Employment: Gilead Sciences; Patent Held/Filed: Gilead Sciences; Stock Shareholder: Gilead Sciences, Pfizer Hongmei Mo – Employment: Gilead Science Inc The following people have nothing to disclose: Viktoria Gontcharova Background: A recently discovered novel surface receptor involved in EX 527 HCV entry, the Niemann-Pick C1-like 1 cholesterol (NPC1L1), is an HCV entry factor amendable to therapeutic intervention. Previously, DNA sequencing studies revealed that nonsynonymous variants in NPC1L1 are collectively common in general population. The aim of the present study was to elucidate whether genetic variants of the NPC1L1 are linked to the antiviral response in a group of patients with Pexidartinib cell line chronic hepatitis C (CHC). Methods: We included 38 patients with CHC genotype 1 treated with pegylated interferon alpha2 and ribavirin (20 patients with SVR and 18 null responders). The whole coding sequence of NPC1L1 gene was amplified by 30 different
PCR reactions. PCR products were barcoded and sequenced in a Junior-454 deep-sequencing platform (Roche). The resulting reads were aligned against the human genome (GRCh37) using BLAT algorithm and the variants were identified using GATK Unified genotyper. The functional consequence of the
sequence variants were determined using SNPEff algorithm and association studies with patient phenotypes were performed using Plink suite. Results: 24 different sequence variants in NPC1L1 gene were identified in total in the 38 patients sequenced. 15 were small insertions and deletions (indels), whereas 9 of them constitute single nucleotide variants (SNV), from which 6 had been already Uroporphyrinogen III synthase reported in dbSNP database. According to a negative selection of deleterious sequence variants in the normal population, most of the identified variants were predicted to play synonymous or regulatory roles in the gene. Nevertheless, even with this limited sample size, association studies shown significative association between two of the sequence variants (a single nucleotide substitution and a single base deletion) with therapy response (rs186726309 and a novel SNV, Chr7: 44575955Del(G)). Additionally, a new SNV has been found which is not present in dbSNP database (Crh7: 44561370) and is present in a low frequency in our cohort; and produces a significant aminoacid change (S919C) in the coding region of the gene. Conclusions.