Skip to main content

Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles

Abstract

The therapeutic promise of small interfering RNAs (siRNAs) for specific gene silencing is dependent on the successful delivery of functional siRNAs to the cytoplasm. Their conjugation to an established delivery platform, such as gold nanoparticles, offers tremendous potential for treating diseases and advancing our understanding of cellular processes. Their success or failure is dependent on both the uptake of the nanoparticles into the cells and subsequent intracellular release of the functional siRNA. In this study, utilizing gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol (PEG) ligand density on knockdown, with a view to determining the optimal nanoparticle design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, as well as the possible effect of steric hindrance of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, while knockdown was determined at the protein and physiological levels (cells in S-phase) by in-cell westerns and BrdU incorporation, respectively.

This is a preview of subscription content, access via your institution.

References

  1. Chiu, Y.-L.; Rana, T. M. siRNA function in RNAi: A chemical modification analysis. RNA 2003, 9, 1034–1048.

    Article  Google Scholar 

  2. Derfus, A. M.; Chen, A. A.; Min, D.-H.; Ruoslahti, E.; Bhatia, S. N. Targeted quantum dot conjugates for siRNA delivery. Bioconjugate Chem. 2007, 18, 1391–1396.

    Article  Google Scholar 

  3. Grigsby, C. L.; Leong, K. W. Balancing protection and release of DNA: Tools to address a bottleneck of non-viral gene delivery. J. R. Soc. Interface 2010, 7, S67–S82.

  4. Lévy, R.; Shaheen, U.; Cesbron, Y.; Sée, V. Gold nanoparticles delivery in mammalian live cells: A critical review. Nano Rev. 2010, 1, 4889.

    Article  Google Scholar 

  5. Karakoti, A. S.; Das, S.; Thevuthasan, S.; Seal, S. PEGylated inorganic nanoparticles. Angew. Chem., Int. Ed. 2011, 50, 1980–1994.

    Article  Google Scholar 

  6. Stefanick, J. F.; Ashley, J. D.; Kiziltepe, T.; Bilgicer, B. A systematic analysis of peptide linker length and liposomal polyethylene glycol coating on cellular uptake of peptidetargeted liposomes. ACS Nano 2013, 7, 2935–2947.

    Article  Google Scholar 

  7. Oishi, M.; Nakaogami, J.; Ishii, T.; Nagasaki, Y. Smart PEGylated gold nanoparticles for the cytoplasmic delivery of siRNA to induce enhanced gene silencing. Chem. Lett. 2001, 35, 1046–1047.

    Article  Google Scholar 

  8. Lushchak, V. I. Glutathione homeostasis and functions: Potential targets for medical interventions. J. Amino Acids 2012, 2012, 736837.

    Article  Google Scholar 

  9. Conde, J.; Tian, F. R.; Hernández, Y.; Bao, C. C.; Cui, D. X.; Janssen, K.-P.; Ibarra, M. R.; Baptista, P. V.; Stoeger, T.; de la Fuente, J. M. In vivo tumor targeting via nanoparticlemediated therapeutic siRNA coupled to inflammatory response in lung cancer mouse models. Biomaterials 2013, 34, 7744–7753.

    Article  Google Scholar 

  10. Cebrián, V.; Martín-Saavedra, F.; Yagüe, C.; Arruebo, M.; Santamaría, J.; Vilaboa, N. Size-dependent transfection efficiency of PEI-coated gold nanoparticles. Acta Biomater. 2011, 7, 3645–3655.

    Article  Google Scholar 

  11. Dreaden, E. C.; Mackey, M. A.; Huang, X. H.; Kang, B.; El-Sayed, M. A. Beating cancer in multiple ways using nanogold. Chem. Soc. Rev. 2011, 40, 3391–3404.

    Article  Google Scholar 

  12. Conde, J.; Ambrosone, A.; Sanz, V.; Hernandez, Y.; Marchesano, V.; Tian, F. R.; Child, H.; Berry, C. C.; Ibarra, M. R.; Baptista, P. V. et al. Design of multifunctional gold nanoparticles for in vitro and in vivo gene silencing. ACS Nano 2012, 6, 8316–8324.

    Article  Google Scholar 

  13. Frens, G. Controlled nucleation for regulation of particle-size in monodisperse gold suspensions. Nat. Phys. Sci. 1973, 241, 20–22.

    Article  Google Scholar 

  14. Turkevich, J.; Stevenson, P. C.; Hillier, J. A study of the nucleation and growth processes in the synthesis of colloidal gold. Discuss. Faraday Soc. 1951, 55–75.

    Google Scholar 

  15. Lee, P. C.; Meisel, D. Adsorption and surface-enhanced Raman of dyes on silver and gold sols. J. Phys. Chem. 1982, 86, 3391–3395.

    Article  Google Scholar 

  16. McBeath, R.; Pirone, D. M.; Nelson, C. M.; Bhadriraju, K.; Chen, C. S. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev. Cell 2004, 6, 483–495.

    Article  Google Scholar 

  17. Conde, J.; Oliva, N.; Artzi, N. Implantable hydrogel embedded dark-gold nanoswitch as a theranostic probe to sense and overcome cancer multidrug resistance. Proc. Natl. Acad. Sci. USA 2015, 112, E1278–E1287.

    Article  Google Scholar 

  18. Ackerson, C. J.; Sykes, M. T.; Kornberg, R. D. Defined DNA/nanoparticle conjugates. Proc. Natl. Acad. Sci. USA 2005, 102, 13383–13385.

    Article  Google Scholar 

  19. Kumar, D.; Meenan, B. J.; Dixon, D. Glutathione-mediated release of Bodipy® from PEG cofunctionalized gold nanoparticles. Int. J. Nanomedicine 2012, 7, 4007–4022.

    Article  Google Scholar 

  20. Chompoosor, A.; Han, G.; Rotello, V. M. Charge dependence of ligand release and monolayer stability of gold nanoparticles by biogenic thiols. Bioconjugate Chem. 2008, 19, 1342–1345.

    Article  Google Scholar 

  21. Hong, R.; Han, G.; Fernández, J. M.; Kim, B.-J.; Forbes, N. S.; Rotello, V. M. Glutathione-mediated delivery and release using monolayer protected nanoparticle carriers. J. Am. Chem. Soc. 2006, 128, 1078–1079.

    Article  Google Scholar 

  22. Han, G.; Chari, N. S.; Verma, A.; Hong, R.; Marin, C. T.; Rotello, V. M. Controlled recovery of the transcription of nanoparticle-bound DNA by intracellular concentrations of glutathione. Bioconjugate Chem. 2005, 16, 1356–1359.

    Article  Google Scholar 

  23. Zhu, Z.-J.; Yeh, Y.-C.; Tang, R.; Yan, B.; Tamayo, J.; Vachet, R. W.; Rotello, V. M. Stability of quantum dots in live cells. Nat. Chem. 2011, 3, 963–968.

    Article  Google Scholar 

  24. Li, D.; Li, G. P.; Guo, W. W.; Li, P. C.; Wang, E. K.; Wang, J. Glutathione-mediated release of functional plasmid DNA from positively charged quantum dots. Biomaterials 2008, 29, 2776–2782.

    Article  Google Scholar 

  25. Meister, A.; Anderson, M. E. Glutathione. Annu. Rev. Biochem. 1983, 52, 711–760.

    Article  Google Scholar 

  26. Wu, G. Y.; Fan, Y.-Z.; Yang, S.; Lupton, J. R.; Turner, N. D. Glutathione metabolism and its implications for health. J. Nutr. 2004, 134, 489–492.

    Google Scholar 

  27. Lei, X. G. In vivo antioxidant role of glutathione peroxidase: Evidence from knockout mice. Methods Enzymol. 2002, 347, 213–225.

    Article  Google Scholar 

  28. Fang, Y.-Z.; Yang, S.; Wu, G. Y. Free radicals, antioxidants, and nutrition. Nutrition 2002, 18, 872–879.

    Article  Google Scholar 

  29. Meister, A. Glutathione metabolism and its selective modification. J. Biol. Chem. 1988, 263, 17205–17208.

    Google Scholar 

  30. Kah, J. C. Y.; Wong, K. Y.; Neoh, K. G.; Song, J. H.; Fu, J. W. P.; Mhaisalkar, S.; Olivo, M.; Richard, C. J. Critical parameters in the pegylation of gold nanoshells for biomedical applications: An in vitro macrophage study. J. Drug Target. 2009, 17, 181–193.

    Article  Google Scholar 

  31. Rana, T. M. Illuminating the silence: Understanding the structure and function of small RNAs. Nat. Rev. Mol. Cell Biol. 2007, 8, 23–36.

    Article  Google Scholar 

  32. Li, Z. H.; Rana, T. M. Molecular mechanisms of RNAtriggered gene silencing machineries. Acc. Chem. Res. 2012, 45, 1122–1131.

    Article  Google Scholar 

  33. Unsworth, L. D.; Sheardown, H.; Brash, J. L. Protein resistance of surfaces prepared by sorption of end-thiolated poly(ethylene glycol) to gold: Effect of surface chain density. Langmuir 2005, 21, 1036–1041.

    Article  Google Scholar 

  34. Knop, K.; Hoogenboom, R.; Fischer, D.; Schubert, U. S. Poly(ethylene glycol) in drug delivery: Pros and cons as well as potential alternatives. Angew. Chem., Int. Ed. 2010, 49, 6288–6308.

    Article  Google Scholar 

  35. Hinterwirth, H.; Kappel, S.; Waitz, T.; Prohaska, T.; Lindner, W.; Lämmerhofer, M. Quantifying thiol ligand density of self-assembled monolayers on gold nanoparticles by inductively coupled plasma-mass spectrometry. ACS Nano 2013, 7, 1129–1136.

    Article  Google Scholar 

  36. Häkkinen, H. The gold-sulfur interface at the nanoscale. Nat. Chem. 2012, 4, 443–455.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mark McCully.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

McCully, M., Hernandez, Y., Conde, J. et al. Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles. Nano Res. 8, 3281–3292 (2015). https://doi.org/10.1007/s12274-015-0828-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12274-015-0828-5

Keywords

  • gold
  • nanoparticles
  • PEG
  • glutathione
  • siRNA
  • drug delivery