Cargando…
ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy
Stimuli-responsive polycations have been developed for improved nucleic acid transfection and enhanced therapeutic efficacy. The most reported mechanisms for controlled release of siRNA are based on polyelectrolyte exchange reactions in the cytoplasm and the degradation of polycations initiated by s...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Ivyspring International Publisher
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6160761/ https://www.ncbi.nlm.nih.gov/pubmed/30279726 http://dx.doi.org/10.7150/thno.26889 |
_version_ | 1783358837653241856 |
---|---|
author | Zhou, Zhanwei Zhang, Qingyan Zhang, Minghua Li, Huipeng Chen, Gang Qian, Chenggen Oupicky, David Sun, Minjie |
author_facet | Zhou, Zhanwei Zhang, Qingyan Zhang, Minghua Li, Huipeng Chen, Gang Qian, Chenggen Oupicky, David Sun, Minjie |
author_sort | Zhou, Zhanwei |
collection | PubMed |
description | Stimuli-responsive polycations have been developed for improved nucleic acid transfection and enhanced therapeutic efficacy. The most reported mechanisms for controlled release of siRNA are based on polyelectrolyte exchange reactions in the cytoplasm and the degradation of polycations initiated by specific triggers. However, the degradation strategy has not always been sufficient due to unsatisfactory kinetics and binding of cationic fragments to siRNA, which limits the gene silencing effect. In this study, a new strategy that combines degradation and charge reversal is proposed. Methods: We prepared a polycation (CrossPPA) by crosslinking of phenylboronic acid (PBA)-grafted 1.8k PEI with alginate. It was compared with 25k PEI, 1.8k PEI and 1.8k PEI-PBA on siRNA encapsulation, ATP-responsive behavior and mechanism, cytotoxicity, cell uptake, siRNA transfection, in vivo biodistribution and in vivo anti-tumor efficacy. The in vitro and in vivo experiments were performed on 4T1 murine breast cancer cells and 4T1 tumor model separately. Results: The crosslinking strategy obviously improve the siRNA loading ability of 1.8k PEI. We validated that intracellular levels of ATP could trigger CrossPPA disassembly and charge reversal, which resulted in efficient and rapid siRNA release due to electrostatic repulsion. Besides, CrossPPA/siRNA showed strong cell uptake in 4T1 cells compared with 1.8k PEI/siRNA. Notably, the cytotoxicity of CrossPPA was pretty low, which was owing to its biodegradability. Furthermore, the crosslinked polyplexes significantly enhanced siRNA transfection and improved tumor accumulation. The high gene silencing ability of CrossPPA polyplex led to strong anti-tumor efficacy when using Bcl2-targeted siRNA. Conclusion: These results indicated that the ATP-triggered disassembly and charge reversal strategy provided a new way for developing stimuli-responsive siRNA carriers and showed potential for nucleic acid delivery in the treatment of cancer. |
format | Online Article Text |
id | pubmed-6160761 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Ivyspring International Publisher |
record_format | MEDLINE/PubMed |
spelling | pubmed-61607612018-10-02 ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy Zhou, Zhanwei Zhang, Qingyan Zhang, Minghua Li, Huipeng Chen, Gang Qian, Chenggen Oupicky, David Sun, Minjie Theranostics Research Paper Stimuli-responsive polycations have been developed for improved nucleic acid transfection and enhanced therapeutic efficacy. The most reported mechanisms for controlled release of siRNA are based on polyelectrolyte exchange reactions in the cytoplasm and the degradation of polycations initiated by specific triggers. However, the degradation strategy has not always been sufficient due to unsatisfactory kinetics and binding of cationic fragments to siRNA, which limits the gene silencing effect. In this study, a new strategy that combines degradation and charge reversal is proposed. Methods: We prepared a polycation (CrossPPA) by crosslinking of phenylboronic acid (PBA)-grafted 1.8k PEI with alginate. It was compared with 25k PEI, 1.8k PEI and 1.8k PEI-PBA on siRNA encapsulation, ATP-responsive behavior and mechanism, cytotoxicity, cell uptake, siRNA transfection, in vivo biodistribution and in vivo anti-tumor efficacy. The in vitro and in vivo experiments were performed on 4T1 murine breast cancer cells and 4T1 tumor model separately. Results: The crosslinking strategy obviously improve the siRNA loading ability of 1.8k PEI. We validated that intracellular levels of ATP could trigger CrossPPA disassembly and charge reversal, which resulted in efficient and rapid siRNA release due to electrostatic repulsion. Besides, CrossPPA/siRNA showed strong cell uptake in 4T1 cells compared with 1.8k PEI/siRNA. Notably, the cytotoxicity of CrossPPA was pretty low, which was owing to its biodegradability. Furthermore, the crosslinked polyplexes significantly enhanced siRNA transfection and improved tumor accumulation. The high gene silencing ability of CrossPPA polyplex led to strong anti-tumor efficacy when using Bcl2-targeted siRNA. Conclusion: These results indicated that the ATP-triggered disassembly and charge reversal strategy provided a new way for developing stimuli-responsive siRNA carriers and showed potential for nucleic acid delivery in the treatment of cancer. Ivyspring International Publisher 2018-09-09 /pmc/articles/PMC6160761/ /pubmed/30279726 http://dx.doi.org/10.7150/thno.26889 Text en © Ivyspring International Publisher This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions. |
spellingShingle | Research Paper Zhou, Zhanwei Zhang, Qingyan Zhang, Minghua Li, Huipeng Chen, Gang Qian, Chenggen Oupicky, David Sun, Minjie ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy |
title | ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy |
title_full | ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy |
title_fullStr | ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy |
title_full_unstemmed | ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy |
title_short | ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy |
title_sort | atp-activated decrosslinking and charge-reversal vectors for sirna delivery and cancer therapy |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6160761/ https://www.ncbi.nlm.nih.gov/pubmed/30279726 http://dx.doi.org/10.7150/thno.26889 |
work_keys_str_mv | AT zhouzhanwei atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT zhangqingyan atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT zhangminghua atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT lihuipeng atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT chengang atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT qianchenggen atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT oupickydavid atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy AT sunminjie atpactivateddecrosslinkingandchargereversalvectorsforsirnadeliveryandcancertherapy |