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Ultrasound enhanced gene therapy: Development of ultrasonic methods of gene transfection for cardiovascular system.

超声增强基因治疗：开发心血管系统基因转染超声方法。

基本信息

批准号：
14570709
负责人：
KOMAMURA Kazuo
金额：
$ 2.11万
依托单位：
National Cardiovascular Center Research Institute
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570709/
关键词：
Ultrasound Gene transfection Microbubble Plasmid Reporter gene Protein 肝細胞増殖因子

项目摘要

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.

超声波触发的微泡破坏已被提议作为针对特定器官的基因治疗的一种手段。据报道，使用这种方法成功表达了报告基因。然而，用这种方法对心血管细胞或组织中的治疗基因产物进行定量尚未得到阐明。我们使用新生大鼠心肌细胞检查了基因量、微泡浓度和孵育时间之间的剂量反应关系。我们采用大肠杆菌β-半乳糖苷酶基因表达质粒、含有CMV-β肌动蛋白杂合启动子的大鼠肝细胞生长因子（HGF）基因表达质粒，并培养大鼠新生心肌细胞进行体外实验设置。由于临床相关性，我们使用频率为 2.5MHz、最大强度为 0.5W/cm^2 的连续波多普勒超声。1.重复暴露于超声触发的半乳糖/棕榈酸微泡破坏可能会产生将基因有效转染到心肌细胞的效果。2.在大鼠左心室腔中使用导管进行心前区超声照射，我们检查了荧光素酶基因转染至心肌的可行性。荧光素酶活性仅在前壁心肌中检测到，其表达水平与体外转染实验中HUVEC细胞中的表达水平相似，表明转染难以突破内皮屏障。 3.在静脉注射基因和微泡的情况下，我们考察了心前区超声照射下荧光素酶基因转染入心肌的可行性。我们在心肌中没有检测到荧光素酶的表达，在肝脏中有一些表达。4.综上所述，我们不妨（1）开发更特异的修饰增强子/启动子的心肌转染基因打靶方法； (2)改进微泡壳，使其对质粒基因和心肌细胞膜具有更高的亲和力。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Treatment of dilated cardiomyopathy with electration of hepatocyte growth factor gene into skeletal muscle

将肝细胞生长因子基因植入骨骼肌治疗扩张型心肌病

DOI：
发表时间：
2004
期刊：
Hypertension 44(3)
影响因子：
0
作者：
Komamura K
通讯作者：
Komamura K

超音波遺伝子治療

超声基因治疗

DOI：
发表时间：
2003
期刊：
超音波TECHNO 15(3)
影响因子：
0
作者：
駒村和雄
通讯作者：
駒村和雄

Differential gene expression in the rat skeletal and heart muscle in glucocorticoid-induced myopathy: Analysis by microarray

DOI：
10.1023/a:1027352703783
发表时间：
2003-07-01
期刊：
CARDIOVASCULAR DRUGS AND THERAPY
影响因子：
3.4
作者：
Komamura, K;Shirotani-Ikejima, H;Miyata, T
通讯作者：
Miyata, T

Improvement of cardiac hypertrophy and ventricular function in a man with Fabry disease by treatment with recombinant α-galactosidase A

通过重组 α-半乳糖苷酶 A 治疗可改善法布里病患者的心脏肥大和心室功能

DOI：
发表时间：
2004
期刊：
Heart 90(6)
影响因子：
0
作者：
Nishikimi T;Mori Y;Kobayashi N;Tadokoro K;Wang X;Akimoto K;Yoshihara F;Kangawa K;Matsuoka H;Komamura K
通讯作者：
Komamura K

Improvement of cardiac hypertrophy and ventricular function in a man with Fabry disease by treatment with recombinant alpha-galactosidase A.

通过重组 α-半乳糖苷酶 A 治疗可改善法布里病患者的心脏肥大和心室功能。