Fitness‐associated substitutions following failure of direct‐acting antivirals assessed by deep sequencing of full‐length hepatitis C virus genomes

• Published in: Alimentary pharmacology & therapeutics 2020-11, Vol.52 (10), p.1583-1591
• Main Authors: Fourati, Slim, Rodriguez, Christophe, Soulier, Alexandre, Donati, Flora, Hamadat, Sabah, Poiteau, Lila, Demontant, Vanessa, Brillet, Rozenn, Ahnou, Nazim, Gricourt, Guillaume, Chevaliez, Stéphane, Ahmed‐Belkacem, Abdelhakim, Pawlotsky, Jean‐Michel
• Format: Article
• Language: English
• Subjects: Adult; Aged; Amino acid sequence; Amino Acid Substitution; Amino Acid Substitution - drug effects; Amino Acid Substitution - genetics; Amino acids; Antiviral Agents; Antiviral Agents - therapeutic use; Cirrhosis; Cohort Studies; DNA Mutational Analysis; DNA Mutational Analysis - methods; Drug Resistance, Viral; Drug Resistance, Viral - genetics; Fitness; Genetic Fitness; Genetic Fitness - drug effects; Genome, Viral; Genome, Viral - drug effects; Genomes; Genotype; Genotype & phenotype; Genotypes; Hepacivirus; Hepacivirus - drug effects; Hepacivirus - genetics; Hepatitis; Hepatitis C; Hepatitis C, Chronic; Hepatitis C, Chronic - drug therapy; Hepatitis C, Chronic - virology; High-Throughput Nucleotide Sequencing; Humans; Life Sciences; Liver cirrhosis; Male; Metagenomics; Middle Aged; Phenotype; Polymorphism, Single Nucleotide; Remission; Replication; Sofosbuvir; Sofosbuvir - therapeutic use; Treatment Failure; Viral Nonstructural Proteins; Viral Nonstructural Proteins - genetics; Virus Replication; Virus Replication - drug effects; Virus Replication - genetics
• ISSN: 0269-2813; 1365-2036; 0269-2813
• DOI: 10.1111/apt.16054

Summary Background In hepatitis C virus (HCV) infection, treatment failure is generally associated with the selection of resistance‐associated substitutions (RAS) conferring reduced susceptibility to direct‐acting antiviral (DAA) drugs. Resistant variants continue to replicate after the end of treatment with potential for transmission. This may result from the selection of “fitness‐associated substitutions”. Aim To characterise potential “fitness‐associated substitutions” in patients infected with genotype 3a failing DAA drugs Methods By means of shotgun metagenomics, we sequenced full‐length HCV genomes at treatment initiation and at virological relapse in eight patients infected with genotype 3a with cirrhosis failing sofosbuvir and an NS5A inhibitor. The impact of amino acid changes occurring outside of DAA target regions selected in at least two patients were assessed on the in vitro susceptibility to an NS5A inhibitor and replication capacity. Results At treatment failure, besides selection of known NS5A RASs, especially Y93H, a large number of amino acid changes was observed outside of DAA target regions. We identified four amino acid positions at which observed changes substantially improved in vitro replication capacity without affecting NS5A inhibitor susceptibility. Conclusions This is the first in vivo observation combined with in vitro confirmation of selection of phenotypically characterised “fitness‐associated substitutions” together with RASs at the time of sofosbuvir‐NS5A inhibitor treatment failure in patients infected with genotype 3a with cirrhosis. Our findings may explain the persistence of resistant HCV variants after treatment in patients who did not achieve sustained virological remission.