A Novel Factor, Tmem176b, Induced by Activin-like Kinase 2 Signal Promotes the Differentiation of Myoblasts into Osteoblasts

Abstract

Previous studies have suggested some interactions between muscle tissues and bone metabolism. The constitutively activating mutation (R206H) of the BMP type I receptor, activin-like-kinase 2 (ALK2), causes fibrodysplasia ossificans progressiva (FOP), which is characterized by extensive ossifications within muscle tissues. In the present study, we revealed that Tmem176b mRNA levels were upregulated by stable transfection of ALK2 (R206H) in mouse myoblastic C2C12 cells. Transient Tmem176b overexpression elevated levels of osteoblast differentiation markers, such as Osterix and alkaline phosphatase, as well as mineralization in C2C12 cells. In addition, Tmem176b overexpression elevated the levels of these markers in mouse osteoblastic MC3T3-E1 cells. On the other hand, Tmem176b overexpression suppressed the levels of myogenic markers, such as MyoD and myogenin in C2C12 cells, although it did not affect the levels of chondrogenic markers, such as type II and X collagens. In conclusion, the present study is the first to demonstrate that Tmem176b induces the differentiation of myoblasts into an osteoblast lineage.

Key words

bone morphogenetic protein - fibrodysplasia ossificans progressiva - osteoblast - myoblast

• References

• 1 Chen G, Deng C, Li YP. TGF-beta and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci 2012; 8: 272-288
• 2 Condamine T, Le Texier L, Howie D et al. Tmem176B and Tmem176A are associated with the immature state of dendritic cells. J Leukoc Biol 2010; 88: 507-515
• 3 Fukuda T, Kohda M, Kanomata K et al. Constitutively activated ALK2 and increased SMAD1/5 cooperatively induce bone morphogenetic protein signaling in fibrodysplasia ossificans progressiva. J Biol Chem 2009; 284: 7149-7156
• 4 Hisa I, Inoue Y, Hendy GN et al. Parathyroid hormone-responsive Smad3-related factor, Tmem119, promotes osteoblast differentiation and interacts with the bone morphogenetic protein-Runx2 pathway. J Biol Chem 2011; 286: 9787-9796
• 5 Jensen ED, Gopalakrishnan R, Westendorf JJ. Regulation of gene expression in osteoblasts. Biofactors 2010; 36: 25-32
• 6 Karsenty G, Wagner EF. Reaching a genetic and molecular understanding of skeletal development. Dev Cell 2002; 2: 389-406
• 7 Komori T. Runx2, a multifunctional transcription factor in skeletal development. J Cell Biochem 2002; 87: 1-8
• 8 Komori T. Regulation of skeletal development by the Runx family of transcription factors. J Cell Biochem 2005; 95: 445-453
• 9 Lang TF. The bone-muscle relationship in men and women. J Osteoporos 2011; 2011: 702-735
• 10 Liu W, Toyosawa S, Furuichi T et al. Overexpression of Cbfa1 in osteoblasts inhibits osteoblast maturation and causes osteopenia with multiple fractures. J Cell Biol 2001; 155: 157-166
• 11 Louvet C, Chiffoleau E, Heslan M et al. Identification of a new member of the CD20/FcepsilonRIbeta family overexpressed in tolerated allografts. Am J Transplant 2005; 5: 2143-2153
• 12 Lurton J, Rose TM, Raghu G et al. Isolation of a gene product expressed by a subpopulation of human lung fibroblasts by differential display. Am J Respir Cell Mol Biol 1999; 20: 327-331
• 13 Mackay AM, Beck SC, Murphy JM et al. Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng 1998; 4: 415-428
• 14 Maeda Y, Fujimura L, O-Wang J et al. Role of Clast1 in development of cerebellar granule cells. Brain Res 2006; 1104: 18-26
• 15 Nakaoka D, Sugimoto T, Kaji H et al. Determinants of bone mineral density and spinal fracture risk in postmenopausal Japanese women. Osteoporos Int 2001; 12: 548-554
• 16 Richards RR, McKee MD, Paitich CB et al. A comparison of the effects of skin coverage and muscle flap coverage on the early strength of union at the site of osteotomy after devascularization of a segment of canine tibia. J Bone Joint Surg Am 1991; 73: 1323-1330
• 17 Sarig R, Baruchi Z, Fuchs O et al. Regeneration and transdifferentiation potential of muscle-derived stem cells propagated as myospheres. Stem Cells 2006; 24: 1769-1778
• 18 Shore EM, Xu M, Feldman GJ et al. A recurrent mutation in the BMP type I receptor ACVR1 causes inherited and sporadic fibrodysplasia ossificans progressiva. Nat Genet 2006; 38: 525-527
• 19 Shore EM, Kaplan FS. Insights from a rare genetic disorder of extra-skeletal bone formation, fibrodysplasia ossificans progressiva (FOP). Bone 2008; 43: 427-433
• 20 Tanaka K, Inoue Y, Hendy GN et al. Interaction of Tmem119 and the bone morphogenetic protein pathway in the commitment of myo­blastic into osteoblastic cells. Bone 2012; 51: 158-167
• 21 Tanaka K, Matsumoto E, Higashimaki Y et al. Role of osteoglycin in the linkage between muscle and bone. J Biol Chem 2012; 287: 11616-11628
• 22 Tanaka K, Matsumoto E, Higashimaki Y et al. FAM5C is a soluble osteo­blast differentiation factor linking muscle to bone. Biochem Biophys Res Commun 2012; 418: 134-139
• 23 van Dinther M, Visser N, de Gorter DJ et al. ALK2 R206H mutation linked to fibrodysplasia ossificans progressiva confers constitutive activity to the BMP type I receptor and sensitizes mesenchymal cells to BMP-induced osteoblast differentiation and bone formation. J Bone Miner Res 2010; 25: 1208-1215
• 24 Zhang D, Schwarz EM, Rosier RN et al. ALK2 functions as a BMP type I receptor and induces Indian hedgehog in chondrocytes during skeletal development. J Bone Miner Res 2003; 18: 1593-1604