KRAS mutant allele-specific expression knockdown in pancreatic cancer model with systemically delivered bi-shRNA KRAS lipoplex

Abstract

The KRAS oncogene, present in over 90% of pancreatic ductal adenocarcinomas, is most frequently the result of one of three gain-of-function substitution mutations of codon 12 glycine. Thus far, RAS mutations have been clinically refractory to both direct and selective inhibition by systemic therapeutics. This report presents the results of pre-clinical assessment of a lipoplex comprising a plasmid-encoded, modular bi-functional shRNA (bi-shRNA), which executes selective and multi-mutant allelic KRASG12mut gene silencing, encased within a fusogenic liposome systemic delivery vehicle. Using both a dual luciferase reporter system and a Restriction Fragment Length Polymorphism (RFLP) assay, selective discrimination of KRASG12mut from KRASwt was confirmed in vitro in PANC1 cells. Subsequently, systemic administration of the bi-shRNAKRAS fusogenic lipoplex into female athymic Nu/Nu mice bearing PANC1 xenografts demonstrated intratumoral plasmid delivery, KRASG12mut knockdown, and inhibition of tumor growth, without adverse effect. Clinical trials with the bi-shRNA lipoplex have been implemented.

Mesoporous Silica Nanoparticle-Based Drug Delivery Systems for the Treatment of Pancreatic Cancer: A Systematic Literature Overview

“…Several alternative targeted delivery strategies have been explored in PDAC over the last decades. These strategies employ-amongst others-liposomes [23][24][25][26], poly(lactic-co-glycolic acid) (PLGA)-based polymeric nanoparticles [27][28][29], solid lipid nanoparticles [30][31][32], and mesoporous silica nanoparticles (MSNs) of which especially the use of MSNs seems an attractive approach. MSNs are nanoscale silica-based particles with a porous structure as implied by their name.…”

RNAi-Based Approaches for Pancreatic Cancer Therapy

“…Rao et al have demonstrated KRAS down-regulation by a lipoplex encoding a short hairpin RNA via systemic injection in a pancreatic cancer model [ 51 ]. The lipoplex contained a plasmid encoding a short hairpin RNA that executes selective silencing of the mutated KRAS gene with tumor cell entry facilitated by facilitated by fusogenic liposomes.…”

A peptide-CpG-DNA-liposome complex vaccine targeting TM4SF5 suppresses growth of pancreatic cancer in a mouse allograft model

“…For example, dioleoyl phosphatidylethanolamine (DOPE) increases the endosomal escape ability of DNA after endocytosis of cationic liposome/DNA complex 23,24. In a pancreatic cancer model, recent studies reported the use of a liposome complex to deliver shRNAs or anti-miRNA oligonucleotides for therapeutic purposes 25,26. Liposomes are also used for efficient antigen delivery and enhanced immune response in developing vaccines 27–29.…”