GENIS:: Gene expression of sodium iodide symporter for noninvasive imaging of gene therapy vectors and quantification of gene expression in vivo
GENIS:: Gene expression of sodium iodide symporter for noninvasive imaging of gene therapy vectors and quantification of gene expression in vivo
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DOI：
10.1016/s1525-0016(03)00153-9
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发表时间：
2003-09-01
影响因子：
12.4
通讯作者：
Brown, SL
中科院分区：
文献类型：
--
作者：
Barton, KN;Tyson, D;Brown, SL
为了优化腺病毒介导的前列腺癌自杀基因治疗，我们开发了一种基于人钠碘同向转运体（hNIS）的方法，该方法允许对腺病毒载体进行非侵入性监测，并定量前列腺内的基因表达量和体积。构建了共表达治疗性酵母胞嘧啶脱氨酶（yCD）/突变型单纯疱疹病毒胸苷激酶（mutTK（SR 39））融合基因和hNIS基因的增殖型腺病毒（Ad 5-yCD/mutTK（SR 39）rep-hNIS）。将Ad 5-yCD/mutTK（SR 39）rep-hNIS和复制缺陷型hNIS腺病毒（rAd-CMV-FLhNIS）注射到狗前列腺的对侧叶中，并在施用（NaTcO 4）-Tc-99 m后使用γ照相机放射照相术监测活动物中的hNIS活性。尽管非常接近膀胱，但使用已安全给予人体的病毒剂量水平（10（10）至10（12）个病毒颗粒），在前列腺中很容易检测到（TcO 4-）-Tc-99 m摄取。由于其快速清除和较短的物理半衰期（6小时），有可能获得每日测量（TcO 4-）-Tc-99 m的体内摄取，允许动态监测报告基因在前列腺内的表达以及整个身体的生物分布。前列腺切片的高分辨率放射自显影结合基因表达的3D重建表明，基因表达的幅度和体积可以用亚毫米分辨率进行量化。在临床中实施GENIS（Na/I同向转运体基因表达）技术将有助于优化未来的人类基因治疗试验。
With the goal of optimizing adenovirus-mediated suicide gene therapy for prostate cancer, we have developed a method based on the human sodium iodide symporter (hNIS) that allows for noninvasive monitoring of adenoviral vectors and quantification of gene expression magnitude and volume within the prostate. A replication-competent adenovirus (Ad5-yCD/mutTK(SR39)rep-hNIS) coexpressing a therapeutic yeast cytosine deaminase (yCD)/mutant herpes simplex virus thymidine kinase (mutTK(SR39)) fusion gene and the hNIS gene was developed. Ad5-yCD/mutTK(SR39)rep-hNIS and a replication-defective hNIS adenovirus (rAd-CMV-FLhNIS) were injected into contralateral lobes of the dog prostate and hNIS activity was monitored in live animals following administration of (NaTcO4)-Tc-99m using gamma camera scintigraphy. Despite the close proximity of the urinary bladder, (TcO4-)-Tc-99m uptake was readily detected in the prostate using viral dose levels (10(10) to 10(12) viral particles) that have been safely administered to humans. Due to its rapid clearance and short physical half-life (6 h), it was possible to obtain daily measurements of (TcO4-)-Tc-99m uptake in vivo, allowing for dynamic monitoring of reporter gene expression within the prostate as well as biodistribution throughout the body. High-resolution autoradiography of prostate sections coupled with 3D reconstruction of gene expression demonstrated that the magnitude and volume of gene expression could be quantified with submillimeter resolution. Implementation of the GENIS (gene expression of Na/I symporter) technology in the clinic will facilitate optimization of future human gene therapy trials.