Siemens D Robert, Crist Scott, Austin J Christopher, Tartaglia James, Ratliff Timothy L
Department of Urology, Cancer Center and Prostate Cancer Research Group, University of Iowa, Iowa City, IA, USA.
J Urol. 2003 Sep;170(3):979-84. doi: 10.1097/01.ju.0000070925.10039.23.
Gene transfer efficiency and specific cell targeting of vectors is a major obstacle in preclinical studies of gene therapy for malignant disease. Previous attempts at gene transfer in bladder cancer models have resulted in variable urothelial and tumor transgene expression after intravesical administration of recombinant viral vectors. In the current study we compared the gene transfer efficiencies of different viral vectors.
We compared the gene transfer efficiencies of the viral vectors replication-deficient adenovirus, attenuated vaccinia virus (NYVAC) and canarypox virus (ALVAC) in vitro and in an orthotopic murine bladder cancer model. We used beta-galactosidase and firefly luciferase reporter gene expression to compare gene transfer efficiency.
Significantly higher transgene expression was observed in vitro when these cells were infected with NYVAC or ALVAC compared with adenovirus vectors. Similarly the efficiency of adenovirus vectors to transfer genetic material into bladder urothelium and orthotopic bladder tumors was inferior to that of ALVAC and NYVAC vectors, which interestingly appeared to have a predilection to infect the orthotopic tumor. Analysis of the expression of coxsackie-adenovirus receptor using reverse transcriptase-polymerase chain reaction revealed the bladder tumor cell lines were lacking this adenovirus receptor. While adenovirus transferred genes poorly to normal bladder, coxsackie-adenovirus receptor expression was high in bladder tissue.
The viral vectors examined in these experiments resulted in significantly different gene transfer in the orthotopic bladder cancer model, underscoring the importance of vector selection in gene therapy protocols.
在恶性疾病基因治疗的临床前研究中，载体的基因转移效率和特异性细胞靶向是一个主要障碍。以往在膀胱癌模型中进行基因转移的尝试，在膀胱内给予重组病毒载体后，导致尿路上皮和肿瘤转基因表达存在差异。在本研究中，我们比较了不同病毒载体的基因转移效率。
我们在体外和原位小鼠膀胱癌模型中比较了复制缺陷型腺病毒、减毒痘苗病毒（NYVAC）和金丝雀痘病毒（ALVAC）的基因转移效率。我们使用β-半乳糖苷酶和萤火虫荧光素酶报告基因表达来比较基因转移效率。
与腺病毒载体相比，当这些细胞用NYVAC或ALVAC感染时，在体外观察到显著更高的转基因表达。同样，腺病毒载体将遗传物质转移到膀胱尿路上皮和原位膀胱肿瘤中的效率低于ALVAC和NYVAC载体，有趣的是，后两者似乎更倾向于感染原位肿瘤。使用逆转录-聚合酶链反应分析柯萨奇腺病毒受体的表达，发现膀胱肿瘤细胞系缺乏这种腺病毒受体。虽然腺病毒向正常膀胱转移基因的能力较差，但膀胱组织中柯萨奇腺病毒受体的表达较高。
在这些实验中检测的病毒载体在原位膀胱癌模型中导致了显著不同的基因转移，强调了在基因治疗方案中载体选择的重要性。
