Molecularly self-assembled nucleic acid nanoparticles for targeted in vivo siRNA delivery

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Abstract

Nanoparticles are used for delivering therapeutics into cells1,2. However, size, shape, surface chemistry and the presentation of targeting ligands on the surface of nanoparticles can affect circulation half-life and biodistribution, cell-specific internalization, excretion, toxicity and efficacy3,4,5,6,7. A variety of materials have been explored for delivering small interfering RNAs (siRNAs)—a therapeutic agent that suppresses the expression of targeted genes8,9. However, conventional delivery nanoparticles such as liposomes and polymeric systems are heterogeneous in size, composition and surface chemistry, and this can lead to suboptimal performance, a lack of tissue specificity and potential toxicity10,11,12. Here, we show that self-assembled DNA tetrahedral nanoparticles with a well-defined size can deliver siRNAs into cells and silence target genes in tumours. Monodisperse nanoparticles are prepared through the self-assembly of complementary DNA strands. Because the DNA strands are easily programmable, the size of the nanoparticles and the spatial orientation and density of cancer-targeting ligands (such as peptides and folate) on the nanoparticle surface can be controlled precisely. We show that at least three folate molecules per nanoparticle are required for optimal delivery of the siRNAs into cells and, gene silencing occurs only when the ligands are in the appropriate spatial orientation. In vivo, these nanoparticles showed a longer blood circulation time (t1/2 ≈ 24.2 min) than the parent siRNA (t1/2 ≈ 6 min).

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Figure 1: Programmable self-assembly of ONPs.
Figure 2: In vitro screening and gene silencing using ONPs.
Figure 3: In vivo pharmacokinetic profile and gene silencing in tumour xenograft mouse model.

References

  1. 1

    Panyam, J. & Labhasetwar, V. Biodegradable nanoparticles for drug and gene delivery to cells and tissue. Adv. Drug Deliv. Rev. 55, 329–347 (2003).

  2. 2

    Peer, D. et al. Nanocarriers as an emerging platform for cancer therapy. Nature Nanotech. 2, 751–760 (2007).

  3. 3

    Petros, R. A. & DeSimone, J. M. Strategies in the design of nanoparticles for therapeutic applications. Nature Rev. Drug Discov. 9, 615–627 (2010).

  4. 4

    Choi, H. S. et al. Design consideration for tumour-targeted nanoparticles. Nature Nanotech. 5, 42–47 (2010).

  5. 5

    Weissleder, R., Kelly, K., Sun, E. Y., Shtatland, T. & Josephson, L. Cell-specific targeting of nanoparticles by multivalent attachment of small molecules. Nature Biotechnol. 23, 1418–1423 (2005).

  6. 6

    De Jong, W. H. & Borm P. Drug delivery and nanoparticles: applications and hazards. Int. J. Nanomed. 3, 133–149 (2008).

  7. 7

    Akinc, A. et al. Targeted delivery of RNAi therapeutics with endogenous and exogenous ligand-based mechanisms. Mol. Ther. 18, 1357–1364 (2010).

  8. 8

    Elbashir, S. M. et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498 (2001).

  9. 9

    Bumcrot, D., Manoharan, M., Koteliansky, V. & Sah, D. W. Y. RNAi therapeutics: a potential new class of pharmaceutical drugs. Nature Chem. Biol. 2, 711–719 (2006).

  10. 10

    Whitehead, K. A., Langer, R. & Anderson, D. G. Knocking down barriers: advances in siRNA delivery. Nature Rev. Drug Discov. 8, 129–138 (2009).

  11. 11

    Oh, Y. K. & Park, T. G. siRNA delivery systems for cancer treatment. Adv. Drug Deliv. Rev. 61, 850–862 (2009).

  12. 12

    Lv, H., Zhang, S., Wang, B., Cui, S. & Yan, J. Toxicity of cationic lipids and cationic polymers in gene delivery. J. Control. Rel. 114, 100–109 (2006).

  13. 13

    Bhatia, D. et al. A synthetic icosahedral DNA-based host–cargo complex for functional in vivo imaging. Nature Commun. 2, 339 (2011).

  14. 14

    Walsh, A. S., Yin, H., Erben, C. M., Wood, M. J. A. & Tuberfield, A. J. DNA cage delivery to mammalian cells. ACS Nano 5, 5427–5432 (2011).

  15. 15

    Keum, J. W., Ahn, J. H. & Bermudez, H. Design, assembly, and activity of antisense DNA nanostructures. Small 7, 3529–3535 (2011).

  16. 16

    Gaglione, M. & Messere, A. Recent progress in chemically modified siRNAs. Mini Rev. Med. Chem. 10, 578–595 (2010).

  17. 17

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

  18. 18

    Allen, T. M. & Cullis, P. R. Drug delivery systems: entering the mainstream. Science 303, 1818–1822 (2004).

  19. 19

    Mok, H., Lee, S. H., Park, J. W. & Park, T. G. Multimeric small interfering ribonucleic acid for highly efficient sequence-specific gene silencing. Nature Mater. 9, 272–278 (2010).

  20. 20

    Song, E. et al. Antibody mediated in vivo delivery of small interfering RNAs via cell-surface receptors. Nature Biotechnol. 23, 709–717 (2005).

  21. 21

    Li, S. D., Chen, Y. C., Hackett, M. J. & Huang, L. Tumor-targeted delivery of siRNA by self-assembled nanoparticles. Mol. Ther. 16, 163–169 (2008).

  22. 22

    Tarapore, P., Shu, Y., Guo, P. & Ho, S. M. Application of Phi29 motor pRNA for targeted therapeutic delivery of siRNA silencing metallothionein-IIA and surviving in ovarian cancers. Mol. Ther. 19, 386–394 (2011).

  23. 23

    Manz, B. N. et al. T-cell triggering thresholds are modulated by the number of antigen within individual T-cell receptor clusters. Proc. Natl Acad. Sci. USA 108, 9089–9094 (2011).

  24. 24

    Nahrendorf, M. et al. Hybrid PET-optical imaging using targeted probes. Proc. Natl Acad. Sci. USA 107, 7910–7915 (2010).

  25. 25

    Leuschner, F. et al. Therapeutic siRNA silencing in inflammatory monocytes in mice. Nature Biotechnol. 29, 1005–1010 (2011).

  26. 26

    Soutschek, J. et al. Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs. Nature 432, 173–178 (2004).

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Acknowledgements

This work was supported by the National Institutes of Health (EB000244), the Center for Cancer Nanotechnology Excellence (U54 CA151884), Alnylam Pharmaceuticals and the National Research Foundation of Korea (NRF-2011-357-D00063). The authors thank J. Hong and C. Hong for figure drawing, and J.B. Lee and A. Schroeder for helpful discussions.

Author information

H.L., A.L.J. and D.G.A planned the experiments. H.L., A.P., K.L., A.S., W.Q., C.Z., J.S.D., A.L.J. and J.T. conducted the experiments. H.L., A.L.J., Y.C., K.L., E.D.K., M.N., R.L. and D.G.A. analysed the data. H.L., A.L.J., M.M. and D.G.A. wrote the paper.

Correspondence to Daniel G. Anderson.

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The authors declare no competing financial interests.

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