Inhibition of Replication of Hepatitis B Virus Using Transcriptional Repressors That Target the Viral DNA

Overview

Journal BMC Infect Dis

Publisher Biomed Central

Specialty Infectious Diseases

Date 2019 Sep 13

PMID 31510934

Citations 9

Authors

Kristie Bloom

Haajira Kaldine

Toni Cathomen

Claudio Mussolino

Abdullah Ely

Patrick Arbuthnot

Affiliations

Soon will be listed here.

Abstract

Background: Chronic infection with hepatitis B virus (HBV) is a serious global health problem. Persistence of the virus occurs as a result of stability of the replication intermediate comprising covalently closed circular DNA (cccDNA). Development of drugs that are capable of disabling this cccDNA is vital.

Methods: To investigate an epigenetic approach to inactivating viral DNA, we engineered transcriptional repressors that comprise an HBV DNA-binding domain of transcription activator like effectors (TALEs) and a fused Krüppel Associated Box (KRAB). These repressor TALEs (rTALEs) targeted the viral surface open reading frame and were placed under transcription control of constitutively active or liver-specific promoters.

Results: Evaluation in cultured cells and following hydrodynamic injection of mice revealed that the rTALEs significantly inhibited production of markers of HBV replication without evidence of hepatotoxicity. Increased methylation of HBV DNA at CpG island II showed that the rTALEs caused intended epigenetic modification.

Conclusions: Epigenetic modification of HBV DNA is a new and effective means of inactivating the virus in vivo. The approach has therapeutic potential and avoids potentially problematic unintended mutagenesis of gene editing.

Citing Articles

Opportunities and challenges for hepatitis B cure.

Roca Suarez A, Zoulim F eGastroenterology. 2025; 1(2):e100021.

PMID: 39944004 PMC: 11731071. DOI: 10.1136/egastro-2023-100021.

Epigenetic regulation and its therapeutic potential in hepatitis B virus covalently closed circular DNA.

Ren J, Cheng S, Ren F, Gu H, Wu D, Yao X Genes Dis. 2024; 12(1):101215.

PMID: 39534573 PMC: 11555349. DOI: 10.1016/j.gendis.2024.101215.

Epigenome editing technologies for discovery and medicine.

McCutcheon S, Rohm D, Iglesias N, Gersbach C Nat Biotechnol. 2024; 42(8):1199-1217.

PMID: 39075148 DOI: 10.1038/s41587-024-02320-1.

The role of HBV cccDNA in occult hepatitis B virus infection.

He P, Zhang P, Fang Y, Han N, Yang W, Xia Z Mol Cell Biochem. 2023; 478(10):2297-2307.

PMID: 36735210 DOI: 10.1007/s11010-023-04660-z.

Gene Editing Technologies to Target HBV cccDNA.

Martinez M, Smekalova E, Combe E, Gregoire F, Zoulim F, Testoni B Viruses. 2022; 14(12).

PMID: 36560658 PMC: 9787400. DOI: 10.3390/v14122654.

References

Sarin S, Kumar M, Lau G, Abbas Z, Chan H, Chen C . Asian-Pacific clinical practice guidelines on the management of hepatitis B: a 2015 update. Hepatol Int. 2015; 10(1):1-98. PMC: 4722087. DOI: 10.1007/s12072-015-9675-4. View

Weinberg M, Ely A, Barichievy S, Crowther C, Mufamadi S, Carmona S . Specific inhibition of HBV replication in vitro and in vivo with expressed long hairpin RNA. Mol Ther. 2007; 15(3):534-41. DOI: 10.1038/sj.mt.6300077. View

Terrault N, Bzowej N, Chang K, Hwang J, Jonas M, Murad M . AASLD guidelines for treatment of chronic hepatitis B. Hepatology. 2015; 63(1):261-83. PMC: 5987259. DOI: 10.1002/hep.28156. View

Guidotti L, Matzke B, Schaller H, Chisari F . High-level hepatitis B virus replication in transgenic mice. J Virol. 1995; 69(10):6158-69. PMC: 189513. DOI: 10.1128/JVI.69.10.6158-6169.1995. View

Kaur P, Paliwal A, Durantel D, Hainaut P, Scoazec J, Zoulim F . DNA methylation of hepatitis B virus (HBV) genome associated with the development of hepatocellular carcinoma and occult HBV infection. J Infect Dis. 2010; 202(5):700-4. DOI: 10.1086/655398. View

Zhang Y, Mao R, Yan R, Cai D, Zhang Y, Zhu H . Transcription of hepatitis B virus covalently closed circular DNA is regulated by CpG methylation during chronic infection. PLoS One. 2014; 9(10):e110442. PMC: 4206413. DOI: 10.1371/journal.pone.0110442. View

Nassal M . The arginine-rich domain of the hepatitis B virus core protein is required for pregenome encapsidation and productive viral positive-strand DNA synthesis but not for virus assembly. J Virol. 1992; 66(7):4107-16. PMC: 241213. DOI: 10.1128/JVI.66.7.4107-4116.1992. View

Guo Y, Li Y, Mu S, Zhang J, Yan Z . Evidence that methylation of hepatitis B virus covalently closed circular DNA in liver tissues of patients with chronic hepatitis B modulates HBV replication. J Med Virol. 2009; 81(7):1177-83. DOI: 10.1002/jmv.21525. View

Brown T, Mackey K, Du T . Analysis of RNA by northern and slot blot hybridization. Curr Protoc Mol Biol. 2008; Chapter 4:Unit 4.9. DOI: 10.1002/0471142727.mb0409s67. View

10.

Liu N, Zhao F, Zhan P, Liu X . Review of small synthetic molecules targeting HBV capsid assembly. Med Chem. 2015; 11(8):710-6. DOI: 10.2174/157340641108151029111243. View

11.

Guo Y, Li Y, Zhao J, Zhang J, Yan Z . HBc binds to the CpG islands of HBV cccDNA and promotes an epigenetic permissive state. Epigenetics. 2011; 6(6):720-6. DOI: 10.4161/epi.6.6.15815. View

12.

Hayashi M, Deng L, Chen M, Gan X, Shinozaki K, Shoji I . Interaction of the hepatitis B virus X protein with the lysine methyltransferase SET and MYND domain-containing 3 induces activator protein 1 activation. Microbiol Immunol. 2015; 60(1):17-25. DOI: 10.1111/1348-0421.12345. View

13.

Costa R, Grayson D . Site-directed mutagenesis of hepatocyte nuclear factor (HNF) binding sites in the mouse transthyretin (TTR) promoter reveal synergistic interactions with its enhancer region. Nucleic Acids Res. 1991; 19(15):4139-45. PMC: 328553. DOI: 10.1093/nar/19.15.4139. View

14.

Blank A, Markert C, Hohmann N, Carls A, Mikus G, Lehr T . First-in-human application of the novel hepatitis B and hepatitis D virus entry inhibitor myrcludex B. J Hepatol. 2016; 65(3):483-9. DOI: 10.1016/j.jhep.2016.04.013. View

15.

Passman M, Weinberg M, Kew M, Arbuthnot P . In situ demonstration of inhibitory effects of hammerhead ribozymes that are targeted to the hepatitis Bx sequence in cultured cells. Biochem Biophys Res Commun. 2000; 268(3):728-33. DOI: 10.1006/bbrc.2000.2209. View

16.

Cohen J . Forgotten no more. Science. 2018; 362(6418):984-987. DOI: 10.1126/science.362.6418.984. View

17.

Bloom K, Ely A, Mussolino C, Cathomen T, Arbuthnot P . Inactivation of hepatitis B virus replication in cultured cells and in vivo with engineered transcription activator-like effector nucleases. Mol Ther. 2013; 21(10):1889-97. PMC: 3808145. DOI: 10.1038/mt.2013.170. View

18.

Thakore P, DIppolito A, Song L, Safi A, Shivakumar N, Kabadi A . Highly specific epigenome editing by CRISPR-Cas9 repressors for silencing of distal regulatory elements. Nat Methods. 2015; 12(12):1143-9. PMC: 4666778. DOI: 10.1038/nmeth.3630. View

19.

Vivekanandan P, Thomas D, Torbenson M . Hepatitis B viral DNA is methylated in liver tissues. J Viral Hepat. 2008; 15(2):103-7. DOI: 10.1111/j.1365-2893.2007.00905.x. View

20.

Marimani M, Ely A, Buff M, Bernhardt S, Engels J, Arbuthnot P . Inhibition of hepatitis B virus replication in cultured cells and in vivo using 2'-O-guanidinopropyl modified siRNAs. Bioorg Med Chem. 2013; 21(20):6145-55. DOI: 10.1016/j.bmc.2013.04.073. View