Department of Pharmaceutics, Rutgers University, Piscataway, NJ 08854, USA.
Department of Pathology and Laboratory Medicine, Rutgers-Robert Wood Johnson Medical School, New Brunswick, NJ 08903, USA.
J Control Release. 2019 Oct;311-312:273-287. doi: 10.1016/j.jconrel.2019.09.006. Epub 2019 Sep 6.
The objective of this study was to develop a stem cell-based system for targeted suicide gene therapy of recurrent, metastatic, and unresectable ovarian cancer. Malignant cells were obtained from the ascites of a patient with advanced recurrent epithelial ovarian cancer (named OVASC-1). Cancer cells were characterized to determine the percentages of drug-resistant ALDH+ cells, MDR-1/ABCG2 overexpressing cells, and cancer stem-like cells. The sensitivity and resistance of the OVASC-1 cells and spheroids to the metabolites of three different enzyme/prodrug systems were assessed, and the most effective one was selected. Adipose-derived stem cells (ASCs) were genetically engineered to express recombinant secretory human carboxylesterase-2 and nanoluciferase genes for simultaneous disease therapy and quantitative imaging. Bioluminescent imaging, magnetic resonance imaging and immuno/histochemistry results show that the engineered ASCs actively targeted and localized at both tumor stroma and necrotic regions. This created the unique opportunity to deliver drugs to not only tumor supporting cells in the stroma, but also to cancer stem-like cells in necrotic/hypoxic regions. The statistical analysis of intraperitoneal OVASC-1 tumor burden and survival rates in mice shows that the administration of the bioengineered ASCs in combination with irinotecan prodrug in the designed sequence and timeline eradicated all intraperitoneal tumors and provided survival benefits. In contrast, treatment of the drug-resistant OVASC-1 tumors with cisplatin/paclitaxel (standard-of-care) did not have any statistically significant benefit. The histopathology and hematology results do not show any toxicity to major peritoneal organs. Our toxicity data in combination with efficacy outcomes delineate a nonsurgical and targeted stem cell-based approach to overcoming drug resistance in recurrent metastatic ovarian cancer.
本研究旨在开发一种基于干细胞的系统，用于复发性、转移性和不可切除的卵巢癌的靶向自杀基因治疗。恶性细胞从晚期复发性上皮性卵巢癌（命名为 OVASC-1）患者的腹水获得。对癌细胞进行特征鉴定，以确定耐药性 ALDH+细胞、MDR-1/ABCG2 过表达细胞和癌症干细胞样细胞的比例。评估 OVASC-1 细胞和球体对三种不同酶/前药系统代谢物的敏感性和耐药性，并选择最有效的一种。脂肪来源的干细胞（ASCs）被遗传工程改造以表达重组分泌人羧酸酯酶-2 和纳米荧光素酶基因，用于同时进行疾病治疗和定量成像。生物发光成像、磁共振成像和免疫/组织化学结果表明，工程化 ASCs 主动靶向并定位于肿瘤基质和坏死区域。这为不仅向基质中的肿瘤支持细胞，而且向坏死/缺氧区域中的癌症干细胞样细胞递药创造了独特的机会。对小鼠腹腔内 OVASC-1 肿瘤负担和存活率的统计学分析表明，以设计的顺序和时间线联合施用生物工程化 ASCs 和伊立替康前药根除了所有腹腔内肿瘤并提供了生存益处。相比之下，用顺铂/紫杉醇（标准护理）治疗耐药性 OVASC-1 肿瘤没有任何统计学上的显著益处。组织病理学和血液学结果显示主要腹膜器官没有任何毒性。我们的毒性数据与疗效结果相结合，描绘了一种非手术和靶向干细胞方法，用于克服复发性转移性卵巢癌的耐药性。
