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Pair correlation microscopy reveals the role of nanoparticle shape in intracellular transport and site of drug release

Nature Nanotechnology volume 12, pages 81–89 (2017)Cite this article

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Abstract

Nanoparticle size, surface charge and material composition are known to affect the uptake of nanoparticles by cells. However, whether nanoparticle shape affects transport across various barriers inside the cell remains unclear. Here we used pair correlation microscopy to show that polymeric nanoparticles with different shapes but identical surface chemistries moved across the various cellular barriers at different rates, ultimately defining the site of drug release. We measured how micelles, vesicles, rods and worms entered the cell and whether they escaped from the endosomal system and had access to the nucleus via the nuclear pore complex. Rods and worms, but not micelles and vesicles, entered the nucleus by passive diffusion. Improving nuclear access, for example with a nuclear localization signal, resulted in more doxorubicin release inside the nucleus and correlated with greater cytotoxicity. Our results therefore demonstrate that drug delivery across the major cellular barrier, the nuclear envelope, is important for doxorubicin efficiency and can be achieved with appropriately shaped nanoparticles.

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Figure 1: Nanoparticles must overcome a number of cellular barriers to deliver and release chemotherapeutics in the nucleus.
Figure 2: Measuring the subcellular distribution of nanoparticles and their mobility across subcellular barriers.
Figure 3: Nanoparticle shape modulates the subcellular distribution of mobile nanoparticles and the mobility across cellular barriers from EX to CYTO and NUC.
Figure 4: Nanoparticle accumulation and escape from early and late endosomes.
Figure 5: Nuclear entry and exit of nanoparticles with and without NLS modification.
Figure 6: High-aspect-ratio nanoparticles increase the efficacy of doxorubicin nuclear delivery.

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  • 30 September 2016

    In the version of this Article originally published online, in the Methods, in the equation for G(τ), in the denominator, the superscript '2' was incorrectly placed, and in the equation for G(τ, δr), in the denominator, there should have been two sets of angled brackets. These errors have been corrected in all versions of the Article.

References

  1. Jiang, W., Kim, B. Y., Rutka, J. T. & Chan, W. C. Nanoparticle-mediated cellular response is size-dependent. Nat. Nanotechnol. 3, 145–150 (2008).

    Article  CAS  Google Scholar 

  2. Davis, M. E., Chen, Z. G. & Shin, D. M. Nanoparticle therapeutics: an emerging treatment modality for cancer. Nat. Rev. Drug. Discov. 7, 771–782 (2008).

    Article  CAS  Google Scholar 

  3. Ahn, S., Seo, E., Kim, K. & Lee, S. J. Controlled cellular uptake and drug efficacy of nanotherapeutics. Sci. Rep. 3, 1997 (2013).

    Article  Google Scholar 

  4. Ruthardt, N., Lamb, D. C. & Brauchle, C. Single-particle tracking as a quantitative microscopy-based approach to unravel cell entry mechanisms of viruses and pharmaceutical nanoparticles. Mol. Ther. 19, 1199–1211 (2011).

    Article  CAS  Google Scholar 

  5. Yarbrough, M. L., Mata, M. A., Sakthivel, R. & Fontoura, B. M. Viral subversion of nucleocytoplasmic trafficking. Traffic 15, 127–140 (2014).

    Article  CAS  Google Scholar 

  6. Dohner, K. & Sodeik, B. The role of the cytoskeleton during viral infection. Curr. Top. Microbiol. Immunol. 285, 67–108 (2005).

    CAS  Google Scholar 

  7. Champion, J. A., Katare, Y. K. & Mitragotri, S. Making polymeric micro- and nanoparticles of complex shapes. Proc. Natl Acad. Sci. USA 104, 11901–11904 (2007).

    Article  CAS  Google Scholar 

  8. Champion, J. A., Katare, Y. K. & Mitragotri, S. Particle shape: a new design parameter for micro- and nanoscale drug delivery carriers. J. Control Release 121, 3–9 (2007).

    Article  CAS  Google Scholar 

  9. Karagoz, B., Boyer, C. & Davis, T. P. Simultaneous polymerization-induced self-assembly (PISA) and guest molecule encapsulation. Macromol. Rapid Commun. 35, 417–421 (2014).

    Article  CAS  Google Scholar 

  10. Barua, S. et al. Particle shape enhances specificity of antibody-displaying nanoparticles. Proc. Natl Acad. Sci. USA 110, 3270–3275 (2013).

    Article  CAS  Google Scholar 

  11. Chu, Z. et al. Unambiguous observation of shape effects on cellular fate of nanoparticles. Sci. Rep. 4, 4495 (2014).

    Article  Google Scholar 

  12. Chithrani, B. D. & Chan, W. C. Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes. Nano. Lett. 7, 1542–1550 (2007).

    Article  CAS  Google Scholar 

  13. Agarwal, R. et al. Mammalian cells preferentially internalize hydrogel nanodiscs over nanorods and use shape-specific uptake mechanisms. Proc. Natl Acad. Sci. USA 110, 17247–17252 (2013).

    Article  CAS  Google Scholar 

  14. Xu, Z. P. et al. Subcellular compartment targeting of layered double hydroxide nanoparticles. J. Control Release 130, 86–94 (2008).

    Article  CAS  Google Scholar 

  15. Hinde, E., Cardarelli, F., Digman, M. A. & Gratton, E. In vivo pair correlation analysis of EGFP intranuclear diffusion reveals DNA-dependent molecular flow. Proc. Natl Acad. Sci. USA 107, 16560–16565 (2010).

    Article  CAS  Google Scholar 

  16. Hinde, E., Cardarelli, F., Digman, M. A. & Gratton, E. Changes in chromatin compaction during the cell cycle revealed by micrometer-scale measurement of molecular flow in the nucleus. Biophys. J. 102, 691–697 (2012).

    Article  CAS  Google Scholar 

  17. Hinde, E. et al. The impact of mitotic versus interphase chromatin architecture on the molecular flow of EGFP by pair correlation analysis. Biophys. J. 100, 1829–1836 (2011).

    Article  CAS  Google Scholar 

  18. Hinde, E., Kong, X., Yokomori, K. & Gratton, E. Chromatin dynamics during DNA repair revealed by pair correlation analysis of molecular flow in the nucleus. Biophys. J. 107, 55–65 (2014).

    Article  CAS  Google Scholar 

  19. Digman, M. A. & Gratton, E. Imaging barriers to diffusion by pair correlation functions. Biophys. J. 97, 665–673 (2009).

    Article  CAS  Google Scholar 

  20. Digman, M. A. & Gratton, E. Lessons in fluctuation correlation spectroscopy. Annu. Rev. Phys. Chem. 62, 645–668 (2011).

    Article  CAS  Google Scholar 

  21. dos Santos, T., Varela, J., Lynch, I., Salvati, A. & Dawson, K. A. Effects of transport inhibitors on the cellular uptake of carboxylated polystyrene nanoparticles in different cell lines. PLoS ONE 6, e24438 (2011).

    Article  CAS  Google Scholar 

  22. Chu, Z. et al. Rapid endosomal escape of prickly nanodiamonds: implications for gene delivery. Sci. Rep. 5, 11661 (2015).

    Article  Google Scholar 

  23. Cardarelli, F. & Gratton, E. In vivo imaging of single-molecule translocation through nuclear pore complexes by pair correlation functions. PLoS ONE 5, e10475 (2010).

    Article  Google Scholar 

  24. Scott, C. C., Vacca, F. & Gruenberg, J. Endosome maturation, transport and functions. Semin. Cell Dev. Biol. 31, 2–10 (2014).

    Article  CAS  Google Scholar 

  25. Huotari, J. & Helenius, A. Endosome maturation. EMBO J. 30, 3481–3500 (2011).

    Article  CAS  Google Scholar 

  26. Kerr, M. & Teasdale, R. D. Live imaging of endosome dynamics. Semin. Cell. Dev. Biol. 31, 11–19 (2014).

    Article  CAS  Google Scholar 

  27. Alber, F. et al. The molecular architecture of the nuclear pore complex. Nature 450, 695–701 (2007).

    Article  CAS  Google Scholar 

  28. Adams, R. L. & Wente, S. R. Uncovering nuclear pore complexity with innovation. Cell 152, 1218–1221 (2013).

    Article  CAS  Google Scholar 

  29. Ghosh, K., Kanapathipillai, M., Korin, N., McCarthy, J. R. & Ingber, D. E. Polymeric nanomaterials for islet targeting and immunotherapeutic delivery. Nano Letters 12, 203–208 (2012).

    Article  CAS  Google Scholar 

  30. Misra, R. & Sahoo, S. K. Intracellular trafficking of nuclear localization signal conjugated nanoparticles for cancer therapy. Eur. J. Pharm. Sci. 39, 152–163 (2010).

    Article  CAS  Google Scholar 

  31. Karagoz, B. et al. Polymerization-induced self-assembly (PISA) - control over the morphology of nanoparticles for drug delivery applications. Poly. Chem. 5, 350–355 (2014).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

E.H. is funded by a Cancer Institute NSW Early Career Fellowship (RG151879) and UNSW Vice Chancellor Research Fellowship. B.K. acknowledges the Scientific and Technological Research Council of Turkey (TUBITAK) for financial support. C.B. is funded by a Future Fellowship (FT1210096) from Australian Research Council (ARC). K.G. acknowledges funding from the ARC Centre of Excellence in Advanced Molecular Imaging (CE140100011) and National Health and Medical Research Council of Australia (1059278, 1037320). J.J.G. acknowledges funding from the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology (CE140100036), the ARC Laureate Fellowship (FL150100060) program and a National Health and Medical Research Council program grant (1091261). The authors thank C. Benzing (UNSW) for discussion on cell uptake and endosomal escape. The authors thank E. Gratton (University of California, Irvine) for discussion on data analysis. The work was supported by the BioMedical Imaging Facility at UNSW.

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Authors and Affiliations

  1. EMBL Australia Node in Single Molecule Science, University of New South Wales, Sydney, 2052, Australia

    Elizabeth Hinde, Kitiphume Thammasiraphop & Katharina Gaus

  2. ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, 2052, Australia

    Elizabeth Hinde, Kitiphume Thammasiraphop & Katharina Gaus

  3. Australian Centre for NanoMedicine, University of New South Wales, Sydney, 2052, Australia

    Elizabeth Hinde, Kitiphume Thammasiraphop, Hien T. T. Duong, Jonathan Yeow, Bunyamin Karagoz, Cyrille Boyer, J. Justin Gooding & Katharina Gaus

  4. Centre for Advanced Macromolecular Design (CAMD), School of Chemical Engineering, University of New South Wales, Sydney, 2052, Australia

    Jonathan Yeow & Cyrille Boyer

  5. School of Chemistry, University of New South Wales, Sydney, 2052, Australia

    Bunyamin Karagoz & J. Justin Gooding

  6. ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, 2052, Australia

    J. Justin Gooding

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Contributions

E.H, C.B, J.J.G and K.G designed the research. E.H and K.T performed imaging experiments. H.T.T.D, B.K and J.Y synthesized the nanoparticles. E.H and K.T. analysed data, and E.H., J.J.G and K.G. wrote the paper. All authors reviewed the manuscript.

Corresponding authors

Correspondence to J. Justin Gooding or Katharina Gaus.

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The authors declare no competing financial interests.

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Hinde, E., Thammasiraphop, K., Duong, H. et al. Pair correlation microscopy reveals the role of nanoparticle shape in intracellular transport and site of drug release. Nature Nanotech 12, 81–89 (2017). https://doi.org/10.1038/nnano.2016.160

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