5-FGR cells (Fig. 4D), consistent with our observations in Huh7 cells (Fig. 2). This down-regulation

in HCV expression correlated with decreased mRNA levels of PPAR-α and ANGPTL3 (Fig. 4E). Our results confirm that miR-27 overexpression selleck chemical inhibits HCV replication. Interestingly, we also observed down-regulation of retinoid X receptor alpha (RXR-α), a previously reported target of miR-27 (Supporting Fig. S9A,B).[31] This protein interacts with several nuclear receptors, including PPAR-α, to regulate liver lipid biosynthesis. Therefore, we examined the functional relevance of miR-27-mediated repression of RXR-α expression on HCV replication and lipid metabolism. We performed CARS imaging on Huh7 cells treated with an RXR-α antagonist, UVI-3003, which inhibits the RXR-α’s interactions with all other nuclear receptors.[32] Huh7 cells treated with this

drug displayed no change in hepatic lipid content (Supporting Fig. S9C). Additionally, RXR-α antagonism in Huh7.5-FGR cells produced no changes in HCV levels. We were also interested in how miR-27′s regulation of PPAR-α signaling would affect viral infectivity. Selleckchem BMN 673 Previous work suggested that increased PPAR-α expression blocks assembly of HCV infectious particles.[33] Huh7.5 cells were cotransfected with JFH-1T RNA and miR-27b mimics and inhibitors, and intracellular HCV RNA levels were measured by qRT-PCR. Neither the miR-27

mimic nor the miR-27 inhibitor had any effect on JFH-1T replication (Supporting Fig. 10), suggesting that miR-27b overexpression has a genotype-specific effect on HCV replication. On the other hand, miR-27b inhibition resulted in a very modest decrease in secretion of infectious HCV, while miR-27b overexpression had no effect on secreted virus’ infectivity, consistent with PPAR-α’s previously reported antiviral role in HCV secretion.[33] Independent of miR-27′s effects on the viral lifecycle, its conserved induction across HCV genotypes manifests globally as a contributor to hepatic steatosis and thus to HCV-associated Meloxicam liver disease. We continued our evaluation of miR-27 expression in a small animal model of acute HCV infection, using the humanized SCID-beige/Alb-uPa mouse model.[34] We infected the chimeric mice with genotype 1a and 2b clinical isolates of HCV (Supporting Fig. S11). qRT-PCR analysis of miR-27b levels revealed a 2.9-fold up-regulation in miR-27b levels 7 weeks postinfection (Fig. 5A). This increase was conserved across both HCV genotypes examined. There was also a 2.0-fold increase in miR-27a levels (Fig. 5B). Oil Red O staining of lipids in the chimeric liver’s human hepatocytes revealed a correlation between cellular lipid levels and miR-27 expression in mice (Fig. 5C), and provides further support for our CARS microscopy results in cell culture experiments.