Ultrasound-Targeted Gene Delivery Induces Angiogenesis After a Myocardial Infarction in Mice

Hiroko Fujii, Zhuo Sun, Shu Hong Li, Jun Wu, Shafie Fazel, Richard D. Weisel, Harry Rakowski, Jonathan Lindner, Ren Ke Li

    Research output: Contribution to journalArticle

    92 Citations (Scopus)

    Abstract

    Objectives: This study evaluated the capacity of ultrasound-targeted microbubble destruction (UTMD) to deliver angiogenic genes, improve perfusion, and recruit progenitor cells after a myocardial infarction (MI) in mice. Background: Angiogenic gene therapy after an MI may become a clinically relevant approach to improve the engraftment of implanted cells if targeted delivery can be accomplished noninvasively. The UTMD technique uses myocardial contrast echocardiography to target plasmid gene delivery to the myocardium and features low toxicity, limited immunogenicity, and the potential for repeated application. Methods: Empty plasmids (control group) or those containing genes for vascular endothelial growth factor (VEGF), stem cell factor (SCF), or green fluorescent protein (to visualize gene delivery) were incubated with perflutren lipid microbubbles. The microbubble-deoxyribonucleic acid mixture was injected intravenously into C57BL/6 mice at 7 days after coronary artery ligation (MI). The UTMD technique facilitated transgene release into the myocardium. Twenty-one days after MI, myocardial perfusion and function were assessed by contrast echocardiography. Protein expression was quantified by Western blot and enzyme-linked immunosorbent assay. Flow cytometry quantified progenitor cell recruitment to the heart. Blood vessel density was evaluated immunohistochemically. Results: Green fluorescent protein expression in the infarcted myocardium demonstrated gene delivery. Myocardial VEGF and SCF levels increased significantly in the respective groups (p <0.05). The physiologic impact of VEGF and SCF gene delivery was confirmed by increased myocardial recruitment of VEGF receptor 2- and SCF receptor (c-kit)-expressing cells, respectively (p <0.05). Consequently, capillary and arteriolar density (Factor VIII and alpha-smooth muscle actin staining), myocardial perfusion, and cardiac function were all enhanced (p <0.01 relative to control group) in recipients of VEGF or SCF. Conclusions: Noninvasive UTMD successfully delivered VEGF and SCF genes into the infarcted heart, increased vascular density, and improved myocardial perfusion and ventricular function. The UTMD technique may be an ideal method for noninvasive, repeated gene delivery after an MI.

    Original languageEnglish (US)
    Pages (from-to)869-879
    Number of pages11
    JournalJACC: Cardiovascular Imaging
    Volume2
    Issue number7
    DOIs
    StatePublished - Jul 2009

    Fingerprint

    Microbubbles
    Stem Cell Factor
    Myocardial Infarction
    Vascular Endothelial Growth Factor A
    Genes
    Perfusion
    Myocardium
    perflutren
    Green Fluorescent Proteins
    Blood Vessels
    Echocardiography
    Plasmids
    Stem Cells
    Proto-Oncogene Proteins c-kit
    Vascular Endothelial Growth Factor Receptor-2
    Control Groups
    Ventricular Function
    Factor VIII
    Transgenes
    Inbred C57BL Mouse

    Keywords

    • angiogenesis
    • echocardiography
    • gene therapy
    • microbubbles
    • myocardial infarction

    ASJC Scopus subject areas

    • Cardiology and Cardiovascular Medicine
    • Radiology Nuclear Medicine and imaging

    Cite this

    Ultrasound-Targeted Gene Delivery Induces Angiogenesis After a Myocardial Infarction in Mice. / Fujii, Hiroko; Sun, Zhuo; Li, Shu Hong; Wu, Jun; Fazel, Shafie; Weisel, Richard D.; Rakowski, Harry; Lindner, Jonathan; Li, Ren Ke.

    In: JACC: Cardiovascular Imaging, Vol. 2, No. 7, 07.2009, p. 869-879.

    Research output: Contribution to journalArticle

    Fujii, H, Sun, Z, Li, SH, Wu, J, Fazel, S, Weisel, RD, Rakowski, H, Lindner, J & Li, RK 2009, 'Ultrasound-Targeted Gene Delivery Induces Angiogenesis After a Myocardial Infarction in Mice', JACC: Cardiovascular Imaging, vol. 2, no. 7, pp. 869-879. https://doi.org/10.1016/j.jcmg.2009.04.008
    Fujii, Hiroko ; Sun, Zhuo ; Li, Shu Hong ; Wu, Jun ; Fazel, Shafie ; Weisel, Richard D. ; Rakowski, Harry ; Lindner, Jonathan ; Li, Ren Ke. / Ultrasound-Targeted Gene Delivery Induces Angiogenesis After a Myocardial Infarction in Mice. In: JACC: Cardiovascular Imaging. 2009 ; Vol. 2, No. 7. pp. 869-879.
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    abstract = "Objectives: This study evaluated the capacity of ultrasound-targeted microbubble destruction (UTMD) to deliver angiogenic genes, improve perfusion, and recruit progenitor cells after a myocardial infarction (MI) in mice. Background: Angiogenic gene therapy after an MI may become a clinically relevant approach to improve the engraftment of implanted cells if targeted delivery can be accomplished noninvasively. The UTMD technique uses myocardial contrast echocardiography to target plasmid gene delivery to the myocardium and features low toxicity, limited immunogenicity, and the potential for repeated application. Methods: Empty plasmids (control group) or those containing genes for vascular endothelial growth factor (VEGF), stem cell factor (SCF), or green fluorescent protein (to visualize gene delivery) were incubated with perflutren lipid microbubbles. The microbubble-deoxyribonucleic acid mixture was injected intravenously into C57BL/6 mice at 7 days after coronary artery ligation (MI). The UTMD technique facilitated transgene release into the myocardium. Twenty-one days after MI, myocardial perfusion and function were assessed by contrast echocardiography. Protein expression was quantified by Western blot and enzyme-linked immunosorbent assay. Flow cytometry quantified progenitor cell recruitment to the heart. Blood vessel density was evaluated immunohistochemically. Results: Green fluorescent protein expression in the infarcted myocardium demonstrated gene delivery. Myocardial VEGF and SCF levels increased significantly in the respective groups (p <0.05). The physiologic impact of VEGF and SCF gene delivery was confirmed by increased myocardial recruitment of VEGF receptor 2- and SCF receptor (c-kit)-expressing cells, respectively (p <0.05). Consequently, capillary and arteriolar density (Factor VIII and alpha-smooth muscle actin staining), myocardial perfusion, and cardiac function were all enhanced (p <0.01 relative to control group) in recipients of VEGF or SCF. Conclusions: Noninvasive UTMD successfully delivered VEGF and SCF genes into the infarcted heart, increased vascular density, and improved myocardial perfusion and ventricular function. The UTMD technique may be an ideal method for noninvasive, repeated gene delivery after an MI.",
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    author = "Hiroko Fujii and Zhuo Sun and Li, {Shu Hong} and Jun Wu and Shafie Fazel and Weisel, {Richard D.} and Harry Rakowski and Jonathan Lindner and Li, {Ren Ke}",
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    AU - Fujii, Hiroko

    AU - Sun, Zhuo

    AU - Li, Shu Hong

    AU - Wu, Jun

    AU - Fazel, Shafie

    AU - Weisel, Richard D.

    AU - Rakowski, Harry

    AU - Lindner, Jonathan

    AU - Li, Ren Ke

    PY - 2009/7

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    N2 - Objectives: This study evaluated the capacity of ultrasound-targeted microbubble destruction (UTMD) to deliver angiogenic genes, improve perfusion, and recruit progenitor cells after a myocardial infarction (MI) in mice. Background: Angiogenic gene therapy after an MI may become a clinically relevant approach to improve the engraftment of implanted cells if targeted delivery can be accomplished noninvasively. The UTMD technique uses myocardial contrast echocardiography to target plasmid gene delivery to the myocardium and features low toxicity, limited immunogenicity, and the potential for repeated application. Methods: Empty plasmids (control group) or those containing genes for vascular endothelial growth factor (VEGF), stem cell factor (SCF), or green fluorescent protein (to visualize gene delivery) were incubated with perflutren lipid microbubbles. The microbubble-deoxyribonucleic acid mixture was injected intravenously into C57BL/6 mice at 7 days after coronary artery ligation (MI). The UTMD technique facilitated transgene release into the myocardium. Twenty-one days after MI, myocardial perfusion and function were assessed by contrast echocardiography. Protein expression was quantified by Western blot and enzyme-linked immunosorbent assay. Flow cytometry quantified progenitor cell recruitment to the heart. Blood vessel density was evaluated immunohistochemically. Results: Green fluorescent protein expression in the infarcted myocardium demonstrated gene delivery. Myocardial VEGF and SCF levels increased significantly in the respective groups (p <0.05). The physiologic impact of VEGF and SCF gene delivery was confirmed by increased myocardial recruitment of VEGF receptor 2- and SCF receptor (c-kit)-expressing cells, respectively (p <0.05). Consequently, capillary and arteriolar density (Factor VIII and alpha-smooth muscle actin staining), myocardial perfusion, and cardiac function were all enhanced (p <0.01 relative to control group) in recipients of VEGF or SCF. Conclusions: Noninvasive UTMD successfully delivered VEGF and SCF genes into the infarcted heart, increased vascular density, and improved myocardial perfusion and ventricular function. The UTMD technique may be an ideal method for noninvasive, repeated gene delivery after an MI.

    AB - Objectives: This study evaluated the capacity of ultrasound-targeted microbubble destruction (UTMD) to deliver angiogenic genes, improve perfusion, and recruit progenitor cells after a myocardial infarction (MI) in mice. Background: Angiogenic gene therapy after an MI may become a clinically relevant approach to improve the engraftment of implanted cells if targeted delivery can be accomplished noninvasively. The UTMD technique uses myocardial contrast echocardiography to target plasmid gene delivery to the myocardium and features low toxicity, limited immunogenicity, and the potential for repeated application. Methods: Empty plasmids (control group) or those containing genes for vascular endothelial growth factor (VEGF), stem cell factor (SCF), or green fluorescent protein (to visualize gene delivery) were incubated with perflutren lipid microbubbles. The microbubble-deoxyribonucleic acid mixture was injected intravenously into C57BL/6 mice at 7 days after coronary artery ligation (MI). The UTMD technique facilitated transgene release into the myocardium. Twenty-one days after MI, myocardial perfusion and function were assessed by contrast echocardiography. Protein expression was quantified by Western blot and enzyme-linked immunosorbent assay. Flow cytometry quantified progenitor cell recruitment to the heart. Blood vessel density was evaluated immunohistochemically. Results: Green fluorescent protein expression in the infarcted myocardium demonstrated gene delivery. Myocardial VEGF and SCF levels increased significantly in the respective groups (p <0.05). The physiologic impact of VEGF and SCF gene delivery was confirmed by increased myocardial recruitment of VEGF receptor 2- and SCF receptor (c-kit)-expressing cells, respectively (p <0.05). Consequently, capillary and arteriolar density (Factor VIII and alpha-smooth muscle actin staining), myocardial perfusion, and cardiac function were all enhanced (p <0.01 relative to control group) in recipients of VEGF or SCF. Conclusions: Noninvasive UTMD successfully delivered VEGF and SCF genes into the infarcted heart, increased vascular density, and improved myocardial perfusion and ventricular function. The UTMD technique may be an ideal method for noninvasive, repeated gene delivery after an MI.

    KW - angiogenesis

    KW - echocardiography

    KW - gene therapy

    KW - microbubbles

    KW - myocardial infarction

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