|
1
|
Belaiba Z, Ayouni K, Gdoura M, Kammoun Rebai W, Touzi H, Sadraoui A, Hammemi W, Yacoubi L, Abdelati S, Hamzaoui L, Msaddak Azzouz M, Chouikha A, Triki H. Whole genome analysis of hepatitis B virus before and during long-term therapy in chronic infected patients: Molecular characterization, impact on treatment and liver disease progression. Front Microbiol 2022; 13:1020147. [PMID: 36325017 PMCID: PMC9618822 DOI: 10.3389/fmicb.2022.1020147] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/15/2022] [Accepted: 09/15/2022] [Indexed: 07/23/2023] Open
Abstract
Hepatitis B virus (HBV) infection remains a serious public health concern worldwide despite the availability of an efficient vaccine and the major improvements in antiviral treatments. The aim of the present study is to analyze the mutational profile of the HBV whole genome in ETV non-responder chronic HBV patients, in order to investigate antiviral drug resistance, immune escape, and liver disease progression to Liver Cirrhosis (LC) or Hepatocellular Carcinoma (HCC). Blood samples were collected from five chronic hepatitis B patients. For each patient, two plasma samples were collected, before and during the treatment. Whole genome sequencing was performed using Sanger technology. Phylogenetic analysis comparing the studied sequences with reference ones was used for genotyping. The mutational profile was analyzed by comparison with the reference sequence M32138. Genotyping showed that the studied strains belong to subgenotypes D1, D7, and D8. The mutational analysis showed high genetic variability. In the RT region of the polymerase gene, 28 amino acid (aa) mutations were detected. The most significant mutations were the pattern rtL180M + rtS202G + rtM204V, which confer treatment resistance. In the S gene, 35 mutations were detected namely sP120T, sT126S, sG130R, sY134F, sS193L, sI195M, and sL216stop were previously described to lead to vaccine, immunotherapy, and/or diagnosis escape. In the C gene, 34 mutations were found. In particular, cG1764A, cC1766G/T, cT1768A, and cC1773T in the BCP; cG1896A and cG1899A in the precore region and cT12S, cE64D, cA80T, and cP130Q in the core region were associated with disease progression to LC and/or HCC. Other mutations were associated with viral replication increase including cT1753V, cG1764A/T, cC1766G/T, cT1768A, and cC1788G in the BCP as well as cG1896A and cG1899A in the precore region. In the X gene, 30 aa substitutions were detected, of which substitutions xT36D, xP46S, xA47T, xI88F, xA102V, xI127T, xK130M, xV131I, and xF132Y were previously described to lead to LC and/or HCC disease progression. In conclusion, our results show high genetic variability in the long-term treatment of chronic HBV patients causing several effects. This could contribute to guiding national efforts to optimize relevant HBV treatment management in order to achieve the global hepatitis elimination goal by 2030.
Collapse
Affiliation(s)
- Zeineb Belaiba
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
- Research Laboratory “Virus, Vectors and Hosts: One Health Approach and Technological Innovation for a Better Health,” LR20IPT02, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Kaouther Ayouni
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
- Research Laboratory “Virus, Vectors and Hosts: One Health Approach and Technological Innovation for a Better Health,” LR20IPT02, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Mariem Gdoura
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
- Research Laboratory “Virus, Vectors and Hosts: One Health Approach and Technological Innovation for a Better Health,” LR20IPT02, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Wafa Kammoun Rebai
- Laboratory of Biomedical Genomics and Oncogenetics (LR16IPT05), Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Henda Touzi
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Amel Sadraoui
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Walid Hammemi
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Lamia Yacoubi
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Salwa Abdelati
- Department of Gastroenterology, Polyclinic of CNSS, Sousse, Tunisia
| | - Lamine Hamzaoui
- Department of Gastroenterology, Hospital of Tahar Maamouri, Nabeul, Tunisia
| | | | - Anissa Chouikha
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
- Research Laboratory “Virus, Vectors and Hosts: One Health Approach and Technological Innovation for a Better Health,” LR20IPT02, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| | - Henda Triki
- Laboratory of Clinical Virology, WHO Reference Laboratory for Poliomyelitis and Measles in the Eastern Mediterranean Region, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
- Research Laboratory “Virus, Vectors and Hosts: One Health Approach and Technological Innovation for a Better Health,” LR20IPT02, Pasteur Institute of Tunis, University Tunis El Manar (UTM), Tunis, Tunisia
| |
Collapse
|
|
2
|
Chen R, Liu Y, Luo D, Si L, Huang B, Wang J, Li X, Cheng F, Xu D, Duan C. Hepatitis B virus mutation pattern rtA181S+T184I+M204I may contribute to multidrug resistance in clinical practice: Analysis of a large cohort of Chinese patients. Antiviral Res 2020; 180:104852. [PMID: 32569703 DOI: 10.1016/j.antiviral.2020.104852] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2019] [Revised: 05/15/2020] [Accepted: 06/09/2020] [Indexed: 02/07/2023]
Abstract
The study aimed to characterize the prevalence and virological features of the rtA181S + T184I + M204I mutant in a large cohort of patients with chronic HBV infection. In total, 22,009 nucleoside/nucleotide analog-treated patients who underwent resistance testing at the Fifth Medical Center of Chinese PLA General Hospital between 2007 and 2016 were enrolled. Serum samples were collected for HBV reverse-transcriptase gene sequencing. Phenotypic analysis of the viral replication capacity and drug susceptibility was performed. The rtA181S mutation was detected in 0.82% (180/22,009) of samples. rtA181S-positive patients had significantly higher lamivudine (LAM), adefovir (ADV), and entecavir (ETV) exposure than rtA181S-negative patients. Of 180 rtA181S-positive patients, 42 had no coexistent resistance mutations, 34 had coexisting LAM-resistance mutation (LAMr), 17 had coexisting ADV-resistance mutation (ADVr), and 86 had coexisting ETV-resistance mutation (ETVr), and one had ADVr + ETVr. rtA181S + T184I + M204I occurred in 79.1% (68/86) of patients with rtA181S + ETVr and 37.8% (68/180) of all rtA181S-positive patients. Longitudinal analysis of the clinical course of resistant mutant evolution for four representative cases showed that rtA181S + T184I + M204I developed in all patients who had received LAM/telbivudine ± ADV and was receiving ETV or ADV + ETV. Compared with wild-type, the rtA181S + T184I + M204I mutant had 53.7% lower replication capacity and >1000-, 3.9-, and 383.3-fold greater LAM, ADV, and ETV resistance, respectively, but remained sensitive to tenofovir. Artificial elimination of rtA181S from the rtA181S + T184I + M204I mutant restored viral susceptibility to ADV but decreased viral replication capacity. Our study presented the first evidence that HBV rtA181S + T184I + M204I mutation had features of multidrug-resistance that contributed to resistance to both nucleoside and nucleotide analogs.
Collapse
Affiliation(s)
- Rongjuan Chen
- Department of Cell Biology and Genetics, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, 400016, China; Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Yan Liu
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Dan Luo
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Lanlan Si
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Bixia Huang
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Jun Wang
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Xiaodong Li
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China
| | - Fengjuan Cheng
- Department of Cell Biology and Genetics, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, 400016, China
| | - Dongping Xu
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100039, China.
| | - Changzhu Duan
- Department of Cell Biology and Genetics, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, 400016, China.
| |
Collapse
|
|
3
|
Hossain MG, Mahmud MM, Nazir KHMNH, Ueda K. PreS1 Mutations Alter the Large HBsAg Antigenicity of a Hepatitis B Virus Strain Isolated in Bangladesh. Int J Mol Sci 2020; 21:ijms21020546. [PMID: 31952213 PMCID: PMC7014173 DOI: 10.3390/ijms21020546] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/12/2019] [Revised: 01/04/2020] [Accepted: 01/13/2020] [Indexed: 02/07/2023] Open
Abstract
Mutations in the hepatitis B virus (HBV) genome can potentially lead to vaccination failure, diagnostic escape, and disease progression. However, there are no reports on viral gene expression and large hepatitis B surface antigen (HBsAg) antigenicity alterations due to mutations in HBV isolated from a Bangladeshi population. Here, we sequenced the full genome of the HBV isolated from a clinically infected patient in Bangladesh. The open reading frames (ORFs) (P, S, C, and X) of the isolated HBV strain were successfully amplified and cloned into a mammalian expression vector. The HBV isolate was identified as genotype C (sub-genotype C2), serotype adr, and evolutionarily related to strains isolated in Indonesia, Malaysia, and China. Clinically significant mutations, such as preS1 C2964A, reverse transcriptase domain I91L, and small HBsAg N3S, were identified. The viral P, S, C, and X genes were expressed in HEK-293T and HepG2 cells by transient transfection with a native subcellular distribution pattern analyzed by immunofluorescence assay. Western blotting of large HBsAg using preS1 antibody showed no staining, and preS1 ELISA showed a significant reduction in reactivity due to amino acid mutations. This mutated preS1 sequence has been identified in several Asian countries. To our knowledge, this is the first report investigating changes in large HBsAg antigenicity due to preS1 mutations.
Collapse
Affiliation(s)
- Md. Golzar Hossain
- Division of Virology, Department of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan
- Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh; (M.M.M.); (K.H.M.N.H.N.)
- Correspondence: (M.G.H.); (K.U.)
| | - Md. Muket Mahmud
- Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh; (M.M.M.); (K.H.M.N.H.N.)
| | - K. H. M. Nazmul Hussain Nazir
- Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh; (M.M.M.); (K.H.M.N.H.N.)
| | - Keiji Ueda
- Division of Virology, Department of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan
- Correspondence: (M.G.H.); (K.U.)
| |
Collapse
|
|
4
|
Luo D, Liu Y, Chen R, Niu M, Liu L, Li X, Li Q, Huang B, Wang J, Xu D, Lin S. Investigation of hepatitis B virus (HBV) rtS78T/sC69* mutation in a large cohort of chronic HBV-infected patients with nucleoside/nucleotide analogue treatment. Antiviral Res 2019; 170:104579. [PMID: 31398372 DOI: 10.1016/j.antiviral.2019.104579] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/27/2019] [Revised: 07/29/2019] [Accepted: 08/05/2019] [Indexed: 02/07/2023]
Abstract
This study aimed to investigate clinical occurrence and significance of the rtS78T/sC69* mutation of hepatitis B virus (HBV). A total of 22,009 consecutive chronic HBV-infected patients who underwent resistance testing at the Fifth Medical Center of Chinese PLA General Hospital (Original name Beijing 302 Hospital) from 2007 to 2016 were enrolled. Serum samples were collected for sequence analysis of HBV reverse-transcriptase (RT) and S regions. Phenotypic analysis was performed to evaluate the viral replication capacity and drug susceptibility. The rtS78T mutation was detected in 0.83% (182/22,009) of the patients' samples. All mutations simultaneously created a stop codon at sC69 (sC69*). The prevalence of rtS78T/sC69* did not differ significantly between the patients with and without entecavir/tenofovir treatment. Of the 182 mutation-positive samples, 41 (22.5%) were detected with signature drug-resistance mutations to adefovir (n = 26), lamivudine (n = 11), entecavir (n = 3), and lamivudine plus adefovir (n = 1). The HBV DNA and RNA levels of the rtS78T/sC69* mutant were significantly increased compared to the wild-type; while the mutant had undetectable secreted and intracellular HBsAg, and its half maximal effective concentration to lamivudine, adefovir, entecavir, and tenofovir were 3.73-, 1.61-, 4.76-, and 3.71-fold of the wild-type, respectively. Artificial elimination of the rtS78T mutation had a limited effect on the drug susceptibilities. The data obtained in the present study suggested that the emergence of the rtS78T/sC69* mutation was not closely related to entecavir/tenofovir treatment and itself appeared insufficient to confer drug resistance unless it coexisted with signature drug-resistance mutations.
Collapse
Affiliation(s)
- Dan Luo
- Department of Infectious Diseases, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China; Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Yan Liu
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Rongjuan Chen
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Ming Niu
- Institute of Chinese Medicine, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Lujie Liu
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Xiaodong Li
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Qi Li
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Bixia Huang
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Jun Wang
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China
| | - Dongping Xu
- Institute of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital (Beijing 302 Hospital), Beijing, 100039, China.
| | - Shumei Lin
- Department of Infectious Diseases, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.
| |
Collapse
|
|
5
|
Liu Y, Zhou Y, Li X, Niu M, Chen R, Shao J, Si L, Luo D, Lin Y, Li L, Zhang K, Xiao X, Xu Z, Liu M, Lu M, Zoulim F, Xu D. Hepatitis B virus mutation pattern rtL180M+A181C+M204V may contribute to entecavir resistance in clinical practice. Emerg Microbes Infect 2019; 8:354-365. [PMID: 30866789 PMCID: PMC6455135 DOI: 10.1080/22221751.2019.1584018] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
Background and Aims: Entecavir (ETV) resistance of hepatitis B virus (HBV) conventionally requires rt184, 202, or 250 mutations plus lamivudine-resistance mutation (rtM204V/I ± L180M). This study aimed to clarify whether rtL180M+A181C+M204V mutations may contribute to HBV ETV resistance. Methods: Serum samples were collected from 22,009 patients who underwent resistance testing in Beijing 302 Hospital from 2007 to 2016. HBV reverse transcriptase (RT) gene was screened by direct sequencing and verified by clonal sequencing. Phenotypic analysis was performed for evaluating replication capacity and drug susceptibility. Results: Classical ETV-resistance mutations of HBV were detected in 1252 patients who were receiving ETV therapy. The rtA181C mutation was detected with rtL180M+M204V mutations in 18 lamivudine-experienced ETV-treated patients, and the emergence of the mutations was associated with virological breakthrough or inadequate virological response to ETV. Patient-derived representative rtA181C-containing mutants, rtL180M+A181C+M204V, rtL180M+A181C+M204V+M250V, and rtL180M+A181C+S202G+M204V, exhibited 45.7%, 25.9%, and 25.0% replication capacity and 85.6-, 356.1-, and 307.1-fold decreased susceptibility to ETV respectively compared to the wild-type strain, while the three mutants remained sensitive to tenofovir (TDF). Artificial elimination of rtA181C largely restored the rtL180M+A181C+M204V mutant’s sensitivity to ETV. Molecular modelling of viral RT binding to ETV showed that the rtL180M+A181C+M204V mutant had a less stable conformation compared to rtL180M+M204V mutant. In clinical practice, undetectable serum HBV DNA was achieved in two of five longitudinally followed rtA181C-positive patients who received switching-to TDF therapy, but not in the other three who received add-on adefovir therapy during observation. Conclusions: Both clinical and experimental data support rtL180M+A181C+M204V as a novel non-classical ETV-resistance mutation pattern.
Collapse
Affiliation(s)
- Yan Liu
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Yi Zhou
- b Department of Infectious Diseases , The First Affiliated Hospital of Xi'an Jiaotong University , Xi'an , People's Republic of China
| | - Xiaodong Li
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Ming Niu
- c Institute of Chinese Medicine , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Rongjuan Chen
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Jinman Shao
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Lanlan Si
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Dan Luo
- b Department of Infectious Diseases , The First Affiliated Hospital of Xi'an Jiaotong University , Xi'an , People's Republic of China
| | - Yayun Lin
- b Department of Infectious Diseases , The First Affiliated Hospital of Xi'an Jiaotong University , Xi'an , People's Republic of China
| | - Le Li
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Kai Zhang
- b Department of Infectious Diseases , The First Affiliated Hospital of Xi'an Jiaotong University , Xi'an , People's Republic of China
| | - Xiaohe Xiao
- c Institute of Chinese Medicine , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Zhihui Xu
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| | - Min Liu
- b Department of Infectious Diseases , The First Affiliated Hospital of Xi'an Jiaotong University , Xi'an , People's Republic of China
| | - Mengji Lu
- d Institute of Virology , University Hospital of Essen, University of Duisburg-Essen , Essen , Germany
| | - Fabien Zoulim
- e Univ Lyon, Universite Claude Bernard Lyon 1, INSERM 1052, CNRS 5286, Centre Leon Berard, Centre de recherche en cancerologie de Lyon , Lyon , France.,f Department of Hepatology, Groupement Hospitalier Nord , Hospices Civils de Lyon , Lyon , France
| | - Dongping Xu
- a Institute of Infectious Diseases , Beijing 302 Hospital , Beijing , People's Republic of China
| |
Collapse
|
|
6
|
Zhang X, Chen X, Wei M, Zhang C, Xu T, Liu L, Xu Z. Potential resistant mutations within HBV reverse transcriptase sequences in nucleos(t)ide analogues-experienced patients with hepatitis B virus infection. Sci Rep 2019; 9:8078. [PMID: 31147594 PMCID: PMC6542804 DOI: 10.1038/s41598-019-44604-6] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2019] [Accepted: 05/16/2019] [Indexed: 12/17/2022] Open
Abstract
This study was performed to analyze the potential resistant mutations within HBV reverse transcriptase (RT) sequences against nucleos(t)ide analogues (NA). HBV DNA RT region spanning from amino acid 169 to 250 was amplified and sequenced from 435 HBV patients who experienced NA treatment. Among study’s cohort, genotypes B and C infected patients were 55.9% and 44.1%, respectively. Mutations were recorded in 54.7% (238/435) patients at 22 positions. Genotype C displayed significant higher frequency of potential NA resistant mutations than genotype B (63.0% vs. 48.1%, P = 0.003). Moreover, eight mutation sites, including 180, 181, 191, 200, 202, 221, 229 and 224, in genotype C showed significant higher frequencies than in genotype B. In contrast, mutation at site 236 was more common in genotype B. Notably, 11 mutations at position 169, 202, 250, 173, 180, 200, 207, 214, 237, 242 and 245 coexisted with M204I or V. Substitutions at nine non-classical mutation sites (191, 207, 213, 218, 221, 224, 229, 238 and 242) were detected in patients with virological breakthrough. Particularly, tenofovir (TDF) resistance was observed in one patient undergoing TDF monotherapy and experienced several NA treatment before. These results might provide clinical useful information under antiviral therapy.
Collapse
Affiliation(s)
- Xiaoman Zhang
- Clinical Liver Center, the 910th hospital of People's Liberation Army, Quanzhou, 362000, China
| | - Xianli Chen
- Department of Infectious and Liver Disease, Xiang'an hospital of Xiamen University, Xiamen, 361000, China
| | - Meijuan Wei
- Clinical Liver Center, the 910th hospital of People's Liberation Army, Quanzhou, 362000, China.,Clinical Liver Center, Decheng hospital of Quanzhou Affiliated of Huaqiao University, Quanzhou, 362000, China
| | - Chunyu Zhang
- Clinical Liver Center, the 910th hospital of People's Liberation Army, Quanzhou, 362000, China
| | - Tao Xu
- Clinical Liver Center, the 910th hospital of People's Liberation Army, Quanzhou, 362000, China
| | - Liguan Liu
- Clinical Liver Center, the 910th hospital of People's Liberation Army, Quanzhou, 362000, China
| | - Zhengju Xu
- Clinical Liver Center, the 910th hospital of People's Liberation Army, Quanzhou, 362000, China.
| |
Collapse
|
|
7
|
Liu L, Liu Y, Chen R, Li X, Luo D, Zhao Y, Li Q, Huang B, Wang FS, Liu X, Xu D. Prevalence of the entecavir-resistance-inducing mutation rtA186T in a large cohort of Chinese hepatitis B virus patients. Antiviral Res 2019; 164:131-138. [PMID: 30796932 DOI: 10.1016/j.antiviral.2019.02.012] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/02/2018] [Revised: 01/29/2019] [Accepted: 02/19/2019] [Indexed: 02/07/2023]
Abstract
This study aimed to clarify whether rtA186T and rtI163V substitutions of hepatitis B virus (HBV) contributed to entecavir (ETV) resistance. A total of 22,009 Chinese patients with chronic HBV infection who received resistance testing at Beijing 302 Hospital from 2007 to 2016 were enrolled. Among them, 6170 patients had been treated with ETV. The HBV reverse transcriptase gene was screened by direct sequencing and verified by clonal sequencing. Phenotypic analysis was performed for evaluating replication capacity and drug susceptibility. Classical ETV-resistance mutations rtT184/S202/M250substitution+rtM204V/I±L180M (LAM-r), rtA186T, and rtI163V were detected in 1252 (5.69%), 14 (0.06%), and 230 (1.05%) of the 22,009 patients, respectively. The rtA186T mutation always coexisted with LAM-r, but not with rtI163V. The 14 rtA186T-positive patients were all treated with LAM and ETV, and the emergence of the rtA186T+LAM-r was closely associated with virological breakthrough or inadequate virological response to ETV. By contrast, the emergence of rtI163V was not related to ETV treatment. Six rtA186T-positive patients were followed up longitudinally, showing that these patients all had received sequential adefovir and LAM monotherapies prior to ETV treatment. Compared to wild-type strain, two patient-derived mutants' rtL180M+A186T+M204V and rtL180M+T184S+A186T+M204V had 86.7% and 89.2% decreased replication capacity, 210- and 555-fold increased ETV resistance, respectively; and artificial elimination of rtA186T largely restored their ETV sensitivity. The rtA186T mutants remained sensitive to tenofovir. In conclusion, our study confirmed that rtA186T plus LAM-r is a novel ETV-resistance mutation pattern which conferred ETV resistance in multiple Chinese patients.
Collapse
Affiliation(s)
- Lujie Liu
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China; Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics/Institute of Aging Research, Guangdong Medical University, Dongguan 523808, Guangdong Province, China
| | - Yan Liu
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China
| | - Rongjuan Chen
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China
| | - Xiaodong Li
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China
| | - Dan Luo
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China
| | - Yangyang Zhao
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China; Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics/Institute of Aging Research, Guangdong Medical University, Dongguan 523808, Guangdong Province, China
| | - Qi Li
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China
| | - Bixia Huang
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China; Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics/Institute of Aging Research, Guangdong Medical University, Dongguan 523808, Guangdong Province, China
| | - Fu-Sheng Wang
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China
| | - Xinguang Liu
- Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics/Institute of Aging Research, Guangdong Medical University, Dongguan 523808, Guangdong Province, China.
| | - Dongping Xu
- Institute of Infectious Diseases, Beijing 302 Hospital/The Fifth Medical Center of PLA General Hospital, Beijing 100039, China; Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics/Institute of Aging Research, Guangdong Medical University, Dongguan 523808, Guangdong Province, China.
| |
Collapse
|
|
8
|
Choi YM, Lee SY, Kim BJ. Naturally occurring hepatitis B virus reverse transcriptase mutations related to potential antiviral drug resistance and liver disease progression. World J Gastroenterol 2018; 24:1708-1724. [PMID: 29713126 PMCID: PMC5922991 DOI: 10.3748/wjg.v24.i16.1708] [Citation(s) in RCA: 31] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/27/2018] [Revised: 04/10/2018] [Accepted: 04/16/2018] [Indexed: 02/06/2023] Open
Abstract
The annual number of deaths caused by hepatitis B virus (HBV)-related disease, including cirrhosis and hepatocellular carcinoma (HCC), is estimated as 887000. The reported prevalence of HBV reverse transcriptase (RT) mutation prior to treatment is varied and the impact of preexisting mutations on the treatment of naïve patients remains controversial, and primarily depends on geographic factors, HBV genotypes, HBeAg serostatus, HBV viral loads, disease progression, intergenotypic recombination and co-infection with HIV. Different sensitivity of detection methodology used could also affect their prevalence results. Several genotype-dependent HBV RT positions that can affect the emergence of drug resistance have also been reported. Eight mutations in RT (rtL80I, rtD134N, rtN139K/T/H, rtY141F, rtM204I/V, rtF221Y, rtI224V, and rtM309K) are significantly associated with HCC progression. HBeAg-negative status, low viral load, and genotype C infection are significantly related to a higher frequency and prevalence of preexisting RT mutations. Preexisting mutations are most frequently found in the A-B interdomain of RT which overlaps with the HBsAg "a" determinant region, mutations of which can lead to simultaneous viral immune escape. In conclusion, the presence of baseline RT mutations can affect drug treatment outcomes and disease progression in HBV-infected populations via modulation of viral fitness and host-immune responses.
Collapse
Affiliation(s)
- Yu-Min Choi
- Department of Microbiology and Immunology, Biomedical Sciences, Liver Research Institute and Cancer Research Institute, Seoul National University, College of Medicine, Seoul 110799, South Korea
| | - So-Young Lee
- Department of Microbiology and Immunology, Biomedical Sciences, Liver Research Institute and Cancer Research Institute, Seoul National University, College of Medicine, Seoul 110799, South Korea
| | - Bum-Joon Kim
- Department of Microbiology and Immunology, Biomedical Sciences, Liver Research Institute and Cancer Research Institute, Seoul National University, College of Medicine, Seoul 110799, South Korea
| |
Collapse
|
|
9
|
Kimberlin DW. Antiviral Agents. PRINCIPLES AND PRACTICE OF PEDIATRIC INFECTIOUS DISEASES 2018:1551-1567.e6. [DOI: 10.1016/b978-0-323-40181-4.00295-4] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
10
|
Fung J, Wong T, Chok K, Chan A, Sin SL, Cheung TT, Dai WC, Ng K, Ng K, Man K, Seto WK, Lai CL, Yuen MF, Lo CM. Oral Nucleos(t)ide Analogs Alone After Liver Transplantation in Chronic Hepatitis B With Preexisting rt204 Mutation. Transplantation 2017; 101:2391-2398. [PMID: 28731907 DOI: 10.1097/tp.0000000000001883] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022]
Abstract
BACKGROUND There is currently limited data regarding the use of oral antiviral therapy alone without hepatitis B immune globulin for chronic hepatitis B patients with preexisting lamivudine (LAM) resistance (LAM-R) undergoing liver transplantation. METHODS This is a cohort study determining the effectiveness and long-term outcome in this group of patients. RESULTS Fifty-seven consecutive chronic hepatitis B patients with preexisting rt204 LAM-R mutations or virological load refractory to LAM undergoing liver transplantation were included, with a median follow-up of 73 months. Fifty-five (96.5%) patients received a regimen that included the use of nucleotide analogs. The cumulative rate of hepatitis B surface antigen seroclearance at 1, 5, and 10 years was 82%, 88%, and 91%, respectively. At the time of transplantation, 39 (72%) patients had detectable hepatitis B virus (HBV) DNA, with a median of 4.5 log copies/mL. The cumulative rate of HBV undetectability was 91% at 1 year, increasing to 100% by 5 years. After 1 year of liver transplantation, over 90% of the patients had undetectable HBV DNA, and from 8 years onward, 100% had undetectable HBV DNA. The overall long-term survival was excellent, with a 12-year survival of 87%. There was no HBV-related graft loss, and no retransplantation or deaths due to HBV reactivation. CONCLUSION Oral antiviral therapy alone without hepatitis B immune globulin is highly effective in preventing HBV reactivation and graft loss from recurrent hepatitis B after liver transplantation in patients with preexisting LAM resistance HBV. The long-term outcome was excellent, with survival of 87% at 12 years after transplantation, without any mortality related to HBV reactivation.
Collapse
Affiliation(s)
- James Fung
- 1 The Liver Transplant Center, Queen Mary Hospital, Pok Fu Lam, Hong Kong. 2 Department of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong. 3 State Key Laboratory for Liver Research, The University of Hong Kong, Pok Fu Lam, Hong Kong. 4 Department of Surgery, The University of Hong Kong, Pok Fu Lam, Hong Kong
| | | | | | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
11
|
Yamada N, Sugiyama R, Nitta S, Murayama A, Kobayashi M, Okuse C, Suzuki M, Yasuda K, Yotsuyanagi H, Moriya K, Koike K, Wakita T, Kato T. Resistance mutations of hepatitis B virus in entecavir-refractory patients. Hepatol Commun 2017; 1:110-121. [PMID: 29404449 PMCID: PMC5721430 DOI: 10.1002/hep4.1022] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/26/2016] [Revised: 12/28/2016] [Accepted: 02/08/2017] [Indexed: 12/17/2022] Open
Abstract
The emergence of resistance mutations in the reverse transcriptase gene of hepatitis B virus (HBV) is associated with treatment failure. Entecavir (ETV) is one of the most potent anti‐HBV reagents; it has a very low resistance rate and is used as the first‐line treatment for chronic hepatitis B. In this study, we isolated HBVs in 4 ETV‐refractory patients (2 with viral breakthrough, 1 with partial virological response, and 1 with flare‐up) and assessed ETV resistance using replication‐competent 1.38‐fold HBV genome‐length molecular clones. The full genome sequences of infected HBVs in ETV‐refractory patients were determined. The HBV molecular clones were generated with the patient‐derived sequences. After transfection of these molecular clones into HepG2 cells, viral replications and ETV susceptibilities were evaluated by measuring the amount of intracellular core‐particle‐associated HBV DNA using Southern blotting and real‐time polymerase chain reaction. Among these cases, ETV‐resistant variants were detected in 2 patients with viral breakthrough and responsible amino acid mutations in reverse transcriptase were successfully identified in these variants. No ETV‐resistant mutation was detected in the other cases. The identified ETV‐resistant mutations did not confer resistance to tenofovir disoproxil fumarate. Conclusion: The HBV replication model with patient‐derived sequences is useful for assessing replication efficiency, susceptibility to anti‐HBV reagents, and responsible resistance mutations and can aid in choosing the appropriate treatment strategy for treatment‐failure cases of chronic hepatitis B. (Hepatology Communications 2017;1:110‐121)
Collapse
Affiliation(s)
- Norie Yamada
- Department of Virology II National Institute of Infectious Diseases Tokyo Japan.,Department of Internal Medicine Center for Liver Diseases, Seizankai Kiyokawa Hospital Tokyo Japan
| | - Ryuichi Sugiyama
- Department of Virology II National Institute of Infectious Diseases Tokyo Japan
| | - Sayuri Nitta
- Department of Virology II National Institute of Infectious Diseases Tokyo Japan.,Department of Gastroenterology and Hepatology Tokyo Medical and Dental University Tokyo Japan
| | - Asako Murayama
- Department of Virology II National Institute of Infectious Diseases Tokyo Japan
| | - Minoru Kobayashi
- Department of Internal Medicine Center for Liver Diseases, Seizankai Kiyokawa Hospital Tokyo Japan
| | - Chiaki Okuse
- Department of Internal Medicine Division of Gastroenterology and Hepatology, St. Marianna University School of Medicine Kanagawa Japan
| | - Michihiro Suzuki
- Department of Internal Medicine Division of Gastroenterology and Hepatology, St. Marianna University School of Medicine Kanagawa Japan
| | - Kiyomi Yasuda
- Department of Internal Medicine Center for Liver Diseases, Seizankai Kiyokawa Hospital Tokyo Japan
| | - Hiroshi Yotsuyanagi
- Division of Infectious Diseases Advanced Clinical Research Center, Institute of Medical Science
| | - Kyoji Moriya
- Department of Infection Control and Prevention Graduate School of Medicine
| | - Kazuhiko Koike
- Department of Gastroenterology Graduate School of Medicine, The University of Tokyo Tokyo Japan
| | - Takaji Wakita
- Department of Virology II National Institute of Infectious Diseases Tokyo Japan
| | - Takanobu Kato
- Department of Virology II National Institute of Infectious Diseases Tokyo Japan
| |
Collapse
|
|
12
|
Abstract
BACKGROUND The long-term use of nucleos(t)ide analogues causes drug resistance and mutations in the HBV reverse transcriptase (RT) region of the polymerase gene. The RT region overlaps the HBV surface gene (S gene) and therefore, the mutations in the RT region simultaneously modify S gene sequence. Certain mutations in the RT region bring about truncated S proteins because the corresponding changed S gene encodes a stop codon which results in the loss of a large portion of the C-terminal hydrophobic region of HBV surface protein. The rtA181T/sW172*, rtM204I/sW196* and rtV191I/sW182* are the most frequently reported drug-resistant mutations with C-terminal truncation, these mutations have oncogenic potential. DATA SOURCES PubMed and Web of Science were searched using terms: "hepatitis B virus", "HBV drug resistance mutation", "HBV surface protein", "HBV truncation", "hepatocellular carcinoma", "rtA181T/sW172*", "rtM204I/sW196*", "rtV191I/sW182*", and relevant articles published in English in the past decades were reviewed. RESULTS The rtA181T/sW172* and rtV191I/sW182* mutants occurred more frequently than the rtM204I/sW196* mutant both in chronic hepatitis B patients and the HBV-related hepatocellular carcinoma tissues. Although these mutations occur naturally, nucleos(t)ide analogues therapy is the main driving force. These mutations may exist alone or coexist with other HBV mutations. All these three mutants impair the virion secretion and result in HBV surface protein retention and serum HBV DNA level reduction. These mutations possess potential carcinogenic properties. The three mutations are resistant to more than one nucleos(t)ide analogue and therefore, it is difficult to treat the patients with the truncated mutations. CONCLUSIONS Nucleos(t)ide analogues induce drug resistance and HBV S gene truncated mutations. These mutations have potential carcinogenesis.
Collapse
Affiliation(s)
- Meng-Lan Wang
- Center of Infectious Diseases, West China Hospital of Sichuan University; Division of Infectious Diseases, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital of Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu 610041, China.
| | | |
Collapse
|
|
13
|
Dong H, Zhou B, Kang H, Jin W, Zhu Y, Shen Y, Sun J, Wang S, Zhao G, Hou J, He Y. Small surface antigen variants of HBV associated with responses to telbivudine treatment in chronic hepatitis B patients. Antivir Ther 2016; 22:43-51. [PMID: 27583985 DOI: 10.3851/imp3078] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 08/22/2016] [Indexed: 10/21/2022]
Abstract
BACKGROUND Nucleoside/nucleotide analogues are widely used to treat chronic HBV infection, but drug resistance is common. The role of HBV surface gene variants in drug resistance to nucleoside/nucleotide analogues is unknown. We are trying to investigate the dynamics of S gene mutations and how they relate to a patient's virological response in this study. METHODS Thirty patients with chronic hepatitis B were enrolled and serum samples were collected at multiple time points during treatment with telbivudine (LdT). The coding regions of the small surface antigen (S-HBsAg) were amplified and sequenced using the 454 GS FLX+ System. RESULTS Sequencing results revealed different dynamics of non-synonymous mutations, such as sL9P, sN40S, sG44E, sW172*, sW182* and sS187F, between patients with a complete virological response and those with a partial virological response. The viral population heterogeneity decreased at week 12 of LdT treatment in patients with a complete virological response, with a concomitant decline in non-synonymous mutations (from an average of 14 to 9.9 per sample) and an increase in the frequencies of major variants (from 14.3% to 40.4%). CONCLUSIONS Our findings suggest that the decrease in viral population heterogeneity at an early stage of LdT treatment was associated with the subsequent optimal virological response, and the early appearance of some specific mutations, such as sG44E, sW172* and sW182*, is a potential indicator of a partial virological response in continuing therapy.
Collapse
Affiliation(s)
- Hui Dong
- Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China
| | - Bin Zhou
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China
| | - Hui Kang
- School of Life Sciences, Fudan University, Shanghai, China
| | - Weirong Jin
- Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China.,Shanghai Shenyou Biotechnology Co., Ltd, Shanghai, China
| | - Yongqiang Zhu
- Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China
| | - Yan Shen
- Shanghai Shenyou Biotechnology Co., Ltd, Shanghai, China
| | - Jian Sun
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China
| | - Shengyue Wang
- Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China
| | - Guoping Zhao
- Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China.,CAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.,Department of Microbiology and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong SAR, China
| | - Jinlin Hou
- State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou, China
| | - Yungang He
- CAS Key Laboratory of Computational Biology, CAS-MPG Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
| |
Collapse
|
|
14
|
Pierra C, Benzaria S, Dukhan D, Loi AG, La Colla P, Bridges E, Mao J, Standring D, Sommadossi JP, Gosselin G. Synthesis, Physicochemical and Pharmacokinetic Studies of Potential Prodrugs of β-L-2′-Deoxycytidine, a Selective and Specific Anti-HBV Agent. ACTA ACUST UNITED AC 2016; 15:269-79. [PMID: 15535049 DOI: 10.1177/095632020401500506] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
β-L-2′-Deoxycytidine (β-L-dC) is a potent, selective and specific anti-hepatitis B virus (HBV) agent. To improve its oral bioavailability, several derivatives involving sugar or base acylation, as well as N4-derivatization with an N,N-(dimethyl-amino)methylene function, were synthesized. The physicochemical characteristics (including chemical stabilities, solubilities and distribution coefficient values) and pharmacokinetics of these compounds were determined and compared with those of the parent drug, β-L-dC. Presented in part at the 14th International Conference on Antiviral Research, Seattle, Washington, USA, 8–13 April 2001. Antiviral Reseach 2001; 50:A79.
Collapse
Affiliation(s)
- Claire Pierra
- Laboratoire Coopératif Idenix-CNRS-Université Montpellier II, Montpellier, France
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
15
|
Zhang BG, Tanaka G, Aihara K, Honda M, Kaneko S, Chen L. Dynamics of an HBV Model with Drug Resistance Under Intermittent Antiviral Therapy. INTERNATIONAL JOURNAL OF BIFURCATION AND CHAOS 2015; 25:1540011. [DOI: 10.1142/s0218127415400118] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/03/2025]
Abstract
This paper studies the dynamics of the hepatitis B virus (HBV) model and the therapy regimens of HBV disease. First, we propose a new mathematical model of HBV with drug resistance, and then analyze its qualitative and dynamical properties. Combining the clinical data and theoretical analysis, we demonstrate that our model is biologically plausible and also computationally viable. Second, we demonstrate that the intermittent antiviral therapy regimen is one of the possible strategies to treat this kind of complex disease. There are two main advantages of this regimen, i.e. it not only may delay the development of drug resistance, but also may reduce the duration of on-treatment time compared with the long-term continuous medication. Moreover, such an intermittent antiviral therapy can reduce the adverse side effects. Our theoretical model and computational results provide qualitative insight into the progression of HBV, and also a possible new therapy for HBV disease.
Collapse
Affiliation(s)
- Ben-Gong Zhang
- School of Mathematics and Computer Science, Wuhan Textile University, Wuhan 430073, P. R. China
| | - Gouhei Tanaka
- Institute of Industrial Science, The University of Tokyo, 153-8505, Japan
| | - Kazuyuki Aihara
- Institute of Industrial Science, The University of Tokyo, 153-8505, Japan
| | - Masao Honda
- Department of Gastroenterology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8641, Japan
| | - Shuichi Kaneko
- Department of Gastroenterology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa 920-8641, Japan
| | - Luonan Chen
- Key Laboratory of Systems Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, P. R. China
| |
Collapse
|
|
16
|
Pondé RAA. Molecular mechanisms underlying HBsAg negativity in occult HBV infection. Eur J Clin Microbiol Infect Dis 2015; 34:1709-31. [PMID: 26105620 DOI: 10.1007/s10096-015-2422-x] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2015] [Accepted: 06/03/2015] [Indexed: 02/06/2023]
Abstract
Although genomic detection is considered the gold standard test on HBV infection identification, the HBsAg investigation is still the most frequent clinical laboratory request to diagnose HBV infection in activity. However, the non-detection of HBsAg in the bloodstream of chronic or acutely infected individuals has been a phenomenon often observed in clinical practice, despite the high sensitivity and specificity of screening assays standardized commercially and adopted in routine. The expansion of knowledge about the hepatitis B virus biology (replication/life cycle, genetic variability/mutability/heterogeneity), their biochemical and immunological properties (antigenicity and immunogenicity), in turn, has allowed to elucidate some mechanisms that may explain the occurrence of this phenomenon. Therefore, the negativity for HBsAg during the acute or chronic infection course may become a fragile or at least questionable result. This manuscript discusses some mechanisms that could explain the negativity for HBsAg in a serological profile of individuals with HBV infection in activity, or factors that could compromise its detection in the bloodstream during HBV infection.
Collapse
Affiliation(s)
- R A A Pondé
- Laboratory of Human Virology, Institute of Tropical Pathology and Public Health, Federal University of Goiás, Goiânia, Goiás, Brazil,
| |
Collapse
|
|
17
|
Koumbi L. Current and future antiviral drug therapies of hepatitis B chronic infection. World J Hepatol 2015; 7:1030-1040. [PMID: 26052392 PMCID: PMC4450180 DOI: 10.4254/wjh.v7.i8.1030] [Citation(s) in RCA: 46] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/28/2014] [Revised: 01/12/2015] [Accepted: 02/04/2015] [Indexed: 02/06/2023] Open
Abstract
Despite significant improvement in the management of chronic hepatitis B virus (HBV) it remains a public health problem, affecting more than 350 million people worldwide. The natural course of the infection is dynamic and involves a complex interplay between the virus and the host's immune system. Currently the approved therapeutic regimens include pegylated-interferon (IFN)-α and monotherapy with five nucleos(t)ide analogues (NAs). Both antiviral treatments are not capable to eliminate the virus and do not establish long-term control of infection after treatment withdrawal. IFN therapy is of finite duration and associates with low response rates, liver decompensating and numerous side effects. NAs are well-tolerated therapies but have a high risk of drug resistance development that limits their prolonged use. The imperative for the development of new approaches for the treatment of chronic HBV infection is a challenging issue that cannot be over-sided. Research efforts are focusing on the identification and evaluation of various viral replication inhibitors that target viral replication and a number of immunomodulators that aim to restore the HBV specific immune hyporesponsiveness without inducing liver damage. This review brings together our current knowledge on the available treatment and discusses potential therapeutic approaches in the battle against chronic HBV infection.
Collapse
Affiliation(s)
- Lemonica Koumbi
- Lemonica Koumbi, Hepatology and Gastroenterology Section, Department of Medicine, Imperial College London, London W2 1PG, United Kingdom
| |
Collapse
|
|
18
|
Xu J, Wu B, Wang JH, Huang L, Wang DY, Zhao L, Zhao GP, Wang Y. Pre-existing mutations in reverse transcriptase of hepatitis B virus in treatment-naive Chinese patients with chronic hepatitis B. PLoS One 2015; 10:e0117429. [PMID: 25821965 PMCID: PMC4379075 DOI: 10.1371/journal.pone.0117429] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/06/2014] [Accepted: 12/23/2014] [Indexed: 02/06/2023] Open
Abstract
High rate of viral replication and lacking of proofreading activity in hepatitis B virus (HBV) polymerase lead to the generation of mutations in HBV virus. Mutations in the reverse transcriptase (RT) region of HBV polymerase are demonstrated to be strongly associated with drug resistance during antiviral treatment. However, the presence of mutations as well as its clinical significance in treatment-naïve hepatitis patients (defined as pre-existing mutations) need to be further investigated. In the present study, a total of 168 serum samples from treatment-naive chronic hepatitis B (CHB) patients were collected, and the RT region of HBV polymerase was sequenced. The results showed that pre-existing mutations in the RT region of HBV polymerase were detected in 43 of 168 (25.6%) treatment-naive CHB patients within which there were no well-characterized primary nucleotide analogs (NAs) resistance sites. Three dominant sites at rt191, rt207 and rt226 were found mutant in 7(16.28%), 8(18.60%), and 14(32.56%) samples respectively among these 43 patients. No significant correlation was found between pre-existing mutations and gender, age, HBV genotype, ALT, HBeAg or HBV DNA loads. However, patients with pre-existing RT mutations under HBeAg sero-negative status exhibited decreased HBV DNA loads, which contributed to the decreased HBV DNA loads in the total HBeAg sero-negative patients. The above investigation indicated that there was a prevalence of pre-existing mutations in RT region of HBV polymerase which might affect the serum HBV DNA level in treatment-naive CHB patients. Its effects on the occurrence of NAs resistance and the prognosis after treatment need to be further investigated.
Collapse
Affiliation(s)
- Jie Xu
- Department of Infectious Diseases, No.3 People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Biao Wu
- Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Shanghai Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai/Shanghai Academy of Science and Technology, Shanghai, China
| | - Jing-Hui Wang
- Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Ling Huang
- Department of Infectious Diseases, No.3 People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Deng-yu Wang
- Shanghai Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai/Shanghai Academy of Science and Technology, Shanghai, China
| | - Ling Zhao
- Department of Infectious Diseases, No.3 People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Guo-ping Zhao
- Shanghai Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai/Shanghai Academy of Science and Technology, Shanghai, China
| | - Ying Wang
- Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Shanghai Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai/Shanghai Academy of Science and Technology, Shanghai, China
- * E-mail:
| |
Collapse
|
|
19
|
Liu Y, Li X, Xin S, Xu Z, Chen R, Yang J, Liu L, Wong VWS, Yang D, Chan HLY, Xu D. The rtA181S mutation of hepatitis B virus primarily confers resistance to adefovir dipivoxil. J Viral Hepat 2015; 22:328-34. [PMID: 25132017 DOI: 10.1111/jvh.12298] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
The study aimed to clarify clinical significance of hepatitis B virus (HBV) rtA181S mutation in Chinese HBV-infected patients. A total of 18 419 patients with chronic HBV infection from Beijing 302 Hospital were investigated. HBV complete reverse transcriptase region of polymerase was screened by direct sequencing, and the results were verified by clonal sequencing. Replication-competent mutant and wild-type HBV genomic amplicons were constructed and transfected into the HepG2 cells and cultured in the presence or absence of serially diluted nucleos(t)ide analogues. Intracellular HBV replicative intermediates were quantitated for calculating the 50% effective concentration of the drug (EC(50)). The rtA181S was detected in 98 patients with 12 kinds of mutational patterns. Genotype C and genotype B HBV infection occupied 91.8% and 8.2% in rtA181S-positive patients, in contrast to 84.6% and 15.4% in rtA181S-negative patients (P < 0.01). All rtA181S-positive patients had received nucleos(t)ide analogues. rtA181S was detected in multiple patients with virologic breakthrough. Phenotypic analysis of patient-derived viral strains showed that rtA181S, rtA181S+N236T, rtN236T and rtA181V strains had 68.5%, 49.9%, 71.4% and 66.2% of natural replication capacity of wild-type strain, and 3.7-fold, 9.8-fold, 7.9-fold and 5.6-fold increased EC(50) to adefovir dipivoxil (ADV). The rtA181S strain remained susceptible to lamivudine, entecavir and tenofovir, and ADV susceptibility was restored after the mutation was eliminated through site-directed mutagenesis. Rescue therapy with entecavir or combination therapy was effective in rtA181S-related ADV-refractory patients. The rtA181S mutation confers moderate resistance to ADV. It could be induced by either lamivudine or ADV and contribute ADV treatment failure.
Collapse
Affiliation(s)
- Y Liu
- Institute of Infectious Diseases/Liver Failure Medical Center, Beijing 302 Hospital of PLA, Beijing, China
| | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
20
|
Liu Y, Xin S, Ye X, Chen R, Xu Z, Li X, Ye H, Cheng S, Xu D. Increased occurrence of mutant rtI233V of HBV in patients with adefovir therapy. Antivir Ther 2015; 21:9-16. [PMID: 26079809 DOI: 10.3851/imp2971] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 05/26/2015] [Indexed: 10/23/2022]
|
|
21
|
Dienstag JL. Antiviral Drugs against Hepatitis Viruses. MANDELL, DOUGLAS, AND BENNETT'S PRINCIPLES AND PRACTICE OF INFECTIOUS DISEASES 2015:563-575.e3. [DOI: 10.1016/b978-1-4557-4801-3.00046-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
22
|
Dienstag JL, Delemos AS. Viral Hepatitis. MANDELL, DOUGLAS, AND BENNETT'S PRINCIPLES AND PRACTICE OF INFECTIOUS DISEASES 2015:1439-1468.e7. [DOI: 10.1016/b978-1-4557-4801-3.00119-3] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
23
|
Bang KB, Kim HJ. Management of antiviral drug resistance in chronic hepatitis B. World J Gastroenterol 2014; 20:11641-11649. [PMID: 25206270 PMCID: PMC4155356 DOI: 10.3748/wjg.v20.i33.11641] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/21/2013] [Revised: 01/10/2014] [Accepted: 05/28/2014] [Indexed: 02/07/2023] Open
Abstract
Rescue antiviral treatment for patients with resistance to preexisting nucleos(t)ide analogues remains a clinical challenge. The correct choice of a first-line treatment of high potency and with a high genetic barrier to achieve sustained long-term suppression of viral replication provides the best chance of preventing treatment failure and the emergence of drug resistance. The management of treatment failure and drug resistance requires a precise and accurate clinical and virologic monitoring. Combination treatment with antiviral drugs that belong to different groups is associated with a lower chance of developing resistance to rescue drugs. To guarantee better control of viral replication in patients with drug resistance, the addition of another drug without a cross resistance profile should be given as early as possible, preferably at the time when genotypic resistance emerges. Long-term surveillance for treatment efficacy and possible emergence of drug resistance should be continued to prevent the emergence of multidrug-resistant strains.
Collapse
|
|
24
|
Li X, Liu Y, Zhao P, Wang Y, Chen L, Xin S, Zhang XX, Xu D. Investigation into drug-resistant mutations of HBV from 845 nucleoside/nucleotide analogue-naive Chinese patients with chronic HBV infection. Antivir Ther 2014; 20:141-147. [PMID: 24992206 DOI: 10.3851/imp2813] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 05/30/2014] [Indexed: 01/04/2023]
Abstract
BACKGROUND This study aimed to clarify the clinical significance of drug-resistant HBV in nucleoside/nucleotide analogue (NA)-naive Chinese patients with chronic HBV infection in real clinical practice. METHODS A total of 845 NA-naive patients who were admitted to Beijing 302 Hospital between July 2007 and March 2012 were included in the study. HBV drug-resistant mutations were examined by direct sequencing of the viral reverse transcriptase gene and verified by clonal sequencing. Phenotypic analysis of viral replication capacity and drug susceptibility were performed by measuring viral replicative intermediate level in 1.1-mer mutant or wild-type HBV amplicon-transfected HepG2 cells in absence or presence of serially diluted drugs. RESULTS Drug-resistant mutations were detected in 2.01% (17/845) of the patients by direct sequencing, including 15 with lamivudine-resistant mutations (rtM204V, rtM204I), one with adefovir-resistant mutation (rtA181V), and one with both lamivudine- and adefovir-resistant mutations (rtA181V, rtM204I). Clonal sequencing identified 13 drug-resistant HBV strains: rtL80I+M204I, rtL80I+M204V, rtL180M+M204I, rtL180M+M204V, rtM204I, rtM204V, rtL80I+L180M+M204I, rtL80I+L180M+M204V, rtA181V, rtA181V+M204I, rtA181T+N236T, rtA181V+N236T and rtN236T. Phenotypic analysis showed that two pre-existing lamivudine-resistant strains (rtL80I+M204I, rtL180M+M204V) had >1,000-fold resistance to lamivudine, and one pre-existing adefovir-resistant strain (rtA181V+N236T) had 15.4-fold resistance to adefovir compared with the wild-type strain. A follow-up study showed that the presence of pre-existing rtM204I strain in one patient increased from 20% at baseline to 85% after 13 months of entecavir treatment with corresponding recession of wild-type strain in the viral pool. CONCLUSIONS The incidence of drug-resistant HBV mutations was low in NA-naive Chinese HBV-infected patients. Pre-existing mutants had similar resistance characteristics to those from NA refractory patients.
Collapse
Affiliation(s)
- Xiaodong Li
- Institute of Infectious Diseases and Liver Failure Research Center, Beijing 302 Hospital of PLA, Beijing, China
| | | | | | | | | | | | | | | |
Collapse
|
|
25
|
Liu Y, Liu W, Li X, Xu Z, Wang X, Li C, Chen L, Xin S, Xu D. Screening and identification of a novel adefovir dipivoxil resistance associated mutation, rtN236V, of HBV from a large cohort of HBV-infected patients. Antivir Ther 2014; 19:551-8. [PMID: 24710668 DOI: 10.3851/imp2775] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 02/23/2014] [Indexed: 10/25/2022]
Abstract
BACKGROUND The study aimed to clarify whether rtN236V mutation of HBV derived from adefovir dipivoxil (ADV)-refractory patients was associated with drug resistance. METHODS A total of 18,419 patients from Beijing 302 Hospital were investigated. HBV complete reverse transcriptase region of polymerase was screened by direct sequencing and verified by clonal sequencing if necessary. Replication-competent wild-type and mutant HBV genomic amplicons were constructed, transfected into HepG2 cells and cultured in the presence or absence of serially diluted nucleoside/nucleotide analogues. Intracellular HBV replicative intermediates were quantitated for calculating the 50% effective concentration of drug. RESULTS rtN236V was detected in six ADV-refractory patients; signature ADV-resistant mutations rtA181V and rtN236T were detected in 1,311 patients. rtN236V mutants emerged predominantly with virological breakthrough in the clinical course of the six patients. Phenotypic analysis of the mutants from two patients was performed. rtN236V mutants from patient 1 and patient 2 exhibited 3.90-fold and 3.10-fold decreased susceptibility to ADV, respectively, compared to the wild-type virus; by contrast, rtN236T mutants from the patients had 4.50-fold and 4.75-fold decreased susceptibility, respectively. Both mutants had a relatively lower viral replication capacity compared to wild-type virus in the absence of antivirals and remained susceptible to lamivudine, entecavir and tenofovir disoproxil fumarate. In clinical practice, switching to entecavir rescue therapy suppressed HBV DNA to an undetectable level and normalized alanine aminotransferase level for both patients. CONCLUSIONS rtN236V was a novel infrequently occurring ADV-resistance-associated mutation. It conferred a moderate resistance to ADV with relatively lower natural replication capacity.
Collapse
Affiliation(s)
- Yan Liu
- Institute of Infectious Diseases/Liver Failure Medical Center, Beijing 302 Hospital, Beijing, China
| | | | | | | | | | | | | | | | | |
Collapse
|
|
26
|
rtM204Q may serve as a novel lamivudine-resistance-associated mutation of hepatitis B virus. PLoS One 2014; 9:e89015. [PMID: 24586482 PMCID: PMC3933355 DOI: 10.1371/journal.pone.0089015] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2013] [Accepted: 01/14/2014] [Indexed: 02/07/2023] Open
Abstract
Background and Aims Lamivudine (LAM) is still widely used for anti-HBV therapy in China. The study aimed to clarify whether a newly-found rtM204Q mutation from patients was associated with the drug resistance. Methods HBV complete reverse-transcriptase region was screened by direct sequencing and verified by clonal sequencing. Replication-competent plasmids containing patient-derived 1.1mer mutant or wild-type viral genome were constructed and transfected into HepG2 cells. After cultured with or without serially-diluted antiviral drugs, intracellular HBV replicative intermediates were quantitated for calculating the 50% effective concentration of drug (EC50). Results A total of 12,000 serum samples of 9,830 patients with chronic HBV infection were screened. rtM204Q mutation was detected in seven LAM-refractory patients. By contrast, rtM204I/rtM204V mutations were detected in 2,502 patients' samples. The rtM204Q emerged either alone or in concomitance with rtM204I/rtM204V, and all were accompanied with virologic breakthrough in clinical course. Clonal sequencing verified that rtM204Q mutant was predominant in viral quasispecies of these samples. Phenotypic analysis showed that rtM204Q mutant had 89.9% of replication capacity and 76-fold increased LAM EC50 of the concomitant wild-type strain. By contrast, rtM204I mutant in the sample had lower replication capacity and higher LAM resistance (46.3% and 1396-fold increased LAM EC50 of the wild-type strain) compared to rtM204Q mutant. rtM204Q mutant was susceptible to adefovir dipivoxil (ADV) in vitro and ADV/ADV+LAM rescue therapy in clinic. Conclusion rtM204Q is suggested to be a novel LAM-resistance-associated mutation. It conferred a moderate resistance with higher competent natural replication capacity compared to rtM204I mutation.
Collapse
|
|
27
|
Amino acid similarities and divergences in the small surface proteins of genotype C hepatitis B viruses between nucleos(t)ide analogue-naïve and lamivudine-treated patients with chronic hepatitis B. Antiviral Res 2013; 102:29-34. [PMID: 24316031 DOI: 10.1016/j.antiviral.2013.11.015] [Citation(s) in RCA: 27] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2013] [Revised: 11/06/2013] [Accepted: 11/28/2013] [Indexed: 02/06/2023]
Abstract
Entire C-genotype small hepatitis B surface (SHBs) sequences were isolated from 139 nucleos(t)ide analogues (NA)-naïve and 74 lamivudine (LMV)-treated chronic hepatitis B (CHB) patients. The conservation and variability of total 226 amino acids (AAs) within the sequences were determined individually, revealing significant higher mutant isolate rate and mutation frequency in LMV-treated cohort than those in the NA-naïve one (P=0.009 and 0.0001, respectively). Three absolutely conserved fragments (s16-s19, s176-s181 and s185-s188) and seven moderately conserved regions (a few AA sites acquiring increased variability after LMV-treatment) were identified. The significant mutation rate increase after LMV-treatment occurred primarily in major hydrophilic region (except 'a' determinant) and transmembrane domain 3/4, but not in other upstream functional regions of SHBs. With little influence on immune escape-associated mutation frequencies within 'a' determinant, LMV-monotherapy significantly induced classical LMVr-associated mirror changes sE164D/rtV173L, sI195M/rtM204V and sW196L/S/rtM204I, as well as non-classical ones sG44E/rtS53N, sT47K/A/rtH55R/Q and sW182stop/rtV191I outside 'a' determinant. Interestingly, another newly-identified truncation mutation sC69stop/rtS78T decreased from 7.91% (11/139) in NA-naïve cohort to 2.70% (2/74) in LMV-treated one. Altogether, the altered AA conservation and diversity in SHBs sequences after LMV-treatment in genotype-C HBV infection might shed new insights into how LMV-therapy affects the SHBs variant evolution and its antigenicity.
Collapse
|
|
28
|
Lv GC, Yao JM, Yang YD, Zheng L, Sheng JF, Chen Y, Li LJ. Efficacy of combined therapy in patients with hepatitis B virus-related decompensated cirrhosis. World J Gastroenterol 2013; 19:3481-3486. [PMID: 23801842 PMCID: PMC3683688 DOI: 10.3748/wjg.v19.i22.3481] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/17/2012] [Revised: 01/24/2013] [Accepted: 03/29/2013] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate the efficacy and safety of combined de novo lamivudine (LAM) and adefovir dipivoxil (ADV) therapy in hepatitis B virus (HBV)-related decompensated liver cirrhosis patients.
METHODS: One hundred and forty patients with HBV-related decompensated cirrhosis were recruited, 70 patients were treated with combined LAM and ADV de novo therapy, and the other 70 patients were treated with LAM alone as controls. The follow-up period was 144 wk. All patients with LAM resistance were shifted to ADV.
RESULTS: The percentage of HBV-related decompensated cirrhosis patients with undetectable HBV DNA in de novo combination group was 51.6% (33/64), 84.2% (48/57), and 92.3% (49/53) by weeks 48, 96, and 144, respectively. In monotherapy group, HBV DNA negativity rate was 46.1% (30/65), 56.1% (32/57), and 39.2% (20/51) by weeks 48, 96 and 144, respectively. There was a significant difference between the two groups by weeks 96 and 144 (P = 0.012 and 0.001). The hepatitis B e antigen seroconversion rate was 28.1% (9/32), 40.0% (12/30), and 53.6% (15/28) in the combination group by weeks 48, 96 and 144, respectively, and 24.2% (8/33), 31.0% (9/29), and 37.0% (10/27) by weeks 48, 96 and 144, respectively, in monotherapy group. A total of 68.6% (44/64), 84.2% (48/57), and 92.5% (49/53) patients achieved alanine aminotransferase (ALT) normalization by weeks 48, 96 and 144, respectively in the combination group. In monotherpy group, the ALT normalization rate was 64.6% (42/65) by week 48, 73.7% (42/57) by week 96, and 80.4% (41/51) by week 144. No patients in the combination group exhibited detectable resistance for at least 144 wk. The cumulative resistance rate in monotherapy group at weeks 48, 96, and 144 was 20.0%, 36.8%, and 56.9%. Both combination group and monotherapy group demonstrated an improvement in Child-Turcotte Pugh and Model for End-Stage Liver Disease scores at weeks 48, 96, and 144. All patients tolerated both combination and monotherapy. The ceratinine levels and glomerular filtration rate remained normal in all patients during the follow-up period.
CONCLUSION: In HBV-related decompensated liver cirrhosis patients, the combined de novo LAM and ADV therapy is more efficacious and safer compared to LAM alone.
Collapse
|
|
29
|
Perrakis A, Förtsch T, Del Medico A, Croner R, Vassos N, Yedibela S, Lohmüller C, Zopf S, Hohenberger W, Müller V. Liver Transplantation for Hepatitis B-Induced Liver Disease: Long-Term Outcome and Effectiveness of Antiviral Therapy for Prevention of Recurrent Hepatitis B Infection. Transplant Proc 2013; 45:1953-6. [DOI: 10.1016/j.transproceed.2012.11.020] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2012] [Accepted: 11/19/2012] [Indexed: 01/11/2023]
|
|
30
|
Zheng J, Zeng Z, Zhang D, Yu Y, Wang F, Pan CQ. Prevalence and significance of Hepatitis B reverse transcriptase mutants in different disease stages of untreated patients. Liver Int 2012; 32:1535-1542. [PMID: 22882650 PMCID: PMC3463715 DOI: 10.1111/j.1478-3231.2012.02859.x] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/10/2012] [Accepted: 07/04/2012] [Indexed: 12/13/2022]
Abstract
AIMS Hepatitis B virus (HBV) reverse transcriptase (RT) mutants, which have not been well characterized according to different disease stages. This study aimed to characterize the profiles of naturally occurring mutations in the HBV RT region and their associated clinical outcomes. METHODS HBV RT region mutations and genotypes were determined by PCR-direct sequencing and compared with p-distance model. RESULTS Among 467 consecutive eligible patients (262 chronic hepatitis B patients, 105 cirrhotic patients and 100 hepatocellular carcinoma patients), the nucleos(t)ide analogues-related mutations (rtI169T, rtV173L, rtL180M, rtA181T, rtS202C, rtM204I/V, rtN236T) were found. The p-distance value reached a peak in the age of 20-30 years in the CHB patients and in the age of 40-45 years in the cirrhotic patients and hepatocellular carcinoma patients. The naturally occurring mutation, rtS106C mutation was higher in chronic hepatitis B patients (14/100, 14.0%) and cirrhotic patients (14/100, 14.0%) than that in hepatocellular carcinoma patients (4/100, 4.0%, P = 0.013). And the rtD134E/G/N/S mutations were also higher in chronic hepatitis B patients (22/100, 22.0%) and cirrhotic patients (21/100, 21.0%) than that in hepatocellular carcinoma patients (10/100, 10.0%, P = 0.021 and P = 0.032 respectively). The mutation frequencies in A-B interdomain were higher in cirrhotic patients (101/1900, 5.3%) than that in hepatocellular carcinoma patients (68/1900, 3.6%) (P = 0.009). CONCLUSIONS The nucleos(t)ide analogues-related mutations do exist in treatment naive patients with different disease stages. rtS106C, rtD134E/G/N/S and A-B interdomain mutations may be associated with necro-inflammation, immune response and cirrhosis development at ages older than 40.
Collapse
Affiliation(s)
- Jinxin Zheng
- Laboratory of Virology, Department of Infectious Diseases, Peking University First Hospital, Beijing, P. R. China
| | - Zheng Zeng
- Laboratory of Virology, Department of Infectious Diseases, Peking University First Hospital, Beijing, P. R. China
| | - Duyi Zhang
- Laboratory of Virology, Department of Infectious Diseases, Peking University First Hospital, Beijing, P. R. China
| | - Yanyan Yu
- Laboratory of Virology, Department of Infectious Diseases, Peking University First Hospital, Beijing, P. R. China
| | - Fang Wang
- Laboratory of Virology, Department of Infectious Diseases, Peking University First Hospital, Beijing, P. R. China
| | - Calvin Q. Pan
- Division of Liver Diseases, Department of Medicine, The Mount Sinai Medical Center, Mount Sinai School of Medicine, New York, USA
- New Discovery, New York, USA
| |
Collapse
|
|
31
|
Ji D, Liu Y, Li L, Xu Z, Si LL, Dai JZ, Li X, Wang L, Yao Z, Xin SJ, Chen GF, Xu D. The rtL229 substitutions in the reverse transcriptase region of hepatitis B virus (HBV) polymerase are potentially associated with lamivudine resistance as a compensatory mutation. J Clin Virol 2012; 54:66-72. [PMID: 22398037 DOI: 10.1016/j.jcv.2012.02.003] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/16/2011] [Revised: 01/23/2012] [Accepted: 02/06/2012] [Indexed: 02/07/2023]
Abstract
BACKGROUND It remains unclear whether hepatitis B virus (HBV) reverse-transcriptase (RT) rtL229 substitutions influence HBV drug resistance. OBJECTIVE The study was to investigate the association of HBV rtL229 substitutions with viral resistance to lamivudine (LAM). STUDY DESIGN Entire HBV RT genes were amplified by nested PCR and sequenced from sera of 6000 nucleos(t)ide analog-experienced patients with chronic HBV infection. The incidence and clinic relevance of rtL229 substitutions were analyzed. Replication-competent viral amplicons which harbored HBV genomes of wild-type, rtM204I, or rtM204I in conjunction with various rtL229 substitutions (rtL229F/W/M/V) were constructed. The amplicons were transfected into HepG2 cells for phenotyping of replication capacity and susceptibility to nucleos(t)ide analogs. RESULTS The rtL229 substitutions were detected in 6.57% (394/6000) of patients. Individual substitution incidences were 2.77%, 0.97%, 0.83% and 0.55% for rtL229V, rtL229F, rtL229M and rtL229W, respectively. The incidence of rtL229 substitutions was significantly higher in LAM-experienced patients (341/4220, 8.1%) than in LAM-naïve patients (53/1780, 3.0%), and were independently associated with genotypic LAM resistance (77.9% vs. 21.2%, OR 8.806, 95%CI 6.345-12.223) and low viral replication (HBV DNA <1000IU/mL) (4.60% vs. 24.2%, OR 0.478, 95%CI 0.254-0.898). Representative cases follow-up showed that rtL229F developed subsequent to rtM204I emergence during LAM treatment and regressed with rtM204I after LAM withdrawal. Functionally, rtL229F did not confer reduced susceptibility to LAM, but could restore replication capacity of rtM204I strain. CONCLUSION The rtL229 substitutions were potentially associated with LAM resistance in Chinese patients and rtL229F had characteristics of a compensatory mutation of rtM204I mutant.
Collapse
Affiliation(s)
- Dong Ji
- Institute of Infectious Diseases and Liver Failure Research Center, Beijing 302 Hospital, Beijing 100039, China
| | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
32
|
Kimberlin DW. Antiviral Agents. PRINCIPLES AND PRACTICE OF PEDIATRIC INFECTIOUS DISEASES 2012:1502-1518.e10. [DOI: 10.1016/b978-1-4377-2702-9.00297-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
33
|
Zhong JH, Li LQ, Wu LC. Lamivudine with or without adefovir dipivoxil for postoperative hepatocellular carcinoma. Cochrane Database Syst Rev 2011:CD008713. [PMID: 22161435 DOI: 10.1002/14651858.cd008713.pub2] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
BACKGROUND Hepatocellular carcinoma (HCC) is a significant cause of death, especially in Asia and sub-Saharan Africa. Removal of the cancer through surgery or other techniques is considered the first-line therapy in early HCC, but relapse of HCC is the main postoperative problem. The main risk factor for HCC is hepatitis B virus (HBV) infection. Lamivudine and adefovir dipivoxil are effective and tolerable for chronic hepatitis B by suppressing the viral load and to reduce fibrosis in the liver. OBJECTIVES To assess the benefits and harms of postoperative administration of lamivudine with or without adefovir dipivoxil in participants with surgically treated HCC and chronic HBV infection or HBV carrier state. SEARCH METHODS A systematic search was performed in The Cochrane Hepato-Biliary Group Controlled Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library, MEDLINE, EMBASE, and Science Citation Index Expanded (SCI Exp) in October 2011. Further trials have been sought through scanning reference lists of relevant articles. SELECTION CRITERIA Randomised clinical trials comparing the administration of lamivudine with and without adefovir dipivoxil for participants with ablation treated HCC (surgical or through other techniques) and chronic HBV infection or HBV carrier state, regardless of publication status, language, blinding, and publication status, were to be included in this review. We planned to extract data on harms from quasi-randomised studies or cohort studies when retrieved with the search results. DATA COLLECTION AND ANALYSIS Two authors independently selected studies for inclusion, and extracted and analysed the data. The type and number of adverse events were reported descriptively. MAIN RESULTS No randomised trials could be included into this systematic review. Thus, we were unable to follow our pre-published protocol and perform meta-analyses.Through our searches for randomised clinical trials, four cohort trials with 230 participants were retrieved. We read them in order to find data on harm, ie, adverse events. Breakthrough hepatitis was a serious adverse event attributable to lamivudine. No other adverse events seemed to be caused by the administration of lamivudine or adefovir dipivoxil were reported in the four cohort studies. AUTHORS' CONCLUSIONS No evidence from randomised trials on the beneficial or harmful effects of lamivudine with or without adefovir dipivoxil for postoperative HCC was found. Randomised clinical trials with large number of participants and long follow-up period should be carried out to direct clinical practice.
Collapse
Affiliation(s)
- Jian Hong Zhong
- Department of Hepato-Biliary Diseases, Tumor Hospital, Guangxi Medical University, Nanning, Guangxi, China, 530021
| | | | | |
Collapse
|
|
34
|
Seto WK, Yuen MF, Fung J, Lai CL. Tenofovir disoproxil fumarate for the treatment of chronic hepatitis B monoinfection. Hepatol Int 2011; 7:327-34. [PMID: 21688182 DOI: 10.1007/s12072-011-9282-y] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/15/2011] [Accepted: 05/30/2011] [Indexed: 02/06/2023]
Abstract
INTRODUCTION Resistance in nucleoside/nucleotide analog (NA) therapy has always been a challenge in the management of chronic hepatitis B (CHB). CLINICAL STUDIES Initially developed for the treatment of HIV infection, early in vitro and clinical observational studies had shown tenofovir disoproxil fumarate (TDF) to be also active against CHB. Recent data from various multicenter phase 3 and 4 clinical trials have confirmed TDF being able to achieve a high viral suppression in both NA-naive and -experienced CHB patients. There are also emerging data on the efficacy of TDF in decompensated CHB. Although there are in vitro studies identifying certain mutation loci associated with a reduced susceptibility to TDF, there have so far been no reports of virologic resistance to TDF in clinical studies. TDF has a favorable safety profile, although more long-term data would be needed. CONCLUSIONS TDF has the makings of an "ideal" first-line drug for the treatment of CHB.
Collapse
Affiliation(s)
- Wai-Kay Seto
- Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam Road, Hong Kong, Hong Kong
| | - Man-Fung Yuen
- Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam Road, Hong Kong, Hong Kong
| | - James Fung
- Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam Road, Hong Kong, Hong Kong
| | - Ching-Lung Lai
- Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam Road, Hong Kong, Hong Kong.
| |
Collapse
|
|
35
|
Stachulski AV, Pidathala C, Row EC, Sharma R, Berry NG, Iqbal M, Bentley J, Allman SA, Edwards G, Helm A, Hellier J, Korba BE, Semple JE, Rossignol JF. Thiazolides as novel antiviral agents. 1. Inhibition of hepatitis B virus replication. J Med Chem 2011; 54:4119-32. [PMID: 21553812 DOI: 10.1021/jm200153p] [Citation(s) in RCA: 44] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
We report the syntheses and activities of a wide range of thiazolides [viz., 2-hydroxyaroyl-N-(thiazol-2-yl)amides] against hepatitis B virus replication, with QSAR analysis of our results. The prototypical thiazolide, nitazoxanide [2-hydroxybenzoyl-N-(5-nitrothiazol-2-yl)amide, NTZ] 1 is a broad spectrum antiinfective agent effective against anaerobic bacteria, viruses, and parasites. By contrast, 2-hydroxybenzoyl-N-(5-chlorothiazol-2-yl)amide 3 is a novel, potent, and selective inhibitor of hepatitis B replication (EC(50) = 0.33 μm) but is inactive against anaerobes. Several 4'- and 5'-substituted thiazolides show good activity against HBV; by contrast, some related salicyloylanilides show a narrower spectrum of activity. The ADME properties of 3 are similar to 1; viz., the O-acetate is an effective prodrug, and the O-aryl glucuronide is a major metabolite. The QSAR study shows a good correlation of observed EC(90) for intracellular virions with thiazolide structural parameters. Finally we discuss the mechanism of action of thiazolides in relation to the present results.
Collapse
Affiliation(s)
- Andrew V Stachulski
- Robert Robinson Laboratories, Department of Chemistry, University of Liverpool, Liverpool L69 7ZD, UK.
| | | | | | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
36
|
Initial virological response and viral mutation with adefovir dipivoxil added to ongoing Lamivudine therapy in Lamivudine-resistant chronic hepatitis B. Dig Dis Sci 2011; 56:1207-14. [PMID: 20927588 DOI: 10.1007/s10620-010-1423-y] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/16/2010] [Accepted: 09/03/2010] [Indexed: 12/24/2022]
Abstract
BACKGROUND Although adefovir dipivoxil (ADV) has been used for antiviral treatment of lamivudine (LAM)-resistant chronic hepatitis B (CHB) patients, the long-term efficacy of this treatment is not well understood. Initial virological response (IVR) has been reported to be an important factor in relation to the development of ADV-resistance. AIMS We therefore examined the factors associated with IVR and ADV mutation in these patients. METHODS Forty-nine LAM-resistant CHB patients with ADV add-on LAM therapy, 47% of whom were hepatitis B e-antigen (HBeAg)-positive with median treatment duration of 23 months, were enrolled in this study. Patients were classified into IVR and non-IVR groups on the basis of viral suppression status. Mutational analysis of the HBV polymerase/reverse transcriptase (rt) domain was performed by PCR-direct sequencing. RESULTS Serum HBV DNA was undetectable (<2.6 log10 copies/mL) in 67, 82, and 84% of patients at 24, 48, and 96 weeks, respectively, after ADV add-on LAM therapy. IVR was achieved in 82% of patients, and ALT normalized at week 24 in 90% of IVR and 78% of non-IVR patients. The lower pretreatment HBV DNA level and virus-containing mutations other than double mutation of rtL180M + rtM204V were significantly associated with IVR (P=0.002 and P=0.014, respectively). ADV-resistant mutations in the RT motif, reported previously, were not detected. CONCLUSION IVR is useful for predicting the antiviral efficacy of ADV and LAM combination therapy in LAM-resistant CHB.
Collapse
|
|
37
|
Outbreak of infections by hepatitis B virus genotype A and transmission of genetic drug resistance in patients coinfected with HIV-1 in Japan. J Clin Microbiol 2011; 49:1017-24. [PMID: 21248087 DOI: 10.1128/jcm.02149-10] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022] Open
Abstract
The major routes of hepatitis B virus (HBV) infection in Japan has been mother-to-child transmission (MTCT) and blood transfusion. However, HBV cases transmitted through sexual contact are increasing, especially among HIV-1-seropositive patients. To understand the molecular epidemiology of HBV in HBV/HIV-1 coinfection, we analyzed HBV genotypes and HIV-1 subtypes in HBV/HIV-1-coinfected patients at Nagoya Medical Center from 2003 to 2007. Among 394 HIV-1-infected Japanese men having sex with men (MSM) who were newly diagnosed during the study period, 31 (7.9%) tested positive for the hepatitis B virus surface antigen. HBV sequence analyses were successful in 26 cases, with 21 (80.7%) and 5 (19.3%) cases determined as genotypes A and C, respectively. Our finding that HBV genotype A was dominant in HIV-1-seropositive patients alerts clinicians to an alternative outbreak of HBV genotype A in the HIV-1-infected MSM population and a shift in HBV genotype from C to A in Japan. The narrow genetic diversity in genotype A cases suggests that genotype A has been recently introduced into the MSM population and that sexual contacts among MSM were more active than speculated from HIV-1 tree analyses. In addition, we found a lamivudine resistance mutation in one naïve case, suggesting a risk of drug-resistant HBV transmission. As genotype A infection has a higher risk than infection with other genotypes for individuals to become HBV carriers, prevention programs are urgently needed for the target population.
Collapse
|
|
38
|
Te HS. Antiviral Therapy: Analysis of Long-term Efficacy and Safety. CURRENT HEPATITIS REPORTS 2010; 9:214-222. [DOI: 10.1007/s11901-010-0052-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
|
|
39
|
Kim KH, Kim ND, Seong BL. Discovery and development of anti-HBV agents and their resistance. Molecules 2010; 15:5878-908. [PMID: 20802402 PMCID: PMC6257723 DOI: 10.3390/molecules15095878] [Citation(s) in RCA: 46] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/04/2010] [Revised: 08/24/2010] [Accepted: 08/26/2010] [Indexed: 02/07/2023] Open
Abstract
Hepatitis B virus (HBV) infection is a prime cause of liver diseases such as hepatitis, cirrhosis and hepatocellular carcinoma. The current drugs clinically available are nucleot(s)ide analogues that inhibit viral reverse transcriptase activity. Most drugs of this class are reported to have viral resistance with breakthrough. Recent advances in methods for in silico virtual screening of chemical libraries, together with a better understanding of the resistance mechanisms of existing drugs have expedited the discovery and development of novel anti-viral drugs. This review summarizes the current status of knowledge about and viral resistance of HBV drugs, approaches for the development of novel drugs as well as new viral and host targets for future drugs.
Collapse
Affiliation(s)
- Kyun-Hwan Kim
- Department of Pharmacology, School of Medicine, and Center for Cancer Research and Diagnostic Medicine, IBST, Konkuk University, Seoul 143-701, Korea
- Research Institute of Medical Sciences, Konkuk University, Seoul 143-701, Korea
- Author to whom correspondence should be addressed; E-Mail: (K.H.K.); Tel.: +82 2 2030 7833; Fax: +82 2 2049 6192; E-Mail: (B.L.S.); Tel.: +82 2 2123 2885; Fax: +82 2 392 3582
| | - Nam Doo Kim
- R&D Center, Equispharm Inc., 11F Gyeonggi Bio-Center, 864-1 Iui-Dong, Yeongtong-gu, Suwon-Shi, Gyeonggi-Do 443-766, Korea
| | - Baik-Lin Seong
- Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Korea
- Translational Research Center for Protein Function Control, Yonsei University, Seoul 120-749, Korea
- Author to whom correspondence should be addressed; E-Mail: (K.H.K.); Tel.: +82 2 2030 7833; Fax: +82 2 2049 6192; E-Mail: (B.L.S.); Tel.: +82 2 2123 2885; Fax: +82 2 392 3582
| |
Collapse
|
|
40
|
Gao Z, Liu FJ, Liu L, Zhou TY, Lei J, Xu L, Liu C, Dai J, Chen EQ, Tang H. Application of hepatitis B virus replication mouse model. World J Gastroenterol 2010; 16:1979-1985. [PMID: 20419834 PMCID: PMC2860074 DOI: 10.3748/wjg.v16.i16.1979] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/07/2010] [Revised: 02/07/2010] [Accepted: 02/14/2010] [Indexed: 02/06/2023] Open
Abstract
AIM To evaluate the value of the hepatitis B virus (HBV) replication mouse model with regard to several aspects of the study of HBV biology. METHODS To evaluate the HBV replication mouse model in detecting the efficacy of anti-HBV agents, the interferon inducer polyinosinic-polytidylin acid (polyIC) and nucleotide analogues adefovir and entecavir were administered to mice injected with wild type pHBV4.1, and the inhibiting effect of these agents on HBV DNA replication was evaluated. To identify the model's value in a replication ability study of HBV drug-resistant mutants and a HBx-minus mutant, telbivudine resistance mutants (rtM204I, ayw subtype), adefovir resistance mutants (rtA181V + rtN236T, ayw subtype) and HBx-minus mutants were injected respectively, and their corresponding HBV DNA replication intermediates in mouse liver were assessed. RESULTS Compared with the wild type HBV replication mouse model without antiviral agent treatment, the HBV DNA replication intermediates of the polyIC-treated group were decreased 1-fold; while in the entecavir- and adefovir-treated groups, the levels of HBV DNA replication intermediates were inhibited 13.6-fold and 1.4-fold, respectively. For the mouse models injected with telbivudine resistance mutant, adefovir resistance mutant and HBx-minus mutant, HBV DNA replication intermediates could still be detected, but the levels of HBV DNA replication intermediates of these mutants decreased 4.5-fold, 5.6-fold and 2.9-fold respectively, compared with the mouse model with wild type HBV plasmid. CONCLUSION The HBV replication mouse model we established was a useful and convenient tool to detect the efficacy of antiviral agents and to study the replication ability of HBV mutants in vivo.
Collapse
|
|
41
|
Liu BM, Li T, Xu J, Li XG, Dong JP, Yan P, Yang JX, Yan L, Gao ZY, Li WP, Sun XW, Wang YH, Jiao XJ, Hou CS, Zhuang H. Characterization of potential antiviral resistance mutations in hepatitis B virus reverse transcriptase sequences in treatment-naïve Chinese patients. Antiviral Res 2009; 85:512-9. [PMID: 20034521 DOI: 10.1016/j.antiviral.2009.12.006] [Citation(s) in RCA: 84] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2009] [Revised: 10/28/2009] [Accepted: 12/15/2009] [Indexed: 02/06/2023]
Abstract
Full-length hepatitis B virus (HBV) reverse transcriptase (RT) sequences were amplified and sequenced among 192 nucleos(t)ide analogue (NA)-naïve Chinese patients with chronic hepatitis B. Deduced amino acids (AAs) at 42 previously reported potential NA resistance (NAr) mutation positions in RT region were analyzed. Patients were found with either B-genotype (28.65%) or C-genotype (71.35%) infections. Rt53, rt91, rt124, rt134, rt221, rt224, rt238 and rt256 were identified as B- and C-genotype-dependent polymorphic AA positions. AA substitutions at 11 classical NAr mutation positions, i.e. rt80, rt169, rt173, rt180, rt181, rt184, rt194, rt202, rt204, rt236 and rt250, were not detected. However, potential NAr mutations were found in 30.73% (59/192) isolates, which involved 18 positions including rt53, rt207, rt229, rt238 and rt256, etc. The concomitant AA changes of HBsAg occurred in 16.67% (32/192) isolates including sG145R mutation. One-third of mutation positions were located in functional RT domains (e.g. rt207 and rt233), A-B interdomains (overlapping HBsAg 'a' determinant and showing most concomitant immune-associated mutations) and non-A-B interdomains (e.g. rt191 and rt213), respectively. Genotypes B and C each showed several preferred positions to mutate. These results might provide insights into understanding the evolution and selection basis of NAr HBV strains under antiviral therapy.
Collapse
Affiliation(s)
- Bao-Ming Liu
- Department of Microbiology, Peking University Health Science Center, Xueyuan Road 38, Haidian District, 100191 Beijing, China
| | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
42
|
Li Y, Zhu M, Guo Y, Chen W, Li G. Full-length hepatitis B virus sequences from naïve patients with fluctuation of viral load during ADV monotherapy. Virus Genes 2009; 40:155-62. [PMID: 20012680 DOI: 10.1007/s11262-009-0429-z] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2009] [Accepted: 11/26/2009] [Indexed: 12/25/2022]
Abstract
The reasons for adefovir dipivoxil (ADV) treatment failures appear diverse. Few studies have reported full-length hepatitis B virus (HBV) genome in patients with ADV treatment failures. The patients were from a phase III clinical trial that investigated the antiviral response to ADV in China. Seven patients had increase in HBV-DNA (>1 log(10) copies/ml above on-treatment nadir) at week 52. The serum HBV-DNA levels were above 10(4)copies/ml at week 92 in four of them. Sixteen full-length HBV genomes from the four patients at four time points were sequenced using cloning sequencing method. The frequency of substitutions at week 52 was higher than at weeks 28(16 wt) and 92(80). HBV-DNA reduction was correlated negatively with the frequency of substitutions at the three time points. No published ADV-resistant mutations were detected. The mutations, including substitutions in immunogenic epitopes and conserved sites of the polymerase gene, were frequent during ADV treatment. Amino acid deletions in X gene and basal core promoter/pre-core mutations appeared before or during ADV treatment. The substitutions in immunogenic epitopes (mainly of the surface gene) and conserved sites of the polymerase gene other than ADV-resistant mutations may have influenced antiviral efficacy in the study. More potent antiviral drugs may be important to rescue individual patients and for public health safety. It is needed to study how these substitutions influence HBV replication, disease progression, and antiviral treatment efficacy.
Collapse
Affiliation(s)
- Yongwei Li
- Department of Infectious Diseases, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
| | | | | | | | | |
Collapse
|
|
43
|
The underlying mechanisms for the 'anti-HBc alone' serological profile. Arch Virol 2009; 155:149-58. [PMID: 20091193 DOI: 10.1007/s00705-009-0559-6] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/23/2009] [Accepted: 10/27/2009] [Indexed: 02/06/2023]
Abstract
The serological pattern, "anti-HBc alone", characterized by the presence of antibodies against the core antigen of hepatitis B virus (anti-HBc) as the only marker of hepatitis B, is not rare in a diagnostic setting. Depending on the prevalence of HBV infection and the patient group investigated, 1-31% of positive anti-HBc results are isolated positive findings. Anti-HBc alone is frequently observed in intravenous drug addicts, HIV-infected individuals, patients who are coinfected with HBV and hepatitis C virus, and pregnant women. However, it is not clear how this profile should be interpreted. Several studies have shown that anti-HBc alone is not only compatible with acute and resolved HBV infection but also with chronic infection. The reasons for the lack of HBsAg and anti-HBs in anti-HBc-alone individuals are not clear, but several mechanisms and possibilities have been suggested that could explain this phenomenon, some of which are delineated in this article.
Collapse
|
|
44
|
Yuen MF, Fung J, Wong DKH, Lai CL. Prevention and management of drug resistance for antihepatitis B treatment. THE LANCET. INFECTIOUS DISEASES 2009; 9:256-64. [PMID: 19324298 DOI: 10.1016/s1473-3099(09)70056-8] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
Emergence of drug resistance in antiviral therapy for chronic hepatitis B negates treatment benefits. There is a lower chance for emergence of resistance for drugs with rapid and potent viral suppression and a high genetic barrier for resistant mutations. Measurement of viral load at 24 weeks' treatment to aid decision making is mandatory for patients receiving drugs that are associated with a higher resistance rate. Combination treatment with drugs that belong to different groups is associated with a lower chance of resistance. To ensure better control of viral replication in patients with drug resistance, the addition of another drug without an overlapping resistance profile should be given as early as possible, preferably at the time when genotypic resistance emerges. With such strategies, most patients can be maintained in clinical remission. However, because of the mechanism of viral persistence, research efforts should continue to anticipate and prevent the emergence of multidrug-resistant strains.
Collapse
Affiliation(s)
- Man-Fung Yuen
- Department of Medicine, University of Hong Kong, Queen Mary Hospital, Hong Kong
| | | | | | | |
Collapse
|
|
45
|
Molecular analysis of an HBsAg-negative hepatitis B virus mutant selected in a tenofovir-treated HIV-hepatitis B virus co-infected patient. AIDS 2009; 23:268-72. [PMID: 19098499 DOI: 10.1097/qad.0b013e3283224316] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/12/2023]
Abstract
The molecular analysis performed in an HIV-hepatitis B virus (HBV) coinfected patient revealed selection of an unusual HBV polymerase mutation (rtV191I) during tenofovir-containing therapy, conferring simultaneously immune escape by HBsAg negativity and resistance to lamivudine but not tenofovir. Phenotypic analysis revealed impaired replicative capacity of mutants, which could be restored by concomitant precore or basal core promoter mutations (HBe-antigen-negativity). HBV mutants carrying drug and vaccine resistance may represent a considerable individual risk and public health concern.
Collapse
|
|
46
|
Pallier C, Rodriguez C, Brillet R, Nordmann P, Hézode C, Pawlotsky JM. Complex dynamics of hepatitis B virus resistance to adefovir. Hepatology 2009; 49:50-9. [PMID: 19065672 PMCID: PMC2956748 DOI: 10.1002/hep.22634] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/19/2022]
Abstract
UNLABELLED In patients with hepatitis B e antigen-negative chronic hepatitis B, adefovir dipivoxil administration selects variants bearing reverse transcriptase rtN236T and/or rtA181V/T substitutions in 29% of cases after 5 years. The aim of this study was to characterize the dynamics of adefovir-resistant variant populations during adefovir monotherapy in order to better understand the molecular mechanisms underlying hepatitis B virus resistance to this class of nucleotide analogues. Patients included in a 240-week clinical trial of adefovir monotherapy who developed adefovir resistance-associated substitutions were studied. The dynamics of hepatitis B virus populations were analyzed over time, after generating nearly 4,000 full-length reverse transcriptase sequences, and compared with the replication kinetics of the virus during therapy. Whatever the viral kinetics pattern, adefovir resistance was characterized by exclusive detection of a dominant wild-type, adefovir-sensitive variant population at baseline and late and gradual selection by adefovir of several coexisting resistant viral populations, defined by the presence of amino acid substitutions at position rt236, position rt181, or both. The gain in fitness of one or the other of these resistant populations during adefovir administration was never associated with the selection of additional amino acid substitutions in the reverse transcriptase. CONCLUSION Our results suggest that adefovir administration selects poorly fit preexisting or emerging viral populations with low-level adefovir resistance, which subsequently compete to fill the replication space. Viral kinetics depends on the initial virological response to adefovir. Lamivudine add-on restores some antiviral efficacy, but adefovir-resistant variants remain predominant. Whether these adefovir resistance-associated substitutions may confer cross-resistance to tenofovir in vivo will need to be determined.
Collapse
Affiliation(s)
- Coralie Pallier
- Centre de référence français des hépatites B, C et delta
AP-HPHôpital Henri MondorUniversité Paris XII Val de MarneDépartement de virologie Créteil,FR,IMRB, Institut Mondor de recherche biomédicale
INSERM : U841Université Paris XII Val de MarneHôpital Henri Mondor 51, av du mal de lattre de tassigny 94010 CRETEIL CEDEX,FR
| | - Christophe Rodriguez
- Centre de référence français des hépatites B, C et delta
AP-HPHôpital Henri MondorUniversité Paris XII Val de MarneDépartement de virologie Créteil,FR,IMRB, Institut Mondor de recherche biomédicale
INSERM : U841Université Paris XII Val de MarneHôpital Henri Mondor 51, av du mal de lattre de tassigny 94010 CRETEIL CEDEX,FR
| | - Rozenn Brillet
- Centre de référence français des hépatites B, C et delta
AP-HPHôpital Henri MondorUniversité Paris XII Val de MarneDépartement de virologie Créteil,FR,IMRB, Institut Mondor de recherche biomédicale
INSERM : U841Université Paris XII Val de MarneHôpital Henri Mondor 51, av du mal de lattre de tassigny 94010 CRETEIL CEDEX,FR
| | - Patrice Nordmann
- Service de bactériologie et virologie
AP-HPHôpital BicêtreUniversité Paris Sud - Paris XILe Kremlin-Bicêtre,FR
| | - Christophe Hézode
- Centre de référence français des hépatites B, C et delta
AP-HPHôpital Henri MondorUniversité Paris XII Val de MarneDépartement de virologie Créteil,FR,IMRB, Institut Mondor de recherche biomédicale
INSERM : U841Université Paris XII Val de MarneHôpital Henri Mondor 51, av du mal de lattre de tassigny 94010 CRETEIL CEDEX,FR,Department of hepatology and gastroenterologyHôpital Henri Mondor, Université Paris 12Creteil,FR
| | - Jean-Michel Pawlotsky
- Centre de référence français des hépatites B, C et delta
AP-HPHôpital Henri MondorUniversité Paris XII Val de MarneDépartement de virologie Créteil,FR,IMRB, Institut Mondor de recherche biomédicale
INSERM : U841Université Paris XII Val de MarneHôpital Henri Mondor 51, av du mal de lattre de tassigny 94010 CRETEIL CEDEX,FR,* Correspondence should be adressed to: Jean-Michel Pawlotsky
| |
Collapse
|
|
47
|
Moon JH, Cho M, Yoon KT, Bae JH, Heo J, Kim GH, Kang DH, Song GA. [The efficacy of adefovir dipivoxil monotherapy and the incidence of genotypic resistance to adefovir dipivoxil in patients with lamivudine-resistant chronic hepatitis B infection]. THE KOREAN JOURNAL OF HEPATOLOGY 2008; 14:503-512. [PMID: 19119245 DOI: 10.3350/kjhep.2008.14.4.503] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
BACKGROUND/AIMS Adefovir dipivoxil (ADV) is a nucleotide analogue that inhibits wild-type hepatitis B virus (HBV) and lamivudine (LMV)-resistant HBV mutants. The aim of this study was to elucidate the efficacy of ADV monotherapy and the incidence of genotypic resistance to ADV in patients with LMV-resistant chronic HBV infection. METHODS This study involved 124 patients with chronic HBV infection who had received ADV monotherapy due to the presence of LMV-resistant HBV mutants. The efficacy of ADV was evaluated by the normalization of serum alanine aminotransferase (ALT) level and by the reduction of serum HBV DNA level (with cutoff levels of 2x10(4) IU/mL and 2x10(2) IU/mL). The cumulative rate of HBeAg loss or seroconversion was assessed in HBeAg-positive patients. The development of mutations in the reverse trancriptase region of HBV DNA polymerase was evaluated by direct sequencing analysis during ADV monotherapy. RESULTS The mean serum HBV DNA level was 5.94 log10IU/mL. At 12 and 24 months after ADV monotherapy, the cumulative rates of serum ALT normalization were 69.4% and 75.5%, respectively, and those of serum HBV DNA reduction were 79.8% and 89.2% for a cutoff level of 2x10(4) IU/mL, and 44.2% and 59.0% for a cutoff of 2x10(2) IU/mL. The mean serum HBV DNA levels at 12 and 24 months were significantly lower than baseline, at 3.24 and 3.04 log10IU/mL, respectively (P<0.001). At 12 months after ADV treatment, the cumulative rates of HBeAg loss and seroconversion were 15.8% and 10.5%, respectively, and the rtN236T and rtA181T/V mutants in HBV DNA polymerase were identified in 25% and 64% of patients, respectively. CONCLUSIONS Although ADV monotherapy is effective, it leads to a high rate of mutations of HBV DNA reverse transcriptase gene in patients with chronic HBV infections who have LMV-resistant HBV mutants.
Collapse
Affiliation(s)
- Jae Hyeon Moon
- Department of Internal Medicine, Pusan National University School of Medicine, Busan, Korea
| | | | | | | | | | | | | | | |
Collapse
|
|
48
|
Antiviral effects of lamivudine, emtricitabine, adefovir dipivoxil, and tenofovir disoproxil fumarate administered orally alone and in combination to woodchucks with chronic woodchuck hepatitis virus infection. Antimicrob Agents Chemother 2008; 52:3617-32. [PMID: 18676881 DOI: 10.1128/aac.00654-08] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023] Open
Abstract
Adefovir dipivoxil (ADV) and tenofovir disoproxil fumarate (TDF) are nucleotide analogs that inhibit the replication of wild-type hepatitis B virus (HBV) and lamivudine (3TC)-resistant virus in HBV-infected patients, including those who are coinfected with human immunodeficiency virus. The combination of ADV or TDF with other nucleoside analogs is a proposed strategy for managing antiviral drug resistance during the treatment of chronic HBV infection. The antiviral effect of oral ADV or TDF, alone or in combination with 3TC or emtricitabine (FTC), against chronic woodchuck hepatitis virus (WHV) infection was evaluated in a placebo-controlled study in the woodchuck, an established and predictive model for antiviral therapy. Once-daily treatment for 48 weeks with ADV plus 3TC or TDF plus FTC significantly reduced serum WHV viremia levels from the pretreatment level by 6.2 log(10) and 6.1 log(10) genome equivalents/ml serum, respectively, followed by TDF plus 3TC (5.6 log(10) genome equivalents/ml), ADV alone (4.8 log(10) genome equivalents/ml), ADV plus FTC (one survivor) (4.4 log(10) genome equivalents/ml), TDF alone (2.9 log(10) genome equivalents/ml), 3TC alone (2.7 log(10) genome equivalents/ml), and FTC alone (2.0 log(10) genome equivalents/ml). Individual woodchucks across all treatment groups also demonstrated pronounced declines in serum WHV surface antigen, characteristically accompanied by declines in hepatic WHV replication and the hepatic expression of WHV antigens. Most woodchucks had prompt recrudescence of WHV replication after drug withdrawal, but individual woodchucks across treatment groups had sustained effects. No signs of toxicity were observed for any of the drugs or drug combinations administered. In conclusion, the oral administration of 3TC, FTC, ADV, and TDF alone and in combination was safe and effective in the woodchuck model of HBV infection.
Collapse
|
|
49
|
Chen EQ, Lei BJ, Tang H. Clinical application and research progress in hepatitis B virus quasispecies. Shijie Huaren Xiaohua Zazhi 2008; 16:1086. [DOI: 10.11569/wcjd.v16.i10.1086] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
|
|
50
|
Curtis M, Zhu Y, Borroto-Esoda K. Hepatitis B virus containing the I233V mutation in the polymerase reverse-transcriptase domain remains sensitive to inhibition by adefovir. J Infect Dis 2007; 196:1483-6. [PMID: 18008227 DOI: 10.1086/522521] [Citation(s) in RCA: 35] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2007] [Accepted: 05/21/2007] [Indexed: 01/12/2023] Open
Abstract
An isoleucine-to-valine change at position 233 (rtI233V) of hepatitis B virus (HBV) polymerase was recently reported to cause decreased in vitro susceptibility to, and treatment failure of, adefovir dipivoxil (ADV). To further evaluate these findings, we screened our ADV clinical-study sequence database of 853 patients and identified 4 who, at baseline, had HBV with this mutation. All 4 patients responded to treatment with ADV, with a median change in HBV DNA levels of 4.0 log(10) copies/mL after 48 weeks of treatment. Phenotypic evaluation of clinical isolates and of a laboratory strain with the rtI233V mutation demonstrated their full susceptibility to adefovir in vitro, and HBV with the rtI233V mutation developed in none of the patients.
Collapse
Affiliation(s)
- Maria Curtis
- Gilead Sciences, 4 University Place, Durham, NC 27707, USA
| | | | | |
Collapse
|