Combined adenovirus-mediated artificial microRNAs targeting mfgl2, mFas, and mTNFR1 protect against fulminant hepatic failure in mice.

Abstract

Hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) has a poor prognosis with high in-hospital mortality. Hepatic and circulating inflammatory cytokines, such as fibrinogen like protein 2 (fgl2), FasL/Fas, and TNFα/TNFR1, play a significant role in the pathophysiology of ACLF. This study aimed to investigate the therapeutic effect of recombinant adenoviral vectors carrying constructed DNA code for non-native microRNA (miRNA) targeting mouse fgl2 (mfgl2) or both mFas and mTNFR1 on murine hepatitis virus (MHV)-3-induced fulminant hepatitis in BALB/cJ mice. Artificial miRNA eukaryotic expression plasmids against mfgl2, mFas, and mTNFR1 were constructed, and their inhibitory effects on the target genes were confirmed in vitro. pcDNA6.2-mFas-mTNFR1- miRNA，which expresses miRNA against both mFas and mTNFR1 simultaneously，was constructed. To construct a miRNA adenovirus expression vector against mfgl2, pcDNA6.2-mfgl2-miRNA was cloned using Gateway technology. Ad-mFas-mTNFR1- miRNA was also constructed by the same procedure. Adenovirus vectors were delivered by tail-vein injection into MHV-3-infected BALB/cJ mice to evaluate the therapeutic effect. 8 of 18 (44.4%) mice recovered from fulminant viral hepatitis in the combined interference group treated with Ad-mfgl2-miRNA and Ad-mFas-mTNFR1-miRNA. But only 4 of 18 (22.2%) mice receiving Ad-mfgl2-miRNA and 3 of 18 (16.7%) mice receiving Ad-mFas-mTNFR1- miRNA survived. These adenovirus vectors significantly ameliorated inflammatory infiltration, fibrin deposition, hepatocyte necrosis and apoptosis, and prolonged survival time. Our data illustrated that combined interference using adenovirus-mediated artificial miRNAs targeting mfgl2, mFas, and mTNFR1 might have significant therapeutic potential for the treatment of fulminant hepatitis.

• Authors: Dong Xi, Ming Wang, Huali Ye, Xiaoping Luo, Qin Ning
• Journal: PloS one (PLoS One) Vol. 8 Issue 11 Pg. e82330 ( 2013) ISSN: 1932-6203 [Electronic] United States
• PMID: 24303082 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Fgl2 protein, mouse
• MicroRNAs
• Receptors, Tumor Necrosis Factor, Type I
• fas Receptor
• Fibrin
• Fibrinogen
• Caspase 3

Topics

• Adenoviridae (genetics)
• Animals
• Apoptosis (genetics)
• CHO Cells
• Caspase 3
• Cricetulus
• Disease Models, Animal
• Female
• Fibrin (metabolism)
• Fibrinogen (genetics, metabolism)
• Gene Expression Regulation
• Genetic Vectors (genetics)
• Liver (metabolism, pathology)
• Liver Failure, Acute (genetics, metabolism)
• Mice
• MicroRNAs (genetics, metabolism)
• Plasmids (genetics)
• RNA Interference
• Receptors, Tumor Necrosis Factor, Type I (genetics)
• fas Receptor (genetics, metabolism)