Effects of TGF-${\beta}1$ Ribbon Antisense on $CCl_4$-induced Liver Fibrosis

  • Doh, Kyung-Oh (Department of Physiology, College of Medicine, Dongguk University)
  • Published : 2008.02.28

Abstract

Ribbon-type antisense oligonucleotide to TGF-${\beta}1$ (TGF-${\beta}1$ RiAS) was designed and tested to prevent or resolve the fibrotic changes induced by $CCl_4$ injection. When Hepa1c1c7 cells were transfected with TGF-${\beta}1$ RiAS, the level of TGF-${\beta}1$ mRNA was effectively reduced. TGF-${\beta}1$ RiAS, mismatched RiAS, and normal saline were each injected to mice via tail veins. When examined for the biochemical effects on the liver, TGF-${\beta}1$ mRNA levels were significantly reduced only in the TGF-${\beta}1$ RiAS-treated group. The results of immunohistochemical studies showed that TGF-${\beta}1$ RiAS prevented the accumulation of collagen and ${\alpha}$-smooth muscle actin, but could not resolve established fibrosis. These results indicate that ribbon antisense to TGF-${\beta}1$ with efficient uptake can effectively prevent fibrosis of the liver.

Keywords

References

  1. Alcolado R, Arthur MJ, Iredale JP. Pathogenesis of liver fibrosis. Clin Sci (Lond) 92: 103-112, 1997 https://doi.org/10.1042/cs0920103
  2. Arias M, Lahme B, Van de Leur E, Gressner AM, Weiskirchen R. Adenoviral delivery of an antisense RNA complementary to the 3' coding sequence of transforming growth factor-beta1 inhibits fibrogenic activities of hepatic stellate cells. Cell Growth Differ 13: 265-273, 2002
  3. Arias M, Sauer-Lehnen S, Treptau J, Janoschek N, Theuerkauf I, Buettner R, Gressner AM, Weiskirchen R. Adenoviral expression of a transforming growth factor-beta1 antisense mRNA is effective in preventing liver fibrosis in bile-duct ligated rats. BMC Gastroenterol 3: 29, 2003 https://doi.org/10.1186/1471-230X-3-29
  4. Arthur MJ, Fibrogenesis II. Metalloproteinases and their inhibitors in liver fibrosis. Am J Physiol Gastrointest Liver Physiol 279: G245-249, 2000 https://doi.org/10.1152/ajpgi.2000.279.2.G245
  5. Bajpai AK, Park JH, Moon IJ, Kang H, Lee YH, Doh KO, Suh SI, Chang BC, Park JG. Rapid blockade of telomerase activity and tumor cell growth by the DPL lipofection of ribbon antisense to hTR. Oncogene 24: 6492-6501, 2005 https://doi.org/10.1038/sj.onc.1208731
  6. Bharath MM, Chandra NR, Rao MRS. Prediction of an HMG-box fold in the C-terminal domain of histone H1: insights into its role in DNA condensation. Proteins 49: 71-81, 2002 https://doi.org/10.1002/prot.10204
  7. Bissell DM, Roulot D, George J. Transforming growth factor beta and the liver. Hepatology 34: 859-867, 2001 https://doi.org/10.1053/jhep.2001.28457
  8. Choi YK, Moon IJ, Jung HK, Jang BC, Seo SI, Park JG. Prevention of tissue injury by ribbon antisense to TGF-beta1 in the kidney. Int J Mol Med 15: 391-399, 2005
  9. Derossi D, Calvet S, Trembleau A, Brunissen A, Chassaing G, Prochiantz A. Cell internalization of the third helix of the Antennapedia homeodomain is receptor-independent. J Biol Chem 271: 18188-18193, 1996 https://doi.org/10.1074/jbc.271.30.18188
  10. Dokka S, Toledo D, Shi X, Ye J, Rojanasakul Y. High-efficiency gene transfection of macrophages by lipoplexes. Int J Pharm 206: 97-104, 2000 https://doi.org/10.1016/S0378-5173(00)00531-7
  11. Eguchi A, Akuta T, Okuyama H, Senda T, Yokoi H, Inokuchi H, Fujita S, Hayakawa T, Takeda K, Hasegawa M, Nakanishi M. Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells. J Biol Chem 276: 26204-26210, 2001 https://doi.org/10.1074/jbc.M010625200
  12. Forbes SJ, Russo FP, Rey V, Burra P, Rugge M, Wright NA, Alison MR. A significant proportion of myofibroblasts are of bone marrow origin in human liver fibrosis. Gastroenterology 126: 955-963, 2004 https://doi.org/10.1053/j.gastro.2004.02.025
  13. Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem 275: 2247-2250, 2000 https://doi.org/10.1074/jbc.275.4.2247
  14. Friedman SL, Roll FJ, Boyles J, Bissell DM. Hepatic lipocytes: the principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci U S A 82: 8681-8685, 1985
  15. Fulda S, Wick W, Weller M, Debatin KM. Smac agonists sensitize for Apo2L/TRAIL- or anticancer drug-induced apoptosis and induce regression of malignant glioma in vivo. Nat Med 8: 808-815, 2002 https://doi.org/10.1038/nm735
  16. Futaki S, Suzuki T, Ohashi W, Yagami T, Tanaka S, Ueda K, Sugiura Y. Arginine-rich peptides. An abundant source of membrane-permeable peptides having potential as carriers for intracellular protein delivery. J Biol Chem 276: 5836-5840, 2001 https://doi.org/10.1074/jbc.M007540200
  17. George J, Roulot D, Koteliansky VE, Bissell DM. In vivo inhibition of rat stellate cell activation by soluble transforming growth factor beta type II receptor: a potential new therapy for hepatic fibrosis. Proc Natl Acad Sci U S A 96: 12719-12724, 1999
  18. Gressner AM, Weiskirchen R, Breitkopf K, Dooley S. Roles of TGF-beta in hepatic fibrosis. Front Biosci 7: d793-807, 2002 https://doi.org/10.2741/gressner
  19. Gryaznov S, Skorski T, Cucco C, Nieborowska-Skorska M, Chiu CY, Lloyd D, Chen JK, Koziolkiewicz M, Calabretta B. Oligonucleotide N3'$\rightarrow$P5' phosphoramidates as antisense agents. Nucleic Acids Res 24: 1508-1514, 1996 https://doi.org/10.1093/nar/24.8.1508
  20. Hammel P, Couvelard A, O'Toole D, Ratouis A, Sauvanet A, Flejou JF, Degott C, Belghiti J, Bernades P, Valla D, Ruszniewski P, Levy P. Regression of liver fibrosis after biliary drainage in patients with chronic pancreatitis and stenosis of the common bile duct. N Engl J Med 344: 418-423, 2001 https://doi.org/10.1056/NEJM200102083440604
  21. Henry SP, Novotny W, Leeds J, Auletta C, Kornbrust DJ. Inhibition of coagulation by a phosphorothioate oligonucleotide. Antisense Nucleic Acid Drug Dev 7: 503-510, 1997 https://doi.org/10.1089/oli.1.1997.7.503
  22. Iredale JP, Benyon RC, Pickering J, McCullen M, Northrop M, Pawley S, Hovell C, Arthur MJ. Mechanisms of spontaneous resolution of rat liver fibrosis. Hepatic stellate cell apoptosis and reduced hepatic expression of metalloproteinase inhibitors. J Clin Invest 102: 538-549, 1998 https://doi.org/10.1172/JCI1018
  23. Issa R, Zhou X, Constandinou CM, Fallowfield J, Millward-Sadler H, Gaca MD, Sands E, Suliman I, Trim N, Knorr A, Arthur MJ, Benyon RC, Iredale JP. Spontaneous recovery from micronodular cirrhosis: evidence for incomplete resolution associated with matrix cross-linking. Gastroenterology 126: 1795-1808, 2004 https://doi.org/10.1053/j.gastro.2004.03.009
  24. Kanzler S, Lohse AW, Keil A, Henninger J, Dienes HP, Schirmacher P, Rose-John S, zum Buschenfelde KH, Blessing M. TGF-beta1 in liver fibrosis: an inducible transgenic mouse model to study liver fibrogenesis. Am J Physiol 276: G1059-1068, 1999
  25. Knittel T, Mehde M, Kobold D, Saile B, Dinter C, Ramadori G. Expression patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: regulation by TNF-alpha and TGF-beta1. J Hepatol 30: 48-60, 1999 https://doi.org/10.1016/S0168-8278(99)80007-5
  26. Ludtke JJ, Zhang G, Sebestyen MG, Wolff JA. A nuclear localization signal can enhance both the nuclear transport and expression of 1 kb DNA. J Cell Sci 112: 2033-2041, 1999
  27. Moon IJ, Choi K, Choi YK, Kim JE, Lee Y, Schreiber AD, Park JG. Potent growth inhibition of leukemic cells by novel ribbon-type antisense oligonucleotides to c-myb1. J Biol Chem 275: 4647-4653, 2000a https://doi.org/10.1074/jbc.275.7.4647
  28. Moon IJ, Kang H, Seu YB, Chang BC, Song DK, Park JG. Marked transfection enhancement by the DPL (DNA/peptide/lipid) complex. Int J Mol Med 20: 429-437, 2007
  29. Moon IJ, Lee Y, Kwak CS, Lee JH, Choi K, Schreiber AD, Park JG. Target site search and effective inhibition of leukaemic cell growth by a covalently closed multiple anti-sense oligonucleotide to c-myb. Biochem J 346 Pt 2: 295-303, 2000b https://doi.org/10.1042/0264-6021:3460295
  30. Nakamura T, Ueno T, Sakamoto M, Sakata R, Torimura T, Hashimoto O, Ueno H, Sata M. Suppression of transforming growth factor-beta results in upregulation of transcription of regeneration factors after chronic liver injury. J Hepatol 41: 974-982, 2004 https://doi.org/10.1016/j.jhep.2004.08.015
  31. Qi Z, Atsuchi N, Ooshima A, Takeshita A, Ueno H. Blockade of type beta transforming growth factor signaling prevents liver fibrosis and dysfunction in the rat. Proc Natl Acad Sci U S A 96: 2345-2349, 1999
  32. Ramadori G, Saile B. Portal tract fibrogenesis in the liver. Lab Invest 84: 153-159, 2004 https://doi.org/10.1038/labinvest.3700030
  33. Schwarze SR, Ho A, Vocero-Akbani A, Dowdy SF. In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285: 1569-1572, 1999 https://doi.org/10.1126/science.285.5433.1569
  34. Sorgi FL, Bhattacharya S, Huang L. Protamine sulfate enhances lipid-mediated gene transfer. Gene Ther 4: 961-968, 1997 https://doi.org/10.1038/sj.gt.3300484
  35. Torchilin VP, Rammohan R, Weissig V, Levchenko TS. TAT peptide on the surface of liposomes affords their efficient intracellular delivery even at low temperature and in the presence of metabolic inhibitors. Proc Natl Acad Sci U S A 98: 8786-8791, 2001
  36. Ueno H, Sakamoto T, Nakamura T, Qi Z, Astuchi N, Takeshita A, Shimizu K, Ohashi H. A soluble transforming growth factor beta receptor expressed in muscle prevents liver fibrogenesis and dysfunction in rats. Hum Gene Ther 11: 33-42, 2000 https://doi.org/10.1089/10430340050016139
  37. Vives E, Brodin P, Lebleu B. A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem 272: 16010-16017, 1997 https://doi.org/10.1074/jbc.272.25.16010
  38. Wagner RW, Matteucci MD, Lewis JG, Gutierrez AJ, Moulds C, Froehler BC. Antisense gene inhibition by oligonucleotides containing C-5 propyne pyrimidines. Science 260: 1510-1513, 1993 https://doi.org/10.1126/science.7684856
  39. Zanta MA, Belguise-Valladier P, Behr JP. Gene delivery: a single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus. Proc Natl Acad Sci U S A 96: 91-96, 1999