Article | Published:

BMP-2 restoration aids in recovery from liver fibrosis by attenuating TGF-β1 signaling

Laboratory Investigationvolume 98pages9991013 (2018) | Download Citation

Abstract

Transforming growth factor-β (TGF-β) plays a central role in hepatic fibrogenesis. This study investigated the function and mechanism of bone morphogenetic protein-2 (BMP-2) in regulation of hepatic fibrogenesis. BMP-2 expression in fibrotic liver was measured in human tissue microarray and mouse models of liver fibrosis induced by bile duct ligation surgery or carbon tetrachloride administration. Adenovirus-mediated BMP-2 gene delivery was used to test the prophylactic effect on liver fibrosis. Primary hepatic stellate cells (HSC), HSC-T6 and clone-9 cell lines were used to study the interplay between BMP-2 and TGF-β1. Hepatic BMP-2 was localized in parenchymal hepatocytes and activated HSCs and significantly decreased in human and mouse fibrotic livers, showing an opposite pattern of hepatic TGF-β1 contents. BMP-2 gene delivery alleviated the elevations of serum hepatic enzymes, cholangiocyte marker CK19, HSC activation markers, and liver fibrosis in both models. Mechanistically, exogenous TGF-β1 dose dependently reduced BMP-2 expression, whereas BMP-2 significantly suppressed expression of TGF-β and its cognate type I and II receptor peptides, as well as the induced Smad3 phosphorylation levels in primary mouse HSCs. Aside from its suppressive effects on cell proliferation and migration, BMP-2 treatment prominently attenuated the TGF-β1-stimulated α-SMA and fibronectin expression, and reversed the TGF-β1-modulated epithelial-to-mesenchymal transition marker expression in mouse HSCs. The mutual regulation between BMP-2 and TGF-β1 signaling axes may constitute the anti-fibrogenic mechanism of BMP-2 in the pathogenesis of liver fibrosis. BMP-2 may potentially serve as a novel therapeutic target for treatment of liver fibrosis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Additional information

These authors contributed equally: Yueh-Hua Chung, Ying-Hsien Huang.

References

  1. 1.

    Lee UE, Friedman SL. Mechanisms of hepatic fibrogenesis. Best Pract Res Clin Gastroenterol. 2011;25:195–206.

  2. 2.

    Friedman SL. Evolving challenges in hepatic fibrosis. Nat Rev Gastroenterol Hepatol. 2010;7:425–36.

  3. 3.

    Bataller R, Brenner DA. Liver fibrosis. J Clin Invest. 2005;115:209–18.

  4. 4.

    Modol T, Brice N, Ruiz de Galarreta M, et al. Fibronectin peptides as potential regulators of hepatic fibrosis through apoptosis of hepatic stellate cells. J Cell Physiol. 2015;230:546–53.

  5. 5.

    Liu XY, Liu RX, Hou F, et al. Fibronectin expression is critical for liver fibrogenesis in vivo and in vitro. Mol Med Rep. 2016;14:3669–75.

  6. 6.

    Bissell DM, Roulot D, George J. Transforming growth factor beta and the liver. Hepatology. 2001;34:859–67.

  7. 7.

    Jonsson JR, Clouston AD, Ando Y, et al. Angiotensin-converting enzyme inhibition attenuates the progression of rat hepatic fibrosis. Gastroenterology. 2001;121:148–55.

  8. 8.

    O’Connor JW, Gomez EW. Biomechanics of TGFbeta-induced epithelial-mesenchymal transition: implications for fibrosis and cancer. Clin Transl Med. 2014;3:23.

  9. 9.

    Bottinger EP. TGF-beta in renal injury and disease. Semin Nephrol. 2007;27:309–20.

  10. 10.

    Liu X, Hu H, Yin JQ. Therapeutic strategies against TGF-beta signaling pathway in hepatic fibrosis. Liver Int. 2006;26:8–22.

  11. 11.

    Zeisberg M, Hanai J, Sugimoto H, et al. BMP-7 counteracts TGF-beta1-induced epithelial-to-mesenchymal transition and reverses chronic renal injury. Nat Med. 2003;9:964–8.

  12. 12.

    Park J, Schwarzbauer JE. Mammary epithelial cell interactions with fibronectin stimulate epithelial-mesenchymal transition. Oncogene. 2014;33:1649–57.

  13. 13.

    Border WA, Noble NA. Transforming growth factor beta in tissue fibrosis. N Engl J Med. 1994;331:1286–92.

  14. 14.

    Wozney JM, Rosen V, Celeste AJ, et al. Novel regulators of bone formation: molecular clones and activities. Science. 1988;242:1528–34.

  15. 15.

    Massague J. The transforming growth factor-beta family. Annu Rev Cell Biol. 1990;6:597–641.

  16. 16.

    Itoh S, Itoh F, Goumans MJ, et al. Signaling of transforming growth factor-beta family members through Smad proteins. Eur J Biochem. 2000;267:6954–67.

  17. 17.

    Feng XH, Derynck R. Specificity and versatility in TGF-beta signaling through Smads. Annu Rev Cell Dev Biol. 2005;21:659–93.

  18. 18.

    Eickelberg O, Morty RE. Transforming growth factor beta/bone morphogenic protein signaling in pulmonary arterial hypertension: remodeling revisited. Trends Cardiovasc Med. 2007;17:263–9.

  19. 19.

    Song JJ, Celeste AJ, Kong FM, et al. Bone morphogenetic protein-9 binds to liver cells and stimulates proliferation. Endocrinology. 1995;136:4293–7.

  20. 20.

    Duncan SA, Watt AJ. BMPs on the road to hepatogenesis. Genes Dev. 2001;15:1879–84.

  21. 21.

    Tsai MS, Suksaweang S, Jiang TX, et al. Proper BMP signaling levels are essential for 3D assembly of hepatic cords from hepatoblasts and mesenchymal cells. Dig Dis Sci. 2015;60:3669–80.

  22. 22.

    Xu CP, Ji WM, van den Brink GR, et al. Bone morphogenetic protein-2 is a negative regulator of hepatocyte proliferation downregulated in the regenerating liver. World J Gastroenterol. 2006;12:7621–5.

  23. 23.

    Yang YL, Liu YS, Chuang LY, et al. Bone morphogenetic protein-2 antagonizes renal interstitial fibrosis by promoting catabolism of type I transforming growth factor-beta receptors. Endocrinology. 2009;150:727–40.

  24. 24.

    Yang YL, Ju HZ, Liu SF, et al. BMP-2 suppresses renal interstitial fibrosis by regulating epithelial-mesenchymal transition. J Cell Biochem. 2011;112:2558–65.

  25. 25.

    Shlyonsky V, Soussia IB, Naeije R, et al. Opposing effects of bone morphogenetic protein-2 and endothelin-1 on lung fibroblast chloride currents. Am J Respir Cell Mol Biol. 2011;45:1154–60.

  26. 26.

    Gao X, Cao Y, Staloch DA, et al. Bone morphogenetic protein signaling protects against cerulein-induced pancreatic fibrosis. PLoS ONE. 2014;9:e89114.

  27. 27.

    Kao YH, Chen CL, Jawan B, et al. Upregulation of hepatoma-derived growth factor is involved in murine hepatic fibrogenesis. J Hepatol. 2010;52:96–105.

  28. 28.

    Massoner P, Kugler KG, Unterberger K, et al. Characterization of transcriptional changes in ERG rearrangement-positive prostate cancer identifies the regulation of metabolic sensors such as neuropeptide Y. PLoS ONE. 2013;8:e55207.

  29. 29.

    Tai MH, Cheng H, Wu JP, et al. Gene transfer of glial cell line-derived neurotrophic factor promotes functional recovery following spinal cord contusion. Exp Neurol. 2003;183:508–15.

  30. 30.

    Huang YH, Tiao MM, Huang LT, et al. Activation of miR-29a in activated hepatic stellate cells modulates its profibrogenic phenotype through inhibition of histone deacetylases 4. PLoS ONE. 2015;10:e0136453.

  31. 31.

    Kao YH, Chen PH, Wu TY, et al. Lipopolysaccharides induce Smad2 phosphorylation through PI3K/Akt and MAPK cascades in HSC-T6 hepatic stellate cells. Life Sci. 2017;184:37–46.

  32. 32.

    Yang YL, Wang FS, Li SC, et al. MicroRNA-29a alleviates bile duct ligation exacerbation of hepatic fibrosis in mice through epigenetic control of methyltransferases. Int J Mol Sci. 2017;18:192.

  33. 33.

    Nakatsuka R, Taniguchi M, Hirata M, et al. Transient expression of bone morphogenic protein-2 in acute liver injury by carbon tetrachloride. J Biochem. 2007;141:113–9.

  34. 34.

    Cai J, Zhao Y, Liu Y, et al. Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology. 2007;45:1229–39.

  35. 35.

    Li X, Yuan J, Li W, et al. Direct differentiation of homogeneous human adipose stem cells into functional hepatocytes by mimicking liver embryogenesis. J Cell Physiol. 2014;229:801–12.

  36. 36.

    Yu Q, Stamenkovic I. Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-beta and promotes tumor invasion and angiogenesis. Genes Dev. 2000;14:163–76.

  37. 37.

    Kao YH, Jawan B, Goto S, et al. Serum factors potentiate hypoxia-induced hepatotoxicity in vitro through increasing transforming growth factor-beta1 activation and release. Cytokine. 2009;47:11–22.

  38. 38.

    Han YP, Yan C, Zhou L, et al. A matrix metalloproteinase-9 activation cascade by hepatic stellate cells in trans-differentiation in the three-dimensional extracellular matrix. J Biol Chem. 2007;282:12928–39.

  39. 39.

    Choi YA, Kang SS, Jin EJ. BMP-2 treatment of C3H10T1/2 mesenchymal cells blocks MMP-9 activity during chondrocyte commitment. Cell Biol Int. 2009;33:887–92.

  40. 40.

    Zhang QD, Xu MY, Cai XB, et al. Myofibroblastic transformation of rat hepatic stellate cells: the role of Notch signaling and epithelial-mesenchymal transition regulation. Eur Rev Med Pharmacol Sci. 2015;19:4130–8.

  41. 41.

    Gao X, Cao Y, Yang W, et al. BMP-2 inhibits TGF-beta-induced pancreatic stellate cell activation and extracellular matrix formation. Am J Physiol Gastrointest Liver Physiol. 2013;304:G804–813.

Download references

Acknowledgements

This study was supported by the grants in part from Kaohsiung Chang Gung Memorial Hospital (Nos. CMRPG8F1561, CMRPG8F1562) and Kaohsiung Armed Forces General Hospital (No. 103-6), Taiwan.

Author information

Author notes

    Affiliations

    1. Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan

      • Yueh-Hua Chung
      • , Shih-Chung Huang
      •  & Ming-Hong Tai
    2. Department of Pediatrics, Kaohsiung Chang Gung Memorial Hospital and Chiayi Chang Gung Memorial Hospital, Puzi City, Taiwan

      • Ying-Hsien Huang
    3. Center for Neuroscience, National Sun Yat-sen University, Kaohsiung, Taiwan

      • Tien-Huei Chu
      •  & Ming-Hong Tai
    4. Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan

      • Chun-Lin Chen
    5. Division of Hepato-Gastroenterology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

      • Pey-Ru Lin
      •  & Tsung-Hui Hu
    6. Department of Internal Medicine, Kaohsiung Armed Forces General Hospital, Kaohsiung, Taiwan

      • Shih-Chung Huang
    7. Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung, Taiwan

      • Deng-Chyang Wu
    8. Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

      • Deng-Chyang Wu
    9. Biobank and Tissue Bank and Department of Pathology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

      • Chao-Cheng Huang
    10. Department of Medical Research, E-Da Hospital, Kaohsiung, Taiwan

      • Ying-Hsien Kao

    Authors

    1. Search for Yueh-Hua Chung in:

    2. Search for Ying-Hsien Huang in:

    3. Search for Tien-Huei Chu in:

    4. Search for Chun-Lin Chen in:

    5. Search for Pey-Ru Lin in:

    6. Search for Shih-Chung Huang in:

    7. Search for Deng-Chyang Wu in:

    8. Search for Chao-Cheng Huang in:

    9. Search for Tsung-Hui Hu in:

    10. Search for Ying-Hsien Kao in:

    11. Search for Ming-Hong Tai in:

    Conflict of interest

    The authors declare that they have no conflict of interest.

    Corresponding authors

    Correspondence to Ying-Hsien Kao or Ming-Hong Tai.

    Electronic supplementary material

    About this article

    Publication history

    Received

    Revised

    Accepted

    Published

    DOI

    https://doi.org/10.1038/s41374-018-0069-9