their transdifferentiation was interrupted in Runx2^fl/flCre mice, leading to lack of primary spongiosa and osteoblasts in the region at E16.5. The osteoblasts appeared in this region at E17.5 in the absence of transdifferentiation, and the number of osteoblasts and the formation of primary spongiosa, but not secondary spongiosa, reached to levels similar those in Runx2^fl/fl mice at birth. The bone structure and volume and all bone histomophometric parameters were similar between Runx2^fl/fl and Runx2^fl/flCre mice after 6 weeks of age. These findings indicate that Runx2 expression in terminal hypertrophic chondrocytes is not required for vascular invasion into the cartilage, but is for their survival and transdifferentiation into osteoblasts, and that the transdifferentiation is necessary for trabecular bone formation in embryonic and neonatal stages, but not for acquiring normal bone structure and volume in young and adult mice.

Klíčová slova:

Cartilage – Bone development – Femur – Chondrocytes – Ossification – Osteoblasts – Safranin staining – Transdifferentiation

Zdroje

1. C KA. Embryonic development of bone and the molecular regulation of intramembranous and endochondral bone formation. Bilezikian JPR, L. G; Rodan G. A, editor. London: Academic Press; 2002.

2. Inada M, Yasui T, Nomura S, Miyake S, Deguchi K, Himeno M, et al. Maturational disturbance of chondrocytes in Cbfa1-deficient mice. Dev Dyn. 1999;214(4):279–90. doi: 10.1002/(SICI)1097-0177(199904)214 : 4<279::AID-AJA1>3.0.CO;2-W 10213384

3. Komori T. Signaling networks in RUNX2-dependent bone development. J Cell Biochem. 2011;112(3):750–5. doi: 10.1002/jcb.22994 21328448

4. Qin X, Jiang Q, Miyazaki T, Komori T. Runx2 regulates cranial suture closure by inducing hedgehog, Fgf, Wnt and Pthlh signaling pathway gene expressions in suture mesenchymal cells. Hum Mol Genet. 2019; 28(6):896–911. doi: 10.1093/hmg/ddy386 30445456

5. Kim IS, Otto F, Zabel B, Mundlos S. Regulation of chondrocyte differentiation by Cbfa1. Mech Dev. 1999;80(2):159–70. doi: 10.1016/s0925-4773(98)00210-x 10072783

6. Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, et al. Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 1997;89(5):755–64. doi: 10.1016/s0092-8674(00)80258-5 9182763

7. Otto F, Thornell AP, Crompton T, Denzel A, Gilmour KC, Rosewell IR, et al. Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell. 1997;89(5):765–71. doi: 10.1016/s0092-8674(00)80259-7 9182764

8. Zelzer E, Glotzer DJ, Hartmann C, Thomas D, Fukai N, Soker S, et al. Tissue specific regulation of VEGF expression during bone development requires Cbfa1/Runx2. Mech Dev. 2001;106(1–2):97–106. doi: 10.1016/s0925-4773(01)00428-2 11472838

9. Himeno M, Enomoto H, Liu W, Ishizeki K, Nomura S, Kitamura Y, et al. Impaired vascular invasion of Cbfa1-deficient cartilage engrafted in the spleen. J Bone Miner Res. 2002;17(7):1297–305. doi: 10.1359/jbmr.2002.17.7.1297 12096844

10. Yoshida CA, Yamamoto H, Fujita T, Furuichi T, Ito K, Inoue K, et al. Runx2 and Runx3 are essential for chondrocyte maturation, and Runx2 regulates limb growth through induction of Indian hedgehog. Genes Dev. 2004;18(8):952–63. doi: 10.1101/gad.1174704 15107406

11. Enomoto H, Enomoto-Iwamoto M, Iwamoto M, Nomura S, Himeno M, Kitamura Y, et al. Cbfa1 is a positive regulatory factor in chondrocyte maturation. J Biol Chem. 2000;275(12):8695–702. doi: 10.1074/jbc.275.12.8695 10722711

12. Takeda S, Bonnamy JP, Owen MJ, Ducy P, Karsenty G. Continuous expression of Cbfa1 in nonhypertrophic chondrocytes uncovers its ability to induce hypertrophic chondrocyte differentiation and partially rescues Cbfa1-deficient mice. Genes Dev. 2001;15(4):467–81. doi: 10.1101/gad.845101 11230154

13. Ueta C, Iwamoto M, Kanatani N, Yoshida C, Liu Y, Enomoto-Iwamoto M, et al. Skeletal malformations caused by overexpression of Cbfa1 or its dominant negative form in chondrocytes. J Cell Biol. 2001;153(1):87–100. doi: 10.1083/jcb.153.1.87 11285276

14. Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM, Tabin CJ. Regulation of rate of cartilage differentiation by Indian hedgehog and PTH-related protein. Science (New York, NY). 1996;273(5275):613–22. doi: 10.1126/science.273.5275.613 8662546

15. St-Jacques B, Hammerschmidt M, McMahon AP. Indian hedgehog signaling regulates proliferation and differentiation of chondrocytes and is essential for bone formation. Genes Dev. 1999;13(16):2072–86. doi: 10.1101/gad.13.16.2072 10465785

16. Yang L, Tsang KY, Tang HC, Chan D, Cheah KS. Hypertrophic chondrocytes can become osteoblasts and osteocytes in endochondral bone formation. Proc Natl Acad Sci U S A. 2014;111(33):12097–102. doi: 10.1073/pnas.1302703111 25092332

17. Zhou X, von der Mark K, Henry S, Norton W, Adams H, de Crombrugghe B. Chondrocytes transdifferentiate into osteoblasts in endochondral bone during development, postnatal growth and fracture healing in mice. PLoS Genet. 2014;10(12):e1004820. doi: 10.1371/journal.pgen.1004820 25474590

18. Jing Y, Zhou X, Han X, Jing J, von der Mark K, Wang J, et al. Chondrocytes Directly Transform into Bone Cells in Mandibular Condyle Growth. J Dent Res. 2015;94(12):1668–75. doi: 10.1177/0022034515598135 26341973

19. Park J, Gebhardt M, Golovchenko S, Perez-Branguli F, Hattori T, Hartmann C, et al. Dual pathways to endochondral osteoblasts: a novel chondrocyte-derived osteoprogenitor cell identified in hypertrophic cartilage. Biology open. 2015;4(5):608–21. doi: 10.1242/bio.201411031 25882555

20. Houben A, Kostanova-Poliakova D, Weissenbock M, Graf J, Teufel S, von der Mark K, et al. beta-catenin activity in late hypertrophic chondrocytes locally orchestrates osteoblastogenesis and osteoclastogenesis. Development. 2016;143(20):3826–38. doi: 10.1242/dev.137489 27621061

21. Gebhard S, Hattori T, Bauer E, Schlund B, Bosl MR, de Crombrugghe B, et al. Specific expression of Cre recombinase in hypertrophic cartilage under the control of a BAC-Col10a1 promoter. Matrix Biol. 2008;27(8):693–9. doi: 10.1016/j.matbio.2008.07.001 18692570

22. Enomoto H, Shiojiri S, Hoshi K, Furuichi T, Fukuyama R, Yoshida CA, et al. Induction of osteoclast differentiation by Runx2 through receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin regulation and partial rescue of osteoclastogenesis in Runx2-/ -⁠ mice by RANKL transgene. J Biol Chem. 2003;278(26):23971–7. doi: 10.1074/jbc.M302457200 12697767

23. Komori T. Cell Death in Chondrocytes, Osteoblasts, and Osteocytes. Int J Mol Sci. 2016;17(12). doi: 10.3390/ijms17122045 27929439

24. Mendonça Gorgulho C, Murthy P, Liotta L, Espina V, Lotze MT. Different measures of HMGB1 location in cancer immunology. Methods Enzymol. 2019;629 : 195–217. doi: 10.1016/bs.mie.2019.10.011 31727241

25. Imayoshi I, Hirano K, Sakamoto M, Miyoshi G, Imura T, Kitano S, et al. A multifunctional teal-fluorescent Rosa26 reporter mouse line for Cre -⁠ and Flp-mediated recombination. Neuroscience research. 2012;73(1):85–91. doi: 10.1016/j.neures.2012.02.003 22343123

26. Komori T. Regulation of bone development and extracellular matrix protein genes by RUNX2. Cell Tissue Res. 2010;339(1):189–95. doi: 10.1007/s00441-009-0832-8 19649655

27. Komori T. Runx2, an inducer of osteoblast and chondrocyte differentiation. Histochem Cell Biol. 2018;149(4):313–23. doi: 10.1007/s00418-018-1640-6 29356961

28. Zelzer E, Mamluk R, Ferrara N, Johnson RS, Schipani E, Olsen BR. VEGFA is necessary for chondrocyte survival during bone development. Development (Cambridge, England). 2004;131(9):2161–71. doi: 10.1242/dev.01053 15073147

29. Duan X, Murata Y, Liu Y, Nicolae C, Olsen BR, Berendsen AD. Vegfa regulates perichondrial vascularity and osteoblast differentiation in bone development. Development. 2015;142(11):1984–91. doi: 10.1242/dev.117952 25977369

30. Colnot C, Lu C, Hu D, Helms JA. Distinguishing the contributions of the perichondrium, cartilage, and vascular endothelium to skeletal development. Dev Biol. 2004;269(1):55–69. doi: 10.1016/j.ydbio.2004.01.011 15081357

31. Cheung WY, Fritton JC, Morgan SA, Seref-Ferlengez Z, Basta-Pljakic J, Thi MM, et al. Pannexin-1 and P2X7-Receptor Are Required for Apoptotic Osteocytes in Fatigued Bone to Trigger RANKL Production in Neighboring Bystander Osteocytes. J Bone Miner Res. 2016;31(4):890–9. doi: 10.1002/jbmr.2740 26553756

32. Mizuhashi K, Ono W, Matsushita Y, Sakagami N, Takahashi A, Saunders TL, et al. Resting zone of the growth plate houses a unique class of skeletal stem cells. Nature. 2018;563(7730):254–8. doi: 10.1038/s41586-018-0662-5 30401834

33. Maes C, Kobayashi T, Selig MK, Torrekens S, Roth SI, Mackem S, et al. Osteoblast precursors, but not mature osteoblasts, move into developing and fractured bones along with invading blood vessels. Dev Cell. 2010;19(2):329–44. doi: 10.1016/j.devcel.2010.07.010 20708594

34. Ono N, Ono W, Nagasawa T, Kronenberg HM. A subset of chondrogenic cells provides early mesenchymal progenitors in growing bones. Nat Cell Biol. 2014;16(12):1157–67. doi: 10.1038/ncb3067 25419849

35. Mizuhashi K, Nagata M, Matsushita Y, Ono W, Ono N. Growth Plate Borderline Chondrocytes Behave as Transient Mesenchymal Precursor Cells. J Bone Miner Res. 2019;34(8):1387–92. doi: 10.1002/jbmr.3719 30888720

36. Dempster DW, Compston JE, Drezner MK, Glorieux FH, Kanis JA, Malluche H, et al. Standardized nomenclature, symbols, and units for bone histomorphometry: a 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res. 2013;28(1):2–17. doi: 10.1002/jbmr.1805 23197339

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