A cell-based, high-throughput screen for small molecule regulators of hepatitis C virus replication

被引:77
作者
Kim, Sun Suk
Peng, Lee F.
Lin, Wenyu
Choe, Won-Hyeok
Sakamoto, Naoya
Schreiber, Stuart L.
Chung, Raymond T.
机构
[1] Massachusetts Gen Hosp, GI Unit, Boston, MA 02114 USA
[2] Gachon Univ, Gil Med Ctr, Dept Gastroenterol & Hepatol, Inchon, South Korea
[3] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA
[4] Harvard Univ, Broad Inst, Cambridge, MA USA
[5] Tokyo Med & Dent Univ, Dept Gastroenterol & Hepatol, Tokyo 113, Japan
[6] Howard Hughes Med Inst, Chevy Chase, MD USA
关键词
D O I
10.1053/j.gastro.2006.10.032
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Background & Aims: 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/RepFeo replicon cell fine 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.
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页码:311 / 320
页数:10
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