TGF-β/BMP signaling and other molecular events: regulation of osteoblastogenesis and bone formation

Abstract

Transforming growth factor-beta (TGF-β)/bone morphogenetic protein (BMP) plays a fundamental role in the regulation of bone organogenesis through the activation of receptor serine/threonine kinases. Perturbations of TGF-β/BMP activity are almost invariably linked to a wide variety of clinical outcomes, i.e., skeletal, extra skeletal anomalies, autoimmune, cancer, and cardiovascular diseases. Phosphorylation of TGF-β (I/II) or BMP receptors activates intracellular downstream Smads, the transducer of TGF-β/BMP signals. This signaling is modulated by various factors and pathways, including transcription factor Runx2. The signaling network in skeletal development and bone formation is overwhelmingly complex and highly time and space specific. Additive, positive, negative, or synergistic effects are observed when TGF-β/BMP interacts with the pathways of MAPK, Wnt, Hedgehog (Hh), Notch, Akt/mTOR, and miRNA to regulate the effects of BMP-induced signaling in bone dynamics. Accumulating evidence indicates that Runx2 is the key integrator, whereas Hh is a possible modulator, miRNAs are regulators, and β-catenin is a mediator/regulator within the extensive intracellular network. This review focuses on the activation of BMP signaling and interaction with other regulatory components and pathways highlighting the molecular mechanisms regarding TGF-β/BMP function and regulation that could allow understanding the complexity of bone tissue dynamics.

Bone formation: Still much to learn about its complexities

How bone morphogenetic protein (BMP), discovered in 1965, interacts within a complex network to form bone is not yet clearly understood. Disturbances in BMP activity are linked to a wide range of autoimmune diseases, cancers and skeletal diseases including fibrodysplasia ossificans progressiva. In their review Md. Shaifur Rahman from Tissue Banking and Biomaterial Research Unit, Atomic Energy Research Establishment, Dahka, Bangladesh, and his colleagues focus on the structure of BMP and its receptors, and the pathways it uses to regulate and stimulate bone growth. A large number of factors and targets have now been identified for the BMP pathways. This has led to the discovery of a complex interactive network that researchers are still trying to understand. Knowing more about how the pathways affect the transformation of bone-forming cells at different developmental stages would help to develop potential therapies for bone-related diseases.

References

[1] Beederman M, Lamplot JD, Nan G et al BMP signaling in mesenchymal stem cell differentiation and bone formation. J Biomed Sci Eng, 2013, 6: 32-52

[2] Botchkarev VA. Bone morphogenetic proteins and their antagonists in skin and hair follicle biology. J Invest Dermatol, 2003, 120: 36-47

[3] Lechleider RJ, Ryan JL, Garrett L et al Targeted mutagenesis of Smad1 reveals an essential role in chorioallantoic fusion. Dev Biol, 2001, 240: 157-167

[4] Zhao GQ. Consequences of knocking out BMP signaling in the mouse. Genesis, 2003, 35: 43-56

[5] Urist MR. Bone morphogenetic protein: the molecularization of skeletal system development. J Bone Miner Res, 1997, 12: 343-346

[6] Carreira AC, Lojudice FH, Halcsik E, Navarro RD, Sogayar MC, Granjeiro JM. Bone morphogenetic proteins: facts, challenges, and future perspectives. J Dent Res, 2014, 93: 335-345

[7] Feng XH, Derynck R. Specificity and versatility in TGF-β signaling through Smads. Annu Rev Cell Dev Biol, 2005, 21: 659-693

[8] Balemans W, Hul WV. Extracellular regulation of BMP signaling in vertebrates: a cocktail of modulators. Dev Biol, 2002, 250: 231-250

[9] Moustakas A, Heldin CH. The regulation of TGF-β signal transduction. Development, 2009, 136: 3699-3714

[10] Schmierer B, Hill CS. TGF-β-SMAD signal transduction: molecular specificity and functional flexibility. Nat Rev Mol Cell Biol, 2007, 8: 970-982

[11] Guo L, Zhao RC, Wu Y. The role of microRNAs in self-renewal and differentiation of mesenchymal stem cells. Exp Hematol, 2011, 39: 608-616

[12] Bessa PC, Casal M, Reis RL. Bone morphogenetic proteins in tissue engineering: the road from the laboratory to the clinic, part I (basic concepts). J Tissue Eng Regen Med, 2008, 2: 1-13

[13] Pan A, Chang L, Nguyen A, James AW. A review of hedgehog signaling in cranial bone development. Front Physiol, 2013, 4: 61

[14] Horikiri Y, Shimo T, Kurio N et al Sonic hedgehog regulates osteoblast function by focal adhesion kinase signaling in the process of fracture healing. PLoS ONE, 2013, 8: e76785

[15] Reichert JC, Schmalzl J, Prager P et al Synergistic effect of Indian hedgehog and bone morphogenetic protein-2 gene transfer to increase the osteogenic potential of human mesenchymal stem cells. Stem Cell Res Ther, 2013, 4: 105

[16] Kim JH, Liu X, Wang J et al Wnt signaling in bone formation and its therapeutic potential for bone diseases. Ther Adv Musculoskelet Dis, 2013, 5: 13-31

[17] Issack PS, Helfet DL, Lane JM. Role of Wnt signaling in bone remodeling and repair. HSS J, 2008, 4: 66-70

[18] Terada K, Misao S, Katase N, Nishimatsu S, Nohno T. Interaction of Wnt Signaling with BMP/Smad Signaling during the Transition from Cell Proliferation to Myogenic Differentiation in Mouse Myoblast-Derived Cells. Int J Cell Biol, 2013, 2013: 616294

[19] Lin GL, Hankenson KD. Integration of BMP, Wnt, and notch signaling pathways in osteoblast differentiation. J Cell Biochem, 2011, 112: 3491-3501

[20] Balboni AL, Hutchinson JA, DeCastro AJ et al ΔNp63α mediated activation of bone morphogenetic protein signaling governs stem cell activity and plasticity in normal and malignant mammary epithelial cells. Cancer Res, 2013, 73: 1020-1030

[21] Cheng H, Jiang W, Phillips FM et al Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs). J Bone Joint Surg Am, 2003, 85-A: 1544-1552

[22] Kang Q, Sun MH, Cheng H et al Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery. Gene Therapy, 2004, 11: 1312-1320

[23] Luther G, Wagner ER, Zhu G et al BMP-9 induced osteogenic differentiation of mesenchymal stem cells: molecular mechanism and therapeutic potential. Curr Gene Ther, 2011, 11: 229-240

[24] Lamplot JD, Qin J, Nan G et al BMP9 signaling in stem cell differentiation and osteogenesis. Am J Stem Cells, 2013, 2: 1-21

[25] Rivera JC, Strohbach CA, Wenke JC, Rathbone CR. Beyond osteogenesis: an in vitro comparison of the potentials of six bone morphogenetic proteins. Front Pharmacol, 2013, 4: 125

[26] Callis TE, Cao D, Wang DZ. Bone morphogenetic protein signaling modulates myocardin transactivation of cardiac genes. Circ Res, 2005, 97: 992-1000

[27] Samartzis D, Khanna N, Shen FH, An HS. Update on bone morphogenetic proteins and their application in spine surgery. J Am Coll Surg, 2005, 200: 236-248

[28] Gamer LW, Cox K, Carlo JM, Rosen V. Overexpression of BMP3 in the developing skeleton alters endochondral bone formation resulting in spontaneous rib fractures. Dev Dyn, 2009, 238: 2374-2381

[29] Knöchel S, Dillinger K, Köster M, Knöchel W. Structure and expression of Xenopus tropicalis BMP-2 and BMP-4 genes. Mech Dev, 2001, 109: 79-82

[30] Zuzarte-Luís V, Montero JA, Rodriguez-León J, Merino R, Rodríguez-Rey JC, Hurlé JM. A new role for BMP5 during limb development acting through the synergic activation of Smad and MAPK pathways. Dev Biol, 2004, 272: 39-52

[31] Paralkar VM, Grgurevic L, Simic P et al A novel role of bone morphogenetic protein-6 (BMP-6) as an endocrine regulator of bone and glucose homeostasis. Bone, 2010, 47: S57

[32] Tsumaki N, Yoshikawa H. The role of bone morphogenetic proteins in endochondral bone formation. Cytokine Growth Factor Rev, 2005, 16: 279-285

[33] Lopez-Coviella I, Mellott TM, Kovacheva VP et al Developmental pattern of expression of BMP receptors and Smads and activation of Smad1 and Smad5 by BMP9 in mouse basal forebrain. Brain Res, 2006, 1088: 49-56

[34] Andersson O, Reissmann E, Ibanez CF. Growth differentiation factor 11 signals through the transforming growth factor-beta receptor ALK5 to regionalize the anterior–posterior axis. EMBO Rep, 2006, 7: 831-837

[35] Shen B, Bhargav D, Wei A et al BMP-13 emerges as a potential inhibitor of bone formation. Int J Biol Sci, 2009, 5: 192-200

[36] Di Pasquale E, Beck-Peccoz P, Persani L. Hypergonadotropic ovarian failure associated with an inherited mutation of human bone morphogenetic protein-15 (BMP15) gene. Am J Hum Genet, 2004, 75: 106-111

[37] Nishimoto Y, Morisaki H, Yamada O, Ichinose Y, Suzuki N. Japanese case of hereditary hemorrhagic telangiectasia type 2 with a novel mutation, c.154A>C (p.Thr52Pro), in the ALK1/ACVRL1 gene. Neurol Clin Neurosci, 2014, 2: 126-128

[38] Fukuda T, Kohda M, Kanomata K et al Constitutively activated ALK2 and increased SMAD1/5 cooperatively induce bone morphogenetic protein signaling in fibrodysplasia ossificans progressive. J Biol Chem, 2009, 284: 7149-7156

[39] Howe JR, Bair JL, Sayed MG et al Germ line mutations of the gene encoding bone morphogenetic protein receptor 1A in juvenile polyposis. Nat Genet, 2001, 28: 184-187

[40] Chen G, Deng C, Li YP. TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci, 2012, 8: 272-288

[41] Fessel JP, Loyd JE, Austin ED. The genetics of pulmonary arterial hypertension in the post-BMPR2 era. Pulm Circ, 2011, 1: 305-319

[42] Pearsall RS, Canalis E, Cornwall-Brady M et al A soluble activin type IIA receptor induces bone formation and improves skeletal integrity. Proc Natl Acad Sci U S A, 2008, 105: 7082-7087

[43] Goncalves MD . The role of the activin receptor type IIb (ActRIIB) in glucose and energy homeostasis. Dissertations available from ProQuest 2010; Paper AAI3447175. http://repository.upenn.edu/dissertations/AAI3447175

[44] Matsunobu T, Torigoe K, Ishikawa M et al Critical roles of the TGF-β type I receptor ALK5 in perichondrial formation and function, cartilage integrity, and osteoblast differentiation during growth plate development. Dev Biol, 2009, 332: 325-338

[45] Miyazono K, Kamiya Y, Morikawa M. Bone morphogenetic protein receptors and signal transduction. J Biochem, 2010, 147: 35-51

[46] Wang T, Xu Z. miR-27 promotes osteoblast differentiation by modulating Wnt signaling. Biochem Biophys Res Commun, 2010, 402: 186-189

[47] Massagué J, Seoane J, Wotton D. Smad transcription factors. Genes Dev, 2005, 19: 2783-2810

[48] Estrada KD, Retting KN, Chin AM, Lyons KM. Smad6 is essential to limit BMP signaling during cartilage development. J Bone Miner Res, 2011, 26: 2498-2510

[49] Song Q, Zhu B, Hu W et al A common SMAD7 variant is associated with risk of colorectal cancer: evidence from a case-control study and a meta-analysis. PLoS ONE, 2012, 7: e33318

[50] Okuda T, van Deursen J, Hiebert SW, Grosveld G, Downing JR. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell, 1996, 84: 321-330

[51] Wu CJ, Lu HK. Smad signal pathway in BMP-2-induced osteogenesis a mini review. J Dent Sci, 2008, 3: 13-21

[52] Inoue K, Ozaki S, Shiga T et al Runx3 controls the axonal projection of proprioceptive dorsal root ganglion neurons. Nat Neurosci, 2002, 5: 946-954

[53] Constam DB, Robertson EJ. Regulation of bone morphogenetic protein activity by prodomains and proprotein convertases. J Cell Biol, 1999, 144: 139-149

[54] Szpalski M, Gunzburg R. Recombinant human bone morphogenetic protein-2: a novel osteoinductive alternative to autogenous bone graft? Acta Orthop Belg, 2005, 71: 133-148

[55] Arrabal PM, Visser R, Santos-Ruiz L, Becerra J, Cifuentes M. Osteogenic molecules for clinical applications: improving the BMP-collagen system. Biol Res, 2013, 46: 421-429

[56] Granjeiro JM, Oliveira RC, Bustos-Valenzuela JC, Sogayar MC, Taga R. Bone morphogenetic proteins: from structure to clinical use. Braz J Med Biol Res, 2005, 38: 1463-1473

[57] Rao SM, Ugale GM, Warad SB. Bone morphogenetic proteins: periodontal regeneration. N Am J Med Sci, 2013, 5: 161-168

[58] Muenster U, Korupolu R, Rastogi R, Read J, Fischer WH. Antagonism of activin by activin chimeras. Vitam Horm, 2011, 85: 105-128

[59] Samad TA, Rebbapragada A, Bell E et al DRAGON, a bone morphogenetic protein co-receptor. J Biol Chem, 2005, 280: 14122-14129

[60] Bonor J, Adams EL, Bragdon B, Moseychuk O, Czymmek KJ, Nohe A. Initiation of BMP2 signaling in domains on the plasma membrane. J Cell Physiol, 2012, 227: 2880-2888

[61] Allendorph GP, , Vale WW, Choe S. Structure of the ternary signaling complex of a TGF-beta superfamily member. Proc Natl Acad Sci U S A, 2006, 103: 7643-7648

[62] Yu PB, Beppu H, Kawai N, Li E, Bloch KD. Bone morphogenetic protein (BMP) type II receptor deletion reveals BMP ligand-specific gain of signaling in pulmonary artery smooth muscle cells. J Biol Chem, 2005, 280: 24443-24450

[63] Miyazono K, Maeda S, Imamur T. BMP receptor signaling: transcriptional targets, regulation of signals, and signaling cross-talk. Cytokine Growth Factor Rev, 2005, 16: 251-263

[64] Shore EM, Xu M, Feldman GJ et al A recurrent mutation in the BMP type I receptor ACVR1 causes inherited and sporadic fibrodysplasia ossificans progressive. Nat Genet, 2006, 38: 525-527

[65] Morovvati Z, SMorovvati S, Alishiri GA, Moosavi SH, Ranjbar R, Moghaddam YB. Mutation detection in activin A receptor, Type I (ACVR1) gene in fibrodysplasia ossificans progressiva in an Iranian family. Cell J, 2014, 16: 91-94

[66] Chalmers J, Gray DH, Rush J. Observations on the induction of bone in soft tissues. J Bone Joint Surg Br, 1975, 57: 36-45

[67] Kaplan FS, Le Merrer M, Glaser DL et al Fibrodysplasia ossificans progressiva. Best Pract Res Clin Rheumatol, 2008, 22: 191-205

[68] Kaplan FS, Groppe J, Shore EM. When one skeleton is enough: approaches and strategies for the treatment of fibrodysplasia ossificans progressiva (FOP). Drug Discov Today Ther Strateg, 2008, 5: 255-262

[69] Yu PB, Deng DY, Lai CS et al BMP type I receptor inhibition reduces heterotopic ossification. Nat Med, 2008, 14: 1363-1369

[70] Gregson CL, Hollingworth P, Williams M et al A novel ACVR1 mutation in the glycine/serine-rich domain found in the most benign case of a fibrodysplasia ossificans progressiva variant reported to date. Bone, 2011, 48: 654-658

[71] Kaplan FS, Xu M, Seemann P et al Classic and atypical fibrodysplasia ossificans progressiva (FOP) phenotypes are caused by mutations in the bone morphogenetic protein (BMP) type I receptor ACVR1. Hum Mutat, 2009, 30: 379-390

[72] Petrie KA, Lee WH, Bullock AN et al Novel mutations in ACVR1 result in atypical features in two fibrodysplasia ossificans progressiva patients. PLoS ONE, 2009, 4: e5005

[73] Ratbi I, Borcciadi R, Regragui A, Ravazzolo R, Sefiani A. Rarely occurring mutation of ACVR1 gene in Moroccan patient with fibrodysplasia ossificans progressiva. Clin Rheumatol, 2010, 29: 119-121

[74] Carvalho DR, Navarro MM, Martins BJ et al Mutational screening of ACVR1 gene in Brazilian fibrodysplasia ossificans progressiva patients. Clin Genet, 2010, 77: 171-176

[75] Furuya H, Ikezoe K, Wang L et al A unique case of fibrodysplasia ossificans progressiva with an ACVR1 mutation, G356D, other than the common mutation (R206H). Am J Med Genet A, 2008, 146A: 459-463

[76] Bocciardi R, Bordo D, Di Duca M, Di Rocco M, Ravazzolo R. Mutational analysis of the ACVR1 gene in Italian patients affected with fibrodysplasia ossificans progressiva: confirmations and advancements. Eur J Hum Genet, 2009, 17: 311-318

[77] Chaikuad A, Alfano I, Kerr G et al Structure of the bone morphogenetic protein receptor ALK2 and implications for fibrodysplasia ossificans progressiva. J Biol Chem, 2012, 287: 36990-36998

[78] Bagarova J, Vonner AJ, Armstrong KA et al Constitutively active ALK2 receptor mutants require Type II receptor cooperation. Mol Cell Biol, 2013, 33: 2413-2424

[79] Lowery JW, Rosen V. Allele-specific RNA interference in FOP silencing the FOP gene. Gene Therapy, 2012, 19: 701-702

[80] Wu JW, Hu M, Chai J et al Crystal structure of a phosphorylated Smad2. Recognition of phosphoserine by the MH2 domain and insights on Smad function in TGF-beta signaling. Mol Cell, 2001, 8: 1277-1289

[81] Makkar P, Metpally RP, Sangadala S, Reddy BV. Modeling and analysis of MH1 domain of Smads and their interaction with promoter DNA sequence motif. J Mol Graph Model, 2009, 27: 803-812

[82] ten Dijke P, Miyazono K, Heldin CH. Signaling inputs converge on nuclear effectors in TGF-beta signaling. Trends Biochem Sci, 2000, 25: 64-70

[83] Zhang J, Fei T, Li Z, Zhu G, Wang L, Chen Y. BMP induces cochlin expression to facilitate self-renewal and suppress neural differentiation of mouse embryonic stem cells. J Biol Chem, 2013, 288: 8053-8060

[84] Lo RS, Chen YG, Shi Y, Pavletich NP, Massague J. The L3 loop: a structural motif determining specific interactions between SMAD proteins and TGF-beta receptors. Embo J, 1998, 17: 996-1005

[85] Liu F, Hata A, Baker JC et al A human Mad protein acting as a BMP-regulated transcriptional activator. Nature, 1996, 381: 620-623

[86] Kusanagi K, Inoue H, Ishidou Y, Mishima HK, Kawabata M, Miyazono K. Characterization of a bone morphogenetic protein-responsive Smad-binding element. Mol Biol Cell, 2000, 11: 555-565

[87] Mukhopadhyay P, Webb CL, Warner DR, Greene RM, Pisano MM. BMP signaling dynamics in embryonic orofacial tissue. J Cell Physiol, 2008, 216: 771-779

[88] Morikawa M, Koinuma D, Tsutsumi S et al ChIP-seq reveals cell type-specific binding patterns of BMP-specific Smads and a novel binding motif. Nucleic Acids Res, 2011, 39: 8712-8727

[89] Ross S, Cheung E, Petrakis TG, Howell M, Kraus WL, Hill CS. Smads orchestrate specific histone modifications and chromatin remodeling to activate transcription. EMBO J, 2006, 25: 4490-4502

[90] James AW. Review of Signaling Pathways Governing MSC Osteogenic and Adipogenic Differentiation. Scientifica (Cairo), 2013, 2013: 684736

[91] Zhao M, Qiao M, Harris SE, Chen D, Oyajobi BO, Mundy GR. The zinc finger transcription factor Gli2 mediates bone morphogenetic protein 2 expression in osteoblasts in response to hedgehog signaling. Mol Cell Biol, 2006, 26: 6197-6208

[92] Edwards PC, Ruggiero S, Fantasia J et al Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. Gene Therapy, 2005, 12: 75-86

[93] Karamboulas K, Dranse HJ, Underhill TM. Regulation of BMP-dependent chondrogenesis in early limb mesenchyme by TGF beta signals. J Cell Sci, 2010, 123: 2068-2076

[94] Yuasa T, Kataoka H, Kinto N et al Sonic hedgehog is involved in osteoblast differentiation by cooperating with BMP-2. J Cell Physiol, 2002, 193: 225-232

[95] Deschaseaux F, Sensebe L, Heymann D. Mechanisms of bone repair and regeneration. Trends Mol Med, 2009, 15: 417-429

[96] Zhang J, Tan X, Li W et al Smad4 is required for the normal organization of the cartilage growth plate. Dev Biol, 2005, 284: 311-322

[97] Kalderon D. Similarities between the Hedgehog and Wnt signaling pathways. Trends Cell Biol, 2002, 12: 523-531

[98] Taipale J, Beachy PA. The Hedgehog and Wnt signaling pathways in cancer. Nature, 2001, 411: 349-354

[99] Komiya Y, Habas R. Wnt signal transduction pathways. Organogenesis, 2008, 4: 68-75

[100] Cadigan KM, Liu YI. Wnt signaling: complexity at the surface. J Cell Sci, 2006, 119: 395-402

[101] Fuentealba LC, Eivers E, Ikeda A et al Integrating patterning signals: Wnt/GSK3 regulates the duration of the BMP/Smad1 signal. Cell, 2007, 131: 980-993

[102] Krishnan V, Bryant HU, MacDougald OA. Regulation of bone mass by Wnt signaling. J Clin Invest, 2006, 116: 1202-1209

[103] Okamoto M, Udagawa N, Uehara S et al Noncanonical Wnt5a enhances Wnt/b-catenin signaling during Osteoblastogenesis. Sci Rep, 2014, 4: 4493

[104] Jin EJ, Lee SY, Choi YA, Jung JC, Bang OS, Kang SS. BMP-2 enhanced chondrogenesis involves p38 MAPK-mediated downregulation of Wnt-7a pathway. Mol Cells, 2006, 22: 353-359

[105] Papathanasiou I, Malizos KN, Tsezou A. Bone morphogenetic protein-2-induced Wnt/β-catenin signaling pathway activation through enhanced low-density-lipoprotein receptor-related protein 5 catabolic activity contributes to hypertrophy in osteoarthritic chondrocytes. Arthritis Res Ther, 2012, 14: R82

[106] Soltanoff CS, Chen W, Yang S, Li YP. Signaling networks that control the lineage commitment and differentiation of bone cells. Crit Rev Eukaryot Gene Expr, 2009, 19: 1-46

[107] McCarthy TL, Centrella M. Novel links among Wnt and TGF-β signaling and Runx2. Mol Endocrinol, 2010, 24: 587-597

[108] Hill TP, Später D, Taketo MM, Birchmeier W, Hartmann C. Canonical Wnt/β-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev Cell, 2005, 8: 727-738

[109] Schramke V, Hairpin AR. RNAs and retrotransposon LTRs effect RNAi and chromatin-based gene silencing. Science, 2003, 301: 1069-1074

[110] Lian JB, Stein GS, van Wijnen AJ et al MicroRNA control of bone formation and homeostasis. Nat Rev Endocrinol, 2012, 8: 212-227

[111] He X, Eberhart JK, Postlethwait JH. MicroRNAs and micromanaging the skeletonin disease, development and evolution. J Cell Mol Med, 2009, 13: 606-618

[112] Jafferji M . Regulation of osteoblast differentiation by microRNAs. Umass Med School 2009; MQP-BIODSA-7214.

[113] Li Z, Hassan MQ, Jafferji M et al Biological functions of miR-29b contribute to positive regulation of osteoblast differentiation. J Biol Chem, 2009, 284: 15676-15684

[114] Hu R, Liu W, Li H et al A Runx2/miR-3960/miR-2861 regulatory feedback loop during mouse osteoblast differentiation. J Biol Chem, 2011, 286: 12328-12339

[115] Kapinas K, Kessler C, Ricks T, Gronowicz G, Delany AM. miR-29 modulates Wnt signaling in human osteoblasts through a positive feedback loop. J Biol Chem, 2010, 285: 25221-25231

[116] Zhang J, Tu Q, Bonewald LF et al Effects of miR-335-5p in modulating osteogenic differentiation by specifically down regulating Wnt antagonist DKK1. J Bone Miner Res, 2011, 26: 1953-1963

[117] Itoh T, Takeda S, Akao Y. MicroRNA-208 modulates BMP-2-stimulated mouse preosteoblast differentiation by directly targeting V-ets erythroblastosis virus E26 oncogene Homolog 1. J Biol Chem, 2010, 285: 27745-27752

[118] Eskildsen T, Taipaleenmäki H, Stenvang J et al MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo. Proc Natl Acad Sci, 2011, 108: 6139-6144

[119] Liu Y, Liu W, Hu C et al MiR-17 modulates osteogenic differentiation through a coherent feed-forward loop in mesenchymal stem cells isolated from periodontal ligaments of patients with periodontitis. Stem Cells, 2011, 29: 1804-1816

[120] Mizoguchi F, Murakami Y, Saito T, Miyasaka N, Kohsaka H. MiR-31 controls osteoclast formation and bone resorption by targeting RhoA. Arthritis Res Ther, 2013, 15: R102

[121] Bae Y, Yang T, Zeng HC et al miRNA-34c regulates Notch signaling during bone development. Hum Mol Genet, 2012, 21: 2991-3000

[122] Mizuno Y, Tokuzawa Y, Ninomiya Y et al miR-210 promotes osteoblastic differentiation through inhibition of AcvR1b. FEBS Lett, 2009, 583: 2263-2268

[123] Mei Y, Bian C, Li J et al miR-21 modulates the ERK-MAPK signaling pathway by regulating SPRY2 expression during human mesenchymal stem cell differentiation. J Cell Biochem, 2013, 114: 1374-1384

[124] Goff LA, Boucher S, Ricupero CL et al Differentiating human multipotent mesenchymal stromal cells regulate microRNAs: prediction of microRNA regulation by PDGF during osteogenesis. Exp Hematol, 2008, 36: 1354-1369

[125] Kim YJ, Bae SW, Yu SS, Bae YC, Jung JS. miR-196a regulates proliferation and osteogenic differentiation in mesenchymal stem cells derived from human adipose tissue. J Bone Miner Res, 2009, 24: 816-825

[126] Schoolmeesters A, Eklund T, Leake D et al Functional profiling reveals critical role for miRNA in differentiation of human mesenchymal stem cells. PLoS ONE, 2009, 4: e5605

[127] Kahai S, Lee SC, Lee DY et al MicroRNA miR-378 regulates nephronectin expression modulating osteoblast differentiation by targeting GalNT-7. PLoS ONE, 2009, 4: e7535

[128] Oskowitz A, Lu J, Penfornis P et al Human multipotent stromal cells from bone marrow and microRNA: regulation of differentiation and leukemia inhibitory factor expression. Proc Natl Acad Sci U S A, 2008, 105: 18372-18377

[129] Kuo PL, Liao SH, Hung JY, Huang MS, Hsu YL. MicroRNA-33a functions as a bone metastasis suppressor in lung cancer by targeting parathyroid hormone related protein. Biochim Biophys Acta, 2013, 1830: 3756-3766

[130] Pollari S, Leivonen SK, Perälä M, Fey V, Käkönen SM, Kallioniemi O. Identification of microRNAs inhibiting TGF-β-induced IL-11 production in bone metastatic breast cancer cells. PLoS ONE, 2012, 7: e37361

[131] Zhao FL, Hu GD, Wang XF, Zhang XH, Zhang YK, Yu ZS. Serum overexpression of microRNA-10b in patients with bone metastatic primary breast cancer. J Int Med Res, 2012, 40: 859-866

[132] Tie J, Pan Y, Zhao L et al MiR-218 inhibits invasion and metastasis of gastric cancer by targeting the Robo1 receptor. PLoS Genet, 2010, 6: e1000879

[133] Octacílio-Silva S, Marques MM, Evangelista AF et al Defining mRNA Targets of MicroRNAs during Osteoblastic Differentiation of Human Mesenchymal Stem Cells by Microarray Transcriptional Interaction Networks, 2010 Resumos do 56° Congresso Brasileiro de Genética, Guarujá-SP, Brasil

[134] Valencia K, Martín-Fernαndez M, Zandueta C et al miR-326 associates with biochemical markers of bone turnover in lung cancer bone metastasis. Bone, 2013, 52: 532-539

[135] Hassan MQ, Gordon JA, Beloti MM et al A network connecting Runx2, SATB2, and the miR-23a∼27a∼24–2 cluster regulates the osteoblast differentiation program. Proc Natl Acad Sci U S A, 2010, 107: 19879-19884

[136] Zeng Y, Qu X, Li H et al MicroRNA-100 regulates osteogenic differentiation of human adipose-derived mesenchymal stem cells by targeting BMPR2. FEBS Lett, 2012, 586: 2375-2381

[137] Vimalraj S, Selvamurugan N. MicroRNAs: Synthesis, gene regulation and osteoblast differentiation. Curr Issues Mol Biol, 2012, 15: 7-18

[138] Inosea H, Ochi H, Kimura A et al A microRNA regulatory mechanism of osteoblast differentiation. Proc Natl Acad Sci U S A, 2009, 106: 20794-20799

[139] Huang G, Nishimoto K, Zhou Z, Hughes D, Kleinerman ES. miR-20a encoded by the miR-17-92 cluster increases the metastatic potential of osteosarcoma cells by regulating Fas expression. Cancer Res, 2012, 72: 908-916

[140] Tomé M, López-Romero P, Albo C et al miR-335 orchestrates cell proliferation, migration and differentiation in human mesenchymal stem cells. Cell Death Diff, 2011, 18: 985-995

[141] Yin Q, Wang X, Fewell C et al MicroRNA miR-155 inhibits bone morphogenetic protein (BMP) signaling and BMP-mediated Epstein–Barr virus reactivation. J Virol, 2010, 84: 6318-6327

[142] Mizuno Y, Yagi K, Tokuzawa Y et al miR-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation. Biochem Biophys Res Commun, 2008, 368: 267-272

[143] Chang L, Karin M. Mammalian MAP kinase signalling cascades. Nature, 2001, 410: 37-40

[144] Kretzschmar M, Doody J, Timokhina I, Massague J. A mechanism of repression of TGF beta/Smad signaling by oncogenic Ras. Genes Dev, 1999, 13: 804-816

[145] Sapkota G, Alarcon C, Spagnoli FM, Brivanlou AH, Massague J. Balancing BMP signaling through integrated inputs into the Smad1 linker. Mol Cell, 2007, 25: 441-454

[146] Matsuura I, Wang G, He D, Liu F. Identification and characterization of ERK MAP kinase phosphorylation sites in Smad3. Biochemistry, 2005, 44: 12546-12553

[147] Herpin A, Cunningham C. Cross-talk between the bone morphogenetic protein pathway and other major signaling pathways results in tightly regulated cell-specific outcomes. FEBS J, 2007, 274: 2977-2985

[148] Leung-Hagesteijn C, Hu MC, Mahendra AS et al Integrin-linked kinase mediates bone morphogenetic protein 7-dependent renal epithelial cell morphogenesis. Mol Cell Biol, 2005, 25: 3648-3657

[149] Eivers E, Fuentealba LC, De Robertis EM. Integrating positional information at the level of Smad1/5/8. Curr Opin Genet Dev, 2008, 18: 304-310

[150] Ghosh-Choudhury N, Abboud SL, Nishimura R, Celeste A, Mahimainathan L, Choudhury GG. Requirement of BMP-2-induced phosphatidylinositol 3-kinase and Akt serine/threonine kinase in osteoblast differentiation and Smad-dependent BMP-2 gene transcription. J Biol Chem, 2002, 277: 33361-33368

[151] Moustakas A, Heldin CH. Non-Smad TGF-beta signals. J Cell Sci, 2005, 118: 3573-3584

[152] Engin F, Lee B. NOTCHing the bone: insights into multi-functionality. Bone, 2010, 46: 274-280

[153] Tezuka K, Yasuda M, Watanabe N et al Stimulation of osteoblastic cell differentiation by Notch. J Bone Miner Res, 2002, 17: 231-239

[154] Turnpenny PD, Alman B, Cornier AS et al Abnormal vertebral segmentation and the notch signaling pathway in man. Dev Dyn, 2007, 236: 1456-1474

[155] Ramasamy SK, Kusumbe AP, Wang L, Adams RH. Endothelial Notch activity promotes angiogenesis and osteogenesis in bone. Nature, 2014, 507: 376-380

[156] Luppen CA, Chandler RL, Noh T, Mortlock DP, Frenkel B. BMP-2 vs. BMP-4 expression and activity in glucocorticoid arrested MC3T3-E1 osteoblasts: Smad signaling, not alkaline phosphatase activity, predicts rescue of mineralization. Growth Factors, 2008, 26: 226-237

[157] Sheng N, Xie Z, Wang C et al Retinoic acid regulates bone morphogenic protein signal duration by promoting the degradation of phosphorylated Smad1. Proc Natl Acad Sci U S A, 2010, 107: 18886-18891

[158] McKeown SJ, Wallace AS, Anderson RB. Expression and function of cell adhesion molecules during neural crest migration. Dev Biol, 2013, 373: 244-257

[159] Zhou J, Lee PL, Lee CI et al BMP receptor-integrin interaction mediates responses of vascular endothelial Smad1/5 and proliferation to disturbed flow. J Thromb Haemost, 2013, 11: 741-755

[160] Lamouille S, Connolly E, Smyth JW, Akhurst RJ, Derynck R. TGF-b-induced activation of mTOR complex 2 drives epithelial–mesenchymal transition and cell invasion. J Cell Sci, 2012, 125: 1259-1273

[161] Xu J, Lamouille S, Derynck R. TGF-b induced epithelial to mesenchymal transition. Cell Res, 2009, 19: 156-172

[162] Song K, Wang H, Krebs TL, Danielpour D. Novel roles of Akt and mTOR in suppressing TGF-b/ALK5-mediated Smad3 activation. EMBO J, 2006, 25: 58-69

[163] Dreieicher E, Beck KF, Lazaroski S et al Nitric oxide inhibits glomerular TGF-b signaling via SMOC-1. J Am Soc Nephrol, 2009, 20: 1963-1974

[164] Saura M, Zaragoza C, Herranz B et al Nitric oxide regulates transforming growth factor-β signaling in endothelial Cells. Circ Res, 2005, 97: 1115-1123

[165] Saito T, Fukai A, Mabuchi A et al Transcriptional regulation of endochondral ossification by HIF-2alpha during skeletal growth and osteoarthritis development. Nat Med, 2010, 16: 678-686

[166] Mariani E, Pulsatelli L, Facchini A. Signaling pathways in cartilage repair. Int J Mol Sci, 2014, 15: 8667-8698

[167] Sagar DR, Ashraf S, Xu L et al Osteoprotegerin reduces the development of pain behaviour and joint pathology in a model of osteoarthritis. Ann Rheum Dis, 2014, 73: 1558-1565

[168] Nakashima T, Takayanagi H. The dynamic interplay between osteoclasts and the immune system. Arch Biochem Biophys, 2008, 473: 166-171

[169] Jimi E, Hirata S, Shin M, Yamazaki M, Fukushima H. Molecular mechanisms of BMP-induced bone formation: cross-talk between BMP and NF-κB signaling pathways in osteoblastogenesis. Jpn Dent Sci Rev, 2010, 46: 33-42

[170] Eliseev RA, Schwarz EM, Zuscik MJ, O'Keefe RJ, Drissi H, Rosier RN. Smad7 mediates inhibition of Saos2 osteosarcoma cell differentiation by NF-κB. Exp Cell Res, 2006, 312: 40-50

[171] Li Y, Li A, Strait K, Zhang H, Nanes MS, Weitzmann MN. Endogenous TNFa lowers maximum peak bone mass and inhibits osteoblastic Smad activation through NF-κB. J Bone Miner Res, 2007, 22: 646-655

[172] Kramer I, Halleux C, Feng JQ, Bonewald LF, Kneissel M. Osteocyte Wnt/β-catenin signaling is required for normal bone homeostasis. Mol Cell Biol, 2010, 30: 3071-3085

[173] Zhang Y, Khan D, Delling J, Tobiasch E. Mechanisms underlying the osteo and adipo differentiation of human mesenchymal stem cells. Sci World J, 2012, 2012: 793823

[174] Das S, Samant RS, Shevde LA. The hedgehog pathway conditions the bone microenvironment for osteolytic metastasis of breast cancer. Int J Breast Cancer, 2012, 2012: 298623

[175] Hughes FJ, Turner W, G Belibasakis G, Martuscelli G. Effects of growth factors and cytokines on osteoblast differentiation. Periodontol 2000, 2006, 41: 48-72

[176] Fuller K, Lawrence KM, Ross JL et al Cathepsin K inhibitors prevent matrix-derived growth factor degradation by human osteoclasts. Bone, 2008, 42: 200-211

[177] Xie H, Cui Z, Wang L et al PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis. Nat Med, 2014, 20: 1270-1278

[178] Gerstenfeld L, Edgar CM, Kakar S, Jacobsen KA, Einhorn TA. Osteogenic growth factors and cytokines and their role in bone repair engineering of functional skeletal tissues. Topics Bone Biol, 2007, 3: 17-45

[179] Li X, Cao X. BMP signaling and skeletogenesis. Ann N Y Acad Sci, 2006, 1068: 26-40

[180] Shimizu T, Tanaka T, Iso T et al Notch signaling pathway enhances bone morphogenetic protein 2 (BMP2) responsiveness of Msx2 gene to induce osteogenic differentiation and mineralization of vascular smooth muscle cells. J Biol Chem, 2011, 286: 19138-19148

[181] Sowa H, Kaji H, Hendy GN et al Menin is required for bone morphogenetic protein 2- and transforming growth factor beta-regulated osteoblastic differentiation through interaction with Smads and Runx2. J Biol Chem, 2004, 279: 40267-40275

[182] Li X, Cao X. BMP signaling and HOX transcription factors in limb development. Front Biosci, 2003, 8: s805-s812