Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape - PubMed (original) (raw)

doi: 10.1038/nbt.2612. Epub 2013 Jun 23.

William Querbes, Anja Zeigerer, Anna Borodovsky, Giovanni Marsico, Undine Schubert, Kevin Manygoats, Sarah Seifert, Cordula Andree, Martin Stöter, Hila Epstein-Barash, Ligang Zhang, Victor Koteliansky, Kevin Fitzgerald, Eugenio Fava, Marc Bickle, Yannis Kalaidzidis, Akin Akinc, Martin Maier, Marino Zerial

Affiliations

• PMID: 23792630
• DOI: 10.1038/nbt.2612

Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape

Jerome Gilleron et al. Nat Biotechnol. 2013 Jul.

Abstract

Delivery of short interfering RNAs (siRNAs) remains a key challenge in the development of RNA interference (RNAi) therapeutics. A better understanding of the mechanisms of siRNA cellular uptake, intracellular transport and endosomal release could critically contribute to the improvement of delivery methods. Here we monitored the uptake of lipid nanoparticles (LNPs) loaded with traceable siRNAs in different cell types in vitro and in mouse liver by quantitative fluorescence imaging and electron microscopy. We found that LNPs enter cells by both constitutive and inducible pathways in a cell type-specific manner using clathrin-mediated endocytosis as well as macropinocytosis. By directly detecting colloidal-gold particles conjugated to siRNAs, we estimated that escape of siRNAs from endosomes into the cytosol occurs at low efficiency (1-2%) and only during a limited window of time when the LNPs reside in a specific compartment sharing early and late endosomal characteristics. Our results provide insights into LNP-mediated siRNA delivery that can guide development of the next generation of delivery systems for RNAi therapeutics.

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Comment in

• A window onto siRNA delivery.
  Wang Y, Huang L. Wang Y, et al. Nat Biotechnol. 2013 Jul;31(7):611-2. doi: 10.1038/nbt.2634. Nat Biotechnol. 2013. PMID: 23839146 No abstract available.
• Lipid nanoparticles as ideal delivery modules for siRNA.
  Friedman H, Holt AT, Pham W. Friedman H, et al. Nanomedicine (Lond). 2013 Dec;8(12):1910-1. Nanomedicine (Lond). 2013. PMID: 24422219 No abstract available.

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