Biodistribution and safety assessment of bladder cancer specific recombinant oncolytic adenovirus in subcutaneous xenografts tumor model in nude mice.
Biodistribution and safety assessment of bladder cancer specific recombinant oncolytic adenovirus in subcutaneous xenografts tumor model in nude mice.
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DOI：
10.2174/156652312800099599
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发表时间：
2012-04-01
影响因子：
3.6
通讯作者：
Zhou L
中科院分区：
文献类型：
--
作者：
Wang F;Wang Z;Tian H;Qi M;Zhai Z;Li S;Li R;Zhang H;Wang W;Fu S;Lu J;Rodriguez R;Guo Y;Zhou L
以往关于构建的膀胱组织特异型腺病毒的安全性评价的工作文献很少。因此，我们研究了膀胱特异性溶瘤腺病毒Ad-PSCAE-UPII-E1A(APU-E1A)和Ad-PSCAE-UPII-E1A-AR(APU-E1A-AR)的生物分布和体内毒性，为开展人体临床试验提供了有意义的信息。用膀胱组织特异性Uroplakin II(UP II)启动子构建条件复制型重组腺病毒(CRADs)APU-E1A、APU-EIA-AR以诱导Ad5E1A基因和E1A-AR融合基因的表达，并在启动子上游插入PSCAE以增强启动子的功能。基于膀胱癌的细胞病变和抗肿瘤作用，将这些CRADs注射到裸鼠皮下移植瘤中。然后，我们通过一般健康和行为评估、肝脏和血液毒性评估、宏观和微观尸检分析来确定毒性。RT-PCR和Western印迹法检测转基因E1a腺病毒的扩散。用APU-Luc给药和荧光素酶试验检测病毒的复制和分布。对动物的总体评估和体重没有显示出一般行为的任何变化。注射5×108pfu或更高剂量(5×109pfu)的APU-E1A和APU-E1A-AR组的血液学改变与PBS组无明显差异，仅在5×109pfu的APU-E1A和APU-E1A-AR组与PBS组相比转氨酶略有升高。E1a转基因未扩散至移植瘤外的器官。荧光素酶分析显示，肿瘤外其他器官未检测到病毒复制。我们的研究表明，重组腺病毒APU-E1A-AR和APU-E1A在小鼠瘤内注射5×107pfu和5×108pfu是安全的，对一般健康和行为没有明显影响。
The previous works about safety evaluation for constructed bladder tissue specific adenovirus are poorly documented. Thus, we investigated the biodistribution and body toxicity of bladder specific oncolytic adenovirus Ad-PSCAE-UPII-E1A (APU-E1A) and Ad-PSCAE-UPII-E1A-AR (APU-E1A-AR), providing meaningful information prior to embarking on human clinical trials. Conditionally replicate recombinant adenovirus (CRADs) APU-E1A, APU-EIA-AR were constructed with bladder tissue specific Uroplakin II (UP II) promoter to induce the expression of Ad5E1A gene and E1A-AR fusing gene, and PSCAE was inserted at upstream of promoter to enhance the function of promoter. Based on the cytopathic and anti-tumor effect of bladder cancer, these CRADs were intratumorally injected into subcutaneous xenografts tumor in nude mice. We then determined the toxicity through general health and behavioral assessment, hepatic and hematological toxicity evaluation, macroscopic and microscopic postmortem analyses. The spread of the transgene E1A of adenovirus was detected with RT-PCR and Western blot. Virus replication and distribution were examined with APU-LUC administration and Luciferase Assay. General assessment and body weight of the animals did not reveal any alteration in general behavior. The hematological alterations of groups which were injected with 5×108 pfu or higher dose (5×109 pfu) of APU-E1A and APU-E1A-AR showed no difference in comparison with PBS group, and only slight increased transaminases in contrast to PBS group at 5×109 pfu of APU-E1A and APU-E1A-AR were observed. E1A transgene did not disseminate to organs outside of xenograft tumor. Virus replication was not detected in other organs beside tumor according to Luciferase Assay. Our study showed that recombinant adenovirus APU-E1A-AR and APU-E1A appear safe with 5×107 pfu and 5×108 pfu intratumorally injection in mice, without any discernable effects on general health and behavior.