Engineering of Bacteria for the Visualization of Targeted Delivery of a Cytolytic Anticancer Agent
Engineering of Bacteria for the Visualization of Targeted Delivery of a Cytolytic Anticancer Agent
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DOI：
10.1038/mt.2013.183
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发表时间：
2013-11-01
影响因子：
12.4
通讯作者：
Min, Jung-Joon
中科院分区：
文献类型：
--
作者：
Jiang, Sheng-Nan;Park, Seung-Hwan;Min, Jung-Joon
最近的一些报道表明，减毒鼠伤寒沙门氏菌能够靶向原发性和转移性肿瘤。使用细菌作为递送抗癌药物的载体需要一种精确调节和可视化基因表达的机制，以确保药物生产的适当时机和位置。为了将这些功能整合到细菌中，我们使用了一种阻遏物调节的四环素外排系统，其中治疗基因和成像报告基因的表达由不同的启动子（tetAP和tetRP）控制，以响应细胞外四环素。减毒鼠伤寒沙门用编码溶细胞素A（一种治疗基因）和海肾荧光素酶变体8（一种成像报道基因）的表达质粒转化鼠伤寒沙门氏菌，并静脉内给予荷瘤小鼠。该基因工程菌成功定位于肿瘤组织，且基因表达依赖于诱导剂的浓度，表明外周调控细菌基因表达的可行性。生物发光信号允许定位细菌的基因表达。工程菌显著抑制了小鼠的原发性和转移性肿瘤，并延长了小鼠的存活时间。因此，携带用于靶向抗癌治疗的治疗和成像报告基因的工程化细菌可以被设计为治疗诊断剂。
A number of recent reports have demonstrated that attenuated Salmonella typhimurium are capable of targeting both primary and metastatic tumors. The use of bacteria as a vehicle for the delivery of anticancer drugs requires a mechanism that precisely regulates and visualizes gene expression to ensure the appropriate timing and location of drug production. To integrate these functions into bacteria, we used a repressor-regulated tetracycline efflux system, in which the expression of a therapeutic gene and an imaging reporter gene were controlled by divergent promoters (tetAP and tetRP) in response to extracellular tetracycline. Attenuated S. typhimurium was transformed with the expression plasmids encoding cytolysin A, a therapeutic gene, and renilla luciferase variant 8, an imaging reporter gene, and administered intravenously to tumor-bearing mice. The engineered Salmonella successfully localized to tumor tissue and gene expression was dependent on the concentration of inducer, indicating the feasibility of peripheral control of bacterial gene expression. The bioluminescence signal permitted the localization of gene expression from the bacteria. The engineered bacteria significantly suppressed both primary and metastatic tumors and prolonged survival in mice. Therefore, engineered bacteria that carry a therapeutic and an imaging reporter gene for targeted anticancer therapy can be designed as a theranostic agent.