De-targeting by miR-143 decreases unwanted transgene expression in non-tumorigenic cells

Abstract

MicroRNA dysregulation often results in the development and progression of cancer. miR-143 is ubiquitously expressed in most human and murine tissues but downregulated in many cancer types. This differential miRNA expression can be utilized for targeted cancer gene therapies. Multiple copies of the miR-143 complementary target sequence were inserted into the 3'UTR of plasmid vectors encoding either for different reporter genes or for the therapeutic gene TNFα. With these transgenes, we analyzed the miR-143-dependent gene expression in cancer cells and normal cells. Moreover, we investigated miR-143-regulated luciferase expression in an NMRI nude/HUH7 xenograft mouse model using a nonviral carrier system for in vivo transfections. We showed low and high levels of miR-143 in cancer cells and normal cells, respectively, leading to a differential gene expression of the reporters and the therapeutic TNFα. According to the miR-143 levels, the luciferase reporter gene expression was silenced in the mouse lungs but not in HUH7 tumors. Thus, we utilized the differential miR-143 expression in healthy and cancerous tissues to de-target the lung by specifically targeting the tumor in an in vivo HUH7 xenograft mouse model. The use of an miR-143-regulated therapeutic transgene may present a promising approach for cancer gene therapy.