Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo
Chronic hepatitis B infection (CHB) is an area of high unmet medical need. Current standard-of-care therapies only rarely lead to a functional cure, defined as durable hepatitis B surface antigen (HBsAg) loss following treatment. The goal for next generation CHB therapies is to achieve a higher rate...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-06-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118300192 |
| _version_ | 1819229033171255296 |
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| author | Hassan Javanbakht Henrik Mueller Johanna Walther Xue Zhou Anaïs Lopez Thushara Pattupara Julie Blaising Lykke Pedersen Nanna Albæk Malene Jackerott Tianlai Shi Corinne Ploix Wouter Driessen Robert Persson Jacob Ravn John A.T. Young Søren Ottosen |
| author_facet | Hassan Javanbakht Henrik Mueller Johanna Walther Xue Zhou Anaïs Lopez Thushara Pattupara Julie Blaising Lykke Pedersen Nanna Albæk Malene Jackerott Tianlai Shi Corinne Ploix Wouter Driessen Robert Persson Jacob Ravn John A.T. Young Søren Ottosen |
| author_sort | Hassan Javanbakht |
| collection | DOAJ |
| description | Chronic hepatitis B infection (CHB) is an area of high unmet medical need. Current standard-of-care therapies only rarely lead to a functional cure, defined as durable hepatitis B surface antigen (HBsAg) loss following treatment. The goal for next generation CHB therapies is to achieve a higher rate of functional cure with finite treatment duration. To address this urgent need, we are developing liver-targeted single-stranded oligonucleotide (SSO) therapeutics for CHB based on the locked nucleic acid (LNA) platform. These LNA-SSOs target hepatitis B virus (HBV) transcripts for RNase-H-mediated degradation. Here, we describe a HBV-specific LNA-SSO that effectively reduces intracellular viral mRNAs and viral antigens (HBsAg and HBeAg) over an extended time period in cultured human hepatoma cell lines that were infected with HBV with mean 50% effective concentration (EC50) values ranging from 1.19 to 1.66 μM. To achieve liver-specific targeting and minimize kidney exposure, this LNA-SSO was conjugated to a cluster of three N-acetylgalactosamine (GalNAc) moieties that direct specific binding to the asialoglycoprotein receptor (ASGPR) expressed specifically on the surface of hepatocytes. The GalNAc-conjugated LNA-SSO showed a strikingly higher level of potency when tested in the AAV-HBV mouse model as compared with its non-conjugated counterpart. Remarkably, higher doses of GalNAc-conjugated LNA-SSO resulted in a rapid and long-lasting reduction of HBsAg to below the detection limit for quantification, i.e., by 3 log10 (p < 0.0003). This antiviral effect depended on a close match between the sequences of the LNA-SSO and its HBV target, indicating that the antiviral effect is not due to non-specific oligonucleotide-driven immune activation. These data support the development of LNA-SSO therapeutics for the treatment of CHB infection. |
| first_indexed | 2024-12-23T11:06:44Z |
| format | Article |
| id | doaj.art-1962d4b211f2443bb9955ca1ed51da8a |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-23T11:06:44Z |
| publishDate | 2018-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-1962d4b211f2443bb9955ca1ed51da8a2022-12-21T17:49:28ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-06-0111C44145410.1016/j.omtn.2018.02.005Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In VivoHassan Javanbakht0Henrik Mueller1Johanna Walther2Xue Zhou3Anaïs Lopez4Thushara Pattupara5Julie Blaising6Lykke Pedersen7Nanna Albæk8Malene Jackerott9Tianlai Shi10Corinne Ploix11Wouter Driessen12Robert Persson13Jacob Ravn14John A.T. Young15Søren Ottosen16Roche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Shanghai, Shanghai 201203, ChinaRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Shanghai, Shanghai 201203, ChinaRoche Pharma Research and Early Development, Roche Innovation Center Copenhagen, 2970 Hørsholm, DenmarkRoche Pharma Research and Early Development, Roche Innovation Center Copenhagen, 2970 Hørsholm, DenmarkRoche Pharma Research and Early Development, Roche Innovation Center Copenhagen, 2970 Hørsholm, DenmarkRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Copenhagen, 2970 Hørsholm, DenmarkRoche Pharma Research and Early Development, Roche Innovation Center Copenhagen, 2970 Hørsholm, DenmarkRoche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Copenhagen, 2970 Hørsholm, DenmarkChronic hepatitis B infection (CHB) is an area of high unmet medical need. Current standard-of-care therapies only rarely lead to a functional cure, defined as durable hepatitis B surface antigen (HBsAg) loss following treatment. The goal for next generation CHB therapies is to achieve a higher rate of functional cure with finite treatment duration. To address this urgent need, we are developing liver-targeted single-stranded oligonucleotide (SSO) therapeutics for CHB based on the locked nucleic acid (LNA) platform. These LNA-SSOs target hepatitis B virus (HBV) transcripts for RNase-H-mediated degradation. Here, we describe a HBV-specific LNA-SSO that effectively reduces intracellular viral mRNAs and viral antigens (HBsAg and HBeAg) over an extended time period in cultured human hepatoma cell lines that were infected with HBV with mean 50% effective concentration (EC50) values ranging from 1.19 to 1.66 μM. To achieve liver-specific targeting and minimize kidney exposure, this LNA-SSO was conjugated to a cluster of three N-acetylgalactosamine (GalNAc) moieties that direct specific binding to the asialoglycoprotein receptor (ASGPR) expressed specifically on the surface of hepatocytes. The GalNAc-conjugated LNA-SSO showed a strikingly higher level of potency when tested in the AAV-HBV mouse model as compared with its non-conjugated counterpart. Remarkably, higher doses of GalNAc-conjugated LNA-SSO resulted in a rapid and long-lasting reduction of HBsAg to below the detection limit for quantification, i.e., by 3 log10 (p < 0.0003). This antiviral effect depended on a close match between the sequences of the LNA-SSO and its HBV target, indicating that the antiviral effect is not due to non-specific oligonucleotide-driven immune activation. These data support the development of LNA-SSO therapeutics for the treatment of CHB infection.http://www.sciencedirect.com/science/article/pii/S2162253118300192HBVHBsAgviral gene expressionAAV-HBV-infected mouse modellocked nucleic acidantisense |
| spellingShingle | Hassan Javanbakht Henrik Mueller Johanna Walther Xue Zhou Anaïs Lopez Thushara Pattupara Julie Blaising Lykke Pedersen Nanna Albæk Malene Jackerott Tianlai Shi Corinne Ploix Wouter Driessen Robert Persson Jacob Ravn John A.T. Young Søren Ottosen Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo Molecular Therapy: Nucleic Acids HBV HBsAg viral gene expression AAV-HBV-infected mouse model locked nucleic acid antisense |
| title | Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo |
| title_full | Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo |
| title_fullStr | Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo |
| title_full_unstemmed | Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo |
| title_short | Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo |
| title_sort | liver targeted anti hbv single stranded oligonucleotides with locked nucleic acid potently reduce hbv gene expression in vivo |
| topic | HBV HBsAg viral gene expression AAV-HBV-infected mouse model locked nucleic acid antisense |
| url | http://www.sciencedirect.com/science/article/pii/S2162253118300192 |
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