Genetically engineered theranostic mesenchymal stem cells for the evaluation of the anticancer efficacy of enzyme/prodrug systems.
Genetically engineered theranostic mesenchymal stem cells for the evaluation of the anticancer efficacy of enzyme/prodrug systems.
复制标题
DOI：
10.1016/j.jconrel.2015.01.003
复制
发表时间：
2015-02-28
期刊：
影响因子：
--
通讯作者：
Hatefi A
中科院分区：
文献类型：
--
作者：
Nouri FS;Wang X;Hatefi A
在过去的十年中，各种酶/前药系统如胸苷激酶/更昔洛韦（TK/GCV）、酵母胞嘧啶脱氨酶/5-氟胞嘧啶（yCD/5-FC）和硝基还原酶/CB 1954（NTR/CB 1954）已用于干细胞介导的癌症自杀基因治疗。然而，还没有进行研究来比较和证明使用一种系统相对于另一种系统的优缺点。知道每种酶/前药系统都有自己的优点和缺点，我们利用间充质干细胞（MSC）作为介质，首次进行比较研究，说明这些系统之间的细微差异对治疗结果的影响。出于治疗目的，我们首先对MSC进行遗传修饰，以稳定表达一组四种自杀基因，包括TK（TK 007和TKSR 39突变体）、酵母胞苷脱氨酶：尿嘧啶磷酸核糖基转移酶（yCD：UPRT）和硝基还原酶（NTR）。然后，我们通过使用对所有四种酶/前药系统敏感的SKOV 3细胞系来评估基因工程MSC的体外和体内抗癌功效。此外，所有MSC都被工程化以稳定表达荧光素酶基因，使其适合于动物中的定量成像和剂量-反应关系研究。考虑到前体药物的旁观者效应所施加的限制，我们的研究结果表明yCD：UPRT/5-FC是测试的酶/前体药物系统中最有效的酶/前体药物系统。我们的研究结果还表明，治疗诊断间充质干细胞是一种可靠的介质，并排评估和筛选的酶/前药系统在临床前水平。这项研究的结果可以帮助利用基于细胞的非病毒或病毒载体进行癌症自杀基因治疗的科学家在选择酶/前药系统时做出更明智的决定。
Over the past decade, various enzyme/prodrug systems such as thymidine kinase/ganciclovir (TK/GCV), yeast cytosine deaminase/5-fluorocytosine (yCD/5-FC) and nitroreductase/CB1954 (NTR/CB1954) have been used for stem cell mediated suicide gene therapy of cancer. Yet, no study has been conducted to compare and demonstrate the advantages and disadvantages of using one system over another. Knowing that each enzyme/prodrug system has its own strengths and weaknesses, we utilized mesenchymal stem cells (MSCs) as a medium to perform for the first time a comparative study that illustrated the impact of subtle differences among these systems on the therapeutic outcome. For therapeutic purposes, we first genetically modified MSCs to stably express a panel of four suicide genes including TK (TK007 and TKSR39 mutants), yeast cytosine deaminase: uracil phosphoribosyltransferase (yCD:UPRT) and nitroreductase (NTR). Then, we evaluated the anticancer efficacies of the genetically engineered MSCs in vitro and in vivo by using SKOV3 cell line which is sensitive to all four enzyme/prodrug systems. In addition, all MSCs were engineered to stably express luciferase gene making them suitable for quantitative imaging and dose-response relationship studies in animals. Considering the limitations imposed by the prodrugs’ bystander effects, our findings show that yCD:UPRT/5-FC is the most effective enzyme/prodrug system among the ones tested. Our findings also demonstrate that theranostic MSCs are a reliable medium for the side-by-side evaluation and screening of the enzyme/prodrug systems at the preclinical level. The results of this study could help scientists who utilize cell-based, non-viral or viral vectors for suicide gene therapy of cancer make more informed decisions when choosing enzyme/prodrug systems.