Inhibition of Hepatitis C Virus Replication by Intracellular Delivery of Multiple SiRNAs by Nanosomes

Overview

Journal Mol Ther

Publisher Cell Press

Specialties Molecular Biology
Pharmacology

Date 2012 May 24

PMID 22617108

Citations 38

Authors

Partha K Chandra

Anup K Kundu

Sidhartha Hazari

Sruti Chandra

Lili Bao

Tara Ooms

Gilbert F Morris

Tong Wu

Tarun K Mandal

Srikanta Dash

Affiliations

Soon will be listed here.

Abstract

Sustained antiviral responses of chronic hepatitis C virus (HCV) infection have improved recently by the use of direct-acting antiviral agents along with interferon (IFN)-α and ribavirin. However, the emergence of drug-resistant variants is expected to be a major problem. We describe here a novel combinatorial small interfering RNA (siRNA) nanosome-based antiviral approach to clear HCV infection. Multiple siRNAs targeted to the highly conserved 5'-untranslated region (UTR) of the HCV genome were synthesized and encapsulated into lipid nanoparticles called nanosomes. We show that siRNA can be repeatedly delivered to 100% of cells in culture using nanosomes without toxicity. Six siRNAs dramatically reduced HCV replication in both the replicon and infectious cell culture model. Repeated treatments with two siRNAs were better than a single siRNA treatment in minimizing the development of an escape mutant, resulting in rapid inhibition of viral replication. Systemic administration of combinatorial siRNA-nanosomes is well tolerated in BALB/c mice without liver injury or histological toxicity. As a proof-of-principle, we showed that systemic injections of siRNA nanosomes significantly reduced HCV replication in a liver tumor-xenotransplant mouse model of HCV. Our results indicate that systemic delivery of combinatorial siRNA nanosomes can be used to minimize the development of escape mutants and inhibition of HCV infection.

Citing Articles

Principle, application and challenges of development siRNA-based therapeutics against bacterial and viral infections: a comprehensive review.

Motamedi H, Mahdizade Ari M, Alvandi A, Abiri R Front Microbiol. 2024; 15:1393646.

PMID: 38939184 PMC: 11208694. DOI: 10.3389/fmicb.2024.1393646.

Nanotechnology Platform for Advancing Vaccine Development against the COVID-19 Virus.

Chowdhury N, Kundu A Diseases. 2023; 11(4).

PMID: 38131983 PMC: 10742622. DOI: 10.3390/diseases11040177.

Developing New Tools to Fight Human Pathogens: A Journey through the Advances in RNA Technologies.

Costa V, Costa S, Saramago M, Cunha M, Arraiano C, Viegas S Microorganisms. 2022; 10(11).

PMID: 36422373 PMC: 9697208. DOI: 10.3390/microorganisms10112303.

In Silico Prediction and Selection of Target Sequences in the SARS-CoV-2 RNA Genome for an Antiviral Attack.

Hussein M, Andrade Dos Ramos Z, Berkhout B, Herrera-Carrillo E Viruses. 2022; 14(2).

PMID: 35215977 PMC: 8880226. DOI: 10.3390/v14020385.

Harnessing Intronic microRNA Structures to Improve Tolerance and Expression of shRNAs in Animal Cells.

Challagulla A, Tizard M, Doran T, Cahill D, Jenkins K Methods Protoc. 2022; 5(1).

PMID: 35200534 PMC: 8879667. DOI: 10.3390/mps5010018.

References

Westerhout E, Ooms M, Vink M, Das A, Berkhout B . HIV-1 can escape from RNA interference by evolving an alternative structure in its RNA genome. Nucleic Acids Res. 2005; 33(2):796-804. PMC: 548362. DOI: 10.1093/nar/gki220. View

Kundu A, Chandra P, Hazari S, Ledet G, Pramar Y, Dash S . Stability of lyophilized siRNA nanosome formulations. Int J Pharm. 2011; 423(2):525-34. PMC: 3298087. DOI: 10.1016/j.ijpharm.2011.11.040. View

Hazari S, Hefler H, Chandra P, Poat B, Gunduz F, Ooms T . Hepatocellular carcinoma xenograft supports HCV replication: a mouse model for evaluating antivirals. World J Gastroenterol. 2011; 17(3):300-12. PMC: 3022289. DOI: 10.3748/wjg.v17.i3.300. View

Fumoto S, Nakadori F, Kawakami S, Nishikawa M, Yamashita F, Hashida M . Analysis of hepatic disposition of galactosylated cationic liposome/plasmid DNA complexes in perfused rat liver. Pharm Res. 2003; 20(9):1452-9. DOI: 10.1023/a:1025766429175. View

Williams R . Global challenges in liver disease. Hepatology. 2006; 44(3):521-6. DOI: 10.1002/hep.21347. View

Kronke J, Kittler R, Buchholz F, Windisch M, Pietschmann T, Bartenschlager R . Alternative approaches for efficient inhibition of hepatitis C virus RNA replication by small interfering RNAs. J Virol. 2004; 78(7):3436-46. PMC: 371081. DOI: 10.1128/jvi.78.7.3436-3446.2004. View

Chevalier C, Saulnier A, Benureau Y, Flechet D, Delgrange D, Colbere-Garapin F . Inhibition of hepatitis C virus infection in cell culture by small interfering RNAs. Mol Ther. 2007; 15(8):1452-62. PMC: 7106008. DOI: 10.1038/sj.mt.6300186. View

Kim M, Shin D, Kim S, Park M . Inhibition of hepatitis C virus gene expression by small interfering RNAs using a tri-cistronic full-length viral replicon and a transient mouse model. Virus Res. 2006; 122(1-2):1-10. DOI: 10.1016/j.virusres.2006.05.003. View

Wood N, Bhattacharya T, Keele B, Giorgi E, Liu M, Gaschen B . HIV evolution in early infection: selection pressures, patterns of insertion and deletion, and the impact of APOBEC. PLoS Pathog. 2009; 5(5):e1000414. PMC: 2671846. DOI: 10.1371/journal.ppat.1000414. View

10.

Westerhout E, Berkhout B . A systematic analysis of the effect of target RNA structure on RNA interference. Nucleic Acids Res. 2007; 35(13):4322-30. PMC: 1934999. DOI: 10.1093/nar/gkm437. View

11.

Howard K . Delivery of RNA interference therapeutics using polycation-based nanoparticles. Adv Drug Deliv Rev. 2009; 61(9):710-20. DOI: 10.1016/j.addr.2009.04.001. View

12.

Jasmijn von Eije K, Brake O, Berkhout B . Human immunodeficiency virus type 1 escape is restricted when conserved genome sequences are targeted by RNA interference. J Virol. 2007; 82(6):2895-903. PMC: 2258968. DOI: 10.1128/JVI.02035-07. View

13.

Aagaard L, Rossi J . RNAi therapeutics: principles, prospects and challenges. Adv Drug Deliv Rev. 2007; 59(2-3):75-86. PMC: 1978219. DOI: 10.1016/j.addr.2007.03.005. View

14.

Boden D, Pusch O, Lee F, Tucker L, Ramratnam B . Human immunodeficiency virus type 1 escape from RNA interference. J Virol. 2003; 77(21):11531-5. PMC: 229317. DOI: 10.1128/jvi.77.21.11531-11535.2003. View

15.

Schubert S, Grunweller A, Erdmann V, Kurreck J . Local RNA target structure influences siRNA efficacy: systematic analysis of intentionally designed binding regions. J Mol Biol. 2005; 348(4):883-93. DOI: 10.1016/j.jmb.2005.03.011. View

16.

Watanabe T, Umehara T, Kohara M . Therapeutic application of RNA interference for hepatitis C virus. Adv Drug Deliv Rev. 2007; 59(12):1263-76. DOI: 10.1016/j.addr.2007.03.022. View

17.

Dash S, Halim A, Tsuji H, Hiramatsu N, Gerber M . Transfection of HepG2 cells with infectious hepatitis C virus genome. Am J Pathol. 1997; 151(2):363-73. PMC: 1858015. View

18.

Tseng Y, Mozumdar S, Huang L . Lipid-based systemic delivery of siRNA. Adv Drug Deliv Rev. 2009; 61(9):721-31. PMC: 3172140. DOI: 10.1016/j.addr.2009.03.003. View

19.

Luo K, Chang D . The gene-silencing efficiency of siRNA is strongly dependent on the local structure of mRNA at the targeted region. Biochem Biophys Res Commun. 2004; 318(1):303-10. DOI: 10.1016/j.bbrc.2004.04.027. View

20.

Yoshinari K, Miyagishi M, Taira K . Effects on RNAi of the tight structure, sequence and position of the targeted region. Nucleic Acids Res. 2004; 32(2):691-9. PMC: 373345. DOI: 10.1093/nar/gkh221. View