Cancer siRNA therapy by tumor selective delivery with ligand-targeted sterically stabilized nanoparticle

TLDR

Potent sequence‑selective gene inhibition by siRNA therapeutics promises ultimate specificity, yet siRNA delivery is limited by poor cellular uptake, short blood half‑life, and non‑specific immune activation. To overcome these barriers, ligand‑targeted, sterically stabilized nanoparticles were engineered for siRNA delivery. The authors constructed self‑assembling PEGylated polyethyleneimine nanoparticles bearing an Arg‑Gly‑Asp (RGD) peptide ligand that targets tumor neovasculature integrins, enabling delivery of siRNA that suppresses VEGFR2 expression and tumor angiogenesis. Cellular uptake and activity of the PEG‑PEI were siRNA‑sequence specific, ligand‑dependent, and competitively inhibited by free peptide; intravenous administration in tumor‑bearing mice produced selective tumor uptake, siRNA‑sequence‑specific VEGFR2 knockdown, reduced angiogenesis and tumor growth, demonstrating dual tissue‑ and gene‑level targeting that could enhance precision therapeutics.

Abstract

Potent sequence selective gene inhibition by siRNA ‘targeted’ therapeutics promises the ultimate level of specificity, but siRNA therapeutics is hindered by poor intracellular uptake, limited blood stability and non-specific immune stimulation. To address these problems, ligand-targeted, sterically stabilized nanoparticles have been adapted for siRNA. Self-assembling nanoparticles with siRNA were constructed with polyethyleneimine (PEI) that is PEGylated with an Arg-Gly-Asp (RGD) peptide ligand attached at the distal end of the polyethylene glycol (PEG), as a means to target tumor neovasculature expressing integrins and used to deliver siRNA inhibiting vascular endothelial growth factor receptor-2 (VEGF R2) expression and thereby tumor angiogenesis. Cell delivery and activity of PEGylated PEI was found to be siRNA sequence specific and depend on the presence of peptide ligand and could be competed by free peptide. Intravenous administration into tumor-bearing mice gave selective tumor uptake, siRNA sequence-specific inhibition of protein expression within the tumor and inhibition of both tumor angiogenesis and growth rate. The results suggest achievement of two levels of targeting: tumor tissue selective delivery via the nanoparticle ligand and gene pathway selectivity via the siRNA oligonucleotide. This opens the door for better targeted therapeutics with both tissue and gene selectivity, also to improve targeted therapies with less than ideal therapeutic targets.