Intracellular siRNA delivery dynamics of integrin-targeted, PEGylated chitosan–poly(ethylene imine) hybrid nanoparticles: A mechanistic insight
| Autor: | Camilla Foged, Kevin Vanvarenberg, Olivier Feron, Gaëlle Vandermeulen, Caroline Bouzin, Héloïse Ragelle, Jacqueline Marchand-Brynaert, Stefano Colombo, Vincent Pourcelle, Véronique Préat |
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| Rok vydání: | 2015 |
| Předmět: |
Intracellular Fluid
Integrins Small interfering RNA Peptidomimetic Endosome media_common.quotation_subject Green Fluorescent Proteins Cell Pharmaceutical Science Polyethylene Glycols Green fluorescent protein Cell Line Tumor medicine Humans Gene silencing RNA Small Interfering Internalization media_common Chitosan Chemistry Gene Transfer Techniques Molecular biology medicine.anatomical_structure Biophysics Nanoparticles Imines Polyethylenes Intracellular |
| Zdroj: | Journal of Controlled Release. 211:1-9 |
| ISSN: | 0168-3659 |
| DOI: | 10.1016/j.jconrel.2015.05.274 |
| Popis: | Integrin-targeted nanoparticles are promising for the delivery of small interfering RNA (siRNA) to tumor cells or tumor endothelium in cancer therapy aiming at silencing genes essential for tumor growth. However, during the process of optimizing and realizing their full potential, it is pertinent to gain a basic mechanistic understanding of the bottlenecks existing for nanoparticle-mediated intracellular delivery. We designed αvβ3 integrin-targeted nanoparticles by coupling arginine-glycine-aspartate (RGD) or RGD peptidomimetic (RGDp) ligands to the surface of poly(ethylene glycol) (PEG) grafted chitosan-poly(ethylene imine) hybrid nanoparticles. The amount of intracellular siRNA delivered by αvβ3-targeted versus non-targeted nanoparticles was quantified in the human non-small cell lung carcinoma cell line H1299 expressing enhanced green fluorescent protein (EGFP) using a stem-loop reverse transcription quantitative polymerase chain reaction (RT-qPCR) approach. Data demonstrated that the internalization of αvβ3-targeted nanoparticles was highly dependent on the surface concentration of the ligand. Above a certain threshold concentration, the use of targeted nanoparticles provided a two-fold increase in the number of siRNA copies/cell, subsequently resulting in as much as 90% silencing of EGFP at well-tolerated carrier concentrations. In contrast, non-targeted nanoparticles mediated low levels of gene silencing, despite relatively high intracellular siRNA concentrations, indicating that these nanoparticles might end up in late endosomes or lysosomes without releasing their cargo to the cell cytoplasm. Thus, the silencing efficiency of the chitosan-based nanoparticles is strongly dependent on the uptake and the intracellular trafficking in H1299 EGFP cells, which is critical information towards a more complete understanding of the delivery mechanism that can facilitate the future design of efficient siRNA delivery systems. |
| Databáze: | OpenAIRE |
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