Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling
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...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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2014
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| Online Access: | http://hdl.handle.net/1721.1/91243 https://orcid.org/0000-0002-2100-1171 https://orcid.org/0000-0003-0131-6552 https://orcid.org/0000-0001-5629-4798 https://orcid.org/0000-0003-4255-0492 |
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| author | Sahay, Gaurav Querbes, William Alabi, Christopher A. Eltoukhy, Ahmed A. Sarkar, Sovan Zurenko, Christopher Karagiannis, Emmanouil Love, Kevin T. Chen, Delai Zoncu, Roberto Buganim, Yosef Schroeder, Avi Langer, Robert Anderson, Daniel Griffith |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Sahay, Gaurav Querbes, William Alabi, Christopher A. Eltoukhy, Ahmed A. Sarkar, Sovan Zurenko, Christopher Karagiannis, Emmanouil Love, Kevin T. Chen, Delai Zoncu, Roberto Buganim, Yosef Schroeder, Avi Langer, Robert Anderson, Daniel Griffith |
| author_sort | Sahay, Gaurav |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:14:03Z |
| format | Article |
| id | mit-1721.1/91243 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:14:03Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | mit-1721.1/912432022-10-01T02:13:22Z Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling Sahay, Gaurav Querbes, William Alabi, Christopher A. Eltoukhy, Ahmed A. Sarkar, Sovan Zurenko, Christopher Karagiannis, Emmanouil Love, Kevin T. Chen, Delai Zoncu, Roberto Buganim, Yosef Schroeder, Avi Langer, Robert Anderson, Daniel Griffith Massachusetts Institute of Technology. Department of Chemical Engineering Massachusetts Institute of Technology. Media Laboratory Koch Institute for Integrative Cancer Research at MIT Sahay, Gaurav Karagiannis, Emmanouil Love, Kevin T. Chen, Delai Alabi, Christopher A. Eltoukhy, Ahmed A. Schroeder, Avi Langer, Robert Anderson, Daniel Griffith 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. Alnylam Pharmaceuticals (Firm) (Control release grant EB000244) National Heart, Lung, and Blood Institute National Institutes of Health (U.S.) (Program of Excellence in Nanotechnology (PEN) Award, Contract #HHSN268201000045C) 2014-10-30T19:26:33Z 2014-10-30T19:26:33Z 2013-06 2013-01 Article http://purl.org/eprint/type/JournalArticle 1087-0156 1546-1696 http://hdl.handle.net/1721.1/91243 Sahay, Gaurav, William Querbes, Christopher Alabi, Ahmed Eltoukhy, Sovan Sarkar, Christopher Zurenko, Emmanouil Karagiannis, et al. “Efficiency of siRNA Delivery by Lipid Nanoparticles Is Limited by Endocytic Recycling.” Nature Biotechnology 31, no. 7 (June 23, 2013): 653–658. https://orcid.org/0000-0002-2100-1171 https://orcid.org/0000-0003-0131-6552 https://orcid.org/0000-0001-5629-4798 https://orcid.org/0000-0003-4255-0492 en_US http://dx.doi.org/10.1038/nbt.2614 Nature Biotechnology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf PMC |
| spellingShingle | Sahay, Gaurav Querbes, William Alabi, Christopher A. Eltoukhy, Ahmed A. Sarkar, Sovan Zurenko, Christopher Karagiannis, Emmanouil Love, Kevin T. Chen, Delai Zoncu, Roberto Buganim, Yosef Schroeder, Avi Langer, Robert Anderson, Daniel Griffith Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling |
| title | Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling |
| title_full | Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling |
| title_fullStr | Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling |
| title_full_unstemmed | Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling |
| title_short | Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling |
| title_sort | efficiency of sirna delivery by lipid nanoparticles is limited by endocytic recycling |
| url | http://hdl.handle.net/1721.1/91243 https://orcid.org/0000-0002-2100-1171 https://orcid.org/0000-0003-0131-6552 https://orcid.org/0000-0001-5629-4798 https://orcid.org/0000-0003-4255-0492 |
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