title: Development and validation of a "capture-fusion" model to study drug sensitivity of patient-derived hepatitis C.
abstract: Emerging therapies for chronic hepatitis C viral (HCV) infection involve inhibition of viral enzymes with drug combinations. Natural, or treatment-induced, enzyme polymorphisms reduce efficacy. We developed a phenotyping assay to aid drug selection based on viral transfer from monocytes to hepatocytes. We studied HCV in monocytes from infected patients and developed a model in which patient-derived HCV is "captured" by the cell line THP-1 and replication assessed after fusion to hepatoma cells. We found that monocytes from HCV-infected patients harbor virus that replicates when cells are fused to hepatocytes. THP-1 cells incubated with infected sera capture HCV, which replicates when fused to hepatocytes. Inhibitable replication of all HCV genotypes was achieved (42 of 52 isolates). We measured sensitivity of telaprevir (TVR) and alisporivir (AVR) in different genotypes, and showed differences in 50% inhibitory concentration (IC50 ) correlating with clinical response (TVR IC50 for genotype (G)1 was 0.042 ± 0.003 vs. 0.117 ± 0.015 μM for G3, whereas AVR IC50 for G1 was 0.139 ± 0.013 vs. 0.044 ± 0.007 μM for G3). We tested TVR-resistant viral isolates and identified changes in IC50 . One patient with a poor clinical response to TVR and wild-type viral sequence showed reduced TVR sensitivity in our assay. We studied samples from a 2-week TVR monotherapy study in which 5 of 8 patients with G3 HCV did not respond whereas 3 of 8 patients did. The "capture-fusion" assay correctly identified responders.
  
  
  CONCLUSIONS
  The capture-fusion model represents a promising new technique that may help identify appropriate treatment strategies for patients with chronic HCV infection.
authors.affiliation: The Liver Unit, Blizard Institute, Queen Mary University of London, London, United Kingdom.
authors.forename: Morven E
authors.initials: ME
authors.lastname: Cunningham
authors.forename: Alia
authors.initials: A
authors.lastname: Javaid
authors.forename: Jenny
authors.initials: J
authors.lastname: Waters
authors.forename: Joseph
authors.initials: J
authors.lastname: Davidson-Wright
authors.forename: Joshua L C
authors.initials: JL
authors.lastname: Wong
authors.forename: Meleri
authors.initials: M
authors.lastname: Jones
authors.forename: Graham R
authors.initials: GR
authors.lastname: Foster
mesh_terms: D002459:Cell Fusion / Q000379:methods; D016572:Cyclosporine / Q000494:pharmacology*; D024882:Drug Resistance, Viral / Q000235:genetics*; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D020128:Inhibitory Concentration 50; D008826:Microbial Sensitivity Tests; D008954:Models, Biological; D009000:Monocytes / Q000821:virology*; D009842:Oligopeptides / Q000494:pharmacology*
chemical_list: D009842:Oligopeptides; C486464:telaprevir; D016572:Cyclosporine; C499715:alisporivir