Therapeutic potential of functionalized siRNA nanoparticles on regression of liver cancer in experimental mice

Mumtaz Jabeen,Mohammad Azam Ansari,Arun Chauhan,Azmat Ali Khan,Amer M Alanazi

doi:10.1038/s41598-019-52142-4

Abstract

Short interfering RNA (siRNA) possesses special ability of silencing specific gene. To increase siRNA stability, transportation and its uptake by tumor cells, effective delivery to the appropriate target cells is a major challenge of siRNA-based therapy. In the present study, an effective, safe and biocompatible survivin siRNA encapsulated, GalNAc decorated PEGylated PLGA nanoconjugates (NCs) viz., GalNAc@PEG@siRNA-PLGA were engineered and their synergistic antitumor efficacy was evaluated for targeted delivery in HCC bearing experimental mice. GalNAc@PEG@siRNA-PLGA NCs were characterized for size, bioavailability, toxicity and biocompatibility. Their antitumor potential was evaluated considering gene silencing, apoptosis, histopathology and survival of treated mice. Exceptional accumulation of hepatocytes, reduction in survivin expression and prominent regression in tumor size confirmed the ASGPR-mediated uptake of ligand-anchored NCs and silencing of survivin gene in a targeted manner. Increased DNA fragmentation and potential modulation of caspase-3, Bax and Bcl-2 factors specified the induction of apoptosis that helped in significant inhibition of HCC progression. The potential synchronous and tumor selective delivery of versatile NCs indicated the effective payloads towards the target site, increased apoptosis in cancer cells and improved survival of treated animals.

Highlights

Short interfering RNA possesses special ability of silencing specific gene
The carboxyl groups present on surface of polylactic acid-co-glycolic acids (PLGAs) of Short interfering RNA (siRNA)-PLGA nanoparticles were increased by adding polyethylene-maleic anhydride (PEMA)
These modified Polyethylene-maleic anhydride (PEMA) treated siRNA-PLGA were incubated with N-hydroxysuccinimide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodimide hydrochloride (EDC)

Summary

Introduction

Short interfering RNA (siRNA) possesses special ability of silencing specific gene. To increase siRNA stability, transportation and its uptake by tumor cells, effective delivery to the appropriate target cells is a major challenge of siRNA-based therapy. GalNAc@PEG@siRNA-PLGA NCs were characterized for size, bioavailability, toxicity and biocompatibility Their antitumor potential was evaluated considering gene silencing, apoptosis, histopathology and survival of treated mice. The potential synchronous and tumor selective delivery of versatile NCs indicated the effective payloads towards the target site, increased apoptosis in cancer cells and improved survival of treated animals. The main advantage of ASGPR is its affinity towards various ligands with as simple as carbohydrates[22,23] In view of this aspect, among various ligands, N-acetylgalactosamine (GalNAc), an oligosaccharide, has high affinity towards ASGPRs and decoration of nanoparticles with GalNAc will be of great importance in selective delivery of gene of interest to HCC tissue via ASGPR receptors. The designed NCs (GalNAc@PEG@siRNA-PLGA) were expected to bind ASGPR receptors of HCC cells, show high transfection efficiency resulting in potential knockdown of survivin gene

Methods

Results

Discussion

Conclusion