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Targeted inhibition of platelet-derived growth factor receptor-β subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats

Gene Therapy volume 15, pages 1424–1435 (2008)Cite this article

Abstract

The activation of hepatic stellate cells (HSCs) is the key event of the pathogenesis of hepatic fibrosis. Platelet-derived growth factor (PDGF) is the most potent mitogen for HSCs, and PDGF receptor-β subunit (PDGFR-β) is required for the proliferation of HSCs induced by PDGF. In this study, a high gene-silencing-efficacy PDGFR-β small interference RNA (siRNA) was synthesized that could suppress the PDGFR-β expression and inhibit the activation and proliferation but could not induce the apoptosis of HSCs in vitro. To avoid the side effect of nonspecific interference of PDGFR-β, we constructed an HSCs-specific short hairpin RNA (shRNA) expression plasmid in which PDGFR-β shRNA was driven by a glial fibrillary acidic protein (GFAP) promoter. The double-staining immunofluorescence examination indicated that GFAP promoter could target the transgene expression into HSCs in carbon tetrachloride induced acute injured rat's liver and bile duct ligation (BDL)-induced chronic injured rat's liver. Furthermore, HSCs-specific PDGFR-β shRNA could relieve liver injury and hepatic fibrosis in the rat's model induced by BDL. This study demonstrates that PDGFR-β siRNA may be presented as an antifibrogenic agent. The application of HSCs-specific RNA interference induced by the GFAP promoter might supply a new powerful tool for cell-specific gene therapy of hepatic fibrogenesis.

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Abbreviations

α-SMA:

α-smooth muscle actin

BDL:

bile duct ligation

CTGF:

connective tissue growth factor

GFAP:

glial fibrillary acidic protein

HSCs:

hepatic stellate cells

PDGF:

platelet-derived growth factor

PDGFR-β:

PDGF receptor-β subunit

RT-PCR:

reverse transcriptional-PCR

shRNA:

short hairpin RNA

siRNA:

small interfering RNA

TIMP-1:

tissue inhibitor of metalloproteinase-1.

References

  1. Safadi R, Friedman SL . Hepatic fibrosis—role of hepatic stellate cell activation. Med Gen Med 2002; 4: 27.

    Article  Google Scholar 

  2. Kinnman N, Francoz C, Barbu V, Wendum D, Rey C, Hultcrantz R et al. The myofibroblastic conversion of peribiliary fibrogenic cells distinct from hepatic stellate cells is stimulated by platelet-derived growth factor during liver fibrogenesis. Lab Invest 2003; 83: 163–173.

    Article  CAS  Google Scholar 

  3. Breitkopf K, Roeyen C, Sawitza I, Wickert L, Floege J, Gressner AM . Expression patterns of PDGF-A, -B, -C and -D and the PDGF- receptors a and b in activated rat hepatic stellate cells (HSC). Cytokine 2005; 31: 349–357.

    Article  CAS  Google Scholar 

  4. Wong L, Yamasaki G, Johnson RJ, Friedman SL . Induction of beta-platelet-derived growth factor receptor in rat hepatic lipocytes during cellular activation in vivo and in culture. J Clin Invest 1994; 94: 1563–1569.

    Article  CAS  Google Scholar 

  5. Pinzani M, Milani S, Herbst H, DeFranco R, Grappone C, Gentilini A et al. Expression of platelet-derived growth factor and its receptors in normal human liver and during active hepatic fibrogenesis. Am J Pathol 1996; 148: 785–800.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Czochra P, Klopcic B, Meyer E, Herkel J, Garcia-Lazaro JF, Thieringer F et al. Liver fibrosis induced by hepatic overexpression of PDGF-B in transgenic mice. J Hepatol 2006; 45: 419–428.

    Article  CAS  Google Scholar 

  7. Prieto J, Qian C, Hernandez-Alcoceba R, Gonzalez-Aseguinolaza G, Mazzolini G, Sangro B et al. Gene therapy of liver diseases. Expert Opin Biol Ther 2004; 4: 1073–1091.

    Article  CAS  Google Scholar 

  8. Pinzani M . PDGF and signal transduction in hepatic stellate cells. Front Biosci 2002; 7: d1720–d1726.

    Article  CAS  Google Scholar 

  9. Marra F, Arrighi MC, Fazi M, Caligiuri A, Pinzani M, Romanelli RG et al. Extracellular signal-regulated kinase activation differentially regulates platelet-derived growth factor's actions in hepatic stellate cells, and is induced by in vivo liver injury in the rat. Hepatology 1999; 30: 951–958.

    Article  CAS  Google Scholar 

  10. Borkham-Kamphorst E, Stoll D, Gressner AM, Weiskirchen R . Antisense strategy against PDGF B-chain proves effective in preventing experimental liver fibrogenesis. Biochem Biophys Res Commun 2004; 321: 413–423.

    Article  CAS  Google Scholar 

  11. Liu XJ, Yang L, Mao YQ, Wang Q, Huang MH, Wang YP et al. Effects of the tyrosine kinase inhibitor genistein on the proliferation, activation of cultured rat hepatic stellate cells. World J Gastroenterol 2002; 8: 739–745.

    Article  CAS  Google Scholar 

  12. Yoshiji H, Noguchi R, Kuriyama S, Ikenaka Y, Yoshii J, Yanase K et al. Imatinib mesylate (STI-571) attenuates liver fibrosis development in rats. Am J Physiol Gastrointest Liver Physiol 2005; 288: G907–G913.

    Article  CAS  Google Scholar 

  13. Di Sario A, Bendia E, Taffetani S, Marzioni M, Candelaresi C, Pigini P et al. Selective Na+/H+ exchange inhibition by cariporide reduces liver fibrosis in the rat. Hepatology 2003; 37: 256–266.

    Article  CAS  Google Scholar 

  14. Iwamoto H, Nakamuta M, Tada S, Sugimoto R, Enjoji M, Nawata H . Platelet-derived growth factor receptor tyrosine kinase inhibitor AG1295 attenuates rat hepatic stellate cell growth. J Lab Clin Med 2000; 135: 406–412.

    Article  CAS  Google Scholar 

  15. Chen YX, Lu CH, Xie WF, Zhang XR, Zhang ZB, Wei LX et al. Effects of ribozyme targeting platelet-derived growth factor receptor beta subunit gene on the proliferation and apoptosis of hepatic stellate cells in vitro. Chin Med J (Engl) 2005; 118: 982–988.

    CAS  Google Scholar 

  16. Borkham-Kamphorst E, Stoll D, Gressner AM, Weiskirchen R . Inhibitory effect of soluble PDGF-beta receptor in culture-activated hepatic stellate cells. Biochem Biophys Res Commun 2004; 317: 451–462.

    Article  CAS  Google Scholar 

  17. Borkham-Kamphorst E, Herrmann J, Stoll D, Treptau J, Gressner AM, Weiskirchen R . Dominant-negative soluble PDGF-beta receptor inhibits hepatic stellate cell activation and attenuates liver fibrosis. Lab Invest 2004; 84: 766–777.

    Article  CAS  Google Scholar 

  18. Vickers TA, Koo S, Bennett CF, Crooke ST, Dean NM, Baker BF . Efficient reduction of target RNAs by small interfering RNA and RNase H-dependent antisense agents. A comparative analysis. J Biol Chem 2003; 278: 7108–7118.

    Article  CAS  Google Scholar 

  19. Grimm D, Kay MA . Therapeutic short hairpin RNA expression in the liver: viral targets and vectors. Gene Ther 2006; 13: 563–575.

    Article  CAS  Google Scholar 

  20. Kinnman N, Francoz C, Barbu V, Wendum D, Rey C, Hultcrantz R et al. The myofibroblastic conversion of peribiliary fibrogenic cells distinct from hepatic stellate cells is stimulated by platelet-derived growth factor during liver fibrogenesis. Lab Invest 2003; 83: 163–173.

    Article  CAS  Google Scholar 

  21. Bedossa P, Paradis V . Liver extracellular matrix in health and disease. J Pathol 2003; 200: 504–515.

    Article  Google Scholar 

  22. Zhang LJ, Chen YX, Chen ZX, Huang YH, Yu JP, Wang XZ . Effect of interleukin-10 and platelet-derived growth factor on expressions of matrix metalloproteinases-2 and tissue inhibitor of metalloproteinases-1 in rat fibrotic liver and cultured hepatic stellate cells. World J Gastroenterol 2004; 10: 2574–2579.

    Article  CAS  Google Scholar 

  23. Rachfal AW, Brigstock DR . Connective tissue growth factor (CTGF/CCN2) in hepatic fibrosis. Hepatol Res 2003; 26: 1–9.

    Article  CAS  Google Scholar 

  24. Gao R, Ball DK, Perbal B, Brigstock DR . Connective tissue growth factor induces c-fos gene activation and cell proliferation through p44/42 MAP kinase in primary rat hepatic stellate cells. J Hepatol 2004; 40: 431–438.

    Article  CAS  Google Scholar 

  25. Garcia-Bañuelos J, Siller-Lopez F, Miranda A, Aguilar LK, Aguilar-Cordova E, Armendariz-Borunda J . Cirrhotic rat livers with extensive fibrosis can be safely transduced with clinical-grade adenoviral vectors. Evidence of cirrhosis reversion. Gene Therapy 2002; 9: 127–134.

    Article  Google Scholar 

  26. Zhang Z, Chen J, Jin D . Platelet-derived growth factor (PDGF)-BB stimulates osteoclastic bone resorption directly: the role of receptor beta. Biochem Biophys Res Commun 1998; 251: 190–194.

    Article  CAS  Google Scholar 

  27. Oreffo RO . Growth factors for skeletal reconstruction and fracture repair. Curr Opin Investig Drugs 2004; 5: 419–423.

    CAS  PubMed  Google Scholar 

  28. Yuantai W, Tiancai W, Qiu Z . PD98059 inhibits expression of pERK1 protein and collagen alpha1(I) mRNA in rat pancreatic stellate cells activated by platelet-derived growth factor. Indian J Gastroenterol 2005; 24: 100–103.

    PubMed  Google Scholar 

  29. Eitner F, Ostendorf T, Van Roeyen C, Kitahara M, Li X, Aase K et al. Expression of a novel PDGF isoform, PDGF-C, in normal and diseased rat kidney. J Am Soc Nephrol 2002; 13: 910–917.

    CAS  PubMed  Google Scholar 

  30. Abdollahi A, Li M, Ping G, Plathow C, Domhan S, Kiessling F et al. Inhibition of platelet-derived growth factor signaling attenuates pulmonary fibrosis. J Exp Med 2005; 201: 925–935.

    Article  CAS  Google Scholar 

  31. Zymek P, Bujak M, Chatila K, Cieslak A, Thakker G, Entman ML et al. The role of platelet-derived growth factor signaling in healing myocardial infarcts. J Am Coll Cardiol 2006; 48: 2315–2323.

    Article  CAS  Google Scholar 

  32. Bahr MJ, Vincent KJ, Arthur MJ, Fowler AV, Smart DE, Wright MC et al. Control of the tissue inhibitor of metalloproteinases-1 promotor in culture-activated rat hepatic stellate cells: regulation by activator protein-1 DNA binding proteins. Hepatology 1999; 29: 839–848.

    Article  CAS  Google Scholar 

  33. Herrmann J, Arias M, Van De Leur E, Gressner AM, Weiskirchen R . CSRP2, TIMP-1, and SM22alpha promotor fragments direct hepatic. stellate cell-specific transgene expression in vitro, but not in vivo. Liver Int 2004; 24: 69–79.

    Article  CAS  Google Scholar 

  34. Inagaki Y, Kushida M, Higashi K, Itoh J, Higashiyama R, Hong YY et al. Cell type-specific intervention of transforming growth factor beta/Smad signaling suppresses collagen gene expression and hepatic fibrosis in mice. Gastroenterology 2005; 129: 259–268.

    Article  CAS  Google Scholar 

  35. Dahl D . The vimentin-GFA protein transition in rat neuroglia cytoskeleton occurs at the time of myelination. J Neurosci Res 1981; 6: 741–748.

    Article  CAS  Google Scholar 

  36. Brenner M, Kisseberth WC, Su Y, Besnard F, Messing A . GFAP promotor directs astrocyte-specific expression in transgenic mice. J Neurosci 1994; 14: 1030–1037.

    Article  CAS  Google Scholar 

  37. Gard AL, White FP, Dutton GR . Extra-neural glial fibrillary acidic protein (GFAP) immunoreactivity in perisinusoidal stellate cells of rat liver. J Neuroimmunol 1985; 8: 359–375.

    Article  CAS  Google Scholar 

  38. Cassiman D, Libbrecht L, Desmet V, Denef C, Roskams T . Hepatic stellate cell/myofibroblast subpopulations in fibrotic human and rat livers. J Hepatol 2002; 36: 200–209.

    Article  Google Scholar 

  39. Maubach G, Lim MC, Zhang CY, Zhuo L . GFAP promotor directs lacZ expression specifically in a rat hepatic stellate cell line. World J Gastroenterol 2006; 12: 723–730.

    Article  CAS  Google Scholar 

  40. Stegmeier F, Hu G, Rickles RJ, Hannon GJ, Elledge SJ . A lentiviral microRNA-based system for single-copy polymerase II-regulated RNA interference in mammalian cells. Proc Natl Acad Sci USA 2005; 102: 13212–13217.

    Article  CAS  Google Scholar 

  41. Zhou H, Xia XG, Xu Z . An RNA polymerase II construct synthesizes short-hairpin RNA with a quantitative indicator and mediates highly efficient RNAi. Nucleic Acids Res 2005; 33: e62.

    Article  Google Scholar 

  42. Vogel S, Piantedosi R, Frank J, Lalazar A, Rockey DC, Friedman SL et al. An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. J Lipid Res 2000; 41: 882–893.

    CAS  PubMed  Google Scholar 

  43. Racine-Samson L, Rockey DC, Bissell DM . The role of alpha1beta1 integrin in wound contraction. A quantitative analysis of liver myofibroblasts in vivo and in primary culture. J Biol Chem 1997; 272: 30911–30917.

    Article  CAS  Google Scholar 

  44. Liu F, Song Y, Liu D . Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Therapy 1999; 6: 1258–1266.

    Article  CAS  Google Scholar 

  45. Maruyama H, Higuchi N, Nishikawa Y, Kameda S, Iino N, Kazama JJ et al. High-level expression of naked DNA delivered to rat liver via tail vein injection. J Gene Med 2002; 4: 333–341.

    Article  CAS  Google Scholar 

  46. Lewis DL, Hagstrom JE, Loomis AG, Wolff JA, Herweijer H . Efficient delivery of siRNA for inhibition of gene expression in postnatal mice. Nat genet 2002; 32: 107–108.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by Grant from National Natural Science Foundation of China (no. 30472049).

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  1. S-W Chen and Y-X Chen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, Changzheng Hospital, Second Military Medical University, Shanghai, China

    S-W Chen, Y-X Chen, X-R Zhang, H Qian, W-Z Chen & W-F Xie

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Correspondence to W-F Xie.

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Chen, SW., Chen, YX., Zhang, XR. et al. Targeted inhibition of platelet-derived growth factor receptor-β subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats. Gene Ther 15, 1424–1435 (2008). https://doi.org/10.1038/gt.2008.93

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