A cell-based, high-throughput screen for small molecule regulators of hepatitis C virus replication.
OBJECTIVE
Only half of patients with chronic hepatitis C virus (HCV) infection experience sustained virologic response to pegylated-interferon and ribavirin, which cause numerous side effects. Thus, the identification of more effective and better tolerated agents is a high priority. We applied chemical biology to screen small molecules that regulate HCV.


METHODS
We first optimized the Huh7/Rep-Feo replicon cell line for the 384-well microplate format and used this line to screen a large library of well-characterized, known biologically active compounds using automated technology. After identifying several molecules capable of either stimulating or inhibiting HCV replication in this primary screen, we then validated our hit compounds using a full-length HCV replicon cell line in secondary screens.


RESULTS
We identified and validated a number of antiviral and proviral agents, including HMG-CoA reductase inhibitors (antiviral) and corticosteroids (proviral). The finding of increased replication associated with corticosteroids suggests that these agents directly promote viral replication independent of their suppressive effects on the immune response. The finding of antiviral activity associated with the HMG-CoA reductase inhibitors implies an important role for lipid metabolism in the viral life cycle.


CONCLUSIONS
We have developed a simple, reproducible, and reliable cell-based high-throughput screening assay system using an HCV replicon model to identify small molecules that regulate HCV replication. This method can be used to identify not only putative antiviral agents, but also cellular regulators of viral replication.
Kim
Sun Suk
SS
Gastrointestinal Unit, Department of Medicine, Massachusetts General Hospital, Boston, MA 02114, USA.
Peng
Lee F
LF
Lin
Wenyu
W
Choe
Won-Hyeok
WH
Sakamoto
Naoya
N
Kato
Nobuyuki
N
Ikeda
Masanori
M
Schreiber
Stuart L
SL
Chung
Raymond T
RT
D000305:Adrenal Cortex Hormones / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D016174:Hepacivirus / Q000187:drug effects* / Q000254:growth & development; D006526:Hepatitis C / Q000188:drug therapy* / Q000821:virology*; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology; D008113:Liver Neoplasms; D011533:Proviruses / Q000187:drug effects / Q000254:growth & development; D012093:Replicon / Q000187:drug effects / Q000502:physiology; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
D000305:Adrenal Cortex Hormones; D000998:Antiviral Agents; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors