TY - JOUR
T1 - Anti-HBV treatment induces novel reverse transcriptase mutations with reflective effect on HBV S antigen
AU - Cento, Valeria
AU - Van Hemert, Formijn
AU - Neumann-Fraune, Maria
AU - Mirabelli, Carmen
AU - Di Maio, Velia Chiara
AU - Salpini, Romina
AU - Bertoli, Ada
AU - Micheli, Valeria
AU - Gubertini, Guido
AU - Romano, Sara
AU - Visca, Michela
AU - De Sanctis, Giuseppe Maria
AU - Berkhout, Ben
AU - Marino, Nicoletta
AU - Mazzotta, Francesco
AU - Cappiello, Giuseppina
AU - Spanò, Alberto
AU - Sarrecchia, Cesare
AU - Ceccherini-Silberstein, Francesca
AU - Andreoni, Massimo
AU - Angelico, Mario
AU - Verheyen, Jens
AU - Perno, Carlo Federico
AU - Svicher, Valentina
PY - 2013/10
Y1 - 2013/10
N2 - Introduction: The identification of novel reverse-transcriptase (RT) drug-resistance mutations is critical in predicting the probability of success to anti-HBV treatment. Furthermore, due to HBV-RT/HBsAg gene-overlap, they can have an impact on HBsAg-detection and quantification. Methods: 356 full-length HBV-RT sequences from 197 drug-naive patients and 159 patients experiencing virological-breakthrough to nucleoside/nucleotide-analogs (NUCs) were analyzed. Mutants and wild-type HBs-antigens were expressed in HuH7-hepatocytes and quantified in cell-supernatants and cell-lysates by Architect HBsAg-assay. Results: Ten novel RT-mutations (rtN53T-rtS78T-rtS85F-rtS135T-rtA181I-rtA200V-rtK212Q-rtL229V/F-rtM309K) correlated with specific NUC-treatments and classical drug-resistance mutations on divergent evolutionary pathways. Some of them reduced RT-binding affinity for anti-HBV drugs and altered S-antigen structure. Indeed, rtS78T (prevalence: 1.1% in drug-naïve and 12.2% in adefovir-failing patients) decreased the RT-affinity for adefovir more than the classical adefovir-resistance mutations rtA181T/V (WT:-9.63kcal/mol, rtA181T:-9.30kcal/mol, rtA181V:-7.96kcal/mol, rtS78T:-7.37kcal/mol). Moreover, rtS78T introduced a stop-codon at HBsAg-position 69, and completely abrogated HBsAg-quantification in both supernatants and cell-lysates, indicating an impaired HBsAg-secretion/production. Furthermore, the HBsAg-mutation sP217L, silent in RT, significantly correlated with M204V/I-related virological-breakthrough and increased HBsAg-quantification in cell-lysate. Conclusions: Mutations beyond those classically known can affect drug-binding affinity of mutated HBV-RT, and may have potential effects on HBsAg. Their cumulative effect on resistance and HBV-pathogenicity indicates the importance of preventing therapeutic failures.
AB - Introduction: The identification of novel reverse-transcriptase (RT) drug-resistance mutations is critical in predicting the probability of success to anti-HBV treatment. Furthermore, due to HBV-RT/HBsAg gene-overlap, they can have an impact on HBsAg-detection and quantification. Methods: 356 full-length HBV-RT sequences from 197 drug-naive patients and 159 patients experiencing virological-breakthrough to nucleoside/nucleotide-analogs (NUCs) were analyzed. Mutants and wild-type HBs-antigens were expressed in HuH7-hepatocytes and quantified in cell-supernatants and cell-lysates by Architect HBsAg-assay. Results: Ten novel RT-mutations (rtN53T-rtS78T-rtS85F-rtS135T-rtA181I-rtA200V-rtK212Q-rtL229V/F-rtM309K) correlated with specific NUC-treatments and classical drug-resistance mutations on divergent evolutionary pathways. Some of them reduced RT-binding affinity for anti-HBV drugs and altered S-antigen structure. Indeed, rtS78T (prevalence: 1.1% in drug-naïve and 12.2% in adefovir-failing patients) decreased the RT-affinity for adefovir more than the classical adefovir-resistance mutations rtA181T/V (WT:-9.63kcal/mol, rtA181T:-9.30kcal/mol, rtA181V:-7.96kcal/mol, rtS78T:-7.37kcal/mol). Moreover, rtS78T introduced a stop-codon at HBsAg-position 69, and completely abrogated HBsAg-quantification in both supernatants and cell-lysates, indicating an impaired HBsAg-secretion/production. Furthermore, the HBsAg-mutation sP217L, silent in RT, significantly correlated with M204V/I-related virological-breakthrough and increased HBsAg-quantification in cell-lysate. Conclusions: Mutations beyond those classically known can affect drug-binding affinity of mutated HBV-RT, and may have potential effects on HBsAg. Their cumulative effect on resistance and HBV-pathogenicity indicates the importance of preventing therapeutic failures.
KW - HBsAg secretion
KW - HBsAg stop codon
KW - HBsAg structure
KW - HBV drug-resistance
KW - Treatment failure
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U2 - 10.1016/j.jinf.2013.05.008
DO - 10.1016/j.jinf.2013.05.008
M3 - Article
C2 - 23796863
AN - SCOPUS:84882856674
VL - 67
SP - 303
EP - 312
JO - Journal of Infection
JF - Journal of Infection
SN - 0163-4453
IS - 4
ER -