Engineered extracellular vesicles for delivering functional Cas9/gRNA to eliminate hepatitis B virus cccDNA and integration

References

• Hepatitis B: World Health Organization. 2022. Available from: https://www.who.int/news-room/fact-sheets/detail/hepatitis-b.
   Google Scholar
• Fanning GC, Zoulim F, Hou J, et al. Therapeutic strategies for hepatitis B virus infection: towards a cure. Nat Rev Drug Discovery. 2019 Nov;18(11):827–844.
   PubMed Web of Science ®Google Scholar
• Pollicino T, Caminiti G. HBV-Integration studies in the clinic: role in the natural history of infection. Viruses. 2021 Feb 26;13(3):368.
   PubMed Web of Science ®Google Scholar
• Terrault NA, Bzowej NH, Chang KM, et al. AASLD guidelines for treatment of chronic hepatitis B. Hepatology (Baltimore, Md). 2016 Jan;63(1):261–283.
   PubMed Web of Science ®Google Scholar
• van den Berg F, Limani SW, Mnyandu N, et al. Advances with RNAi-Based Therapy for Hepatitis B Virus Infection. Viruses. 2020 Aug 4;12(8):851.
   PubMed Web of Science ®Google Scholar
• Penna A, Laccabue D, Libri I, et al. Peginterferon-α does not improve early peripheral blood HBV-specific T-cell responses in HBeAg-negative chronic hepatitis. J Hepatol. 2012 Jun;56(6):1239–1246.
   PubMed Web of Science ®Google Scholar
• Makarova KS, Wolf YI, Iranzo J, et al. Evolutionary classification of CRISPR-Cas systems: a burst of class 2 and derived variants. Nat Rev Microbiol. 2020 Feb;18(2):67–83.
   PubMed Web of Science ®Google Scholar
• Lin SR, Yang HC, Kuo YT, et al. The CRISPR/Cas9 system facilitates clearance of the Intrahepatic HBV Templates In Vivo. Mol Ther Nucleic Acids. 2014 Aug 19;3(8):e186.
   PubMedGoogle Scholar
• Seeger C, Sohn JA. Targeting hepatitis B virus with CRISPR/Cas9. Mol Ther Nucleic Acids. 2014 Dec 16;3(12):e216.
   PubMedGoogle Scholar
• Wang J, Xu ZW, Liu S, et al. Dual gRNAs guided CRISPR/Cas9 system inhibits hepatitis B virus replication. World J Gastroenterol. 2015 Aug 28;21(32):9554–9565.
   PubMed Web of Science ®Google Scholar
• Wang J, Chen R, Zhang R, et al. The gRNA-miRNA-gRNA Ternary Cassette Combining CRISPR/Cas9 with RNAi Approach Strongly Inhibits Hepatitis B virus replication. Theranostics. 2017;7(12):3090–3105.
   PubMed Web of Science ®Google Scholar
• Zhen S, Hua L, Liu YH, et al. Harnessing the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated Cas9 system to disrupt the hepatitis B virus. Gene Ther. 2015 May;22(5):404–412.
   PubMed Web of Science ®Google Scholar
• Liu Y, Zhao M, Gong M, et al. Inhibition of hepatitis B virus replication via HBV DNA cleavage by Cas9 from Staphylococcus aureus. Antiviral Res. 2018 Apr;152:58–67.
   PubMed Web of Science ®Google Scholar
• Nelson CE, Wu Y, Gemberling MP, et al. Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy. Nat Med. 2019 Mar;25(3):427–432.
   PubMed Web of Science ®Google Scholar
• Hanlon KS, Kleinstiver BP, Garcia SP, et al. High levels of AAV vector integration into CRISPR-induced DNA breaks. Nat Commun. 2019 Sep 30;10(1):4439.
   PubMedGoogle Scholar
• Tong S, Moyo B, Lee CM, et al. Engineered materials for in vivo delivery of genome-editing machinery. Nat Rev Mater. 2019 Nov;4:726–737.
   PubMed Web of Science ®Google Scholar
• Chen R, Huang H, Liu H, et al. Friend or Foe? Evidence indicates endogenous exosomes can deliver functional gRNA and Cas9 protein. Small (Weinheim an der Bergstrasse, Germany). 2019 Sep;15(38):e1902686.
   PubMed Web of Science ®Google Scholar
• Rädler J, Gupta D, Zickler A, et al. Exploiting the biogenesis of extracellular vesicles for bioengineering and therapeutic cargo loading [published online ahead of print, 2023 Feb 20]. Mol Ther: The J Am Soc Gene Ther. 2023 Feb 20: S1525-0016(23)00076-X.
   Google Scholar
• Kennedy MJ, Hughes RM, Peteya LA, et al. Rapid blue-light-mediated induction of protein interactions in living cells. Nat Methods. 2010 Dec;7(12):973–975.
   PubMed Web of Science ®Google Scholar
• Shen B, Wu N, Yang JM, et al. Protein targeting to exosomes/microvesicles by plasma membrane anchors. J Biol Chem. 2011 Apr 22;286(16):14383–14395.
   PubMed Web of Science ®Google Scholar
• Sato I, Obata Y, Kasahara K, et al. Differential trafficking of Src, Lyn, Yes and Fyn is specified by the state of palmitoylation in the SH4 domain. J Cell Sci. 2009 Apr 1;122(Pt 7):965–975.
   PubMed Web of Science ®Google Scholar
• Michailidis E, Pabon J, Xiang K, et al. A robust cell culture system supporting the complete life cycle of hepatitis B virus. Sci Rep. 2017 Nov 30;7(1):16616.
   PubMedGoogle Scholar
• 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. Antimicrob Agents Chemother. 1997 Aug;41(8):1715–1720.
   PubMed Web of Science ®Google Scholar
• Gupta D, Liang X, Pavlova S, et al. Quantification of extracellular vesicles in vitro and in vivo using sensitive bioluminescence imaging. J Extracell Vesicles. 2020 Aug 21;9(1):1800222.
   PubMed Web of Science ®Google Scholar
• Xia Y, Stadler D, Ko C, et al. Analyses of HBV cccDNA Quantification and Modification. Methods Mol Biol (Clifton, NJ). 2017;1540:59–72.
   PubMedGoogle Scholar
• Luangsay S, Gruffaz M, Isorce N, et al. Early inhibition of hepatocyte innate responses by hepatitis B virus. J Hepatol. 2015 Dec;63(6):1314–1322.
   PubMed Web of Science ®Google Scholar
• Martinez MG, Combe E, Inchauspe A, et al. CRISPR-Cas9 targeting of hepatitis B virus covalently closed circular DNA generates transcriptionally active episomal variants. mBio. 2022 Apr 26;13(2):e0288821.
   PubMed Web of Science ®Google Scholar
• Rohner E, Yang R, Foo KS, et al. Unlocking the promise of mRNA therapeutics. Nat Biotechnol. 2022 Nov;40(11):1586–1600.
   PubMed Web of Science ®Google Scholar
• Wiklander OPB, Brennan M, Lötvall J, et al. Advances in therapeutic applications of extracellular vesicles. Sci Transl Med. 2019 May 15;11(492):eaav8521.
   PubMed Web of Science ®Google Scholar
• El Andaloussi S, Lakhal S, Mäger I, et al. Exosomes for targeted siRNA delivery across biological barriers. Adv Drug Delivery Rev. 2013 Mar;65(3):391–397.
   PubMed Web of Science ®Google Scholar
• Hung ME, Leonard JN. A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery. J Extracell Vesicles. 2016;5:31027.
   PubMed Web of Science ®Google Scholar
• Schrezenmeier E, Dörner T. Mechanisms of action of hydroxychloroquine and chloroquine: implications for rheumatology. Nat Rev Rheumatol. 2020 Mar;16(3):155–166.
   PubMed Web of Science ®Google Scholar
• Li Y, Cho MH, Lee SS, et al. Hydroxychloroquine-loaded hollow mesoporous silica nanoparticles for enhanced autophagy inhibition and radiation therapy. J Controlled Release: Official J Controlled Release Soc. 2020 Sep 10;325:100–110.
   PubMed Web of Science ®Google Scholar
• Sung WK, Zheng H, Li S, et al. Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat Genet. 2012 May 27;44(7):765–769.
   PubMed Web of Science ®Google Scholar
• Li X, Zhang J, Yang Z, et al. The function of targeted host genes determines the oncogenicity of HBV integration in hepatocellular carcinoma. J Hepatol. 2014 May;60(5):975–984.
   PubMed Web of Science ®Google Scholar
• Yang YC, Yang HC. Recent progress and future prospective in HBV Cure by CRISPR/Cas. Viruses. 2021 Dec 21;14(1):4.
   PubMed Web of Science ®Google Scholar
• Whitley JA, Kim S, Lou L, et al. Encapsulating Cas9 into extracellular vesicles by protein myristoylation. J Extracell Vesicles. 2022 Apr;11(4):e12196.
   PubMed Web of Science ®Google Scholar
• Rädler J, Gupta D, Zickler A, et al. Exploiting the biogenesis of extracellular vesicles for bioengineering and therapeutic cargo loading. Mol Ther: The J Am Soc Gene Ther. 2023 May 3;31(5):1231–1250.
   PubMed Web of Science ®Google Scholar
• Yim N, Ryu SW, Choi K, et al. Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein-protein interaction module. Nat Commun. 2016 Jul 22;7:12277.
   PubMed Web of Science ®Google Scholar
• Osteikoetxea X, Silva A, Lázaro-Ibáñez E, et al. Engineered Cas9 extracellular vesicles as a novel gene editing tool. J Extracell Vesicles. 2022 May;11(5):e12225.
   PubMed Web of Science ®Google Scholar
• Gee P, Lung MSY, Okuzaki Y, et al. Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping. Nat Commun. 2020 Mar 13;11(1):1334.
   PubMed Web of Science ®Google Scholar
• Ilahibaks NF, Ardisasmita AI, Xie S, et al. TOP-EVs: technology of protein delivery through extracellular vesicles is a versatile platform for intracellular protein delivery. J Controlled Release: Official J Controlled Release Soc. 2023 Mar;355:579–592.
   PubMed Web of Science ®Google Scholar
• Heath N, Osteikoetxea X, de Oliveria TM, et al. Endosomal escape enhancing compounds facilitate functional delivery of extracellular vesicle cargo. Nanomedicine (London, England). 2019 Nov;14(21):2799–2814.
   PubMed Web of Science ®Google Scholar
• Zhou C, Tan L, Sun Y, et al. Exosomes carry microRNAs into neighboring cells to promote diffusive infection of newcastle disease virus. Viruses. 2019 Jun 6;11(6):527.
   PubMed Web of Science ®Google Scholar
• Li J, Liu K, Liu Y, et al. Exosomes mediate the cell-to-cell transmission of IFN-α-induced antiviral activity. Nat Immunol. 2013 Aug;14(8):793–803.
   PubMed Web of Science ®Google Scholar
• Hamilton JR, Tsuchida CA, Nguyen DN, et al. Targeted delivery of CRISPR-Cas9 and transgenes enables complex immune cell engineering. Cell Rep. 2021 Jun 1;35(9):109207.
   PubMed Web of Science ®Google Scholar