Cardiac-specific gene expression facilitated by an enhanced myosin light chain promoter

Wolfgang Boecker; Oliver Y. Bernecker; Joseph C. Wu; Xinsheng Zhu; Tomohiro Sawa; Luanda Grazette; Anthony Rosenzweig; Federica Del Monte; Ulrich Schmidt; Roger J. Hajjar

doi:10.1162/1535350041464847

Cardiac-specific gene expression facilitated by an enhanced myosin light chain promoter

Wolfgang Boecker, Oliver Y. Bernecker, Joseph C. Wu, Xinsheng Zhu, Tomohiro Sawa, Luanda Grazette, Anthony Rosenzweig, Federica Del Monte, Ulrich Schmidt, Roger J. Hajjar

Medical Information Systems Research Center

Research output: Contribution to journal › Article › peer-review

40 Scopus citations

Abstract

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 (lue 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 lue 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.

Original language	English
Pages (from-to)	69-75
Number of pages	7
Journal	Molecular Imaging
Volume	3
Issue number	2
DOIs	https://doi.org/10.1162/1535350041464847
State	Published - Apr 2004

Keywords

Adenovirus
Cardiac promoters
Gene therapy

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1162/1535350041464847

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title = "Cardiac-specific gene expression facilitated by an enhanced myosin light chain promoter",

abstract = "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 (lue 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 lue 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.",

keywords = "Adenovirus, Cardiac promoters, Gene therapy",

author = "Wolfgang Boecker and Bernecker, {Oliver Y.} and Wu, {Joseph C.} and Xinsheng Zhu and Tomohiro Sawa and Luanda Grazette and Anthony Rosenzweig and {Del Monte}, Federica and Ulrich Schmidt and Hajjar, {Roger J.}",

year = "2004",

month = apr,

doi = "10.1162/1535350041464847",

language = "英語",

volume = "3",

pages = "69--75",

journal = "Molecular Imaging",

issn = "1535-3508",

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TY - JOUR

T1 - Cardiac-specific gene expression facilitated by an enhanced myosin light chain promoter

AU - Boecker, Wolfgang

AU - Bernecker, Oliver Y.

AU - Wu, Joseph C.

AU - Zhu, Xinsheng

AU - Sawa, Tomohiro

AU - Grazette, Luanda

AU - Rosenzweig, Anthony

AU - Del Monte, Federica

AU - Schmidt, Ulrich

AU - Hajjar, Roger J.

PY - 2004/4

Y1 - 2004/4

N2 - 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 (lue 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 lue 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.

AB - 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 (lue 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 lue 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.

KW - Adenovirus

KW - Cardiac promoters

KW - Gene therapy

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DO - 10.1162/1535350041464847

M3 - 記事

C2 - 15296671

AN - SCOPUS:4243061120

SN - 1535-3508

VL - 3

SP - 69

EP - 75

JO - Molecular Imaging

JF - Molecular Imaging

IS - 2

ER -

Cardiac-specific gene expression facilitated by an enhanced myosin light chain promoter

Abstract

Keywords

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