A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA
Protein-based methods of targeted short-interfering RNA (siRNA) delivery have the potential to solve some of the problems faced by nanoparticle-based methods, such as poor pharmacokinetics and biodistribution, low tumor penetration, and polydispersity. However, protein-based targeted delivery has be...
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| Format: | Article |
| Language: | en_US |
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2014
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| Online Access: | http://hdl.handle.net/1721.1/91602 https://orcid.org/0000-0003-2398-5896 https://orcid.org/0000-0002-0882-7761 |
| _version_ | 1826193307749842944 |
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| author | Liu, David V. Yang, Nicole Jie Yeon Wittrup, Karl Dane |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Liu, David V. Yang, Nicole Jie Yeon Wittrup, Karl Dane |
| author_sort | Liu, David V. |
| collection | MIT |
| description | Protein-based methods of targeted short-interfering RNA (siRNA) delivery have the potential to solve some of the problems faced by nanoparticle-based methods, such as poor pharmacokinetics and biodistribution, low tumor penetration, and polydispersity. However, protein-based targeted delivery has been limited to fusion proteins with polycationic peptides as siRNA carriers, whose high charge density in some cases results in undesirable biophysical and in vivo properties. Here, we present a fully proteinaceous, multiagent approach for targeted siRNA delivery to epidermal growth factor receptor (EGFR), using a nonpolycationic carrier for siRNA. Each agent contributes a fundamentally different mechanism of action that work together for potent targeted RNA interference. The first agent is an EGFR-targeted fusion protein that uses a double-stranded RNA-binding domain as a nonpolycationic siRNA carrier. This double-stranded RNA-binding domain fusion protein can deliver siRNA to the endosomes of an EGFR-expressing cell line. A second agent delivers the cholesterol-dependent cytolysin, perfringolysin O, in a targeted manner, which enhances the endosomal escape of siRNA and induces gene silencing. A third agent that clusters EGFR increases gene-silencing potency and decreases cytolysin toxicity. Altogether, this system is potent, with only 16 nmol/l siRNA required for gene silencing and a therapeutic window that spans two orders of magnitude of targeted cytolysin concentrations. |
| first_indexed | 2024-09-23T09:37:13Z |
| format | Article |
| id | mit-1721.1/91602 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:37:13Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | mit-1721.1/916022022-09-26T12:40:26Z A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA Liu, David V. Yang, Nicole Jie Yeon Wittrup, Karl Dane Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemical Engineering Koch Institute for Integrative Cancer Research at MIT Liu, David V. Yang, Nicole Jie Yeon Wittrup, Karl Dane Protein-based methods of targeted short-interfering RNA (siRNA) delivery have the potential to solve some of the problems faced by nanoparticle-based methods, such as poor pharmacokinetics and biodistribution, low tumor penetration, and polydispersity. However, protein-based targeted delivery has been limited to fusion proteins with polycationic peptides as siRNA carriers, whose high charge density in some cases results in undesirable biophysical and in vivo properties. Here, we present a fully proteinaceous, multiagent approach for targeted siRNA delivery to epidermal growth factor receptor (EGFR), using a nonpolycationic carrier for siRNA. Each agent contributes a fundamentally different mechanism of action that work together for potent targeted RNA interference. The first agent is an EGFR-targeted fusion protein that uses a double-stranded RNA-binding domain as a nonpolycationic siRNA carrier. This double-stranded RNA-binding domain fusion protein can deliver siRNA to the endosomes of an EGFR-expressing cell line. A second agent delivers the cholesterol-dependent cytolysin, perfringolysin O, in a targeted manner, which enhances the endosomal escape of siRNA and induces gene silencing. A third agent that clusters EGFR increases gene-silencing potency and decreases cytolysin toxicity. Altogether, this system is potent, with only 16 nmol/l siRNA required for gene silencing and a therapeutic window that spans two orders of magnitude of targeted cytolysin concentrations. National Institutes of Health (U.S.) (Grant AI065824) National Institutes of Health (U.S.) (Grant CA101830) 2014-11-17T19:52:04Z 2014-11-17T19:52:04Z 2014-05 2014-01 Article http://purl.org/eprint/type/JournalArticle 2162-2531 http://hdl.handle.net/1721.1/91602 Liu, David V, Nicole J Yang, and K Dane Wittrup. “A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA.” Mol Ther Nucleic Acids 3, no. 5 (May 13, 2014): e162. © 2014 The American Society of Gene & Cell Therapy https://orcid.org/0000-0003-2398-5896 https://orcid.org/0000-0002-0882-7761 en_US http://dx.doi.org/10.1038/mtna.2014.14 Molecular Therapy—Nucleic Acids Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License http://creativecommons.org/licenses/by-nc-nd/3.0/ application/pdf Molecular Therapy Nucleic Acids |
| spellingShingle | Liu, David V. Yang, Nicole Jie Yeon Wittrup, Karl Dane A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA |
| title | A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA |
| title_full | A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA |
| title_fullStr | A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA |
| title_full_unstemmed | A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA |
| title_short | A Nonpolycationic Fully Proteinaceous Multiagent System for Potent Targeted Delivery of siRNA |
| title_sort | nonpolycationic fully proteinaceous multiagent system for potent targeted delivery of sirna |
| url | http://hdl.handle.net/1721.1/91602 https://orcid.org/0000-0003-2398-5896 https://orcid.org/0000-0002-0882-7761 |
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