1887

Abstract

Hepatitis B virus (HBV) is a major human pathogen that causes liver diseases. The main HBV RNAs are unspliced transcripts that encode the key viral proteins. Recent studies have shown that some of the HBV spliced transcript isoforms are predictive of liver cancer, yet the roles of these spliced transcripts remain elusive. Furthermore, there are nine major HBV genotypes common in different regions of the world, these genotypes may express different spliced transcript isoforms. To systematically study the HBV splice variants, we transfected human hepatoma cells, Huh7, with four HBV genotypes (A2, B2, C2 and D3), followed by deep RNA-sequencing. We found that 13–28 % of HBV RNAs were splice variants, which were reproducibly detected across independent biological replicates. These comprised 6 novel and 10 previously identified splice variants. In particular, a novel, singly spliced transcript was detected in genotypes A2 and D3 at high levels. The biological relevance of these splice variants was supported by their identification in HBV-positive liver biopsy and serum samples, and in HBV-infected primary human hepatocytes. Interestingly the levels of HBV splice variants varied across the genotypes, but the spliced pregenomic RNA SP1 and SP9 were the two most abundant splice variants. Counterintuitively, these singly spliced SP1 and SP9 variants had a suboptimal 5′ splice site, supporting the idea that splicing of HBV RNAs is tightly controlled by the viral post-transcriptional regulatory RNA element.

Funding
This study was supported by the:
  • National Health and Medical Research Council (Award APP1145977)
    • Principle Award Recipient: PeterA. Revill
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2021-01-13
2024-03-29
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References

  1. Betz-Stablein BD, Töpfer A, Littlejohn M, Yuen L, Colledge D et al. Single-molecule sequencing reveals complex genome variation of hepatitis B virus during 15 years of chronic infection following liver transplantation. J Virol 2016; 90:7171–7183 [View Article][PubMed]
    [Google Scholar]
  2. Hou J, Lin L, Zhou W, Wang Z, Ding G et al. Identification of miRNomes in human liver and hepatocellular carcinoma reveals miR-199a/b-3p as therapeutic target for hepatocellular carcinoma. Cancer Cell 2011; 19:232–243 [View Article][PubMed]
    [Google Scholar]
  3. Huang Q, Lin B, Liu H, Ma X, Mo F et al. RNA-Seq analyses generate comprehensive transcriptomic landscape and reveal complex transcript patterns in hepatocellular carcinoma. PLoS One 2011; 6:e26168 [View Article][PubMed]
    [Google Scholar]
  4. Chen J, Li Y, Lai F, Wang Y, Sutter K et al. Functional comparison of IFN‐α subtypes reveals potent HBV suppression by a concerted action of IFN‐α and ‐γ signaling. Hepatology 2020 [View Article][PubMed]
    [Google Scholar]
  5. Sato A, Ono C, Tamura T, Mori H, Izumi T et al. Rimonabant suppresses RNA transcription of hepatitis B virus by inhibiting hepatocyte nuclear factor 4α. Microbiol Immunol 2020; 64:345–355 [View Article][PubMed]
    [Google Scholar]
  6. Liu PJ, Harris JM, Marchi E, D'Arienzo V, Michler T et al. Hypoxic gene expression in chronic hepatitis B virus infected patients is not observed in state-of-the-art in vitro and mouse infection models. Sci Rep 2020; 10:14101 [View Article][PubMed]
    [Google Scholar]
  7. Yang Y, Chen L, Gu J, Zhang H, Yuan J et al. Recurrently deregulated lncRNAs in hepatocellular carcinoma. Nat Commun 2017; 8:14421 [View Article][PubMed]
    [Google Scholar]
  8. Niu C, Livingston CM, Li L, Beran RK, Daffis S et al. The Smc5/6 complex restricts HBV when localized to ND10 without inducing an innate immune response and is counteracted by the HBV X protein shortly after infection. PLoS One 2017; 12:e0169648 [View Article][PubMed]
    [Google Scholar]
  9. Yoo S, Wang W, Wang Q, Fiel MI, Lee E et al. A pilot systematic genomic comparison of recurrence risks of hepatitis B virus-associated hepatocellular carcinoma with low- and high-degree liver fibrosis. BMC Med 2017; 15:214 [View Article][PubMed]
    [Google Scholar]
  10. Hensel KO, Cantner F, Bangert F, Wirth S, Postberg J. Episomal HBV persistence within transcribed host nuclear chromatin compartments involves HBx. Epigenetics Chromatin 2018; 11:34 [View Article][PubMed]
    [Google Scholar]
  11. Niu C, Li L, Daffis S, Lucifora J, Bonnin M et al. Toll-like receptor 7 agonist GS-9620 induces prolonged inhibition of HBV via a type I interferon-dependent mechanism. J Hepatol 2018; 68:922–931 [View Article][PubMed]
    [Google Scholar]
  12. Song M, Sun Y, Tian J, He W, Xu G et al. Silencing retinoid X receptor alpha expression enhances early-stage hepatitis B virus infection in cell cultures. J Virol 2018; 92:e01771-17 [View Article][PubMed]
    [Google Scholar]
  13. Winer BY, Gaska JM, Lipkowitz G, Bram Y, Parekh A et al. Analysis of host responses to hepatitis B and delta viral infections in a micro-scalable hepatic co-culture system. Hepatology 2020; 71:14–30 [View Article][PubMed]
    [Google Scholar]
  14. Moreau P, Cournac A, Palumbo GA, Marbouty M, Mortaza S et al. Tridimensional infiltration of DNA viruses into the host genome shows preferential contact with active chromatin. Nat Commun 2018; 9:4268 [View Article][PubMed]
    [Google Scholar]
  15. De Crignis E, Romal S, Carofiglio F, Moulos P, Verstegen MMA. Human liver organoids; a patient-derived primary model for HBV infection and related hepatocellular carcinoma. bioRxiv 2020568147
    [Google Scholar]
  16. Li C, Hou Y, Xu J, Zhang A, Liu Z et al. A direct test of selection in cell populations using the diversity in gene expression within tumors. Mol Biol Evol 2017; 34:1730–1742 [View Article][PubMed]
    [Google Scholar]
  17. Liu L, He C, Liu H, Wang G, Lv Z et al. Transcriptomic profiling of long non-coding RNAs in non-virus associated hepatocellular carcinoma. Cell Biochem Biophys 2020; 78:465–474 [View Article][PubMed]
    [Google Scholar]
  18. Wang H, Zhang CZ, Lu S-X, Zhang M-F, Liu L-L et al. A coiled-coil domain containing 50 splice variant is modulated by serine/arginine-rich splicing factor 3 and promotes hepatocellular carcinoma in mice by the Ras signaling pathway. Hepatology 2019; 69:179–195 [View Article][PubMed]
    [Google Scholar]
  19. Lim CS, Brown CM. Hepatitis B virus nuclear export elements: RNA stem-loop α and β, key parts of the HBV post-transcriptional regulatory element. RNA Biol 2016; 13:743–747 [View Article][PubMed]
    [Google Scholar]
  20. Stadelmayer B, Diederichs A, Chapus F, Rivoire M, Neveu G et al. Full-length 5'RACE identifies all major HBV transcripts in HBV-infected hepatocytes and patient serum. J Hepatol 2020; 73:40–51 [View Article][PubMed]
    [Google Scholar]
  21. Krause-Kyora B, Susat J, Key FM, Kühnert D, Bosse E et al. Neolithic and medieval virus genomes reveal complex evolution of hepatitis B. eLife 2018; 7:e36666 [View Article][PubMed]
    [Google Scholar]
  22. Yuen LKW, Littlejohn M, Duchêne S, Edwards R, Bukulatjpi S et al. Tracing ancient human migrations into Sahul using hepatitis B virus genomes. Mol Biol Evol 2019; 36:942–954 [View Article][PubMed]
    [Google Scholar]
  23. Revill PA, Tu T, Netter HJ, Yuen LKW, Locarnini SA et al. The evolution and clinical impact of hepatitis B virus genome diversity. Nat Rev Gastroenterol Hepatol 2020; 17:618–634 [View Article][PubMed]
    [Google Scholar]
  24. McNaughton AL, Revill PA, Littlejohn M, Matthews PC, Ansari MA. Analysis of genomic-length HBV sequences to determine genotype and subgenotype reference sequences. J Gen Virol 2020; 101:271–283 [View Article][PubMed]
    [Google Scholar]
  25. Sozzi V, Walsh R, Littlejohn M, Colledge D, Jackson K et al. In vitro studies show that sequence variability contributes to marked variation in hepatitis B virus replication, protein expression, and function observed across genotypes. J Virol 2016; 90:10054–10064 [View Article][PubMed]
    [Google Scholar]
  26. Kramvis A. Genotypes and genetic variability of hepatitis B virus. Intervirology 2014; 57:141–150 [View Article][PubMed]
    [Google Scholar]
  27. Kao J-H. Molecular epidemiology of hepatitis B virus. Korean J Intern Med 2011; 26:255–261 [View Article][PubMed]
    [Google Scholar]
  28. Kramvis A, Kostaki E-G, Hatzakis A, Paraskevis D. Immunomodulatory function of HBeAg related to short-sighted evolution, transmissibility, and clinical manifestation of hepatitis B virus. Front Microbiol 2018; 9:2521 [View Article][PubMed]
    [Google Scholar]
  29. Chotiyaputta W, Lok ASF. Hepatitis B virus variants. Nat Rev Gastroenterol Hepatol 2009; 6:453–462 [View Article][PubMed]
    [Google Scholar]
  30. Lee GH, Wasser S, Lim SG. Hepatitis B pregenomic RNA splicing – the products, the regulatory mechanisms and its biological significance. Virus Res 2008; 136:1–7 [View Article][PubMed]
    [Google Scholar]
  31. Candotti D, Allain J-P. Biological and clinical significance of hepatitis B virus RNA splicing: an update. Ann Blood 2016; 2:6 [View Article]
    [Google Scholar]
  32. Huang C-C, Kuo T-M, Yeh C-T, Hu C, Chen Y-L et al. One single nucleotide difference alters the differential expression of spliced RNAs between HBV genotypes A and D. Virus Res 2013; 174:18–26 [View Article][PubMed]
    [Google Scholar]
  33. Ito N, Nakashima K, Sun S, Ito M, Suzuki T. Cell type diversity in hepatitis B virus RNA splicing and its regulation. Front Microbiol 2019; 10:207 [View Article][PubMed]
    [Google Scholar]
  34. Sommer G, van Bömmel F, Will H. Genotype-specific synthesis and secretion of spliced hepatitis B virus genomes in hepatoma cells. Virology 2000; 271:371–381 [View Article][PubMed]
    [Google Scholar]
  35. Chen PJ, Chen CR, Sung JL, Chen DS. Identification of a doubly spliced viral transcript joining the separated domains for putative protease and reverse transcriptase of hepatitis B virus. J Virol 1989; 63:4165–4171 [View Article][PubMed]
    [Google Scholar]
  36. Su TS, Lai CJ, Huang JL, Lin LH, Yauk YK et al. Hepatitis B virus transcript produced by RNA splicing. J Virol 1989; 63:4011–4018 [View Article][PubMed]
    [Google Scholar]
  37. Terré S, Petit MA, Bréchot C. Defective hepatitis B virus particles are generated by packaging and reverse transcription of spliced viral RNAs in vivo . J Virol 1991; 65:5539–5543 [View Article][PubMed]
    [Google Scholar]
  38. Wu HL, Chen PJ, Tu SJ, Lin MH, Lai MY et al. Characterization and genetic analysis of alternatively spliced transcripts of hepatitis B virus in infected human liver tissues and transfected HepG2 cells. J Virol 1991; 65:1680–1686 [View Article][PubMed]
    [Google Scholar]
  39. Rosmorduc O, Petit MA, Pol S, Capel F, Bortolotti F et al. In vivo and in vitro expression of defective hepatitis B virus particles generated by spliced hepatitis B virus RNA. Hepatology 1995; 22:10–19[PubMed]
    [Google Scholar]
  40. Günther S, Sommer G, Iwanska A, Will H. Heterogeneity and common features of defective hepatitis B virus genomes derived from spliced pregenomic RNA. Virology 1997; 238:363–371 [View Article][PubMed]
    [Google Scholar]
  41. Abraham TM, Lewellyn EB, Haines KM, Loeb DD. Characterization of the contribution of spliced RNAs of hepatitis B virus to DNA synthesis in transfected cultures of Huh7 and HepG2 cells. Virology 2008; 379:30–37 [View Article][PubMed]
    [Google Scholar]
  42. El Chaar M, El Jisr T, Allain J-P. Hepatitis B virus DNA splicing in Lebanese blood donors and genotype A to E strains: implications for hepatitis B virus DNA quantification and infectivity. J Clin Microbiol 2012; 50:3159–3167 [View Article][PubMed]
    [Google Scholar]
  43. Chen J, Wu M, Wang F, Zhang W, Wang W et al. Hepatitis B virus spliced variants are associated with an impaired response to interferon therapy. Sci Rep 2015; 5:16459 [View Article][PubMed]
    [Google Scholar]
  44. Lam AM, Ren S, Espiritu C, Kelly M, Lau V et al. Hepatitis B virus capsid assembly modulators, but not nucleoside analogs, inhibit the production of extracellular pregenomic RNA and spliced RNA variants. Antimicrob Agents Chemother 2017; 61:e00680-17 [View Article][PubMed]
    [Google Scholar]
  45. Hass M, Hannoun C, Kalinina T, Sommer G, Manegold C et al. Functional analysis of hepatitis B virus reactivating in hepatitis B surface antigen-negative individuals. Hepatology 2005; 42:93–103 [View Article][PubMed]
    [Google Scholar]
  46. Candotti D, Lin CK, Belkhiri D, Sakuldamrongpanich T, Biswas S et al. Occult hepatitis B infection in blood donors from South East Asia: molecular characterisation and potential mechanisms of occurrence. Gut 2012; 61:1744–1753 [View Article][PubMed]
    [Google Scholar]
  47. Soussan P, Tuveri R, Nalpas B, Garreau F, Zavala F et al. The expression of hepatitis B spliced protein (HBSP) encoded by a spliced hepatitis B virus RNA is associated with viral replication and liver fibrosis. J Hepatol 2003; 38:343–348 [View Article][PubMed]
    [Google Scholar]
  48. Suzuki T, Kajino K, Masui N, Saito I, Miyamura T. Alternative splicing of hepatitis B virus RNAs in HepG2 cells transfected with the viral DNA. Virology 1990; 179:881–885 [View Article][PubMed]
    [Google Scholar]
  49. Soussan P, Pol J, Garreau F, Schneider V, Le Pendeven C et al. Expression of defective hepatitis B virus particles derived from singly spliced RNA is related to liver disease. J Infect Dis 2008; 198:218–225 [View Article][PubMed]
    [Google Scholar]
  50. Duriez M, Mandouri Y, Lekbaby B, Wang H, Schnuriger A et al. Alternative splicing of hepatitis B virus: a novel virus/host interaction altering liver immunity. J Hepatol 2017; 67:687–699 [View Article][PubMed]
    [Google Scholar]
  51. Köck J, Nassal M, Deres K, Blum HE, von Weizsäcker F. Hepatitis B virus nucleocapsids formed by carboxy-terminally mutated core proteins contain spliced viral genomes but lack full-size DNA. J Virol 2004; 78:13812–13818 [View Article][PubMed]
    [Google Scholar]
  52. Wang Y-L, Liou G-G, Lin C-H, Chen M-L, Kuo T-M et al. The inhibitory effect of the hepatitis B virus singly-spliced RNA-encoded p21.5 protein on HBV nucleocapsid formation. PLoS One 2015; 10:e0119625 [View Article][PubMed]
    [Google Scholar]
  53. Chen W-N, Chen J-Y, Lin W-S, Lin J-Y, Lin X. Hepatitis B doubly spliced protein, generated by a 2.2 kb doubly spliced hepatitis B virus RNA, is a pleiotropic activator protein mediating its effects via activator protein-1- and CCAAT/enhancer-binding protein-binding sites. J Gen Virol 2010; 91:2592–2600 [View Article][PubMed]
    [Google Scholar]
  54. Huang HL, Jeng KS, Hu CP, Tsai CH, Lo SJ et al. Identification and characterization of a structural protein of hepatitis B virus: a polymerase and surface fusion protein encoded by a spliced RNA. Virology 2000; 275:398–410 [View Article][PubMed]
    [Google Scholar]
  55. Tsai K-N, Chong C-L, Chou Y-C, Huang C-C, Wang Y-L et al. Doubly spliced RNA of hepatitis B virus suppresses viral transcription via TATA-binding protein and induces stress granule assembly. J Virol 2015; 89:11406–11419 [View Article][PubMed]
    [Google Scholar]
  56. Bayliss J, Lim L, Thompson AJV, Desmond P, Angus P et al. Hepatitis B virus splicing is enhanced prior to development of hepatocellular carcinoma. J Hepatol 2013; 59:1022–1028 [View Article][PubMed]
    [Google Scholar]
  57. Pan M-H, Hu H-H, Mason H, Bayliss J, Littlejohn M et al. Hepatitis B splice variants are strongly associated with and are indeed predictive of hepatocellular carcinoma. J Hepatol 2018; 68:S474–S475 [View Article]
    [Google Scholar]
  58. Andrews S. FastQC: a Quality Control Tool for High Throughput Sequence Data Cambridge: Babraham Institute; 2016
  59. Sultan M, Amstislavskiy V, Risch T, Schuette M, Dökel S et al. Influence of RNA extraction methods and library selection schemes on RNA-seq data. BMC Genomics 2014; 15:675 [View Article][PubMed]
    [Google Scholar]
  60. Jiang H, Lei R, Ding S-W, Zhu S. Skewer: a fast and accurate adapter trimmer for next-generation sequencing paired-end reads. BMC Bioinformatics 2014; 15:182 [View Article][PubMed]
    [Google Scholar]
  61. Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics 2013; 29:15–21 [View Article][PubMed]
    [Google Scholar]
  62. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J et al. The Sequence Alignment/Map format and SAMtools. Bioinformatics 2009; 25:2078–2079 [View Article][PubMed]
    [Google Scholar]
  63. Li H. Minimap2: pairwise alignment for nucleotide sequences. Bioinformatics 2018; 34:3094–3100 [View Article][PubMed]
    [Google Scholar]
  64. Parker MT, Barton GJ, Simpson GG. Two-pass alignment using machine-learning-filtered splice junctions increases the accuracy of intron detection in long-read RNA sequencing. bioRxiv 2020118679
    [Google Scholar]
  65. Tang AD, Soulette CM, van Baren MJ, Hart K, Hrabeta-Robinson E et al. Full-length transcript characterization of SF3B1 mutation in chronic lymphocytic leukemia reveals downregulation of retained introns. Nat Commun 2020; 11:1438 [View Article][PubMed]
    [Google Scholar]
  66. Lee CM, Barber GP, Casper J, Clawson H, Diekhans M et al. UCSC Genome Browser enters 20th year. Nucleic Acids Res 2020; 48:D756–D761 [View Article][PubMed]
    [Google Scholar]
  67. Hayer J, Jadeau F, Deléage G, Kay A, Zoulim F et al. HBVdb: a knowledge database for hepatitis B virus. Nucleic Acids Res 2013; 41:D566–D570 [View Article][PubMed]
    [Google Scholar]
  68. Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 2004; 32:1792–1797 [View Article][PubMed]
    [Google Scholar]
  69. Eddy SR. Accelerated profile HMM searches. PLoS Comput Biol 2011; 7:e1002195 [View Article][PubMed]
    [Google Scholar]
  70. Wheeler TJ, Eddy SR. nhmmer: DNA homology search with profile HMMs. Bioinformatics 2013; 29:2487–2489 [View Article][PubMed]
    [Google Scholar]
  71. Pertea M, Pertea GM, Antonescu CM, Chang T-C, Mendell JT et al. StringTie enables improved reconstruction of a transcriptome from RNA-seq reads. Nat Biotechnol 2015; 33:290–295 [View Article][PubMed]
    [Google Scholar]
  72. Pertea G, Pertea M. GFF utilities: GffRead and GffCompare. F1000Res 2020; 9:304 [View Article][PubMed]
    [Google Scholar]
  73. Hahne F, Ivanek R. Visualizing genomic data using Gviz and Bioconductor. Methods Mol Biol 2016; 1418:335–351 [View Article][PubMed]
    [Google Scholar]
  74. Lawrence M, Huber W, Pagès H, Aboyoun P, Carlson M et al. Software for computing and annotating genomic ranges. PLoS Comput Biol 2013; 9:e1003118 [View Article][PubMed]
    [Google Scholar]
  75. Altinel K, Hashimoto K, Wei Y, Neuveut C, Gupta I et al. Single-nucleotide resolution mapping of hepatitis B virus promoters in infected human livers and hepatocellular carcinoma. J Virol 2016; 90:10811–10822 [View Article][PubMed]
    [Google Scholar]
  76. Yeo G, Burge CB. Maximum entropy modeling of short sequence motifs with applications to RNA splicing signals. J Comput Biol 2004; 11:377–394 [View Article][PubMed]
    [Google Scholar]
  77. McLaren W, Gil L, Hunt SE, Riat HS, Ritchie GRS et al. The Ensembl variant effect predictor. Genome Biol 2016; 17:122 [View Article][PubMed]
    [Google Scholar]
  78. Jian X, Boerwinkle E, Liu X. In silico prediction of splice-altering single nucleotide variants in the human genome. Nucleic Acids Res 2014; 42:13534–13544 [View Article][PubMed]
    [Google Scholar]
  79. Pervouchine DD, Knowles DG, Guigó R. Intron-centric estimation of alternative splicing from RNA-seq data. Bioinformatics 2013; 29:273–274 [View Article][PubMed]
    [Google Scholar]
  80. Crooks GE, Hon G, Chandonia J-M, Brenner SE. WebLogo: a sequence logo generator. Genome Res 2004; 14:1188–1190 [View Article][PubMed]
    [Google Scholar]
  81. Zytnicki M. mmquant: how to count multi-mapping reads?. BMC Bioinformatics 2017; 18:411 [View Article][PubMed]
    [Google Scholar]
  82. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 2014; 15:550 [View Article][PubMed]
    [Google Scholar]
  83. Ritchie ME, Phipson B, Wu D, Hu Y, Law CW et al. Limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res 2015; 43:e47 [View Article][PubMed]
    [Google Scholar]
  84. Huang DW, Sherman BT, Lempicki RA. Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 2009; 37:1–13 [View Article][PubMed]
    [Google Scholar]
  85. Huang DW, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 2009; 4:44–57 [View Article][PubMed]
    [Google Scholar]
  86. R Core Team R: a Language and Environment for Statistical Computing Vienna: R Foundation for Statistical Computing; 2020
    [Google Scholar]
  87. Hothorn T, Hornik K. exactRankTests: Exact Distributions for Rank and Permutation Tests Vienna: Comprehensive R Archive Network; 2019
  88. Wickham H. ggplot2: Elegant Graphics for Data Analysis New York: Springer; 2016
    [Google Scholar]
  89. Heise T, Sommer G, Reumann K, Meyer I, Will H et al. The hepatitis B virus PRE contains a splicing regulatory element. Nucleic Acids Res 2006; 34:353–363 [View Article][PubMed]
    [Google Scholar]
  90. Halgand B, Desterke C, Rivière L, Fallot G, Sebagh M et al. Hepatitis B virus pregenomic RNA in hepatocellular carcinoma: a nosological and prognostic determinant. Hepatology 2018; 67:86–96 [View Article][PubMed]
    [Google Scholar]
  91. Jin Y, Lee WY, Toh ST, Tennakoon C, Toh HC et al. Comprehensive analysis of transcriptome profiles in hepatocellular carcinoma. J Transl Med 2019; 17:273 [View Article][PubMed]
    [Google Scholar]
  92. Marchio A, Cerapio JP, Ruiz E, Cano L, Casavilca S et al. Early-onset liver cancer in South America associates with low hepatitis B virus DNA burden. Sci Rep 2018; 8:12031 [View Article][PubMed]
    [Google Scholar]
  93. Fang Y, Teng X, Xu W-Z, Li D, Zhao H-W et al. Molecular characterization and functional analysis of occult hepatitis B virus infection in Chinese patients infected with genotype C. J Med Virol 2009; 81:826–835 [View Article][PubMed]
    [Google Scholar]
  94. Zhou T-C, Li X, Li L, Li X-F, Zhang L et al. Evolution of full-length genomes of HBV quasispecies in sera of patients with a coexistence of HBsAg and anti-HBs antibodies. Sci Rep 2017; 7:661 [View Article][PubMed]
    [Google Scholar]
  95. Hao R, Xiang K, Peng Y, Hou J, Sun J et al. Naturally occurring deletion/insertion mutations within HBV whole genome sequences in HBeAg-positive chronic hepatitis B patients are correlated with baseline serum HBsAg and HBeAg levels and might predict a shorter interval to HBeAg loss and seroconversion during antiviral treatment. Infect Genet Evol 2015; 33:261–268 [View Article][PubMed]
    [Google Scholar]
  96. Niller HH, Ay E, Banati F, Demcsák A, Takacs M et al. Wild type HBx and truncated HBx: pleiotropic regulators driving sequential genetic and epigenetic steps of hepatocarcinogenesis and progression of HBV-associated neoplasms. Rev Med Virol 2016; 26:57–73 [View Article][PubMed]
    [Google Scholar]
  97. Chauhan K, Kalam H, Dutt R, Kumar D. RNA splicing: a new paradigm in host-pathogen interactions. J Mol Biol 2019; 431:1565–1575 [View Article][PubMed]
    [Google Scholar]
  98. Padgett RA. New connections between splicing and human disease. Trends Genet 2012; 28:147–154 [View Article][PubMed]
    [Google Scholar]
  99. Li X, Pan E, Zhu J, Xu L, Chen X et al. Cisplatin enhances hepatitis B virus replication and PGC-1α expression through endoplasmic reticulum stress. Sci Rep 2018; 8:3496 [View Article][PubMed]
    [Google Scholar]
  100. Liu X, Green RM. Endoplasmic reticulum stress and liver diseases. Liver Res 2019; 3:55–64 [View Article][PubMed]
    [Google Scholar]
  101. Montalbano R, Honrath B, Wissniowski TT, Elxnat M, Roth S et al. Exogenous hepatitis B virus envelope proteins induce endoplasmic reticulum stress: involvement of cannabinoid axis in liver cancer cells. Oncotarget 2016; 7:20312–20323 [View Article][PubMed]
    [Google Scholar]
  102. Sasaki R, Kanda T, Nakamura M, Nakamoto S, Haga Y et al. Possible involvement of hepatitis B virus infection of hepatocytes in the attenuation of apoptosis in hepatic stellate cells. PLoS One 2016; 11:e0146314 [View Article][PubMed]
    [Google Scholar]
  103. Wang Q, Lin L, Yoo S, Wang W, Blank S et al. Impact of non-neoplastic vs intratumoural hepatitis B viral DNA and replication on hepatocellular carcinoma recurrence. Br J Cancer 2016; 115:841–847 [View Article][PubMed]
    [Google Scholar]
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