Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies
Herpes simplex virus-1 (HSV-1) is highly contagious, and there is a need for a therapeutic means to eradicate it. We have identified an siRNA (siHSV) that knocks down gene expression of the infected cell protein 0 (ICP0), which is important in the regulation of HSV infection. The selected siHSV was...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/3/633 |
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| author | Doaa Jbara-Agbaria Saskia Blondzik Anke Burger-Kentischer Majd Agbaria Mirjam M. Nordling-David Anna Giterman Gil Aizik Steffen Rupp Gershon Golomb |
| author_facet | Doaa Jbara-Agbaria Saskia Blondzik Anke Burger-Kentischer Majd Agbaria Mirjam M. Nordling-David Anna Giterman Gil Aizik Steffen Rupp Gershon Golomb |
| author_sort | Doaa Jbara-Agbaria |
| collection | DOAJ |
| description | Herpes simplex virus-1 (HSV-1) is highly contagious, and there is a need for a therapeutic means to eradicate it. We have identified an siRNA (siHSV) that knocks down gene expression of the infected cell protein 0 (ICP0), which is important in the regulation of HSV infection. The selected siHSV was encapsulated in liposomes to overcome its poor stability, increase cell permeability, and prolonging siRNA circulation time. Several siRNAs against ICP0 have been designed and identified. We examined the role of various parameters, including formulation technique, lipids composition, and ratio. An optimal liposomal siHSV formulation (Lip<sup>DOPE</sup>-siHSV) was characterized with desirable physiochemical properties, in terms of nano-size, low polydispersity index (PDI), neutral surface charge, high siHSV loading, spherical shape, high stability in physiologic conditions in vitro, and long-term shelf-life stability (>1 year, 4 °C). The liposomes exhibited profound internalization by human keratinocytes, no cytotoxicity in cell cultures, no detrimental effect on mice liver enzymes, and a gradual endo-lysosomal escape. Mice biodistribution studies in intact mice revealed accumulation, mainly in visceral organs but also in the trigeminal ganglion. The therapeutic potential of siHSV liposomes was demonstrated by significant antiviral activity both in the plaque reduction assay and in the 3D epidermis model, and the mechanism of action was validated by the reduction of ICP0 expression levels. |
| first_indexed | 2024-03-09T12:58:06Z |
| format | Article |
| id | doaj.art-a89dfc1974d145ccb2aba1113f8ec524 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T12:58:06Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-a89dfc1974d145ccb2aba1113f8ec5242023-11-30T21:57:43ZengMDPI AGPharmaceutics1999-49232022-03-0114363310.3390/pharmaceutics14030633Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity StudiesDoaa Jbara-Agbaria0Saskia Blondzik1Anke Burger-Kentischer2Majd Agbaria3Mirjam M. Nordling-David4Anna Giterman5Gil Aizik6Steffen Rupp7Gershon Golomb8Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, IsraelFraunhofer Institute for Interfacial Engineering and Biotechnology, 70569 Stuttgart, GermanyFraunhofer Institute for Interfacial Engineering and Biotechnology, 70569 Stuttgart, GermanyInstitute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, IsraelInstitute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, IsraelInstitute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, IsraelInstitute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, IsraelFraunhofer Institute for Interfacial Engineering and Biotechnology, 70569 Stuttgart, GermanyInstitute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, IsraelHerpes simplex virus-1 (HSV-1) is highly contagious, and there is a need for a therapeutic means to eradicate it. We have identified an siRNA (siHSV) that knocks down gene expression of the infected cell protein 0 (ICP0), which is important in the regulation of HSV infection. The selected siHSV was encapsulated in liposomes to overcome its poor stability, increase cell permeability, and prolonging siRNA circulation time. Several siRNAs against ICP0 have been designed and identified. We examined the role of various parameters, including formulation technique, lipids composition, and ratio. An optimal liposomal siHSV formulation (Lip<sup>DOPE</sup>-siHSV) was characterized with desirable physiochemical properties, in terms of nano-size, low polydispersity index (PDI), neutral surface charge, high siHSV loading, spherical shape, high stability in physiologic conditions in vitro, and long-term shelf-life stability (>1 year, 4 °C). The liposomes exhibited profound internalization by human keratinocytes, no cytotoxicity in cell cultures, no detrimental effect on mice liver enzymes, and a gradual endo-lysosomal escape. Mice biodistribution studies in intact mice revealed accumulation, mainly in visceral organs but also in the trigeminal ganglion. The therapeutic potential of siHSV liposomes was demonstrated by significant antiviral activity both in the plaque reduction assay and in the 3D epidermis model, and the mechanism of action was validated by the reduction of ICP0 expression levels.https://www.mdpi.com/1999-4923/14/3/633nanomedicineliposomesgene deliveryHSV-13D epidermis model |
| spellingShingle | Doaa Jbara-Agbaria Saskia Blondzik Anke Burger-Kentischer Majd Agbaria Mirjam M. Nordling-David Anna Giterman Gil Aizik Steffen Rupp Gershon Golomb Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies Pharmaceutics nanomedicine liposomes gene delivery HSV-1 3D epidermis model |
| title | Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies |
| title_full | Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies |
| title_fullStr | Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies |
| title_full_unstemmed | Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies |
| title_short | Liposomal siRNA Formulations for the Treatment of Herpes Simplex Virus-1: In Vitro Characterization of Physicochemical Properties and Activity, and In Vivo Biodistribution and Toxicity Studies |
| title_sort | liposomal sirna formulations for the treatment of herpes simplex virus 1 in vitro characterization of physicochemical properties and activity and in vivo biodistribution and toxicity studies |
| topic | nanomedicine liposomes gene delivery HSV-1 3D epidermis model |
| url | https://www.mdpi.com/1999-4923/14/3/633 |
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