Positron emission tomography imaging of cardiac reporter gene expression in living rats
Positron emission tomography imaging of cardiac reporter gene expression in living rats
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DOI：
10.1161/01.cir.0000023620.59633.53
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发表时间：
2002-07-09
期刊：
影响因子：
37.8
通讯作者：
Gambhir, SS
中科院分区：
文献类型：
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作者：
Wu, JC;Inubushi, M;Gambhir, SS
背景-成像报告基因的表达对于基因治疗的非侵入性监测是有用的。方法与结果：10只大鼠心肌内注射携带巨细胞病毒启动子驱动的单纯疱疹病毒I型突变胸苷激酶(Ad-CMV-HSV1-sr39tk)的腺病毒1×10(9)pfu作为PET报告基因。对照组(n=4)注射携带巨细胞病毒启动子驱动的萤火虫荧光素酶的腺病毒(Ad-CMV-Fluc)1×10(9)pfu。在第2~4天，尾静脉注射氮-13氨([N-13]-NH3)作为心肌灌注示踪剂，然后用9-(4-[F-18]-氟-3-羟甲基丁基)鸟嘌呤([F-18]-FHBG)检测HSV1-sr39tk的表达。成像后，取出心脏进行体外[F-18]伽马计数和胸腺嘧啶核苷酶测定。结果显示，在所有的microPET图像上，[N-13]-NH3在心肌中分布均匀。与对照组相比，注射Ad-CMV-HSV1-sr39tk的大鼠在前外侧壁有显著的[F-18]-FHBG摄取。伽马计数显示，注射Ad-CMV-HSV1-sr39tk大鼠的放射性增加了20.0+/-4.4倍，而酶试验显示胸苷激酶活性增加了22.1+/-6.1倍(P
Background-Imaging reporter gene expression is useful for noninvasive monitoring of gene therapy. In this study, we imaged cardiac reporter gene expression in living rats using micro positron emission tomography (microPET).Methods and Results-Rats (n=10) underwent intramyocardial injection with 1x10(9) pfu of adenovirus carrying cytomegalovirus promoter-driving herpes simplex virus type I mutant thymidine kinase (Ad-CMV-HSV1-sr39tk) as PET reporter gene. Control rats (n=4) received 1x10(9) pfu of adenovirus carrying cytomegalovirus promoter-driving, firefly luciferase (Ad-CMV-Fluc). On days 2 to 4, microPET images were obtained after a tail vein injection of nitrogen-13 ammonia ([N-13]-NH3) as myocardial perfusion tracer, followed by 9-(4-[F-18]-fluoro-3 hydroxymethylbutyl) guanine ([F-18]-FHBG) to assess HSV1-sr39tk expression. After imaging, hearts were removed for ex vivo [F-18] gamma counting and thymidine kinase enzyme assay. Results show homogenous myocardial distribution of [N-13]-NH3 on all microPET images. Rats injected with Ad-CMV-HSV1-sr39tk have significant [F-18]-FHBG uptake in the anterolateral wall compared with background signal in controls. Gamma counting shows 20.0 +/- 4.4-fold increase of radioactivity, whereas enzyme assay shows 22.1 +/- 6.1-fold increase of thymidine kinase activity in Ad-CMV-HSV1-sr39tk injected rats (P