Abstract
Although human heme oxygenase-1 (hHO-1) could provide a useful approach for cellular protection in the ischemic heart, constitutive overexpression of hHO-1 may lead to unwanted side effects. To avoid this, we designed a hypoxia-regulated hHO-1 gene therapy system that can be switched on and off. This vigilant plasmid system is composed of myosin light chain-2v promoter and a gene switch that is based on an oxygen-dependent degradation domain from the hypoxia inducible factor-1-alpha. The vector can sense ischemia and switch on the hHO-1 gene system, specifically in the heart. In an in vivo experiment, the vigilant hHO-1 plasmid or saline was injected intramyocardially into myocardial infarction mice or sham operation mice. After gene transfer, expression of hHO-1 was only detected in the ischemic heart treated with vigilant hHO-1 plasmids. Masson trichrome staining showed significantly fewer fibrotic areas in vigilant hHO-1 plasmids-treated mice compared with saline control (43.0%+/-4.8% versus 62.5%+/-3.3%, P<0.01). The reduction of interstitial fibrosis is accompanied by an increase in myocardial hHO-1 expression in peri-infarct border areas, concomitant with higher Bcl-2 levels and lower Bax, Bak, and caspase 3 levels in the ischemic myocardium compared with saline control. By use of a cardiac catheter, heart from vigilant hHO-1 plasmids-treated mice showed improved recovery of contractile and diastolic performance after myocardial infarction compared with saline control. This study documents the beneficial regulation and therapeutic potential of vigilant plasmid-mediated hHO-1 gene transfer. This novel gene transfer strategy can provide cardiac-specific protection from future repeated bouts of ischemic injury.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Apoptosis / genetics
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Binding Sites
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Biosensing Techniques
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Caspase 3
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Caspases / biosynthesis
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Caspases / genetics
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Cell Hypoxia* / genetics
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DNA-Binding Proteins
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Endomyocardial Fibrosis / etiology
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Endomyocardial Fibrosis / prevention & control*
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Gene Expression Regulation / genetics*
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Genes, Synthetic*
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Genetic Therapy*
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Genetic Vectors / genetics
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Heme Oxygenase (Decyclizing) / biosynthesis
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Heme Oxygenase (Decyclizing) / genetics*
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Heme Oxygenase (Decyclizing) / physiology
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Heme Oxygenase-1
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit
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Membrane Proteins / biosynthesis
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Membrane Proteins / genetics
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Myocardial Infarction / complications
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Myocardial Infarction / enzymology
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Myocardial Infarction / therapy*
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Myosin Light Chains / genetics
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Plasmids / genetics
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Protein Structure, Tertiary
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Proto-Oncogene Proteins / biosynthesis
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins c-bcl-2 / biosynthesis
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Regulatory Sequences, Nucleic Acid / genetics*
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Saccharomyces cerevisiae Proteins / genetics
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TATA Box
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Transcription Factors / chemistry
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Transcription Factors / genetics
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Transcriptional Activation
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bcl-2 Homologous Antagonist-Killer Protein
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bcl-2-Associated X Protein
Substances
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BAK1 protein, human
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BAX protein, human
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DNA-Binding Proteins
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GAL4 protein, S cerevisiae
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HIF1A protein, human
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Hypoxia-Inducible Factor 1, alpha Subunit
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Membrane Proteins
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Myosin Light Chains
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Proto-Oncogene Proteins
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Proto-Oncogene Proteins c-bcl-2
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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bcl-2 Homologous Antagonist-Killer Protein
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bcl-2-Associated X Protein
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HMOX1 protein, human
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Heme Oxygenase (Decyclizing)
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Heme Oxygenase-1
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CASP3 protein, human
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Caspase 3
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Caspases