Experimental Efficacy of Adenovirus-mediated Gene Therapy for Gallbladder Cancer

新的治疗方法，如基因治疗，是必要的晚期胆囊癌（GBC），但很少有研究。最近的研究已经引入了E1突变腺病毒（Ads），其显示肿瘤特异性复制和有希望的临床结果。为了提高这种方法的安全性，我们新构建了AxdAdB-3，这是一种具有突变E1 A和E1 B-55 kD缺失的双限制性Ad。我们研究了该Ad在体外和体内对GBC的作用以及与野生型Ad或E1 B-55 kD缺失的Ad AxE 1AdB相比其对正常人细胞的安全性。AxdAdB-3在几种GBC细胞系中复制并引起溶瘤，其体外效率与野生型Ad或AxE 1AdB一样。相比之下，AxdAdB-3在原代正常细胞（例如，上皮细胞、内皮细胞和肝细胞）中的细胞病变效应，并且与野生型Ad.此外，AxdAdB-3对正常细胞的细胞毒性比AxE 1AdB轻。AxdAdB-3显著（p<0.01）抑制GBC异种移植物的生长。AxdAdB-3还显示出对患有腹膜播散性GBC的小鼠的治疗功效，显示出肿瘤选择性复制和肿瘤溶解，导致显著（p<0.05）延长的存活。本研究表明，E1双限制性Ad在体外和体内有效地和选择性地复制并引起GBC的溶瘤，对正常细胞的负面影响减少，这表明这可能是一种有前途的GBC基因治疗工具。

New treatments, such as gene therapy, are necessary for advanced gallbladder cancer (GBC) but little has been studied. Recent studies have introduced E1 mutant adenoviruses (Ads) which show tumor-specific replication and promising clinical results. To enhance the safety of this approach, we newly constructed AxdAdB-3, a double-restricted Ad with a mutant E1A and E1B-55kD deletion. We studied the in vitro and in vivo effects of this Ad on GBC as well as its safety for normal human cells in comparison with wild-type Ad or an E1B-55kD deleted Ad, AxE1AdB. AxdAdB-3 replicated in and caused oncolysis of several GBC cell lines as efficiently as wild-type Ad or AxE1AdB in vitro. By contrast, AxdAdB-3, replicated much less effectively in primary normal cells (e.g., epithelial cells, endothelial cells and hepatocytes) than in GBC cells and had only a mild cytopathic effects, unlike wild-type Ad. Furthermore, cytotoxicity of AxdAdB-3 in normal cells was milder than AxE1AdB. AxdAdB-3 significantly (p<0.01) suppressed the growth of GBC xenografts. AxdAdB-3 also showed therapeutic efficacy for mice with peritoneally disseminated GBC, showing tumor-selective replication and oncolysis that resulted in significantly (p<0.05) prolonged survival. The present study showed that- the El double-restricted Ad effectively and selectively replicates in and causes oncolysis of GBC in vitro and in vivo with reduced negative effects on normal cells, suggesting that this could be a promising gene therapy tool for GBC.