Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites
Lipid nanoparticles (LNPs) are often liver tropic, presenting challenges for LNP-delivered mRNA therapeutics intended for other tissues, as off-target expression in the liver may increase side effects and modulate immune responses. To avoid off-target expression in the liver, miR-122 binding sites h...
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Elsevier
2025-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050124002183 |
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| author | Brian J. Parrett Satoko Yamaoka Michael A. Barry |
| author_facet | Brian J. Parrett Satoko Yamaoka Michael A. Barry |
| author_sort | Brian J. Parrett |
| collection | DOAJ |
| description | Lipid nanoparticles (LNPs) are often liver tropic, presenting challenges for LNP-delivered mRNA therapeutics intended for other tissues, as off-target expression in the liver may increase side effects and modulate immune responses. To avoid off-target expression in the liver, miR-122 binding sites have been used by others in viral and non-viral therapeutics. Here, we use a luciferase reporter system to compare different copy numbers and insertion locations of miR-122 binding sequences to restrict liver expression. We inserted one to five miR-122 binding sites into the 5′ or 3′ untranslated regions (UTRs) of luciferase mRNAs and tested them in LNPs in vitro and in vivo via systemic intravenous and local intramuscular injections in mice. Our results showed no significant differences in de-targeting efficacy between mRNAs harboring one or multiple miR-122 binding sites or between those with 5′ or 3′ UTR placements. To test the impact of miR-122 binding sites on antibody response to a mRNA vaccine, Ebola virus matrix protein VP40 mRNAs were modified with or without miR-122 binding sites and injected in mice intramuscularly. This work reinforces the utility of miR-122 binding sites while providing a comparison of these sites to aid the future development of LNP-mRNA therapies for non-hepatic tissues. |
| format | Article |
| id | doaj-art-4c6be28b12f14253a700c0629ad401b0 |
| institution | Kabale University |
| issn | 2329-0501 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
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| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj-art-4c6be28b12f14253a700c0629ad401b02025-01-05T04:28:16ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-03-01331101402Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sitesBrian J. Parrett0Satoko Yamaoka1Michael A. Barry2Virology and Gene Therapy (VGT) Graduate Program, Mayo Clinic, Rochester, MN 55902, USADepartment of Medicine, Division of Infectious Diseases, Mayo Clinic, Rochester, MN 55902, USADepartment of Medicine, Division of Infectious Diseases, Mayo Clinic, Rochester, MN 55902, USA; Department of Immunology, Mayo Clinic, Rochester, MN 55902, USA; Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55902, USA; Corresponding author: Michael A. Barry, Department of Medicine, Division of Infectious Diseases, Mayo Clinic, 200 First Street SW, Rochester, MN 55902, USA.Lipid nanoparticles (LNPs) are often liver tropic, presenting challenges for LNP-delivered mRNA therapeutics intended for other tissues, as off-target expression in the liver may increase side effects and modulate immune responses. To avoid off-target expression in the liver, miR-122 binding sites have been used by others in viral and non-viral therapeutics. Here, we use a luciferase reporter system to compare different copy numbers and insertion locations of miR-122 binding sequences to restrict liver expression. We inserted one to five miR-122 binding sites into the 5′ or 3′ untranslated regions (UTRs) of luciferase mRNAs and tested them in LNPs in vitro and in vivo via systemic intravenous and local intramuscular injections in mice. Our results showed no significant differences in de-targeting efficacy between mRNAs harboring one or multiple miR-122 binding sites or between those with 5′ or 3′ UTR placements. To test the impact of miR-122 binding sites on antibody response to a mRNA vaccine, Ebola virus matrix protein VP40 mRNAs were modified with or without miR-122 binding sites and injected in mice intramuscularly. This work reinforces the utility of miR-122 binding sites while providing a comparison of these sites to aid the future development of LNP-mRNA therapies for non-hepatic tissues.http://www.sciencedirect.com/science/article/pii/S2329050124002183lipid nanoparticlemRNAmicroRNAvaccinede-targetingmiR-122 |
| spellingShingle | Brian J. Parrett Satoko Yamaoka Michael A. Barry Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites Molecular Therapy: Methods & Clinical Development lipid nanoparticle mRNA microRNA vaccine de-targeting miR-122 |
| title | Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites |
| title_full | Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites |
| title_fullStr | Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites |
| title_full_unstemmed | Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites |
| title_short | Reducing off-target expression of mRNA therapeutics and vaccines in the liver with microRNA binding sites |
| title_sort | reducing off target expression of mrna therapeutics and vaccines in the liver with microrna binding sites |
| topic | lipid nanoparticle mRNA microRNA vaccine de-targeting miR-122 |
| url | http://www.sciencedirect.com/science/article/pii/S2329050124002183 |
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