2003 Fiscal Year Final Research Report Summary
Ultrasound enhanced gene therapy: Development of ultrasonic methods of gene transfection for cardiovascular system.
| Project/Area Number |
14570709
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | National Cardiovascular Center Research Institute |
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Principal Investigator |
KOMAMURA Kazuo National Cardiovascular Center Research Institute, Department of Cardiovascular Dynamics, Head of Lab, 循環動態機能部, 室長 (90311448)
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| Co-Investigator(Kenkyū-buntansha) |
BEPPU Shintaro Osaka University School of Allied Health Sciences, Dept. of Functional Diagnostic Science, Professor, 医学部保健学科・医用物理学講座, 教授 (40113500)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Ultrasound / Gene transfection / Microbubble / Plasmid / Reporter gene / Protein |
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| Research Abstract |
Ultrasound-triggered microbubble destruction has been proposed as a means of targeting gene therapy to specific organs. Successful reporter gene expression using this approach has been reported. However, quantification of therapeutic gene products with this method has not been elucidated yet in cardiovascular cells or tissues. We examined the dose-response relations among the amount of gene, microbubble concentration and the duration of incubation using neonatal rat cardiomyocytes. We employed expression plasmid of Escheririchia Coli β-Galactosidase gene, expression plasmid of rat hepatocyte growth factor (HGF) gene containing CMV-β actin hybrid promoter, and cultured rat neonatal cardiomyocytes for in vitro experimental setups. We used continuous-wave Doppler ultrasound with frequency of 2.5MHz and maximal intensity of 0.5W/cm^2 because of clinical relevance.
1.Repetitive exposure to ultrasound-triggered galactose/palmitic acid microbubble destruction may yield efficient transfection of genes to cardiomyocytes.
2.Using catheter in the rat left ventricular cavity with precordial ultrasound irradiation, we examined the feasibility of luciferase gene transfection into the myocardium. Luciferase activity was detected only in the anterior myocardium, and the level of expression was similar to that in HUVEC cells for in vitro experiments for transfection, suggesting difficulty to transfect beyond the endothelial barrier.
3.We examined the feasibility of luciferase gene transfection into the myocardium with precordial ultrasound irradiation in the setting of intravenous administration of gene and microbubble. We detected no expression of luciferase in the myocardium, and some expression in the liver.
4.Taken together, we might as well (1)develop more specific gene targeting method for myocardial transfection with modifying enhancer/promoter ; and (2)improve microbubble shells with higher affinity for both plasmid-gene and myocardial cell membrane.
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[Journal Article] The role of a common TNNT2 polymorphism in cardiac hypertrophy.2004
Author(s)
Komamura, K., Iwai, N., Kokame, K., Yasumura, Y., Kim, J., Yamagishi, M., Morisaki, T., Kimura, A., Tomoike, H., Kitakaze, M., Miyatake, K.
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Journal Title
J Hum Genet. 49(3)
Pages: 129-133
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Treatment of dilated cardiomyopathy with electroporation of hepatocyte growth factor gene into skeletal muscle.2004
Author(s)
Komamura, K., Tatsumi, R., Miyazaki, J., Matsumoto, K., Yamato, E., Nakamura, T., Shimizu, Y., Nakatani, T., Kitamura, S., Tomoike, H., Kitakaze, M., Kangawa, K., Miyatake, K.
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Journal Title
Hypertension. 44(3)
Pages: 365-371
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Differential gene expression in the rat skeletal and heart muscle in glucocorticoid-induced myopathy: analysis by microarray.2003
Author(s)
Komamura, K., Shirotani-Ikejima, H., Tatsumi, R., Tsujita-Kuroda, Y., Kitakaze, M., Miyatake, K., Sunagawa, K., Miyata, T.
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Journal Title
Cardiovasc Drugs Ther. 17(4)
Pages: 303-310
Description
「研究成果報告書概要(欧文)」より
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