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


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.
Institute of Clinical Chemistry and Pathobiochemistry, RWTH-University Hospital Aachen, Aachen, Germany. ekamphorst@ukaachen.de
Erawan
E
Borkham-Kamphorst
Sebastian
S
Huss
Eddy
E
Van de Leur
Ute
U
Haas
Ralf
R
Weiskirchen
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002478:Cells, Cultured; D001651:Cholestasis, Extrahepatic / Q000150:complications / Q000235:genetics / Q000473:pathology; D003135:Common Bile Duct / Q000601:surgery; D004195:Disease Models, Animal; D018450:Disease Progression; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology*; D008026:Ligation; D008103:Liver Cirrhosis / Q000209:etiology / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D055514:Nephroblastoma Overexpressed Protein / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D055514:Nephroblastoma Overexpressed Protein; D012333:RNA, Messenger; D017382:Reactive Oxygen Species