Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA.
OBJECTIVE
We have reported previously that synthetic small interfering RNA (siRNA) and DNA-based siRNA expression vectors efficiently and specifically suppress hepatitis C virus (HCV) replication in vitro. In this study, we investigated the effects of the siRNA targeting HCV-RNA in vivo.


METHODS
We constructed recombinant retrovirus and adenovirus expressing short hairpin RNA (shRNA), and transfected into replicon-expressing cells in vitro and transgenic mice in vivo.


RESULTS
Retroviral transduction of Huh7 cells to express shRNA and subsequent transfection of an HCV replicon into the cells showed that the cells had acquired resistance to HCV replication. Infection of cells expressing the HCV replicon with an adenovirus expressing shRNA resulted in efficient vector delivery and expression of shRNA, leading to suppression of the replicon in the cells by approximately 10(-3). Intravenous delivery of the adenovirus expressing shRNA into transgenic mice that can be induced to express HCV structural proteins by the Cre/loxP switching system resulted in specific suppression of virus protein synthesis in the liver.


CONCLUSIONS
Taken together, our results support the feasibility of utilizing gene targeting therapy based on siRNA and/or shRNA expression to counteract HCV replication, which might prove valuable in the treatment of hepatitis C.
Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University, Tokyo, Japan. nsakamoto.gast@tmd.ac.jp
Naoya
N
Sakamoto
Yoko
Y
Tanabe
Takanori
T
Yokota
Kenichi
K
Satoh
Yuko
Y
Sekine-Osajima
Mina
M
Nakagawa
Yasuhiro
Y
Itsui
Megumi
M
Tasaka
Yuki
Y
Sakurai
Chen
C
Cheng-Hsin
Masahiko
M
Yano
Shogo
S
Ohkoshi
Yutaka
Y
Aoyagi
Shinya
S
Maekawa
Nobuyuki
N
Enomoto
Michinori
M
Kohara
Mamoru
M
Watanabe
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005240:Feasibility Studies; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D006526:Hepatitis C / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control*; D006801:Humans; D050835:Interferon Regulatory Factors / Q000235:genetics; D008099:Liver / Q000378:metabolism / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D012367:RNA, Viral; D012190:Retroviridae / Q000235:genetics; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014779:Virus Replication
D050835:Interferon Regulatory Factors; D034741:RNA, Small Interfering; D012367:RNA, Viral; D014764:Viral Proteins