Adenoviral CCN3/NOV gene transfer fails to mitigate liver fibrosis in an experimental bile duct ligation model because of hepatocyte apoptosis

Liver Int. 2012 Oct;32(9):1342-53. doi: 10.1111/j.1478-3231.2012.02837.x. Epub 2012 Jun 15.

Abstract

Background and aims: CCN3/NOV, a matricellular protein of the CYR61-CTGF-NOV (CCN) family, comprises six secreted proteins that associate specifically with the extracellular matrix. CCN proteins lack specific high-affinity receptors; instead, they regulate crucial biological processes, such as fibrosis, by signalling via integrins and proteoglycans. Recent studies have linked overexpression of CCN3/NOV to mitigate kidney fibrosis. This study aims to investigate CCN3/NOV overexpression in liver fibrogenesis in vivo.

Methods: The biological efficacy of adenoviral expressed CCN3/NOV directed under transcriptional control of the constitutively active Cytomegalovirus promoter (Ad-NOV) was analysed in a bile duct ligation model and in cultured primary hepatocytes.

Results and conclusions: Even though Ad-NOV gene transfer in a 3-week bile duct ligation mouse model showed the expected high levels of CCN3/NOV in both mRNA and protein, it failed to reduce liver fibrogenesis, but instead enhanced hepatocyte apoptosis. Furthermore, overexpressed CCN3/NOV in cultured primary hepatocytes resulted in decreased levels of CCN2/CTGF, the profibrotic marker protein in liver fibrosis. Both Ad-NOV and Ad-CTGF induced reactive oxygen species production, enhanced p38 and JNK activation. Therefore, we conclude that CCN3/NOV overexpression in vivo is insufficient to mitigate liver fibrogenesis because of the induction of hepatocyte injury and apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Apoptosis / physiology*
  • Cells, Cultured
  • Cholestasis, Extrahepatic / complications
  • Cholestasis, Extrahepatic / genetics
  • Cholestasis, Extrahepatic / pathology
  • Common Bile Duct / surgery
  • Disease Models, Animal
  • Disease Progression
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Vectors / genetics
  • Hepatocytes / metabolism
  • Hepatocytes / pathology*
  • Ligation
  • Liver Cirrhosis / etiology
  • Liver Cirrhosis / genetics
  • Liver Cirrhosis / pathology*
  • Mice
  • Mice, Inbred C57BL
  • Nephroblastoma Overexpressed Protein / genetics*
  • Nephroblastoma Overexpressed Protein / metabolism
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / metabolism

Substances

  • Nephroblastoma Overexpressed Protein
  • RNA, Messenger
  • Reactive Oxygen Species