Noninvasive radiological imaging of pulmonary gene transfer and expression using the human sodium iodide symporter
Noninvasive radiological imaging of pulmonary gene transfer and expression using the human sodium iodide symporter
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DOI：
10.1007/s00259-004-1715-6
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发表时间：
2005-04-01
影响因子：
9.1
通讯作者：
Domann, FE
中科院分区：
文献类型：
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作者：
Niu, G;Krager, KJ;Domann, FE
目的：在本研究中，我们研究了人类碘化钠同向转运体（hNIS）作为报告基因的应用，以无创方式实时成像体内基因转移和肺组织中的表达。方法。将表达人NIS的腺病毒（Ad-hNIS）或空腺病毒（Ad-Bgl II）通过鼻孔注入Cotton大鼠的肺部。感染后3、10和17天后，用(TcO4-)-Tc-99m进行伽马相机闪烁扫描，观察基因转移的分布和持续时间。感染后20天，进行逆转录聚合酶链反应检测hNIS基因表达。使用双表达载体 Ad-hNIS-eGFP 通过荧光显微术检测感染肺组织中的转基因表达。使用I-124-作为示踪剂对肺部基因转移进行正电子发射断层扫描（PET）成像。最后，通过磷光成像评估 hNIS 向极化人气道上皮细胞层的转移。结果：感染 Ad-hNIS 的动物的肺部在闪烁扫描和 PET 扫描中清晰可见，而感染 Ad-Bgl II 的动物则检测不到。 Ad-hNIS 感染动物的肺部在 17 天时仍然可见，但在 20 天时不再可检测到。荧光显微镜显示，Ad-hNIS-eGFP感染的肺组织的GFP信号强度明显高于Ad-Bgl II感染的肺组织。结论：使用hNIS基因作为报告基因来监测肺部基因治疗过程中传递的基因表达的位置、幅度和时间是可行的。实时无创地通过断层扫描可视化基因表达的能力对人类基因治疗具有重大的转化意义。
Purpose: In this study we investigated the application of the human sodium iodide symporter (hNIS) as a reporter gene to noninvasively image in vivo gene transfer and expression in lung tissue in real time.Methods. Human NIS-expressing adenoviruses (Ad-hNIS) or empty adenoviruses (Ad-Bgl II) were instilled into the lungs of Cotton rats via the nostrils. After 3, 10, and 17 days post infection, gamma camera scintigraphy with (TcO4-)-Tc-99m was performed to observe the distribution and duration of gene transfer. At 20 days after infection, reverse transcription polymerase chain reaction was performed to detect hNIS gene expression. Dual expressing vector Ad-hNIS-eGFP was used to detect transgene expression by fluorescence photomicroscopy in infected lung tissue. Positron emission tomography ( PET) imaging of gene transfer to the lungs was performed using I-124- as tracer. Finally, hNIS transfer to a polarized human airway epithelial cell layer was evaluated by phosphorimaging.Results: Lungs in animals infected with Ad-hNIS were clearly visible on scintigraphy and PET scans, while those infected with Ad-Bgl II were undetectable. Lungs in Ad-hNIS infected animals could still be visualized at 17 days but were no longer detectable at 20 days. Fluorescence microscopy showed that lung tissue infected with Ad-hNIS-eGFP had significantly higher GFP signal intensity than that infected with Ad-Bgl II.Conclusion: It is feasible to use the hNIS gene as a reporter gene to monitor the location, magnitude, and timing of expression of genes delivered during pulmonary gene therapy. The ability to noninvasively visualize gene expression tomographically in real time has significant translational implications in human gene therapy.