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 R.
AU - Leong-Poi, Howard
N1 - Funding Information:
Supported by an operating grant (FRN 62763) from the Canadian Institutes of Health Research, Ottawa, Ontario, Canada, and an equipment grant from the Canadian Foundation for Innovation, Ottawa, Ontario, Canada. Dr. Lindner is supported by grants (R01-HL-074443, R01-HL-078610, R01-DK-063508) from the National Institutes of Health, Bethesda, Maryland, and has served on the scientific advisory board for Visual Sonics. Dr. Leong-Poi is supported by a New Investigator Award from the Canadian Institutes of Health Research, Ottawa, Ontario, Canada, and an Early Researcher Award from the Ministry of Research and Innovation, Ontario, Canada.
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
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U2 - 10.1016/j.jacc.2009.07.023
DO - 10.1016/j.jacc.2009.07.023
M3 - Article
C2 - 19850216
AN - SCOPUS:70349986673
SN - 0735-1097
VL - 54
SP - 1735
EP - 1742
JO - Journal of the American College of Cardiology
JF - Journal of the American College of Cardiology
IS - 18
ER -