TGF-β2 silencing to target biliary-derived liver diseases

Gut. 2020 Sep;69(9):1677-1690. doi: 10.1136/gutjnl-2019-319091. Epub 2020 Jan 28.

Abstract

Objective: TGF-β2 (TGF-β, transforming growth factor beta), the less-investigated sibling of TGF-β1, is deregulated in rodent and human liver diseases. Former data from bile duct ligated and MDR2 knockout (KO) mouse models for human cholestatic liver disease suggested an involvement of TGF-β2 in biliary-derived liver diseases.

Design: As we also found upregulated TGFB2 in liver tissue of patients with primary sclerosing cholangitis (PSC) and primary biliary cholangitis (PBC), we now fathomed the positive prospects of targeting TGF-β2 in early stage biliary liver disease using the MDR2-KO mice. Specifically, the influence of TgfB2 silencing on the fibrotic and inflammatory niche was analysed on molecular, cellular and tissue levels.

Results: TgfB2-induced expression of fibrotic genes in cholangiocytes and hepatic stellate cellswas detected. TgfB2 expression in MDR2-KO mice was blunted using TgfB2-directed antisense oligonucleotides (AON). Upon AON treatment, reduced collagen deposition, hydroxyproline content and αSMA expression as well as induced PparG expression reflected a significant reduction of fibrogenesis without adverse effects on healthy livers. Expression analyses of fibrotic and inflammatory genes revealed AON-specific regulatory effects on Ccl3, Ccl4, Ccl5, Mki67 and Notch3 expression. Further, AON treatment of MDR2-KO mice increased tissue infiltration by F4/80-positive cells including eosinophils, whereas the number of CD45-positive inflammatory cells decreased. In line, TGFB2 and CD45 expression correlated positively in PSC/PBC patients and localised in similar areas of the diseased liver tissue.

Conclusions: Taken together, our data suggest a new mechanistic explanation for amelioration of fibrogenesis by TGF-β2 silencing and provide a direct rationale for TGF-β2-directed drug development.

Keywords: TGF-beta; cholestasis; fibrosis; primary biliary cirrhosis; primary sclerosing cholangitis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B / genetics
  • ATP-Binding Cassette Sub-Family B Member 4
  • Animals
  • Cholangitis, Sclerosing* / metabolism
  • Cholangitis, Sclerosing* / pathology
  • Disease Models, Animal
  • Drug Discovery
  • Gene Expression Regulation
  • Gene Silencing*
  • Hepatic Stellate Cells / metabolism
  • Humans
  • Liver Cirrhosis* / metabolism
  • Liver Cirrhosis* / pathology
  • Liver Cirrhosis* / prevention & control
  • Liver Cirrhosis, Biliary* / metabolism
  • Liver Cirrhosis, Biliary* / pathology
  • Mice
  • Mice, Knockout
  • Oligonucleotides, Antisense*
  • Transforming Growth Factor beta2 / genetics*
  • Transforming Growth Factor beta2 / metabolism*
  • Up-Regulation

Substances

  • ATP Binding Cassette Transporter, Subfamily B
  • Oligonucleotides, Antisense
  • TGFB2 protein, human
  • Tgfb2 protein, mouse
  • Transforming Growth Factor beta2