Enhancement of gene therapy specificity for diffuse colon carcinoma liver metastases with recombinant herpes simplex virus
Enhancement of gene therapy specificity for diffuse colon carcinoma liver metastases with recombinant herpes simplex virus
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DOI：
10.1097/00000658-199609000-00008
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发表时间：
1996-09-01
影响因子：
9
通讯作者：
Tanabe, KK
中科院分区：
文献类型：
--
作者：
Carroll, NM;Chiocca, EA;Tanabe, KK
作者确定了一种重组单纯疱疹病毒（HSV）载体是否能在体外破坏人结肠癌细胞，以及该载体是否能在体内选择性地在结肠癌肝转移灶中复制，而不是在周围的肝细胞中复制。核糖核苷酸还原酶在活跃分裂的细胞中表达，并产生用于DNA合成的脱氧核糖核苷酸。hrR 3仅在能提供核糖核苷酸还原酶互补的活跃分裂细胞中复制，而在静止细胞如正常肝细胞中不复制。对于体内研究，无胸腺BALB/c裸鼠在7天后脾内注射HT 29和脾内注射hrR 3，并在病毒注射后7天处死。他们的肝脏进行了组织化学检查lacZ expression.ResultsAll的HT 29细胞在体外被破坏时，hrR 3加入在每10个肿瘤细胞1空斑形成单位的滴度。在通过脾内注射hrR 3处理的小鼠的肝切片中检查的105个肿瘤结节中有101个显示lacZ表达。最小的β-半乳糖苷酶活性存在于正常liver.ConclusionsHRR3 HSV载体有效地破坏HT 29人结肠癌细胞在非常低的感染复数。肝转移瘤和正常肝脏之间核糖核苷酸还原酶的差异表达允许hrR 3选择性地在肿瘤细胞中复制，而在周围正常肝脏中复制最少。进一步研究基于HSV的载体作为肝转移瘤的溶瘤剂是必要的。
ObjectiveThe authors determined whether a recombinant herpes simplex virus (HSV) vector could destroy human colon carcinoma cells in vitro and whether the vector would selectively replicate in colon carcinoma liver metastases but not surrounding hepatocytes in vivo.BackgroundThe HSV vector hrR3 is defective in the gene encoding ribonucleotide reductase and contains the lacZ reporter gene. Ribonucleotide reductase is expressed in actively dividing cells and generates deoxyribonucleotides for DNA synthesis. hrR3 replicates only in actively dividing cells that can provide ribonucleotide reductase in complementation, but not in quiescent cells such as normal hepatocytes.MethodshrR3- mediated lysis of HT29 human colon carcinoma cells was first determined in vitro. For in vivo studies, athymic BALB/c nude mice underwent intrasplenic injection of HT29 and intrasplenic injection of hrR3 7 days later, and were killed 7 days after viral injection. Their livers were examined histochemically for lacZ expression.ResultsAll the HT29 cells were destroyed in vitro when hrR3 was added at a titer of 1 plaque-forming unit per 10 tumor cells. One hundred one of 105 tumor nodules examined in liver sections from mice treated by intrasplenic injection of hrR3 demonstrated lacZ expression. Minimal beta-galactosidase activity was present in normal liver.ConclusionsThe hrR3 HSV vector effectively destroys HT29 human colon carcinoma cells at very low multiplicities of infection. Differential expression of ribonucleotide reductase between liver metastases and normal liver allows hrR3 to selectively replicate in tumor cells with minimal replication in surrounding normal liver. Further investigation of HSV-based vectors as oncolytic agents for liver metastases is warranted.