Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling

By G. Sahay, W. Querbes, Christopher A. Alabi, Ahmed A. Eltoukhy, Sovan Sarkar, Christopher Zurenko, Emannouil Karagiannis, Kevin T. Love, Delai Chen, Roberto Zoncu and 4 more. Nature Biotechnology, 2013

867 citations recorded by the source corpus.

Despite efforts to understand the interactions between nanoparticles and cells, the cellular processes that determine the efficiency of intracellular drug delivery remain unclear. Here we examine cellular uptake of short interfering RNA (siRNA) delivered in lipid nanoparticles (LNPs) using cellular trafficking probes in combination with automated high-throughput confocal microscopy. We also employed defined perturbations of cellular pathways paired with systems biology approaches to uncover protein-protein and protein–small molecule interactions. We show that multiple cell signaling effectors are required for initial cellular entry of LNPs through macropinocytosis, including proton pumps, mTOR and cathepsins. siRNA delivery is substantially reduced as ≅70% of the internalized siRNA undergoes exocytosis through egress of LNPs from late endosomes/lysosomes. Niemann-Pick type C1 (NPC1) is shown to be an important regulator of the major recycling pathways of LNP-delivered siRNAs. NPC1-deficient cells show enhanced cellular retention of LNPs inside late endosomes and lysosomes, and increased gene silencing of the target gene. Our data suggest that siRNA delivery efficiency might be improved by designing delivery vehicles that can escape the recycling pathways.

Subfields: Biomaterials, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/be7e521b76f3337cb7ab60e96af235f0fe528234

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Citations in the corpus

Delivering the Messenger: Advances in Technologies for Therapeutic mRNA Delivery. (2019, positive): “22,23 Once inside the cell, LNPs are routed into early endosomes, followed by late endosomes, and ﬁ nally the lysosomes where the mRNA contents are enzymatically degraded.”
The role of lipid components in lipid nanoparticles for vaccines and gene therapy (2022, positive): “That said, due to endocytic recycling, the proportion of nucleic acid cargo that escapes the endosomes is reportedly less than 5% [41].”
The delivery of therapeutic oligonucleotides (2016, background): “Another interesting report demonstrated that the NPC1 lipid transport system, previously shown to be involved in LNP processing (158), is also involved in the processing of certain types of polymeric nanoparticles (227).”
Brief update on endocytosis of nanomedicines (2019, background): “Adapted from [6] (A), [7,222,229,247,253] (B), [282] (C), [9,289] (D), and [293] (E) with permission.”