ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy

Zhanwei Zhou, Qingyan Zhang, Minghua Zhang, Huipeng Li, Gang Chen, Chenggen Qian, David Oupicky, Minjie Sun

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)4604-4619
Number of pages16
JournalTheranostics
Volume8
Issue number17
DOIs
StatePublished - Jan 1 2018

Fingerprint

Small Interfering RNA
Adenosine Triphosphate
Neoplasms
Therapeutics
Transfection
Gene Silencing
Nucleic Acids
Static Electricity
Cytoplasm
Breast Neoplasms

Keywords

  • ATP-responsive
  • Cancer therapy
  • Charge reversal
  • Phenylboronic acid
  • SiRNA delivery

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Pharmacology, Toxicology and Pharmaceutics (miscellaneous)

Cite this

Zhou, Z., Zhang, Q., Zhang, M., Li, H., Chen, G., Qian, C., ... Sun, M. (2018). ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy. Theranostics, 8(17), 4604-4619. https://doi.org/10.7150/thno.26889

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.

In: Theranostics, Vol. 8, No. 17, 01.01.2018, p. 4604-4619.

Research output: Contribution to journalArticle

Zhou, Z, Zhang, Q, Zhang, M, Li, H, Chen, G, Qian, C, Oupicky, D & Sun, M 2018, 'ATP-activated decrosslinking and charge-reversal vectors for siRNA delivery and cancer therapy', Theranostics, vol. 8, no. 17, pp. 4604-4619. https://doi.org/10.7150/thno.26889
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. In: Theranostics. 2018 ; Vol. 8, No. 17. pp. 4604-4619.
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AU - Zhou, Zhanwei

AU - Zhang, Qingyan

AU - Zhang, Minghua

AU - Li, Huipeng

AU - Chen, Gang

AU - Qian, Chenggen

AU - Oupicky, David

AU - Sun, Minjie

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AB - 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.

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