Fig 1.
Humanized-liver mouse model for HCV infection.
A. Experimental design for the creation and validation of the HCV infection model in humanized-liver chimeric MUP-uPA-SCID/Beige mice. B. Chemical structures of the NS5Ai Velpatasvir and Ledipasvir, and the CypI CRV431 and Alisporivir.
Fig 2.
Individual analysis of anti-HCV efficacy of Velpatasvir, Ledipasvir, CRV431 and Alisporivir in humanized-liver chimeric mice.
MUP-uPA-SCID-Beige mice implanted with human hepatocytes were infected i.v. with plasma from HCV-infected chimpanzee (100 infectious doses (CID50)/mL of GT1a, GT2a, GT3a and GT4a). Drugs (50 mg/kg) were given orally 9 weeks post-infection. Blood was collected retro-orbitally every week until week 15 post-HCV infection. Ten mice per treatment. Viral replication (HCV RNA copies/ml of serum) was quantified by real-time reverse transcription PCR. Error bars corresponds to the standard deviation (SD) between 10 animals. At week 12 and 15 post-infection, SD are small due to the minimal viral loads due to the drug treatment efficacy. Data are representative of two independent experiments.
Fig 3.
Combinational analysis of anti-HCV efficacy of Velpatasvir, Ledipasvir, Alisporivir and CRV431 in humanized-liver chimeric mice.
A. MUP-uPA-SCID-Beige mice implanted with human hepatocytes were infected i.v. with plasma from HCV-infected chimpanzee (100 infectious doses (CID50)/mL of GT1a, GT2a, GT3a and GT4a). Drugs were orally given alone (50 mg/kg) or in combination (50/50 mg/kg) 9 weeks post-infection. Blood was collected until week 15 post-HCV infection. Ten mice per treatment. Viral replication (HCV RNA copies/ml of serum) was quantified by real-time reverse transcription PCR. Error bars corresponds to the SD between 10 animals. At week 12 and 15 post-infection, SD are small due to the minimal viral loads due to the drug treatment efficacy. B. Statistical analyses for drug combination effect between HCV genotypes. C. Statistical analyses for Ledipasvir combination effect on HCV GT1a, GT2a, GT3a and GT4a. D. Statistical analyses for Velpatasvir combination effect on HCV GT1a, GT2a, GT3a and GT4a. For each treatment/genotype combination, we simulated 1,000 draws using the mean and standard deviation from the experimental results to account for the uncertainty in estimating the Area Under the Curve for the level of viremia. Area Under the Curve was estimated using the trapezoidal method. Groups were compared using t-tests and the permutation test. P-values were adjusted for multiple comparisons using the Bonferroni correction. Data are representative of two independent experiments.
Fig 4.
HCV rebound analysis in humanized-liver chimeric mice.
MUP-uPA-SCID-Beige mice implanted with human hepatocytes were infected i.v. with plasma from HCV-infected chimpanzee (100 infectious doses (CID50)/mL of GT1a, GT2a, GT3a and GT4a). Drugs—Velpatasvir, Ledipasvir, CRV431, Alisporivir—(50 mg/kg) were orally given alone or in combination 2 months post-infection. Ten mice per treatment. Blood was collected until month 9 post-HCV infection. Viral replication (HCV RNA copies/ml of serum) was quantified by real-time reverse transcription PCR. At week 12 and 15 post-infection, SD are small due to the minimal viral loads due to the drug treatment efficacy. Data are representative of two independent experiments.