1. Academic Validation
  2. TGF-β2 silencing to target biliary-derived liver diseases

TGF-β2 silencing to target biliary-derived liver diseases

  • Gut. 2020 Sep;69(9):1677-1690. doi: 10.1136/gutjnl-2019-319091.
Anne Dropmann 1 Steven Dooley 2 Bedair Dewidar 1 3 Seddik Hammad 1 4 Tatjana Dediulia 1 Julia Werle 1 Vanessa Hartwig 1 Shahrouz Ghafoory 5 Stefan Woelfl 5 Hanna Korhonen 6 Michel Janicot 6 Katja Wosikowski 6 Timo Itzel 7 Andreas Teufel 7 Detlef Schuppan 8 9 10 Ana Stojanovic 11 Adelheid Cerwenka 11 Stefanie Nittka 12 Albrecht Piiper 13 Timo Gaiser 14 Naiara Beraza 15 16 Malgorzata Milkiewicz 17 Piotr Milkiewicz 18 John G Brain 19 David E J Jones 19 Thomas S Weiss 20 Ulrich M Zanger 21 22 Matthias Ebert 23 Nadja M Meindl-Beinker 2
Affiliations

Affiliations

  • 1 Molecular Hepatology-Alcohol Associated Diseases, Department of Medicine II, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
  • 2 Molecular Hepatology-Alcohol Associated Diseases, Department of Medicine II, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany steven.dooley@medma.uni-heidelberg.de Nadja.Meindl-Beinker@medma.uni-heidelberg.de.
  • 3 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tanta University, Tanta, Egypt.
  • 4 Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt.
  • 5 Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany.
  • 6 Isarna Therapeutics GmbH, Munchen, Germany.
  • 7 Hepatology and Clinical Bioinformatics, Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.
  • 8 Research Center for Immunotherapy, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
  • 9 Institute of Translational Immunology, University Medical Center, Johannes Gutenberg University Mainz, Mainz, Germany.
  • 10 Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
  • 11 Department of Immunobiochemistry, Centre for Biomedicine and Medical Technology (CBTM) and European Center for Angioscience (ECAS), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
  • 12 Institute for Clinical Chemistry, Medical Faculty Mannheim of the University of Heidelberg, University Hospital Mannheim, Mannheim, Germany.
  • 13 Medizinische Klinik 1, Klinikum der Goethe-Universität, Frankfurt am Main, Germany.
  • 14 Institute of Pathology, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
  • 15 Gut Microbes and Health Institute Strategic Programme, Quadram Institute, Norwich, UK.
  • 16 CIC bioGUNE, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Derio, Spain.
  • 17 Department of Medical Biology, Pomeranian Medical University, Szczecin, Poland.
  • 18 Liver and Internal Medicine Unit, Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
  • 19 NIHR Applied Immunobiology and Transplant Research Group, Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, UK.
  • 20 Department of Pediatrics and Juvenile Medicine, Center for Liver Cell Research, University of Regensburg Hospital, Regensburg, Germany.
  • 21 Department of Molecular and Cell Biology, Dr Margarete Fischer-Bosch-Institute of Clinical Pharmacology, Stuttgart, Germany.
  • 22 Eberhard-Karls-University Tübingen, Tübingen, Germany.
  • 23 Department of Medicine II, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
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.

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