Li Hongwei, Li Jin Zhong, Helm Gregory A, Pan Dongfeng
Department of Radiology, University of Virginia, Charlottesville, VA 22908, USA.
Biotechnol Appl Biochem. 2007 Apr;46(Pt 4):179-84. doi: 10.1042/BA20060081.
Monitoring the expression of therapeutic genes in targeted tissues in disease models is important to assess the effectiveness and safety of systems of gene therapy delivery. In the present study, we employed a CCD (charge-coupled-device) imaging system to monitor how a prostate-specific adenovirus vector (AdPSA-Luc) mediated the long-term, sustained expression of firefly luciferase (Luc) in living human prostate cancer mouse models. The in vivo bioluminescence imaging revealed significantly high levels of luciferase expression up to 1 month, not only in prostate tumours, but also in lungs after intratumoural injection. Systemic tail vein injection of AdPSA-Luc revealed significant luciferase expression in lungs of both human prostate cancer mouse models and naïve mice, but significantly higher in the former, while the control virus, AdCMV-Luc, containing CMV (cytomegalovirus) promoter and luciferase gene, just restricted expression in the livers. Our findings demonstrate the ability of the cooled CCD camera to sensitively and non-invasively track the location, magnitude and persistence of luciferase gene expression in human prostate cancer mouse models. Monitoring of gene therapy studies in small animals may be aided considerably with further extensions of this technique.
在疾病模型中监测靶向组织中治疗性基因的表达对于评估基因治疗递送系统的有效性和安全性至关重要。在本研究中,我们采用电荷耦合器件(CCD)成像系统来监测前列腺特异性腺病毒载体(AdPSA-Luc)如何在活体人前列腺癌小鼠模型中介导萤火虫荧光素酶(Luc)的长期、持续表达。体内生物发光成像显示,瘤内注射后长达1个月,不仅在前列腺肿瘤中,而且在肺部,荧光素酶表达水平都显著升高。对人前列腺癌小鼠模型和未处理小鼠进行AdPSA-Luc的全身尾静脉注射后,肺部均出现显著的荧光素酶表达,但在前者中表达水平显著更高,而含有巨细胞病毒(CMV)启动子和荧光素酶基因的对照病毒AdCMV-Luc仅在肝脏中表达。我们的研究结果表明,冷却的CCD相机能够灵敏且无创地追踪人前列腺癌小鼠模型中荧光素酶基因表达的位置、强度和持续时间。随着该技术的进一步拓展,有望极大地助力小动物基因治疗研究的监测工作。