Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors
Systemically delivered adeno-associated viral vector serotype 9 (AAV9) effectively transduces murine heart, but provides transgene expression also in liver and skeletal muscles. Improvement of the selectivity of transgene expression can be achieved through incorporation of target sites (TSs) for miR...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050120300930 |
| _version_ | 1828758932374421504 |
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| author | Izabela Kraszewska Mateusz Tomczyk Kalina Andrysiak Monika Biniecka Anja Geisler Henry Fechner Michał Zembala Jacek Stępniewski Józef Dulak Agnieszka Jaźwa-Kusior |
| author_facet | Izabela Kraszewska Mateusz Tomczyk Kalina Andrysiak Monika Biniecka Anja Geisler Henry Fechner Michał Zembala Jacek Stępniewski Józef Dulak Agnieszka Jaźwa-Kusior |
| author_sort | Izabela Kraszewska |
| collection | DOAJ |
| description | Systemically delivered adeno-associated viral vector serotype 9 (AAV9) effectively transduces murine heart, but provides transgene expression also in liver and skeletal muscles. Improvement of the selectivity of transgene expression can be achieved through incorporation of target sites (TSs) for miRNA-122 and miRNA-206 into the 3′ untranslated region (3′ UTR) of the expression cassette. Here, we aimed to generate such miRNA-122- and miRNA-206-regulated AAV9 vector for a therapeutic, heart-specific overexpression of heme oxygenase-1 (HO-1). We successfully validated the vector functionality in murine cell lines corresponding to tissues targeted by AAV9. Next, we evaluated biodistribution of transgene expression following systemic vector delivery to HO-1-deficient mice of mixed C57BL/6J × FVB genetic background. Although AAV genomes were present in the hearts of these animals, HO-1 protein expression was either absent or significantly impaired. We found that miRNA-122, earlier described as liver specific, was present also in the hearts of C57BL/6J × FVB mice. Various levels of miRNA-122 expression were observed in the hearts of other mouse strains, in heart tissues of patients with cardiomyopathy, and in human induced pluripotent stem cell-derived cardiomyocytes in which we also confirmed such posttranscriptional regulation of transgene expression. Our data clearly indicate that therapeutic utilization of miRNA-based regulation strategy needs to consider inter-individual variability. |
| first_indexed | 2024-12-11T00:50:42Z |
| format | Article |
| id | doaj.art-e31943af4dc24b31b50d9af45a05ad0e |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-11T00:50:42Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-e31943af4dc24b31b50d9af45a05ad0e2022-12-22T01:26:39ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-06-011711901201Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV VectorsIzabela Kraszewska0Mateusz Tomczyk1Kalina Andrysiak2Monika Biniecka3Anja Geisler4Henry Fechner5Michał Zembala6Jacek Stępniewski7Józef Dulak8Agnieszka Jaźwa-Kusior9Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, PolandDepartment of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, PolandDepartment of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, PolandKardio-Med Silesia, 41-800 Zabrze, PolandDepartment of Applied Biochemistry, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, GermanyDepartment of Applied Biochemistry, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, GermanyDepartment of Cardiac Surgery, Heart and Lung Transplantation and Mechanical Circulatory Support, Silesian Center for Heart Diseases, 41-800 Zabrze, PolandDepartment of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, PolandDepartment of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, Poland; Kardio-Med Silesia, 41-800 Zabrze, PolandDepartment of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, Poland; Corresponding author: Agnieszka Jaźwa-Kusior, PhD, Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa Str. 7, 30-387 Kraków, Poland.Systemically delivered adeno-associated viral vector serotype 9 (AAV9) effectively transduces murine heart, but provides transgene expression also in liver and skeletal muscles. Improvement of the selectivity of transgene expression can be achieved through incorporation of target sites (TSs) for miRNA-122 and miRNA-206 into the 3′ untranslated region (3′ UTR) of the expression cassette. Here, we aimed to generate such miRNA-122- and miRNA-206-regulated AAV9 vector for a therapeutic, heart-specific overexpression of heme oxygenase-1 (HO-1). We successfully validated the vector functionality in murine cell lines corresponding to tissues targeted by AAV9. Next, we evaluated biodistribution of transgene expression following systemic vector delivery to HO-1-deficient mice of mixed C57BL/6J × FVB genetic background. Although AAV genomes were present in the hearts of these animals, HO-1 protein expression was either absent or significantly impaired. We found that miRNA-122, earlier described as liver specific, was present also in the hearts of C57BL/6J × FVB mice. Various levels of miRNA-122 expression were observed in the hearts of other mouse strains, in heart tissues of patients with cardiomyopathy, and in human induced pluripotent stem cell-derived cardiomyocytes in which we also confirmed such posttranscriptional regulation of transgene expression. Our data clearly indicate that therapeutic utilization of miRNA-based regulation strategy needs to consider inter-individual variability.http://www.sciencedirect.com/science/article/pii/S2329050120300930 |
| spellingShingle | Izabela Kraszewska Mateusz Tomczyk Kalina Andrysiak Monika Biniecka Anja Geisler Henry Fechner Michał Zembala Jacek Stępniewski Józef Dulak Agnieszka Jaźwa-Kusior Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors Molecular Therapy: Methods & Clinical Development |
| title | Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors |
| title_full | Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors |
| title_fullStr | Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors |
| title_full_unstemmed | Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors |
| title_short | Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors |
| title_sort | variability in cardiac mirna 122 level determines therapeutic potential of mirna regulated aav vectors |
| url | http://www.sciencedirect.com/science/article/pii/S2329050120300930 |
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