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Cited by 22Bone Tissue Engineering MaterialsBone Metabolism and DiseasesTGF-β signaling in diseases

Small molecule inhibition of transforming growth factor beta signaling enables the endogenous regenerative potential of the mammalian calvarium

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Senarath-Yapa, Kshemendra, Li, Shuli, Walmsley, Graham G., Zielins, Elizabeth, Paik, Kevin, Britto, Jonathan A., Grigoriadis, Agamemnon E., Wan, Derrick C. +3 more·2016·Tissue Engineering Part A
DOI 10.1089/ten.tea.2015.0527PMC PMC4876548
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Abstract

Current approaches for the treatment of skeletal defects are suboptimal, principally because the ability of bone to repair and regenerate is poor. Although the promise of effective cellular therapies for skeletal repair is encouraging, these approaches are limited by the risks of infection, cellular contamination, and tumorigenicity. Development of a pharmacological approach would therefore help avoid some of these potential risks. This study identifies transforming growth factor beta (TGFβ) signaling as a potential pathway for pharmacological modulation in vivo. We demonstrate that inhibition of TGFβ signaling by the small molecule SB431542 potentiates calvarial skeletal repair through activation of bone morphogenetic protein (BMP) signaling on osteoblasts and dura mater cells participating in healing of calvarial defects. Cells respond to inhibition of TGFβ signaling by producing higher levels of BMP2 that upregulates inhibitory Smad6 expression, thus providing a negative feedback loop to contain excessive BMP signaling. Importantly, study on human osteoblasts indicates that molecular mechanism(s) triggered by SB431542 are conserved. Collectively, these data provide insights into the use of small molecules to modulate key signaling pathways for repairing skeletal defects.

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  • Autologous and allogeneic bone grafts are the most frequently employed clinical techniques for bone repair but have limitations including donor-site defects, immune rejection, and surgical pain.

    “Autologous and allogeneic bone grafts are the most frequently employed clinical techniques [129,130]. However, there are still some limitations to these treatments, such as the risk of creating new defects at the donor site, potential immune rejection, and the pain associated with surgical procedures [131,132].”

    From Mussel-Inspired Injectable Adhesive Hydrogels for Biomedical Applications (2024) · cites this paper as [129]

    The Introduction identifies autologous and allogeneic grafts as current techniques for craniofacial bone reconstruction and describes donor-site morbidity and chronic pain for autologous grafts, plus immunologic rejection for allogeneic bone. It does not say these are the most frequently employed clinical techniques. (LLM, conf=0.86)

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Research works printed but not linked: 55

  1. [1]Amini A.R., Laurencin C.T., and Nukavarapu S.P. Bone tissue engineering: recent advances and challenges. Crit Rev Biomed Eng 40, 363, 2012 [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. [2]Baroli B. From natural bone grafts to tissue engineering therapeutics: brainstorming on pharmaceutical formulative requirements and challenges. J Pharm Sci 98, 1317, 2009 [DOI] [PubMed] [Google Scholar]
  3. [3]Steiner C., Elixhauser A., and Schnaier J. The healthcare cost and utilization project: an overview. Eff Clin Pract 5, 143, 2002 [PubMed] [Google Scholar]
  4. [4]Shenaq S.M. Reconstruction of complex cranial and craniofacial defects utilizing iliac crest-internal oblique microsurgical free flap. Microsurgery 9, 154, 1988 [DOI] [PubMed] [Google Scholar]
  5. [5]Rah D.K. Art of replacing craniofacial bone defects. Yonsei Med J 41, 756, 2000 [DOI] [PubMed] [Google Scholar]
  6. [6]Bruens M.L., Pieterman H., de Wijn J.R., and Vaandrager J.M. Porous polymethylmethacrylate as bone substitute in the craniofacial area. J Craniofac Surg 14, 63, 2003 [DOI] [PubMed] [Google Scholar]
  7. [7]St John T.A., Vaccaro A.R., Sah A.P., Schaefer M., Berta S.C., Albert T., and Hilibrand A. Physical and monetary costs associated with autogenous bone graft harvesting. Am J Orthop (Belle Mead NJ) 32, 18, 2003 [PubMed] [Google Scholar]
  8. [8]Karsenty G. Minireview: transcriptional control of osteoblast differentiation. Endocrinology 142, 2731, 2001 [DOI] [PubMed] [Google Scholar]
  9. [9]Karsenty G. Transcriptional control of skeletogenesis. Annu Rev Genomics Hum Genet 9, 183, 2008 [DOI] [PubMed] [Google Scholar]
  10. [10]Jiang X., Iseki S., Maxson R.E., Sucov H.M., and Morriss-Kay G.M. Tissue origins and interactions in the mammalian skull vault. Dev Biol 241, 106, 2002 [DOI] [PubMed] [Google Scholar]
  11. [11]Quarto N., Wan D.C., Kwan M.D., Panetta N.J., Li S., and Longaker M.T. Origin matters: differences in embryonic tissue origin and Wnt signaling determine the osteogenic potential and healing capacity of frontal and parietal calvarial bones. J Bone Miner Res 25, 1680, 2010 [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. [12]Li S., Quarto N., and Longaker M.T. Activation of FGF signaling mediates proliferative and osteogenic differences between neural crest derived frontal and mesoderm parietal derived bone. PLoS One 5, e14033, 2010 [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. [13]Quarto N., Behr B., Li S., and Longaker M.T. Differential FGF ligands and FGF receptors expression pattern in frontal and parietal calvarial bones. Cells Tissues Organs 190, 158, 2009 [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. [14]Behr B., Panetta N.J., Longaker M.T., and Quarto N. Different endogenous threshold levels of Fibroblast Growth Factor-ligands determine the healing potential of frontal and parietal bones. Bone 47, 281, 2010 [DOI] [PubMed] [Google Scholar]
  15. [15]Senarath-Yapa K., Li S., Meyer N.P., Longaker M.T., and Quarto N. Integration of multiple signaling pathways determines differences in the osteogenic potential and tissue regeneration of neural crest-derived and mesoderm-derived calvarial bones. Int J Mol Sci 14, 5978, 2013 [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. [16]Li S., Meyer N.P., Quarto N., and Longaker M.T. Integration of multiple signaling regulates through apoptosis the differential osteogenic potential of neural crest-derived and mesoderm-derived Osteoblasts. PLoS One 8, e58610, 2013 [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. [17]Inman G.J., Nicolas F.J., Callahan J.F., Harling J.D., Gaster L.M., Reith A.D., Laping N.J., and Hill C.S. SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol 62, 65, 2002 [DOI] [PubMed] [Google Scholar]
  18. [18]Bonewald L.F., and Dallas S.L. Role of active and latent transforming growth factor beta in bone formation. J Cell Biochem 55, 350, 1994 [DOI] [PubMed] [Google Scholar]
  19. [19]Robey P.G., Young M.F., Flanders K.C., Roche N.S., Kondaiah P., Reddi A.H., Termine J.D., Sporn M.B., and Roberts A.B. Osteoblasts synthesize and respond to transforming growth factor-type beta (TGF-beta) in vitro. J Cell Biol 105, 457, 1987 [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. [20]Dallas S.L., Sivakumar P., Jones C.J., Chen Q., Peters D.M., Mosher D.F., Humphries M.J., and Kielty C.M. Fibronectin regulates latent transforming growth factor-beta (TGF beta) by controlling matrix assembly of latent TGF beta-binding protein-1. J Biol Chem 280, 18871, 2005 [DOI] [PubMed] [Google Scholar]
  21. [22]Noda M., and Camilliere J.J. In vivo stimulation of bone formation by transforming growth factor-beta. Endocrinology 124, 2991, 1989 [DOI] [PubMed] [Google Scholar]
  22. [23]Zhang H., Ahmad M., and Gronowicz G. Effects of transforming growth factor-beta 1 (TGF-beta1) on in vitro mineralization of human osteoblasts on implant materials. Biomaterials 24, 2013, 2003 [DOI] [PubMed] [Google Scholar]
  23. [24]Srouji S., Blumenfeld I., Rachmiel A., and Livne E. Bone defect repair in rat tibia by TGF-beta1 and IGF-1 released from hydrogel scaffold. Cell Tissue Bank 5, 223, 2004 [DOI] [PubMed] [Google Scholar]
  24. [25]Ripamonti U., Ferretti C., Teare J., and Blann L. Transforming growth factor-beta isoforms and the induction of bone formation: implications for reconstructive craniofacial surgery. J Craniofac Surg 20, 1544, 2009 [DOI] [PubMed] [Google Scholar]
  25. [26]Tachi K., Takami M., Sato H., Mochizuki A., Zhao B., Miyamoto Y., Tsukasaki H., Inoue T., Shintani S., Koike T., Honda Y., Suzuki O., Baba K., and Kamijo R. Enhancement of bone morphogenetic protein-2-induced ectopic bone formation by transforming growth factor-beta1. Tissue Eng Part A 17, 597, 2011 [DOI] [PubMed] [Google Scholar]
  26. [27]Lind M. Growth factor stimulation of bone healing. Effects on osteoblasts, osteomies, and implants fixation. Acta Orthop Scand Suppl 283, 2, 1998 [PubMed] [Google Scholar]
  27. [28]Alliston T., Choy L., Ducy P., Karsenty G., and Derynck R. TGF-beta-induced repression of CBFA1 by Smad3 decreases cbfa1 and osteocalcin expression and inhibits osteoblast differentiation. EMBO J 20, 2254, 2001 [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. [29]Erlebacher A., and Derynck R. Increased expression of TGF-beta 2 in osteoblasts results in an osteoporosis-like phenotype. J Cell Biol 132, 195, 1996 [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. [30]Heldin C.H., Miyazono K., and ten Dijke P. TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature 390, 465, 1997 [DOI] [PubMed] [Google Scholar]
  30. [31]Derynck R., and Zhang Y.E. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature 425, 577, 2003 [DOI] [PubMed] [Google Scholar]
  31. [32]Shi Y., and Massague J. Mechanisms of TGF-beta signaling from cell membrane to the nucleus. Cell 113, 685, 2003 [DOI] [PubMed] [Google Scholar]
  32. [33]Piek E., Heldin C.H., and Ten Dijke P. Specificity, diversity, and regulation in TGF-beta superfamily signaling. FASEB J 13, 2105, 1999 [PubMed] [Google Scholar]
  33. [34]Wallace E.M., Lyssikatos J.P., Yeh T., Winkler J.D., and Koch K. Progress towards therapeutic small molecule MEK inhibitors for use in cancer therapy. Curr Top Med Chem 5, 215, 2005 [DOI] [PubMed] [Google Scholar]
  34. [35]Blake R.A., Broome M.A., Liu X., Wu J., Gishizky M., Sun L., and Courtneidge S.A. SU6656, a selective src family kinase inhibitor, used to probe growth factor signaling. Mol Cell Biol 20, 9018, 2000 [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. [36]Hong C.C., and Yu P.B. Applications of small molecule BMP inhibitors in physiology and disease. Cytokine Growth Factor Rev 20, 409, 2009 [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. [37]Tanaka H., Shinto O., Yashiro M., Yamazoe S., Iwauchi T., Muguruma K., Kubo N., Ohira M., and Hirakawa K. Transforming growth factor beta signaling inhibitor, SB-431542, induces maturation of dendritic cells and enhances anti-tumor activity. Oncol Rep 24, 1637, 2010 [DOI] [PubMed] [Google Scholar]
  37. [38]Takeuchi K., Abe M., Hiasa M., Oda A., Amou H., Kido S., Harada T., Tanaka O., Miki H., Nakamura S., Nakano A., Kagawa K., Yata K., Ozaki S., and Matsumoto T. Tgf-Beta inhibition restores terminal osteoblast differentiation to suppress myeloma growth. PLoS One 5, e9870, 2010 [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. [39]Waghabi M.C., de Souza E.M., de Oliveira G.M., Keramidas M., Feige J.J., Araujo-Jorge T.C., and Bailly S. Pharmacological inhibition of transforming growth factor beta signaling decreases infection and prevents heart damage in acute Chagas' disease. Antimicrob Agents Chemother 53, 4694, 2009 [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. [40]Cabello-Verrugio C., Cordova G., Vial C., Zuniga L.M., and Brandan E. Connective tissue growth factor induction by lysophosphatidic acid requires transactivation of transforming growth factor type beta receptors and the JNK pathway. Cell Signal 23, 449, 2011 [DOI] [PubMed] [Google Scholar]
  40. [41]Sugiyama K., Ishii G., Ochiai A., and Esumi H. Improvement of the breaking strength of wound by combined treatment with recombinant human G-CSF, recombinant human M-CSF, and a TGF-beta1 receptor kinase inhibitor in rat skin. Cancer Sci 99, 1021, 2008 [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. [42]Nakamura R., Kayamori K., Oue E., Sakamoto K., Harada K., and Yamaguchi A. Transforming growth factor-beta synthesized by stromal cells and cancer cells participates in bone resorption induced by oral squamous cell carcinoma. Biochem Biophys Res Commun 458, 777, 2015 [DOI] [PubMed] [Google Scholar]
  42. [43]Cowan C.M., Shi Y.Y., Aalami O.O., Chou Y.F., Mari C., Thomas R., Quarto N., Contag C.H., Wu B., and Longaker M.T. Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol 22, 560, 2004 [DOI] [PubMed] [Google Scholar]
  43. [44]Quarto N., Wan D.C., and Longaker M.T. Molecular mechanisms of FGF-2 inhibitory activity in the osteogenic context of mouse adipose-derived stem cells (mASCs). Bone 42, 1040, 2008 [DOI] [PubMed] [Google Scholar]
  44. [45]Levi B., Hyun J.S., Nelson E.R., Li S., Montoro D.T., Wan D.C., Jia F.J., Glotzbach J.C., James A.W., Lee M., Huang M., Quarto N., Gurtner G.C., Wu J.C., and Longaker M.T. Nonintegrating knockdown and customized scaffold design enhances human adipose-derived stem cells in skeletal repair. Stem Cells 29, 2018, 2011 [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. [46]Quarto N., Senarath-Yapa K., Renda A., and Longaker M.T. TWIST1 silencing enhances in vitro and in vivo osteogenic differentiation of human adipose-derived stem cells by triggering activation of BMP-ERK/FGF signaling and TAZ upregulation. Stem Cells 33, 833, 2015 [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. [47]Quarto N., Li S., Renda A., and Longaker M.T. Exogenous activation of BMP-2 signaling overcomes TGFbeta-mediated inhibition of osteogenesis in Marfan embryonic stem cells and Marfan patient-specific induced pluripotent stem cells. Stem Cells 30, 2709, 2012 [DOI] [PubMed] [Google Scholar]
  47. [48]Russell H.K., Jr A modification of Movat's pentachrome stain. Arch Pathol 94, 187, 1972 [PubMed] [Google Scholar]
  48. [49]Behr B., Sorkin M., Lehnhardt M., Renda A., Longaker M.T., and Quarto N. A comparative analysis of the osteogenic effects of BMP-2, FGF-2, and VEGFA in a calvarial defect model. Tissue Eng Part A 18, 1079, 2012 [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. [50]Gosain A.K., Santoro T.D., Song L.S., Capel C.C., Sudhakar P.V., and Matloub H.S. Osteogenesis in calvarial defects: contribution of the dura, the pericranium, and the surrounding bone in adult versus infant animals. Plast Reconstr Surg 112, 515, 2003 [DOI] [PubMed] [Google Scholar]
  50. [51]Wang J., and Glimcher M.J. Characterization of matrix-induced osteogenesis in rat calvarial bone defects: II. Origins of bone-forming cells. Calcif Tissue Int 65, 486, 1999 [DOI] [PubMed] [Google Scholar]
  51. [52]Maeda S., Hayashi M., Komiya S., Imamura T., and Miyazono K. Endogenous TGF-beta signaling suppresses maturation of osteoblastic mesenchymal cells. EMBO J 23, 552, 2004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. [53]Miyazono K., and Miyazawa K. Id: a target of BMP signaling. Sci STKE 2002, pe40, 2002 [DOI] [PubMed] [Google Scholar]
  53. [54]Ishida W., Hamamoto T., Kusanagi K., Yagi K., Kawabata M., Takehara K., Sampath T.K., Kato M., and Miyazono K. Smad6 is a Smad1/5-induced smad inhibitor. Characterization of bone morphogenetic protein-responsive element in the mouse Smad6 promoter. J Biol Chem 275, 6075, 2000 [DOI] [PubMed] [Google Scholar]
  54. [55]Wang Q., Wei X., Zhu T., Zhang M., Shen R., Xing L., O'Keefe R.J., and Chen D. Bone morphogenetic protein 2 activates Smad6 gene transcription through bone-specific transcription factor Runx2. J Biol Chem 282, 10742, 2007 [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. [56]Schindeler A., Morse A., Peacock L., Mikulec K., Yu N.Y., Liu R., Kijumnuayporn S., McDonald M.M., Baldock P.A., Ruys A.J., and Little D.G. Rapid cell culture and pre-clinical screening of a transforming growth factor-beta (TGF-beta) inhibitor for orthopaedics. BMC Musculoskelet Disord 11, 105, 2010 [DOI] [PMC free article] [PubMed] [Google Scholar]

Cited by

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Authors
Senarath-Yapa, K., Li, S., Walmsley, G. G., Zielins, E., Paik, K., Britto, J. A., Grigoriadis, A. E., Wan, D. C., Liu, K. J., Longaker, M. T., & Quarto, N.
Affiliations
  • King's College London (GB)
  • Stanford Medicine (US)
  • Great Ormond Street Hospital (GB)
Corresponding author
  • Longaker, Michael T.
  • Quarto, Natalina
ORCID
Venue
Tissue Engineering Part A
Publisher
SAGE Publications
Published
2016-05
Volume
22
Issue
9-10
Pages
707-720
ISSN
  • 1937-3341 (ISSN-L)
  • 1937-335X
DOI
10.1089/ten.tea.2015.0527
PMC
PMC4876548
OpenAlex
W2395501646
Cited by
22
References
1
Citations by year
2026: 1 · 2025: 2 · 2024: 2 · 2022: 5 · 2021: 4 · 2020: 3 · 2019: 1 · 2018: 1 · 2017: 3
FWCI
0.80
Type
journal article
Language
en
Open access
Licence
www.liebertpub.com/nv/resources-tools/text-and-data-mining-policy/121/ (tdm)
Locations
  • Tissue Engineering Part A (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Research Portal (King's College London) (submittedVersion)
Funders
  • Medical Research Council (MR/L017237/1)
  • National Heart, Lung, and Blood Institute (U01HL099776)
  • National Institute of Dental and Craniofacial Research (R01DE021683)
Topics
  • Physical Sciences › Engineering › Biomedical Engineering › Bone Tissue Engineering Materials
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › TGF-β signaling in diseases
Keywords
TGF-beta inhibition, SB431542, calvarial bone regeneration, BMP signaling, BMP-2, Smad6, osteoblasts, calvarial defect, skeletal repair, small molecule therapy, negative feedback loop
MeSH terms
Animals, Benzamides, Bone Regeneration, Dioxoles, Gene Expression Regulation, Humans, Osteoblasts, Skull, Signal Transduction, Transforming Growth Factor beta, Mice, Smad6 Protein, Bone Morphogenetic Protein 2

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