Bioluminescence imaging allows monitoring hepatitis C virus core protein inhibitors in mice.
BACKGROUND
The development of small molecule inhibitors of hepatitis C virus (HCV) core protein as antiviral agents has been intensively pursued as a viable strategy to eradicate HCV infection. However, lack of a robust and convenient small animal model has hampered the assessment of in vivo efficacy of any antiviral compound.


RESULTS
The objective of this work was to develop a novel method to screen anti-core protein siRNA in the mouse liver by bioluminescence imaging. The inhibitory effect of two shRNAs targeting the highly conserved core region of the HCV genome, shRNA452 and shRNA523, was examined using this method. In the transient mouse model, the effect of shRNA-523 was detectable at as early as 24 h and became even more pronounced at later time points. The effect of shRNA-452 was not detectable until 48 h post-transduction. In a stable mouse model, shRNA523 reduced luciferase levels by up to 76.4±26.0% and 91.8±8.0% at 6 h and 12 h after injection respectively, and the inhibitory effect persisted for 1 day after a single injection while shRNA-Scramble did not seem to have an effect on the luciferase activity in vivo.


CONCLUSIONS
Thus, we developed a simple and quantitative assay for real-time monitoring of HCV core protein inhibitors in mice.
Beijing Institute of Transfusion Medicine, Beijing, China.
Juan
J
Du
Fang
F
Zhao
Yong
Y
Zhou
Hu
H
Yan
Xiang-guo
XG
Duan
Sheng-qiang
SQ
Liang
Ying-li
YL
Wang
Qiu-xia
QX
Fu
Xiao-hui
XH
Wang
Jian-chun
JC
Peng
Lin-sheng
LS
Zhan
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D053583:Interleukin-1beta / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D008099:Liver / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D013997:Time Factors; D014162:Transfection / Q000379:methods; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*
D053583:Interleukin-1beta; D015850:Interleukin-6; D034741:RNA, Small Interfering; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D008156:Luciferases; D000410:Alanine Transaminase