Monitoring adenoviral DNA delivery, using a mutant herpes simplex virus type 1 thymidine kinase gene as a PET reporter gene
Monitoring adenoviral DNA delivery, using a mutant herpes simplex virus type 1 thymidine kinase gene as a PET reporter gene
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DOI：
10.1038/sj.gt.3301899
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发表时间：
2002-12-01
期刊：
影响因子：
5.1
通讯作者：
Herschman, HR
中科院分区：
文献类型：
--
作者：
Liang, Q;Nguyen, K;Herschman, HR
目前的基因治疗方案通常无法监测体细胞DNA递送后基因表达的位点、水平和持续性。单纯疱疹病毒1型胸苷激酶（HSV 1-tk）目前正在作为报告基因进行深入研究，用于报告基因表达的体内成像。通过全身注射正电子发射的、放射性核素标记的胸苷类似物或无环鸟苷HSV 1-TK底物，并随后通过正电子发射断层扫描检测捕获的磷酸化产物，重复和非侵入性地监测HSV 1-tk报告基因的存在。为了提高HSV 1-tk PET报告基因系统的功效，已经描述了HSV 1-tk基因中的替代底物和突变。我们使用复制缺陷型腺病毒将HSV 1-sr 39 tk突变酶和野生型HSV 1-tk酶递送至小鼠。使用9-[（4-[F-18]fluoro-3-hydroxymethylbutyl）guanine作为探针，在腺病毒介导的小鼠肝脏表达后，HSV 1-sr 39 TK在体内表现出比野生型HSV 1-TK酶更高的灵敏度。使用这种腺病毒递送系统，可以通过microPET非侵入性地、定量地和重复地监测HSV 1-sr 39 tk PET报告基因表达的位置、幅度和持续时间超过3个月。
Current gene therapy protocols often suffer from an inability to monitor the site, level and persistence of gene expression following somatic DNA delivery. Herpes simplex virus 1 thymidine kinase (HSV1-tk) is currently under intensive investigation as a reporter gene for in vivo imaging of reporter gene expression. The presence of the HSV1-tk reporter gene is repetitively and non-invasively monitored by systemic injection of positron-emitting, radionuclide-labeled thymidine analogues or acycloguanosine HSV1-TK substrates and subsequent detection, by positron emission tomography, of trapped, phosphorylated product. To improve the efficacy of the HSV1-tk PET reporter gene system, both alternative substrates and mutations in the HSV1-tk gene have been described. We used a replication defective adenovirus to deliver the HSV1-sr39tk mutant enzyme and the wild-type HSV1-tk enzyme to mice. HSV1-sr39TK demonstrates greater sensitivity than wild-type HSV1-TK enzyme in vivo, using 9-[(4-[F-18]fluoro-3-hydroxymethylbutyl)guanine as probe, following adenovirus-mediated hepatic expression in mice. Using this adenoviral delivery system, the location, magnitude and duration of HSV1-sr39tk PET reporter gene expression could be non-invasively, quantitatively and repetitively monitored for over 3 months by microPET.