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Prospects of viral vector-mediated delivery of sequences encoding anti-HBV designer endonucleases

Gene Therapy (2022)Cite this article

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Abstract

Available treatment for chronic hepatitis B virus (HBV) infection offers modest functional curative efficacy. The viral replicative intermediate comprising covalently closed circular DNA (cccDNA) is responsible for persistent chronic HBV infection. Hence, current efforts have focused on developing therapies that disable cccDNA. Employing gene editing tools has emerged as an attractive strategy, with the end goal of establishing permanently inactivated cccDNA. Although anti-HBV designer nucleases are effective in vivo, none has yet progressed to clinical trial. Lack of safe and efficient delivery systems remains the limiting factor. Several vectors may be used to deliver anti-HBV gene editor-encoding sequences, with viral vectors being at the forefront. Despite the challenges associated with packaging large gene editor-encoding sequences into viral vectors, advancement in the field is overcoming such limitations. Translation of viral vector-mediated gene editing against HBV to clinical application is within reach. This review discusses the prospects of delivering HBV targeted designer nucleases using viral vectors.

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Fig. 1: Viral vectors expressing anti-HBV gene editing nucleases.

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References

  1. WHO. Interim guidance for country validation of viral hepatitis elimination. WHO; 2021.

  2. Xia Y, Guo H. Hepatitis B virus cccDNA: Formation, regulation and therapeutic potential. Antiviral Res. 2020;180:e104824.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Hu J, Protzer U, Siddiqui A. Revisiting hepatitis B virus: challenges of curative therapies. J Virol. 2019;93:e01032–19.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Dandri M, Petersen J. cccDNA maintenance in chronic hepatitis B–targeting the matrix of viral replication. Infect Drug Resist. 2020;13:e3873.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Seeger C, Mason WS. Molecular biology of hepatitis B virus infection. Virology. 2015;479:672–86.

    Article  PubMed  CAS  Google Scholar 

  6. Yan H, Zhong G, Xu G, He W, Jing Z, Gao Z, et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife. 2012;1:1–14. https://doi.org/10.7554/eLife.00049.

    Article  CAS  Google Scholar 

  7. Iwamoto M, Saso W, Sugiyama R, Ishii K, Ohki M, Nagamori S, et al. Epidermal growth factor receptor is a host-entry cofactor triggering hepatitis B virus internalization. Proc Natl Acad Sci USA. 2019;116:8487–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Huang H-C, Chen C-C, Chang W-C, Tao M-H, Huang C. Entry of hepatitis B virus into immortalized human primary hepatocytes by clathrin-dependent endocytosis. J Virol. 2012;86:9443–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Blondot M-L, Bruss V, Kann M. Intracellular transport and egress of hepatitis B virus. J Hepatol. 2016;64:S49–S59.

    Article  CAS  PubMed  Google Scholar 

  10. Gallucci L, Kann M. Nuclear import of hepatitis B virus capsids and genome. Viruses. 2017;9:e21.

    Article  PubMed Central  CAS  Google Scholar 

  11. Schreiner S, Nassal M. A role for the host DNA damage response in hepatitis B virus cccDNA formation—and beyond? Viruses. 2017;9:e125.

    Article  PubMed Central  CAS  Google Scholar 

  12. Zhang Z-Q, Shi B-S, Lu W, Liu D-P, Huang D, Feng Y-L. Quantitative HBcrAg and HBcAb versus HBsAg and HBV DNA in predicting liver fibrosis levels of chronic hepatitis B patients. Gastroenterol Hepatol. 2020;43:526–36.

    Article  CAS  PubMed  Google Scholar 

  13. Hadziyannis E, Laras A. Viral biomarkers in chronic HBeAg negative HBV infection. Genes. 2018;9:e469.

    Article  PubMed Central  CAS  Google Scholar 

  14. Demirören K, Kocamaz H, Doğan Y. The importance of the serum quantitative levels of hepatitis B surface antigen and hepatitis B e antigen in children with chronic hepatitis B. Turk J Gastroenterol. 2015;26:36–41.

    Article  PubMed  Google Scholar 

  15. Li J, Sun X, Fang J, Wang C, Han G, Ren W. Analysis of intrahepatic total HBV DNA, cccDNA and serum HBsAg level in chronic hepatitis B patients with undetectable serum HBV DNA during oral antiviral therapy. Clin Res Hepatol Gastroenterol. 2017;41:635–43.

    Article  CAS  PubMed  Google Scholar 

  16. Qu B, Ni Y, Lempp FA, Vondran FWR, Urban S. T5 exonuclease hydrolysis of hepatitis B virus replicative intermediates allows reliable quantification and fast drug efficacy testing of covalently closed circular DNA by PCR. J Virol. 2018;92:e01117–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Nassal M. HBV cccDNA: viral persistence reservoir and key obstacle for a cure of chronic hepatitis B. Gut. 2015;64:1972–84.

    Article  CAS  PubMed  Google Scholar 

  18. Li X, Zhao J, Yuan Q, Xia N. Detection of HBV covalently closed circular DNA. Viruses. 2017;9:139–57.

    Article  PubMed Central  CAS  Google Scholar 

  19. Zeisel MB, Lucifora J, Mason WS, Sureau C, Beck J, Levrero M, et al. Towards an HBV cure: state-of-the-art and unresolved questions—report of the ANRS workshop on HBV cure. Gut. 2015;64:1314–26.

    Article  CAS  PubMed  Google Scholar 

  20. Ligat G, Verrier ER, Nassal M, Baumert TF. Hepatitis B virus–host interactions and novel targets for viral cure. Curr Opin Virol. 2021;49:41–51.

    Article  CAS  PubMed  Google Scholar 

  21. Liver EAFTSOT. EASL 2017 Clinical Practice Guidelines on the management of hepatitis B virus infection. J Hepatol. 2017;67:370–98.

    Article  Google Scholar 

  22. Prifti G-M, Moianos D, Giannakopoulou E, Pardali V, Tavis JE, Zoidis G. Recent advances in hepatitis B treatment. Pharmaceuticals. 2021;14:417.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Lam AM, Espiritu C, Vogel R, Ren S, Lau V, Kelly M, et al. Preclinical characterization of NVR 3-778, a first-in-class capsid assembly modulator against hepatitis B virus. Antimicrobial Agents Chemother. 2019;63:e01734–18.

    Article  CAS  Google Scholar 

  24. Maepa MB, Bloom K, Ely A, Arbuthnot P. Hepatitis B virus: promising drug targets and therapeutic implications. Expert Opin Ther Targets. 2021;25:451–66.

    Article  CAS  PubMed  Google Scholar 

  25. 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;21:1889–97.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Smith T, Singh P, Chmielewski KO, Bloom K, Cathomen T, Arbuthnot P, et al. Improved specificity and safety of anti-hepatitis B virus TALENs using obligate heterodimeric FokI nuclease domains. Viruses. 2021;13:e1344.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Weber ND, Stone D, Sedlak RH, De Silva Feelixge HS, Roychoudhury P, Schiffer JT, et al. AAV-mediated delivery of zinc finger nucleases targeting hepatitis B virus inhibits active replication. PloS One. 2014;9:e97579.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Schiwon M, Ehrke-Schulz E, Oswald A, Bergmann T, Michler T, Protzer U, et al. One-vector system for multiplexed CRISPR/Cas9 against hepatitis B virus cccDNA utilizing high-capacity adenoviral vectors. Mol Ther Nucleic Acids. 2018;12:242–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Rothstein RJ. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–11.

    Article  CAS  PubMed  Google Scholar 

  30. Lieber MR. The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway. Ann Rev Biochem. 2010;79:181–211.

    Article  CAS  PubMed  Google Scholar 

  31. Mao Z, Bozzella M, Seluanov A, Gorbunova V. DNA repair by nonhomologous end joining and homologous recombination during cell cycle in human cells. Cell Cycle. 2008;7:2902–6.

    Article  CAS  PubMed  Google Scholar 

  32. Ely A, Moyo B, Arbuthnot P. Progress with developing use of gene editing to cure chronic infection with hepatitis B virus. Mol Ther. 2016;24:671–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Kim Y-G, Cha J, Chandrasegaran S. Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain. Proc Natl Acad Sci. 1996;93:1156–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Miller J, McLachlan A, Klug A. Repetitive zinc‐binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO J. 1985;4:1609–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Cornu TI, Thibodeau-Beganny S, Guhl E, Alwin S, Eichtinger M, Joung JK, et al. DNA-binding specificity is a major determinant of the activity and toxicity of zinc-finger nucleases. Mol Ther. 2008;16:352–8.

    Article  CAS  PubMed  Google Scholar 

  36. Cradick TJ, Keck K, Bradshaw S, Jamieson AC, McCaffrey AP. Zinc-finger nucleases as a novel therapeutic strategy for targeting hepatitis B virus DNAs. Mol Ther. 2010;18:947–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Tebas P, Stein D, Tang WW, Frank I, Wang SQ, Lee G, et al. Gene editing of CCR5 in autologous CD4 T cells of persons infected with HIV. N Engl J Med. 2014;370:901–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Miller JC, Tan S, Qiao G, Barlow KA, Wang J, Xia DF, et al. A TALE nuclease architecture for efficient genome editing. Nat Biotechnol. 2011;29:143–8.

    Article  CAS  PubMed  Google Scholar 

  39. Fujikawa T, Ishihara H, Leach JE, Tsuyumu S. Suppression of defense response in plants by the avrBs3/pthA gene family of Xanthomonas spp. Mol Plant Microbe Interact. 2006;19:342–9.

    Article  CAS  PubMed  Google Scholar 

  40. Morbitzer R, Elsaesser J, Hausner J, Lahaye T. Assembly of custom TALE-type DNA binding domains by modular cloning. Nucleic Acids Res. 2011;39:5790–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, et al. Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res. 2011;39:e82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Reyon D, Tsai SQ, Khayter C, Foden JA, Sander JD, Joung JK. FLASH assembly of TALENs for high-throughput genome editing. Nat Biotechnol. 2012;30:460–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Ashmore-Harris C, Fruhwirth GO. The clinical potential of gene editing as a tool to engineer cell-based therapeutics. Clin Transl Med. 2020;9:1–22.

    Article  Google Scholar 

  44. Chen J, Zhang W, Lin J, Wang F, Wu M, Chen C, et al. An efficient antiviral strategy for targeting hepatitis B virus genome using transcription activator-like effector nucleases. Mol Ther. 2014;22:303–11.

    Article  PubMed  CAS  Google Scholar 

  45. Marraffini LA. The CRISPR-Cas system of Streptococcus pyogenes: function and applications. Streptococcus pyogenes: Basic Biology to Clinical Manifestations [Internet]. 2016.

  46. Yourik P, Fuchs RT, Mabuchi M, Curcuru JL, Robb GB. Staphylococcus aureus Cas9 is a multiple-turnover enzyme. RNA. 2019;25:35–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012;337:816–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Lin SR, Yang HC, Kuo YT, Liu CJ, Yang TY, Sung KC, et al. The CRISPR/Cas9 system facilitates clearance of the intrahepatic HBV templates in vivo. Mol Ther Nucleic Acids. 2014;3:e186.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Dong C, Qu L, Wang H, Wei L, Dong Y, Xiong S. Targeting hepatitis B virus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication. Antiviral Res. 2015;118:110–7.

    Article  CAS  PubMed  Google Scholar 

  50. Zhen S, Hua L, Liu Y, Gao L, Fu J, Wan D, et al. Harnessing the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated Cas9 system to disrupt the hepatitis B virus. Gene Ther. 2015;22:404–12.

    Article  CAS  PubMed  Google Scholar 

  51. Kennedy EM, Bassit LC, Mueller H, Kornepati AV, Bogerd HP, Nie T, et al. Suppression of hepatitis B virus DNA accumulation in chronically infected cells using a bacterial CRISPR/Cas RNA-guided DNA endonuclease. Virology. 2015;476:196–205.

    Article  CAS  PubMed  Google Scholar 

  52. Li H, Sheng C, Wang S, Yang L, Liang Y, Huang Y, et al. Removal of integrated hepatitis B virus DNA using CRISPR-Cas9. Front Cell Infect Microbiol. 2017;7:91.

    Article  PubMed  PubMed Central  Google Scholar 

  53. Seeger C, Sohn JA. Targeting hepatitis B virus with CRISPR/Cas9. Mol Ther Nucleic Acids. 2014;3:e216.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Belloni L, Pollicino T, De Nicola F, Guerrieri F, Raffa G, Fanciulli M, et al. Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function. Proc Natl Acad Sci. 2009;106:19975–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Wang J, Xu Z-W, Liu S, Zhang R-Y, Ding S-L, Xie X-M, et al. Dual gRNAs guided CRISPR/Cas9 system inhibits hepatitis B virus replication. World J Gastroenterol WJG. 2015;21:e9554.

    Article  CAS  PubMed  Google Scholar 

  56. Liu X, Hao R, Chen S, Guo D, Chen Y. Inhibition of hepatitis B virus by the CRISPR/Cas9 system via targeting the conserved regions of the viral genome. J Gen Virol. 2015;96:2252–61.

    Article  CAS  PubMed  Google Scholar 

  57. Ramanan V, Shlomai A, Cox DB, Schwartz RE, Michailidis E, Bhatta A, et al. CRISPR/Cas9 cleavage of viral DNA efficiently suppresses hepatitis B virus. Sci Rep. 2015;5:1–9.

    Article  CAS  Google Scholar 

  58. Jiang C, Mei M, Li B, Zhu X, Zu W, Tian Y, et al. A non-viral CRISPR/Cas9 delivery system for therapeutically targeting HBV DNA and pcsk9 in vivo. Cell Res. 2017;27:440–3.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Rees HA, Liu DR. Base editing: precision chemistry on the genome and transcriptome of living cells. Nat Rev Genet. 2018;19:770–88.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Yang Y-C, Chen Y-H, Kao J-H, Ching C, Liu I-J, Wang C-C, et al. Permanent inactivation of HBV genomes by CRISPR/Cas9-mediated non-cleavage base editing. Mol Ther Nucleic Acids. 2020;20:480–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nat Biotechnol. 2013;31:822–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Pattanayak V, Lin S, Guilinger JP, Ma E, Doudna JA, Liu DR. High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. Nat Biotechnol. 2013;31:839–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Kang S-H, Lee W-J, An J-H, Lee J-H, Kim Y-H, Kim H, et al. Prediction-based highly sensitive CRISPR off-target validation using target-specific DNA enrichment. Nat Commun. 2020;11:1–11.

    Article  CAS  Google Scholar 

  64. Ran FA, Hsu PD, Lin C-Y, Gootenberg JS, Konermann S, Trevino AE, et al. Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell. 2013;154:1380–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Karimova M, Beschorner N, Dammermann W, Chemnitz J, Indenbirken D, Bockmann J-H, et al. CRISPR/Cas9 nickase-mediated disruption of hepatitis B virus open reading frame S and X. Sci Rep. 2015;5:1–16.

    Article  Google Scholar 

  66. Sakuma T, Masaki K, Abe‐Chayama H, Mochida K, Yamamoto T, Chayama K. Highly multiplexed CRISPR‐Cas9‐nuclease and Cas9‐nickase vectors for inactivation of hepatitis B virus. Genes Cells. 2016;21:1253–62.

    Article  CAS  PubMed  Google Scholar 

  67. Kurihara T, Fukuhara T, Ono C, Yamamoto S, Uemura K, Okamoto T, et al. Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9. Sci Rep. 2017;7:1–13.

    Article  CAS  Google Scholar 

  68. Watts JM, Dang KK, Gorelick RJ, Leonard CW, Bess JW Jr, Swanstrom R, et al. Architecture and secondary structure of an entire HIV-1 RNA genome. Nature. 2009;460:711–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Escors D, Breckpot K. Lentiviral vectors in gene therapy: their current status and future potential. Arch Immunol Ther Exp (Warsz). 2010;58:107–19.

    Article  CAS  Google Scholar 

  70. Ramanan V, Shlomai A, Cox DB, Schwartz RE, Michailidis E, Bhatta A, et al. CRISPR/Cas9 cleavage of viral DNA efficiently suppresses hepatitis B virus. Sci Rep. 2015;5:e10833.

    Article  CAS  Google Scholar 

  71. Crowther C, Mowa MB, Ely A, Arbuthnot PB. Inhibition of HBV replication in vivo using helper-dependent adenovirus vectors to deliver antiviral RNA interference expression cassettes. Antivir Ther. 2014;19:363–73.

    Article  CAS  PubMed  Google Scholar 

  72. Mowa MB, Crowther C, Ely A, Arbuthnot P. Inhibition of hepatitis B virus replication by helper dependent adenoviral vectors expressing artificial anti-HBV pri-miRs from a liver-specific promoter. Biomed Res Int. 2014;2014:e718743.

    Article  Google Scholar 

  73. Tasca F, Wang Q, Goncalves M. Adenoviral vectors meet gene editing: a rising partnership for the genomic engineering of human stem cells and their progeny. Cells. 2020;9:e953.

    Article  PubMed  CAS  Google Scholar 

  74. Nelson CE, Gersbach CA. Engineering delivery vehicles for genome editing. Annu Rev Chem Biomol Eng. 2016;7:637–62.

    Article  CAS  PubMed  Google Scholar 

  75. Cui YR, Wang SJ, Chen J, Li J, Chen W, Wang S, et al. Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides. Genome Biol. 2020;21:51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  76. Li C, Psatha N, Gil S, Wang H, Papayannopoulou T, Lieber A. HDAd5/35(++) adenovirus vector expressing anti-crispr peptides decreases CRISPR/Cas9 toxicity in human hematopoietic stem cells. Mol Ther Methods Clin Dev. 2018;9:390–401.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Schiffer JT, Swan DA, Stone D, Jerome KR. Predictors of hepatitis B cure using gene therapy to deliver DNA cleavage enzymes: a mathematical modeling approach. PLoS Comput Biol. 2013;9:e1003131.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Maggio I, Stefanucci L, Janssen JM, Liu J, Chen X, Mouly V, et al. Selection-free gene repair after adenoviral vector transduction of designer nucleases: rescue of dystrophin synthesis in DMD muscle cell populations. Nucleic Acids Res. 2016;44:1449–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Saydaminova K, Ye X, Wang H, Richter M, Ho M, Chen H, et al. Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation. Mol Ther Methods Clin Dev. 2015;1:e14057.

    Article  CAS  Google Scholar 

  80. Holkers M, Maggio I, Liu J, Janssen JM, Miselli F, Mussolino C, et al. Differential integrity of TALE nuclease genes following adenoviral and lentiviral vector gene transfer into human cells. Nucleic Acids Res. 2013;41:e63.

    Article  CAS  PubMed  Google Scholar 

  81. Brescia M, Janssen JM, Liu J, Goncalves M. High-capacity adenoviral vectors permit robust and versatile testing of DMD gene repair tools and strategies in human cells. Cells. 2020;9:e869.

    Article  PubMed  CAS  Google Scholar 

  82. Li C, Wang H, Georgakopoulou A, Gil S, Yannaki E, Lieber A. In vivo HSC gene therapy using a bi-modular HDAd5/35++ vector cures sickle cell disease in a mouse model. Mol Ther. 2021;29:822–37.

    Article  CAS  PubMed  Google Scholar 

  83. Kato Y, Tabata H, Sato K, Nakamura M, Saito I, Nakanishi T. Adenovirus vectors expressing eight multiplex guide RNAs of CRISPR/Cas9 efficiently disrupted diverse hepatitis B virus gene derived from heterogeneous patient. Int J Mol Sci. 2021;22:e10570.

    Article  CAS  Google Scholar 

  84. Yuan J, Wang J, Crain K, Fearns C, Kim KA, Hua KL, et al. Zinc-finger nuclease editing of human cxcr4 promotes HIV-1 CD4(+) T cell resistance and enrichment. Mol Ther. 2012;20:849–59.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Provasi E, Genovese P, Lombardo A, Magnani Z, Liu PQ, Reik A, et al. Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer. Nat Med. 2012;18:807–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Li L, Krymskaya L, Wang J, Henley J, Rao A, Cao LF, et al. Genomic editing of the HIV-1 coreceptor CCR5 in adult hematopoietic stem and progenitor cells using zinc finger nucleases. Mol Ther. 2013;21:1259–69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  87. Darrow JJ. Luxturna: FDA documents reveal the value of a costly gene therapy. Drug Discovery Today. 2019;24:949–54.

    Article  PubMed  Google Scholar 

  88. Mendell JR, Al-Zaidy S, Shell R, Arnold WD, Rodino-Klapac LR, Prior TW, et al. Single-dose gene-replacement therapy for spinal muscular atrophy. N Engl J Med. 2017;377:1713–22.

    Article  CAS  PubMed  Google Scholar 

  89. Li C, Samulski RJ. Engineering adeno-associated virus vectors for gene therapy. Nat Rev Genet. 2020;21:255–72.

    Article  CAS  PubMed  Google Scholar 

  90. Naso MF, Tomkowicz B, Perry WL 3rd, Strohl WR. Adeno-associated virus (AAV) as a vector for gene therapy. BioDrugs. 2017;31:317–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  91. Anguela XM, Sharma R, Doyon Y, Miller JC, Li H, Haurigot V, et al. Robust ZFN-mediated genome editing in adult hemophilic mice. Blood. 2013;122:3283–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Scott T, Moyo B, Nicholson S, Maepa MB, Watashi K, Ely A, et al. ssAAVs containing cassettes encoding SaCas9 and guides targeting hepatitis B virus inactivate replication of the virus in cultured cells. Sci Rep. 2017;7:e7401.

    Article  CAS  Google Scholar 

  93. Liu Y, Zhao M, Gong M, Xu Y, Xie C, Deng H, et al. Inhibition of hepatitis B virus replication via HBV DNA cleavage by Cas9 from Staphylococcus aureus. Antiviral Res. 2018;152:58–67.

    Article  CAS  PubMed  Google Scholar 

  94. Yan K, Feng J, Liu X, Wang H, Li Q, Li J, et al. Inhibition of hepatitis B virus by AAV8-derived CRISPR/SaCas9 expressed from liver-specific promoters. Front Microbiol. 2021;12:1–14.

    Article  Google Scholar 

  95. Kayesh MEH, Amako Y, Hashem MA, Murakami S, Ogawa S, Yamamoto N, et al. Development of an in vivo delivery system for CRISPR/Cas9-mediated targeting of hepatitis B virus cccDNA. Virus Res. 2020;290:e198191.

    Article  CAS  Google Scholar 

  96. Li H, Sheng C, Liu H, Wang S, Zhao J, Yang L, et al. Inhibition of HBV expression in HBV transgenic mice using AAV-delivered CRISPR-SaCas9. Front Immunol. 2018;9:e2080.

    Article  CAS  Google Scholar 

  97. Stone D, Long KR, Loprieno MA, Feelixge HSDS, Kenkel EJ, Liley RM, et al. CRISPR-Cas9 gene editing of hepatitis B virus in chronically infected humanized mice. Mol Ther Methods Clin Dev 2021;20:258–75.

    Article  CAS  PubMed  Google Scholar 

  98. Charlesworth CT, Deshpande PS, Dever DP, Camarena J, Lemgart VT, Cromer MK, et al. Identification of preexisting adaptive immunity to Cas9 proteins in humans. Nat Med. 2019;25:249–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  99. Gough V, Gersbach CA. Immunity to Cas9 as an obstacle to persistent genome editing. Mol Ther. 2020;28:1389–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Gallinaro A, Borghi M, Pirillo MF, Cecchetti S, Bona R, Canitano A, et al. Development and preclinical evaluation of an integrase defective lentiviral vector vaccine expressing the HIVACAT T cell immunogen in mice. Mol Ther Methods Clin Dev. 2020;17:418–28.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  101. Morante V, Borghi M, Farina I, Michelini Z, Grasso F, Gallinaro A, et al. Integrase-defective lentiviral vector is an efficient vaccine platform for cancer immunotherapy. Viruses. 2021;13:e355.

    Article  CAS  Google Scholar 

  102. Iwakuma T, Cui Y, Chang LJ. Self-inactivating lentiviral vectors with U3 and U5 modifications. Virology. 1999;261:120–32.

    Article  CAS  PubMed  Google Scholar 

  103. Palmer DJ, Turner DL, Ng P. Production of CRISPR/Cas9-mediated self-cleaving helper-dependent adenoviruses. Mol Ther Methods Clin Dev. 2019;13:432–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  104. Palmer DJ, Turner DL, Ng P. A single “all-in-one” helper-dependent adenovirus to deliver donor DNA and CRISPR/Cas9 for efficient homology-directed repair. Mol Ther Methods Clin Dev. 2020;17:441–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  105. Mendonca SA, Lorincz R, Boucher P, Curiel DT. Adenoviral vector vaccine platforms in the SARS-CoV-2 pandemic. NPJ Vaccines. 2021;6:e97.

    Article  CAS  Google Scholar 

  106. Bockstal V, Gaddah A, Goldstein N, Shukarev G, Bart S, Luhn K, et al. Assessments of different batches and dose levels of a two-dose Ad26.ZEBOV and MVA-BN-Filo vaccine regimen. NPJ Vaccines. 2021;6:e157.

    Article  CAS  Google Scholar 

  107. Raper SE, Chirmule N, Lee FS, Wivel NA, Bagg A, Gao GP, et al. Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. Mol Genet Metab. 2003;80:148–58.

    Article  CAS  PubMed  Google Scholar 

  108. Fontanellas A, Hervas-Stubbs S, Mauleon I, Dubrot J, Mancheno U, Collantes M, et al. Intensive pharmacological immunosuppression allows for repetitive liver gene transfer with recombinant adenovirus in nonhuman primates. Mol Ther. 2010;18:754–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  109. Ahi YS, Bangari DS, Mittal SK. Adenoviral vector immunity: its implications and circumvention strategies. Curr Gene Ther. 2011;11:307–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  110. Mnyandu N, Limani SW, Arbuthnot P, Maepa MB. Advances in designing Adeno-associated viral vectors for development of anti-HBV gene therapeutics. Virol J. 2021;18:e247.

    Article  CAS  Google Scholar 

  111. Ellis B, Hirsch M, Porter S, Samulski R, Porteus M. Zinc-finger nuclease-mediated gene correction using single AAV vector transduction and enhancement by Food and Drug Administration-approved drugs. Gene Ther. 2013;20:35–42.

    Article  CAS  PubMed  Google Scholar 

  112. Metzger MJ, McConnell-Smith A, Stoddard BL, Miller AD. Single-strand nicks induce homologous recombination with less toxicity than double-strand breaks using an AAV vector template. Nucleic Acids Res. 2011;39:926–35.

    Article  CAS  PubMed  Google Scholar 

  113. Mingozzi F, Maus MV, Sabatino DE, Hui DJ, Rasko JE, Ragni MV, et al. AAV-2 capsid-specific CD8+ T cells limit the duration of gene therapy in humans and cross-react with AAV-8 capsid. Blood. 2006;108:e455.

    Article  Google Scholar 

  114. Velazquez VM, Meadows AS, Pineda RJ, Camboni M, McCarty DM, Fu H. Effective depletion of pre-existing anti-AAV antibodies requires broad immune targeting. Molecular Therapy-Methods & Clinical. Development. 2017;4:159–68.

    CAS  Google Scholar 

  115. Vig K, Herzog R, Martin D, Moore EG, Dennis VA, Pillai S, et al. Recombinant adeno-associated virus as vaccine delivery vehicles. Gene Ther Mol Biol. 2008;12:277–92.

    Google Scholar 

  116. Herzog RW. Immune responses to AAV capsid: are mice not humans after all? Mol Ther. 2007;15:649–50.

    Article  CAS  PubMed  Google Scholar 

  117. Crowther C, Ely A, Hornby J, Mufamadi S, Salazar F, Marion P, et al. Efficient inhibition of hepatitis B virus replication in vivo, using polyethylene glycol-modified adenovirus vectors. Hum Gene Ther. 2008;19:1325–31.

    Article  CAS  PubMed  Google Scholar 

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Funding

Financial support from the South African National Research Foundation (Unique Grant Numbers: 118022 and 120383) and South African Medical Research Council of South Africa is gratefully acknowledged.

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Authors and Affiliations

  1. Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa

    Ridhwaanah Jacobs, Prashika Singh, Tiffany Smith, Patrick Arbuthnot & Mohube Betty Maepa

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  1. Ridhwaanah Jacobs
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  2. Prashika Singh
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  3. Tiffany Smith
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  4. Patrick Arbuthnot
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  5. Mohube Betty Maepa
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Contributions

RJ wrote the sections on LVs and AAVs, put the first draft together and revised all the drafts. PS wrote the introduction section on HBV biology and revised all drafts. TS wrote the introduction section on anti-HBV designer nucleases and revised all drafts. PA revised the final draft. MBM conceptualized the idea, wrote the section on Ads, revised all the drafts and finalized the manuscript for submission.

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Correspondence to Mohube Betty Maepa.

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Jacobs, R., Singh, P., Smith, T. et al. Prospects of viral vector-mediated delivery of sequences encoding anti-HBV designer endonucleases. Gene Ther (2022). https://doi.org/10.1038/s41434-022-00342-5

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  • DOI: https://doi.org/10.1038/s41434-022-00342-5

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