Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter
Background: Adenoviral gene transfer has been shown to be effective in cardiac myocytes in vitro and in vivo. A major limitation of myocardial gene therapy is the extracardiac transgene expression. Methods: To minimize extracardiac gene expression, we have constructed a tissue-specific promoter for...
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2004-04-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.1162/15353500200404103 |
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| author | Wolfgang Boecker Oliver Y. Bernecker Joseph C. Wu Xinsheng Zhu Tomohiro Sawa Luanda Grazette Anthony Rosenzweig Federica del Monte Ulrich Schmidt Roger J. Hajjar |
| author_facet | Wolfgang Boecker Oliver Y. Bernecker Joseph C. Wu Xinsheng Zhu Tomohiro Sawa Luanda Grazette Anthony Rosenzweig Federica del Monte Ulrich Schmidt Roger J. Hajjar |
| author_sort | Wolfgang Boecker |
| collection | DOAJ |
| description | Background: Adenoviral gene transfer has been shown to be effective in cardiac myocytes in vitro and in vivo. A major limitation of myocardial gene therapy is the extracardiac transgene expression. Methods: To minimize extracardiac gene expression, we have constructed a tissue-specific promoter for cardiac gene transfer, namely, the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene, which is known to be expressed in a tissue-specific manner in ventricular myocardium followed by a luciferase (luc) reporter gene (Ad.4 × MLC 250 .Luc). Rat cardiomyocytes, liver and kidney cells were infected with Ad.4 × MLC.Luc or control vectors. For in vivo testing, Ad.4 × MLC 250 .Luc was injected into the myocardium or in the liver of rats. Kinetics of promoter activity were monitored over 8 days using a cooled CCD camera. Results: In vitro: By infecting hepatic versus cardiomyocyte cells, we found that the promoter specificity ratio (luc activity in cardiomyocytes per liver cells) was 20.4 versus 0.9 (Ad.4 × MLC 250 .Luc vs. Ad.CMV). In vivo: Ad.4 × MLC 250 .Luc significantly reduced luc activity in liver (38.4-fold), lung (16.1-fold), and kidney (21.8-fold) versus Ad.CMV ( p = .01); whereas activity in the heart was only 3.8-fold decreased. The gene expression rate of cardiomyocytes versus hepatocytes was 7:1 (Ad.4 × MLC.Luc) versus 1:1.4 (Ad.CMV.Luc). Discussion: This new vector may be useful to validate therapeutic approaches in animal disease models and offers the perspective for selective expression of therapeutic genes in the diseased heart. |
| format | Article |
| id | doaj-art-8bb6a274c6324e4e9b8bd5f3f56ae6c5 |
| institution | Kabale University |
| issn | 1536-0121 |
| language | English |
| publishDate | 2004-04-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-8bb6a274c6324e4e9b8bd5f3f56ae6c52025-01-03T01:20:02ZengSAGE PublishingMolecular Imaging1536-01212004-04-01310.1162/1535350020040410310.1162_15353500200404103Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain PromoterWolfgang Boecker0Oliver Y. Bernecker1Joseph C. Wu2Xinsheng Zhu3Tomohiro Sawa4Luanda Grazette5Anthony Rosenzweig6Federica del Monte7Ulrich Schmidt8Roger J. Hajjar9Klinikum GrosshadernMassachusetts General HospitalUCLA School of MedicineHarvard Medical SchoolTeikyo University School of MedicineMassachusetts General HospitalMassachusetts General HospitalMassachusetts General HospitalHarvard Medical SchoolHarvard Medical SchoolBackground: Adenoviral gene transfer has been shown to be effective in cardiac myocytes in vitro and in vivo. A major limitation of myocardial gene therapy is the extracardiac transgene expression. Methods: To minimize extracardiac gene expression, we have constructed a tissue-specific promoter for cardiac gene transfer, namely, the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene, which is known to be expressed in a tissue-specific manner in ventricular myocardium followed by a luciferase (luc) reporter gene (Ad.4 × MLC 250 .Luc). Rat cardiomyocytes, liver and kidney cells were infected with Ad.4 × MLC.Luc or control vectors. For in vivo testing, Ad.4 × MLC 250 .Luc was injected into the myocardium or in the liver of rats. Kinetics of promoter activity were monitored over 8 days using a cooled CCD camera. Results: In vitro: By infecting hepatic versus cardiomyocyte cells, we found that the promoter specificity ratio (luc activity in cardiomyocytes per liver cells) was 20.4 versus 0.9 (Ad.4 × MLC 250 .Luc vs. Ad.CMV). In vivo: Ad.4 × MLC 250 .Luc significantly reduced luc activity in liver (38.4-fold), lung (16.1-fold), and kidney (21.8-fold) versus Ad.CMV ( p = .01); whereas activity in the heart was only 3.8-fold decreased. The gene expression rate of cardiomyocytes versus hepatocytes was 7:1 (Ad.4 × MLC.Luc) versus 1:1.4 (Ad.CMV.Luc). Discussion: This new vector may be useful to validate therapeutic approaches in animal disease models and offers the perspective for selective expression of therapeutic genes in the diseased heart.https://doi.org/10.1162/15353500200404103 |
| spellingShingle | Wolfgang Boecker Oliver Y. Bernecker Joseph C. Wu Xinsheng Zhu Tomohiro Sawa Luanda Grazette Anthony Rosenzweig Federica del Monte Ulrich Schmidt Roger J. Hajjar Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter Molecular Imaging |
| title | Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter |
| title_full | Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter |
| title_fullStr | Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter |
| title_full_unstemmed | Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter |
| title_short | Cardiac-Specific Gene Expression Facilitated by an Enhanced Myosin Light Chain Promoter |
| title_sort | cardiac specific gene expression facilitated by an enhanced myosin light chain promoter |
| url | https://doi.org/10.1162/15353500200404103 |
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