Investigation of multidrug-resistance mutations of hepatitis B virus (HBV) in a large cohort of chronic HBV-infected patients with treatment of nucleoside/nucleotide analogs
Yan Liu 1 , Rongjuan Chen 1 , Wenhui Liu 2 , Lanlan Si 1 , Le Li 1 , Xiaodong Li 1 , ZengtaoYao 1 , Hao Liao 1 , Jun Wang 1 , Yuanhua Li 1 , Jun Zhao 3 , Dongping Xu 4
Affiliations
Affiliations
1
Department of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China.
2
Department of Gastroenterology, The Second Medical Center of Chinese PLA General Hospital, National Clinical Research Center for Geriatric Diseases, Beijing, 100853, China.
3
Department of Liver Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China. Electronic address: [email protected].
4
Department of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China. Electronic address: [email protected].
Multidrug-resistance hepatitis B virus (MDR HBV), defined as those with mutations resistant to both nucleoside analogs lamivudine/telbivudine/entecavir (LAM/LdT/ETV) and nucleotide analog adefovir (ADV), has potential to cause treatment difficulty. To clarify clinical prevalence and virological features of MDR HBV, we investigated serum samples from 28,236 chronic HBV-infected patients with treatment of nucleoside/nucleotide analogs. All patients underwent resistance testing in the Fifth Medical Center of Chinese PLA General Hospital between 2007 and 2019. MDR mutations were screened by direct sequencing; MDR strains (with mutations co-located on the same viral genome) were verified by clonal sequencing (≥20 clones/sample) and subjected to phenotypic analysis if necessary. MDR mutations were detected in 0.81% (229/28,236) patients. MDR strains were verified in 83.0% (190/229) of MDR mutation-positive patients. As ETV-resistance mutation (ETVr) had additional mutation(s) on LAMr conferring more resistance, MDR mutations fell into LAMr+ADVr and ETVr+ADVr subsets. Sixteen mutation patterns of MDR strains were verified, including eight with LAMr+ADVr and eight with ETVr+ADVr. Refractory to sequential therapies of LAM/LdT/ETV and ADV were closely linked with MDR HBV development. Ten representative MDR strains (five LAMr+ADVr and five ETVr+ADVr) tested all had decrease in replication capacity compared to wild-type strains and decrease extent was positively related with the number of primary resistance on viral genome. Compared to ADV+ETV, TDF/TDF+ETV showed higher inhibitory rates on MDR HBV, especially for the five ETVr+ADVr strains (74.5%-97.6% vs. 60.2%-79.5%, all P < 0.05). This study significantly extends the knowledge on MDR HBV and has clinical implications for resistance management.
Keywords: Hepatitis B virus; entecavir; inhibitory rates; multidrug-resistance; mutation.