Comparison of Gene Delivery Techniques for Therapeutic Angiogenesis. Ultrasound-Mediated Destruction of Carrier Microbubbles Versus Direct Intramuscular Injection

Jeremy Kobulnik, Michael A. Kuliszewski, Duncan J. Stewart, Jonathan Lindner, Howard Leong-Poi

    Research output: Contribution to journalArticle

    49 Citations (Scopus)

    Abstract

    Objectives: This study was designed to compare the efficacy of angiogenic gene delivery by ultrasound-mediated (UM) destruction of intravenous carrier microbubbles to direct intramuscular (IM) injections. Background: Current trials of gene therapy for angiogenesis remain limited by suboptimal, invasive delivery techniques. Methods: Hind-limb ischemia was produced by iliac artery ligation in 99 rats. In 32 rats, UM delivery of green fluorescent protein (GFP)/vascular endothelial growth factor-165 (VEGF165) plasmid deoxyribonucleic acid was performed. Thirty-five animals received IM injections of VEGF165/GFP plasmid. Remaining rats received no treatment. Before delivery (day 14 after ligation) and at days 17, 21, and 28 and week 8 after ligation, microvascular blood volume and microvascular blood flow to the proximal hind limbs were assessed by contrast-enhanced ultrasound (n = 8 per group). Total transfection was assessed by reverse transcriptase-polymerase chain reaction, and localization of transfection was determined by immunohistochemistry. Results: By day 28, both IM and UM delivery of VEGF165 produced significant increases in microvascular blood volume and microvascular blood flow. Whereas increases in microvascular blood volume were similar between treatment groups, microvascular blood flow was greater (p <0.005) in UM-treated animals as compared with IM-treated animals, persisting to week 8. The VEGF165/GFP messenger ribonucleic acid expression was greater (p <0.05) for IM-treated animals. A strong GFP signal was detected for both groups and was localized to focal perivascular regions and myocytes around injection sites for IM and to the vascular endothelium of arterioles/capillaries in a wider distribution for UM delivery. Conclusions: Despite lower transfection levels, UM delivery of VEGF165 is as effective as IM injections. The UM delivery results in directed vascular transfection over a wider distribution, which may account for the more efficient angiogenesis.

    Original languageEnglish (US)
    Pages (from-to)1735-1742
    Number of pages8
    JournalJournal of the American College of Cardiology
    Volume54
    Issue number18
    DOIs
    StatePublished - Oct 27 2009

    Fingerprint

    Microbubbles
    Intramuscular Injections
    Green Fluorescent Proteins
    Transfection
    Blood Volume
    Ligation
    Genes
    Plasmids
    Extremities
    Therapeutics
    Iliac Artery
    Vascular Endothelium
    Arterioles
    Blood Group Antigens
    Reverse Transcriptase Polymerase Chain Reaction
    Genetic Therapy
    Muscle Cells
    Blood Vessels
    Ischemia
    Immunohistochemistry

    Keywords

    • angiogenesis
    • chronic ischemia
    • gene therapy

    ASJC Scopus subject areas

    • Cardiology and Cardiovascular Medicine

    Cite this

    Comparison of Gene Delivery Techniques for Therapeutic Angiogenesis. Ultrasound-Mediated Destruction of Carrier Microbubbles Versus Direct Intramuscular Injection. / Kobulnik, Jeremy; Kuliszewski, Michael A.; Stewart, Duncan J.; Lindner, Jonathan; Leong-Poi, Howard.

    In: Journal of the American College of Cardiology, Vol. 54, No. 18, 27.10.2009, p. 1735-1742.

    Research output: Contribution to journalArticle

    @article{78535c510e5448a18e7628c8c38d2d1a,
    title = "Comparison of Gene Delivery Techniques for Therapeutic Angiogenesis. Ultrasound-Mediated Destruction of Carrier Microbubbles Versus Direct Intramuscular Injection",
    abstract = "Objectives: This study was designed to compare the efficacy of angiogenic gene delivery by ultrasound-mediated (UM) destruction of intravenous carrier microbubbles to direct intramuscular (IM) injections. Background: Current trials of gene therapy for angiogenesis remain limited by suboptimal, invasive delivery techniques. Methods: Hind-limb ischemia was produced by iliac artery ligation in 99 rats. In 32 rats, UM delivery of green fluorescent protein (GFP)/vascular endothelial growth factor-165 (VEGF165) plasmid deoxyribonucleic acid was performed. Thirty-five animals received IM injections of VEGF165/GFP plasmid. Remaining rats received no treatment. Before delivery (day 14 after ligation) and at days 17, 21, and 28 and week 8 after ligation, microvascular blood volume and microvascular blood flow to the proximal hind limbs were assessed by contrast-enhanced ultrasound (n = 8 per group). Total transfection was assessed by reverse transcriptase-polymerase chain reaction, and localization of transfection was determined by immunohistochemistry. Results: By day 28, both IM and UM delivery of VEGF165 produced significant increases in microvascular blood volume and microvascular blood flow. Whereas increases in microvascular blood volume were similar between treatment groups, microvascular blood flow was greater (p <0.005) in UM-treated animals as compared with IM-treated animals, persisting to week 8. The VEGF165/GFP messenger ribonucleic acid expression was greater (p <0.05) for IM-treated animals. A strong GFP signal was detected for both groups and was localized to focal perivascular regions and myocytes around injection sites for IM and to the vascular endothelium of arterioles/capillaries in a wider distribution for UM delivery. Conclusions: Despite lower transfection levels, UM delivery of VEGF165 is as effective as IM injections. The UM delivery results in directed vascular transfection over a wider distribution, which may account for the more efficient angiogenesis.",
    keywords = "angiogenesis, chronic ischemia, gene therapy",
    author = "Jeremy Kobulnik and Kuliszewski, {Michael A.} and Stewart, {Duncan J.} and Jonathan Lindner and Howard Leong-Poi",
    year = "2009",
    month = "10",
    day = "27",
    doi = "10.1016/j.jacc.2009.07.023",
    language = "English (US)",
    volume = "54",
    pages = "1735--1742",
    journal = "Journal of the American College of Cardiology",
    issn = "0735-1097",
    publisher = "Elsevier USA",
    number = "18",

    }

    TY - JOUR

    T1 - Comparison of Gene Delivery Techniques for Therapeutic Angiogenesis. Ultrasound-Mediated Destruction of Carrier Microbubbles Versus Direct Intramuscular Injection

    AU - Kobulnik, Jeremy

    AU - Kuliszewski, Michael A.

    AU - Stewart, Duncan J.

    AU - Lindner, Jonathan

    AU - Leong-Poi, Howard

    PY - 2009/10/27

    Y1 - 2009/10/27

    N2 - Objectives: This study was designed to compare the efficacy of angiogenic gene delivery by ultrasound-mediated (UM) destruction of intravenous carrier microbubbles to direct intramuscular (IM) injections. Background: Current trials of gene therapy for angiogenesis remain limited by suboptimal, invasive delivery techniques. Methods: Hind-limb ischemia was produced by iliac artery ligation in 99 rats. In 32 rats, UM delivery of green fluorescent protein (GFP)/vascular endothelial growth factor-165 (VEGF165) plasmid deoxyribonucleic acid was performed. Thirty-five animals received IM injections of VEGF165/GFP plasmid. Remaining rats received no treatment. Before delivery (day 14 after ligation) and at days 17, 21, and 28 and week 8 after ligation, microvascular blood volume and microvascular blood flow to the proximal hind limbs were assessed by contrast-enhanced ultrasound (n = 8 per group). Total transfection was assessed by reverse transcriptase-polymerase chain reaction, and localization of transfection was determined by immunohistochemistry. Results: By day 28, both IM and UM delivery of VEGF165 produced significant increases in microvascular blood volume and microvascular blood flow. Whereas increases in microvascular blood volume were similar between treatment groups, microvascular blood flow was greater (p <0.005) in UM-treated animals as compared with IM-treated animals, persisting to week 8. The VEGF165/GFP messenger ribonucleic acid expression was greater (p <0.05) for IM-treated animals. A strong GFP signal was detected for both groups and was localized to focal perivascular regions and myocytes around injection sites for IM and to the vascular endothelium of arterioles/capillaries in a wider distribution for UM delivery. Conclusions: Despite lower transfection levels, UM delivery of VEGF165 is as effective as IM injections. The UM delivery results in directed vascular transfection over a wider distribution, which may account for the more efficient angiogenesis.

    AB - Objectives: This study was designed to compare the efficacy of angiogenic gene delivery by ultrasound-mediated (UM) destruction of intravenous carrier microbubbles to direct intramuscular (IM) injections. Background: Current trials of gene therapy for angiogenesis remain limited by suboptimal, invasive delivery techniques. Methods: Hind-limb ischemia was produced by iliac artery ligation in 99 rats. In 32 rats, UM delivery of green fluorescent protein (GFP)/vascular endothelial growth factor-165 (VEGF165) plasmid deoxyribonucleic acid was performed. Thirty-five animals received IM injections of VEGF165/GFP plasmid. Remaining rats received no treatment. Before delivery (day 14 after ligation) and at days 17, 21, and 28 and week 8 after ligation, microvascular blood volume and microvascular blood flow to the proximal hind limbs were assessed by contrast-enhanced ultrasound (n = 8 per group). Total transfection was assessed by reverse transcriptase-polymerase chain reaction, and localization of transfection was determined by immunohistochemistry. Results: By day 28, both IM and UM delivery of VEGF165 produced significant increases in microvascular blood volume and microvascular blood flow. Whereas increases in microvascular blood volume were similar between treatment groups, microvascular blood flow was greater (p <0.005) in UM-treated animals as compared with IM-treated animals, persisting to week 8. The VEGF165/GFP messenger ribonucleic acid expression was greater (p <0.05) for IM-treated animals. A strong GFP signal was detected for both groups and was localized to focal perivascular regions and myocytes around injection sites for IM and to the vascular endothelium of arterioles/capillaries in a wider distribution for UM delivery. Conclusions: Despite lower transfection levels, UM delivery of VEGF165 is as effective as IM injections. The UM delivery results in directed vascular transfection over a wider distribution, which may account for the more efficient angiogenesis.

    KW - angiogenesis

    KW - chronic ischemia

    KW - gene therapy

    UR - http://www.scopus.com/inward/record.url?scp=70349986673&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=70349986673&partnerID=8YFLogxK

    U2 - 10.1016/j.jacc.2009.07.023

    DO - 10.1016/j.jacc.2009.07.023

    M3 - Article

    C2 - 19850216

    AN - SCOPUS:70349986673

    VL - 54

    SP - 1735

    EP - 1742

    JO - Journal of the American College of Cardiology

    JF - Journal of the American College of Cardiology

    SN - 0735-1097

    IS - 18

    ER -