In vitro cell model and mouse model of HBV cccDNA
-
摘要: HBV慢性感染仍然是世界性公共健康问题。现有抗病毒药物能够有效抑制HBV复制,却无法使之完全治愈。共价闭合环状DNA(cccDNA)是HBV复制的原始模版,具有显著的稳定性,是HBV持续感染的分子基础,也是实现感染治愈的关键靶点。由于较低的细胞内拷贝数,以及缺乏灵敏可靠的检测方法,限制了对HBV cccDNA生物合成、代谢和调控等方面的相关研究。建立合适的cccDNA研究模型有助于了解这些生物学过程。综述了近年来HBV cccDNA细胞培养模型和实验小鼠模型等方面的主要进展,以期为进一步研究HBV病毒学和抗病毒药物研发提供帮助。Abstract: Chronic hepatitis B virus (HBV) infection remains a major public health threat worldwide. Current antiviral drugs can effectively inhibit the replication of HBV, but they fail to achieve a complete cure. As the original template for HBV replication, covalently closed circular DNA (cccDNA) is intrinsically stable in vivo and is regarded as the molecular basis for persistent HBV infection and the key target for the cure of HBV infection. Studies on biosynthesis, metabolism, and regulation of HBV cccDNA have been limited by the low copy number of cccDNA within cells and the lack of sensitive and reliable detection methods. Establishment of appropriate research models of cccDNA helps to understand related biological processes. This article reviews the latest research advances in cell models and mouse models of HBV cccDNA, in order to facilitate future studies on HBV virology and development of antiviral drugs against HBV.
-
Key words:
- hepatitis B virus /
- cccDNA /
- cell culture model /
- models, animal
-
[1]BECK J, NASSAL M.Hepatitis B virus replication[J].World JGastroenterol, 2007, 13 (1) :48-64. [2]YANG W, SUMMERS J.Illegitimate replication of linear hepadnavirus DNA through nonhomologous recombination[J].J Virol, 1995, 69 (7) :4029-4036. [3]GAO W, HU J.Formation of hepatitis B virus covalently closed circular DNA:Removal of genome-linked protein[J].J Virol, 2007, 81 (12) :6164-6174. [4]GUO H, JIANG D, ZHOU T, et al.Characterization of the intracellular deproteinized relaxed circular DNA of hepatitis B virus:An intermediate of covalently closed circular DNA formation[J].J Virol, 2007, 81 (22) :12472-12484. [5]BOCK CT, SCHWINN S, LOCARNINI S, et al.Structural organization of the hepatitis B virus minichromosome[J].J Mol Biol, 2001, 307 (1) :183-196. [6]HONG X, KIM ES, GUO H.Epigenetic regulation of hepatitis B virus covalently closed circular DNA:Implications for epigenetic therapy against chronic hepatitis B[J].Hepatology, 2017, 66 (6) :2066-2077. [7]WERLE-LAPOSTOLLE B, BOWDEN S, LOCARNINI S, et al.Persistence of cccDNA during the natural history of chronic hepatitis Band decline during adefovir dipivoxil therapy[J].Gastroenterology, 2004, 126 (7) :1750-1758. [8]BOURNE EJ, DIENSTAG JL, LOPEZ VA, et al.Quantitative analysis of HBV cccDNA from clinical specimens:Correlation with clinical and virological response during antiviral therapy[J].J Viral Hepat, 2007, 14 (1) :55-63. [9]ALLWEISS L, DANDRI M.The role of cccDNA in HBV maintenance[J].Viruses, 2017, 9 (6) :e156. [10]CHEN X, OIDOVSAMBUU O, LIU P, et al.A novel quantitative microarray antibody capture assay identifies an extremely high hepatitis delta virus prevalence among hepatitis B virusinfected mongolians[J].Hepatology, 2017, 66 (6) :1739-1749. [11]WIELAND S, THIMME R, PURCELL RH, et al.Genomic analysis of the host response to hepatitis B virus infection[J].Proc Natl Acad Sci U S A, 2004, 101 (17) :6669-6674. [12]LUCIFORA J, XIA Y, REISINGER F, et al.Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA[J].Science, 2014, 343 (6176) :1221-1228. [13]SHEN F, LI Y, WANG Y, et al.Hepatitis B virus sensitivity to interferon-alpha in hepatocytes is more associated with cellular interferon response than with viral genotype[J].Hepatology, 2018, 67 (4) :1237-1252. [14]GRIPON P, DIOT C, THEZE N, et al.Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide[J].J Virol, 1988, 62 (11) :4136-4143. [15]ORTEGA-PRIETO AM, SKELTON JK, WAI SN, et al.3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B virus infection[J].Nat Commun, 2018, 9 (1) :682. [16]ZHANG K, ZHANG L, LIU W, et al.In vitro expansion of primary human hepatocytes with efficient liver repopulation capacity[J].Cell Stem Cell, 2018, 23 (6) :806-819, e804. [17]FU GB, HUANG WJ, ZENG M, et al.Expansion and differentiation of human hepatocyte-derived liver progenitor-like cells and their use for the study of hepatotropic pathogens[J].Cell Res, 2019, 29 (1) :8-22. [18]GRIPON P, RUMIN S, URBAN S, et al.Infection of a human hepatoma cell line by hepatitis B virus[J].Proc Natl Acad Sci U S A, 2002, 99 (24) :15655-15660. [19]HANTZ O, PARENT R, DURANTEL D, et al.Persistence of the hepatitis B virus covalently closed circular DNA in HepaRGhuman hepatocyte-like cells[J].J Gen Virol, 2009, 90 (Pt1) :127-135. [20]LUCIFORA J, DURANTEL D, TESTONI B, et al.Control of hepatitis B virus replication by innate response of HepaRG cells[J].Hepatology, 2010, 51 (1) :63-72. [21]ZHONG G, YAN H, WANG H, et al.Sodium taurocholate cotransporting polypeptide mediates woolly monkey hepatitis Bvirus infection of Tupaia hepatocytes[J].J Virol, 2013, 87 (12) :7176-7184. [22]YAN H, LI W.Sodium taurocholate cotransporting polypeptide acts as a receptor for hepatitis B and D virus[J].Dig Dis, 2015, 33 (3) :388-396. [23]NI Y, LEMPP FA, MEHRLE S, et al.Hepatitis B and D viruses exploit sodium taurocholate co-transporting polypeptide for species-specific entry into hepatocytes[J].Gastroenterology, 2014, 146 (4) :1070-1083. [24]QI Y, GAO Z, XU G, et al.DNA polymerase kappa is a key cellular factor for the formation of covalently closed circular DNA of hepatitis B virus[J].PLo S Pathog, 2016, 12 (10) :e1005893. [25]LONG Q, YAN R, HU J, et al.The role of host DNA ligases in hepadnavirus covalently closed circular DNA formation[J].PLo S Pathog, 2017, 13 (12) :e1006784. [26]GUIDOTTI LG, MATZKE B, SCHALLER H, et al.High-level hepatitis B virus replication in transgenic mice[J].J Virol, 1995, 69 (10) :6158-6169. [27]RANEY AK, EGGERS CM, KLINE EF, et al.Nuclear covalently closed circular viral genomic DNA in the liver of hepatocyte nuclear factor 1 alpha-null hepatitis B virus transgenic mice[J].J Virol, 2001, 75 (6) :2900-2911. [28]LI H, ZHUANG Q, WANG Y, et al.HBV life cycle is restricted in mouse hepatocytes expressing human NTCP[J].Cell Mol Immunol, 2014, 11 (2) :175-183. [29]BOGOMOLOV P, ALEXANDROV A, VORONKOVA N, et al.Treatment of chronic hepatitis D with the entry inhibitor myrcludex B:First results of a phase Ib/IIa study[J].J Hepatol, 2016, 65 (3) :490-498. [30]HE W, REN B, MAO F, et al.Hepatitis D virus infection of mice expressing human sodium taurocholate co-transporting polypeptide[J].PLo S Pathog, 2015, 11 (4) :e1004840. [31]CUI X, CLARK DN, LIU K, et al.Viral DNA-dependent induction of innate immune response to hepatitis B virus in immortalized mouse hepatocytes[J].J Virol, 2016, 90 (1) :486-496. [32]SUMMERS J, SMITH PM, HORWICH AL.Hepadnavirus envelope proteins regulate covalently closed circular DNA amplification[J].J Virol, 1990, 64 (6) :2819-2824. [33]SUMMERS J, SMITH PM, HUANG MJ, et al.Morphogenetic and regulatory effects of mutations in the envelope proteins of an avian hepadnavirus[J].J Virol, 1991, 65 (3) :1310-1317. [34]KOCK J, ROSLER C, ZHANG JJ, et al.Generation of covalently closed circular DNA of hepatitis B viruses via intracellular recycling is regulated in a virus specific manner[J].PLo SPathog, 2010, 6 (9) :e1001082. [35]SELLS MA, CHEN ML, ACS G.Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA[J].Proc Natl Acad Sci U S A, 1987, 84 (4) :1005-1009. [36]LADNER SK, OTTO MJ, BARKER CS, et al.Inducible expression of human hepatitis B virus (HBV) in stably transfected hepatoblastoma cells:A novel system for screening potential inhibitors of HBV replication[J].Antimicrob Agents Chemother, 1997, 41 (8) :1715-1720. [37]GUO JT, GUO H.Metabolism and function of hepatitis B virus cccDNA:Implications for the development of cccDNA-targeting antiviral therapeutics[J].Antiviral Res, 2015, 122:91-100. [38]CAI D, WANG X, YAN R, et al.Establishment of an inducible HBV stable cell line that expresses cccDNA-dependent epitope-tagged HBe Ag for screening of cccDNA modulators[J].Antiviral Res, 2016, 132:26-37. [39]GNTHER S, LI BC, MISKA S, et al.A novel method for efficient amplification of whole hepatitis B virus genomes permits rapid functional analysis and reveals deletion mutants in immunosuppressed patients[J].J Virol, 1995, 69 (9) :5437-5444. [40]POLLICINO T, BELLONI L, RAFFA G, et al.Hepatitis B virus replication is regulated by the acetylation status of hepatitis Bvirus cccDNA-bound H3 and H4 histones[J].Gastroenterology, 2006, 130 (3) :823-837. [41]BELLONI L, POLLICINO T, de NICOLA F, et al.Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function[J].Proc Natl Acad Sci U S A, 2009, 106 (47) :19975-19979. [42]BELLONI L, ALLWEISS L, GUERRIERI F, et al.IFN-alpha inhibits HBV transcription and replication in cell culture and in humanized mice by targeting the epigenetic regulation of the nuclear cccDNA minichromosome[J].J Clin Invest, 2012, 122 (2) :529-537. [43]QI Z, LI G, HU H, et al.Recombinant covalently closed circular hepatitis B virus DNA induces prolonged viral persistence in immunocompetent mice[J].J Virol, 2014, 88 (14) :8045-8056. [44]ZHU YF, LI GY, CHANG H, et al.Preparing recombinant cccDNA of hepatitis B virus in vitro using minicircle DNA vector technogy[J].J Microbes Infect, 2017, 12 (4) :229-234. (in Chinese) 朱园飞, 李改云, 常豪, 等.应用微环DNA技术体外诱导和制备重组的乙型肝炎病毒共价闭合环状DNA[J].微生物与感染, 2017, 12 (4) :229-234. [45]YAN Z, ZENG J, YU Y, et al.HBVcircle:A novel tool to investigate hepatitis B virus covalently closed circular DNA[J].JHepatol, 2017, 66 (6) :1149-1157. [46]LI F, CHENG L, MURPHY CM, et al.Minicircle HBV cccDNAwith a Gaussia luciferase reporter for investigating HBV cccD-NA biology and developing cccDNA-targeting drugs[J].Sci Rep, 2016, (6) :36483. [47]RHIM JA, SANDGREN EP, PALMITER RD, et al.Complete reconstitution of mouse liver with xenogeneic hepatocytes[J].Proc Natl Acad Sci U S A, 1995, 92 (11) :4942-4946. [48]DANDRI M, BURDA MR, BURKLE A, et al.Increase in de novo HBV DNA integrations in response to oxidative DNA damage or inhibition of poly (ADP-ribosyl) ation[J].Hepatology, 2002, 35 (1) :217-223. [49]BISSIG KD, WIELAND SF, TRAN P, et al.Human liver chimeric mice provide a model for hepatitis B and C virus infection and treatment[J].J Clin Invest, 2010, 120 (3) :924-930. [50]VOLZ T, ALLWEISS L, BEN MM, et al.The entry inhibitor Myrcludex-B efficiently blocks intrahepatic virus spreading in humanized mice previously infected with hepatitis B virus[J].J Hepatol, 2013, 58 (5) :861-867. [51]ALLWEISS L, VOLZ T, GIERSCH K, et al.Proliferation of primary human hepatocytes and prevention of hepatitis B virus reinfection efficiently deplete nuclear cccDNA in vivo[J].Gut, 2018, 67 (3) :542-552. [52]YANG PL, ALTHAGE A, CHUNG J, et al.Hydrodynamic injection of viral DNA:A mouse model of acute hepatitis B virus infection[J].Proc Natl Acad Sci U S A, 2002, 99 (21) :13825-13830. [53]HUANG LR, WU HL, CHEN PJ, et al.An immunocompetent mouse model for the tolerance of human chronic hepatitis B virus infection[J].Proc Natl Acad Sci U S A, 2006, 103 (47) :17862-17867. [54]LI G, ZHU Y, SHAO D, et al.Recombinant covalently closed circular DNA of hepatitis B virus induces long-term viral persistence with chronic hepatitis in a mouse model[J].Hepatology, 2018, 67 (1) :56-70. [55]DONG XY, YUCHI J, WANG G, et al.Establishment of hepatitis B virus (HBV) chronic infection mouse model by in vivo transduction with a recombinant adeno-associated virus 8carrying 1.3 copies of HBV genome (r AAN8-1.3HBV) [J].Chin J Virol, 2010, 26 (6) :425-431. (in Chinese) 董小岩, 尉迟捷, 王刚, 等.高嗜肝性8型重组腺相关病毒体内转导法制备乙型肝炎病毒持续感染小鼠模型[J].病毒学报, 2010, 26 (6) :425-431. [56]DION S, BOURGINE M, GODON O, et al.Adeno-associated virus-mediated gene transfer leads to persistent hepatitis B virus replication in mice expressing HLA-A2 and HLA-DR1 molecules[J].J Virol, 2013, 87 (10) :5554-5563. [57]LUCIFORA J, PROTZER U.Attacking hepatitis B virus cccD-NA--The holy grail to hepatitis B cure[J].J Hepatol, 2016, 64 (1 Suppl) :s41-s48. [58]HUANG LR, GABEL YA, GRAF S, et al.Transfer of HBV genomes using low doses of adenovirus vectors leads to persistent infection in immune competent mice[J].Gastroenterology, 2012, 142 (7) :1447-1450, e1443.
本文二维码
计量
- 文章访问数: 2073
- HTML全文浏览量: 62
- PDF下载量: 534
- 被引次数: 0