Aptamer-functionalized peptide H₃CR₅C as a novel nanovehicle for codelivery of fasudil and miRNA-195 targeting hepatocellular carcinoma

Abstract

Liver cancer is the fifth most commonly diagnosed malignancy, of which hepatocellular carcinoma (HCC) represents the dominating histological subtype. Antiangiogenic therapy aimed at vascular endothelial growth factor (VEGF) has shown promising but deficient clinical prospects on account of vasculogenic mimicry, a highly patterned vascular channel distinguished from the endothelium-dependent blood vessel, which may function as blood supply networks occurring in aggressive tumors including HCC. In this study, we used a new cationic peptide, disulfide cross-linked stearylated polyarginine peptide modified with histidine (H₃R₅), as a reducible vector, cell penetrating peptide-modified aptamer (ST21) with specific binding to HCC cells to conjugate to peptide H₃R₅ as the targeting probe, miRNA-195 (miR195) as a powerful gene drug to inhibit VEGF, and fasudil to suppress vasculogenic mimicry by blocking ROCK2, all of which were simultaneously encapsulated in the same nanoparticles. Fasudil was loaded by ammonium sulfate-induced transmembrane electrochemical gradient and miR195 was condensed through electrostatic interaction. ST21-H₃R₅-polyethylene glycol (PEG) exhibited excellent loading capacities for both fasudil and miR195 with adjustable dosing ratios. Western blot analysis showed that ^FasudilST21-H₃R₅-PEG_miR195 had strong silencing activity of ROCK2 and VEGF, as compared with ^FasudilH₃R₅-PEG_miR195. In vitro and in vivo experiments confirmed that ST21-modified nanoparticles showed significantly higher cellular uptake and therapeutic efficacy in tumor cells or tumor tissues than the unmodified counterparts. These findings suggest that aptamer-conjugated peptide holds great promise for delivering chemical drugs and gene drugs simultaneously to overcome HCC.

Keywords:

• aptamer
• fasudil
• miR195
• combined therapy
• hepatocellular carcinoma

Acknowledgments

The authors acknowledge Professor Shen Gao from Changhai Hospital affiliated to Second Military Medical University for his kind guidance in this research. This research work was supported by the funds provided by the National Natural Science Foundation of China (number 81302212), the Natural Science Foundation of Shanghai (16ZR1428000), and the Crossing Program between Medicine and Industry supported by Shanghai Jiaotong University (numbers YG2014MS32, YG2015QN14).

Disclosure

The authors report no conflicts of interest in this work.