Synthetic switch-based baculovirus for transgene expression control and selective killing of hepatocellular carcinoma cells

• Published in: Nucleic acids research Vol. 46; no. 15; p. e93
• Main Authors: Lin, Mei-Wei, Tseng, Yen-Wen, Shen, Chih-Che, Hsu, Mu-Nung, Hwu, Jih-Ru, Chang, Chin-Wei, Yeh, Chung-Ju, Chou, Min-Yuan, Wu, Jaw-Ching, Hu, Yu-Chen
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 06.09.2018
• Subjects: Animals; Baculoviridae - genetics; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Carcinoma, Hepatocellular - therapy; Cell Line, Tumor; Cell Survival - genetics; Gene Expression Regulation, Neoplastic; Genetic Vectors - genetics; HEK293 Cells; Hep G2 Cells; Humans; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Liver Neoplasms - therapy; Methods Online; MicroRNAs - genetics; MicroRNAs - metabolism; Reproducibility of Results; Sf9 Cells; Spodoptera; Synthetic Biology - methods; Transgenes - genetics
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/gky447

Abstract Baculovirus (BV) holds promise as a vector for anticancer gene delivery to combat the most common liver cancer-hepatocellular carcinoma (HCC). However, in vivo BV administration inevitably results in BV entry into non-HCC normal cells, leaky anticancer gene expression and possible toxicity. To improve the safety, we employed synthetic biology to engineer BV for transgene expression regulation. We first uncovered that miR-196a and miR-126 are exclusively expressed in HCC and normal cells, respectively, which allowed us to engineer a sensor based on distinct miRNA expression signature. We next assembled a synthetic switch by coupling the miRNA sensor and RNA binding protein L7Ae for translational repression, and incorporated the entire device into a single BV. The recombinant BV efficiently entered HCC and normal cells and enabled cis-acting transgene expression control, by turning OFF transgene expression in normal cells while switching ON transgene expression in HCC cells. Using pro-apoptotic hBax as the transgene, the switch-based BV selectively killed HCC cells in separate culture and mixed culture of HCC and normal cells. These data demonstrate the potential of synthetic switch-based BV to distinguish HCC and non-HCC normal cells for selective transgene expression control and killing of HCC cells.