Nanobubble-mediated co-delivery of Ce6 and miR-195 for synergized sonodynamic and checkpoint blockade combination therapy with elicitation of robust immune response in hepatocellular carcinoma.

MicroRNAs (miRNAs) is the tiny and highly conserved noncoding RNAs, regulate gene expression at the post-transcriptional level by binding to the 3'-UTR of target mRNAs. Several studies found that miR-195 plays an unavoidable role in the regulation of cell proliferation, cycle and apoptosis in hepatocellular carcinoma (HCC). Here, we constructed miR-195 and Chlorine e6 (Ce6) co-loading NBs (nanobubbles), making use of NBs as carriers to deliver miR-195 and Ce6 to mouse tumor models. Our results showed that the binding between PD-1 and PD-L1 was blocked by upregulating miR-195 expression. The analysis of CTL (Cytotoxic T Cell) immune activity in the treatment group was higher than the control group. Simultaneously, Ce6 was used as sonosensitizer to induce SDT (sonodynamic therapy) and trigger ICD (immunogenic cell death) of tumor cell via generation of ROS. Recent studies have found that ICD may further enhance anti-tumor immunity against PD-L1. Results indicated that combination treatment effectively stimulated infiltration of T cell and the activation of natural killer (NK) cells as well as the maturation of dendritic cells (DCs), and the combination treatment group exibited the highest CTL killing activity. These results indicate that a stronger antitumor immunity was triggered via combination of SDT-induced tumor cell ICD and immune checkpoint blockade of PD-1/PD-L1 mediated by upregulation of miR-195. In conclusion, we have successfully constructed an efficient delivery system with great potential to provide a new strategy for synergistic immunotherapy.