Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

A DNA nanorobot functions as a cancer therapeutic in response to a molecular trigger in vivo

Abstract

Nanoscale robots have potential as intelligent drug delivery systems that respond to molecular triggers1,2,3,4. Using DNA origami we constructed an autonomous DNA robot programmed to transport payloads and present them specifically in tumors. Our nanorobot is functionalized on the outside with a DNA aptamer that binds nucleolin, a protein specifically expressed on tumor-associated endothelial cells5, and the blood coagulation protease thrombin within its inner cavity. The nucleolin-targeting aptamer serves both as a targeting domain and as a molecular trigger for the mechanical opening of the DNA nanorobot. The thrombin inside is thus exposed and activates coagulation at the tumor site. Using tumor-bearing mouse models, we demonstrate that intravenously injected DNA nanorobots deliver thrombin specifically to tumor-associated blood vessels and induce intravascular thrombosis, resulting in tumor necrosis and inhibition of tumor growth. The nanorobot proved safe and immunologically inert in mice and Bama miniature pigs. Our data show that DNA nanorobots represent a promising strategy for precise drug delivery in cancer therapy.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Design and characterization of thrombin-functionalized DNA nanorobot.
Figure 2: Analysis of DNA nanorobot-triggered activation and endothelial cell targeting.
Figure 3: DNA nanorobots target tumors, induce thrombosis in tumor vessels and inhibit tumor growth in vivo.
Figure 4: Treatment with nanorobot-Th inhibits the growth of melanoma and poorly vascularized SK-OV3 ovarian cancer.

References

  1. Seeman, N.C. DNA in a material world. Nature 421, 427–431 (2003).

    Article  Google Scholar 

  2. Modi, S., Nizak, C., Surana, S., Halder, S. & Krishnan, Y. Two DNA nanomachines map pH changes along intersecting endocytic pathways inside the same cell. Nat. Nanotechnol. 8, 459–467 (2013).

    Article  CAS  Google Scholar 

  3. Douglas, S.M., Bachelet, I. & Church, G.M. A logic-gated nanorobot for targeted transport of molecular payloads. Science 335, 831–834 (2012).

    Article  CAS  Google Scholar 

  4. Amir, Y. et al. Universal computing by DNA origami robots in a living animal. Nat. Nanotechnol. 9, 353–357 (2014).

    Article  CAS  Google Scholar 

  5. Huang, Y. et al. The angiogenic function of nucleolin is mediated by vascular endothelial growth factor and nonmuscle myosin. Blood 107, 3564–3571 (2006).

    Article  CAS  Google Scholar 

  6. Jungmann, R. et al. Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT. Nat. Methods 11, 313–318 (2014).

    Article  CAS  Google Scholar 

  7. Bhatia, D., Surana, S., Chakraborty, S., Koushika, S.P. & Krishnan, Y. A synthetic icosahedral DNA-based host-cargo complex for functional in vivo imaging. Nat. Commun. 2, 339 (2011).

    Article  Google Scholar 

  8. Lee, H. et al. Molecularly self-assembled nucleic acid nanoparticles for targeted in vivo siRNA delivery. Nat. Nanotechnol. 7, 389–393 (2012).

    Article  CAS  Google Scholar 

  9. Chauhan, V.P. & Jain, R.K. Strategies for advancing cancer nanomedicine. Nat. Mater. 12, 958–962 (2013).

    Article  CAS  Google Scholar 

  10. Huang, X. et al. Tumor infarction in mice by antibody-directed targeting of tissue factor to tumor vasculature. Science 275, 547–550 (1997).

    Article  CAS  Google Scholar 

  11. Hu, P. et al. Comparison of three different targeted tissue factor fusion proteins for inducing tumor vessel thrombosis. Cancer Res. 63, 5046–5053 (2003).

    CAS  PubMed  Google Scholar 

  12. Jain, R.K. Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307, 58–62 (2005).

    Article  CAS  Google Scholar 

  13. Agemy, L. et al. Nanoparticle-induced vascular blockade in human prostate cancer. Blood 116, 2847–2856 (2010).

    Article  CAS  Google Scholar 

  14. Sambrano, G.R., Weiss, E.J., Zheng, Y.W., Huang, W. & Coughlin, S.R. Role of thrombin signalling in platelets in haemostasis and thrombosis. Nature 413, 74–78 (2001).

    Article  CAS  Google Scholar 

  15. Pinheiro, A.V., Han, D., Shih, W.M. & Yan, H. Challenges and opportunities for structural DNA nanotechnology. Nat. Nanotechnol. 6, 763–772 (2011).

    Article  CAS  Google Scholar 

  16. Rothemund, P.W.K. Folding DNA to create nanoscale shapes and patterns. Nature 440, 297–302 (2006).

    Article  CAS  Google Scholar 

  17. Gerling, T., Wagenbauer, K.F., Neuner, A.M. & Dietz, H. Dynamic DNA devices and assemblies formed by shape-complementary, non-base pairing 3D components. Science 347, 1446–1452 (2015).

    Article  CAS  Google Scholar 

  18. Benson, E. et al. DNA rendering of polyhedral meshes at the nanoscale. Nature 523, 441–444 (2015).

    Article  CAS  Google Scholar 

  19. Schüller, V.J. et al. Cellular immunostimulation by CpG-sequence-coated DNA origami structures. ACS Nano 5, 9696–9702 (2011).

    Article  Google Scholar 

  20. Jiang, Q. et al. DNA origami as a carrier for circumvention of drug resistance. J. Am. Chem. Soc. 134, 13396–13403 (2012).

    Article  CAS  Google Scholar 

  21. Zhang, Q. et al. DNA origami as an in vivo drug delivery vehicle for cancer therapy. ACS Nano 8, 6633–6643 (2014).

    Article  CAS  Google Scholar 

  22. Chen, Y.J., Groves, B., Muscat, R.A. & Seelig, G. DNA nanotechnology from the test tube to the cell. Nat. Nanotechnol. 10, 748–760 (2015).

    Article  CAS  Google Scholar 

  23. Soundararajan, S., Chen, W., Spicer, E.K., Courtenay-Luck, N. & Fernandes, D.J. The nucleolin targeting aptamer AS1411 destabilizes Bcl-2 messenger RNA in human breast cancer cells. Cancer Res. 68, 2358–2365 (2008).

    Article  CAS  Google Scholar 

  24. Nutiu, R. & Li, Y. Structure-switching signaling aptamers. J. Am. Chem. Soc. 125, 4771–4778 (2003).

    Article  CAS  Google Scholar 

  25. Blanco, E., Shen, H. & Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 33, 941–951 (2015).

    Article  CAS  Google Scholar 

  26. Kong, G., Braun, R.D. & Dewhirst, M.W. Hyperthermia enables tumor-specific nanoparticle delivery: effect of particle size. Cancer Res. 60, 4440–4445 (2000).

    CAS  PubMed  Google Scholar 

  27. Zhang, H. et al. Identification of urine protein biomarkers with the potential for early detection of lung cancer. Sci. Rep. 5, 11805 (2015).

    Article  Google Scholar 

  28. Surana, S., Shenoy, A.R. & Krishnan, Y. Designing DNA nanodevices for compatibility with the immune system of higher organisms. Nat. Nanotechnol. 10, 741–747 (2015).

    Article  CAS  Google Scholar 

  29. Liu, Y., Zeng, B.H., Shang, H.T., Cen, Y.Y. & Wei, H. Bama miniature pigs (Sus scrofa domestica) as a model for drug evaluation for humans: comparison of in vitro metabolism and in vivo pharmacokinetics of lovastatin. Comp. Med. 58, 580–587 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Goldberg, S.N. et al. Image-guided tumor ablation: proposal for standardization of terms and reporting criteria. Radiology 228, 335–345 (2003).

    Article  Google Scholar 

  31. Miniard, A.C., Middleton, L.M., Budiman, M.E., Gerber, C.A. & Driscoll, D.M. Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression. Nucleic Acids Res. 38, 4807–4820 (2010).

    Article  CAS  Google Scholar 

  32. Thompson, J.S. et al. BAFF binds to the tumor necrosis factor receptor-like molecule B cell maturation antigen and is important for maintaining the peripheral B cell population. J. Exp. Med. 192, 129–135 (2000).

    Article  CAS  Google Scholar 

  33. Douglas, S.M., Chou, J.J. & Shih, W.M. DNA-nanotube-induced alignment of membrane proteins for NMR structure determination. Proc. Natl. Acad. Sci. USA 104, 6644–6648 (2007).

    Article  CAS  Google Scholar 

  34. Alvarez-Erviti, L. et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 29, 341–345 (2011).

    Article  CAS  Google Scholar 

  35. Guyer, R.A. & Macara, I.G. Loss of the polarity protein PAR3 activates STAT3 signaling via an atypical protein kinase C (aPKC)/NF-κB/interleukin-6 (IL-6) axis in mouse mammary cells. J. Biol. Chem. 290, 8457–8468 (2015).

    Article  CAS  Google Scholar 

  36. Gottfries, J., Melgar, S. & Michaëlsson, E. Modelling of mouse experimental colitis by global property screens: a holistic approach to assess drug effects in inflammatory bowel disease. PLoS One 7, e30005 (2012).

    Article  CAS  Google Scholar 

  37. O'Callaghan, P., Li, J.P., Lannfelt, L., Lindahl, U. & Zhang, X. Microglial heparan sulfate proteoglycans facilitate the cluster-of-differentiation 14 (CD14)/Toll-like receptor 4 (TLR4)-dependent inflammatory response. J. Biol. Chem. 290, 14904–14914 (2015).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank L.Z. Xu (Medical and Health Analysis Center of Peking University) for animal imaging and G. Z. Shi (Laboratory Animal Center of Institute of Biophysics, Chinese Academy of Sciences) for histological examination of minipigs. We also thank A. Sheftel from High Impact Editing for improving the English of the manuscript. This work was supported by grants from National Basic Research Plan of China (MoST Program 2016YFA0201601 to G.N. and B.D.), the National Natural Science Foundation of China (31730032 to G.N., 21222311, 21573051, 91127021 to B.D., the National Distinguished Young Scientists program 31325010 to G.N.), Innovation Research Group of National Natural Science Foundation (11621505 to G.N. and Yuliang Z., 21721002 to B.D.), Beijing Municipal Science & Technology Commission (Z161100000116035 to G.N., Z161100000116036 to B.D.), CAS Interdisciplinary Innovation Team to B.D., G.N. & Yuliang Z., Key Research Program of Frontier Sciences, CAS, Grant No. QYZDB-SSW-SLH029 to B.D. and US National Institute of Health Director's Transformative Research Award (R01GM104960-01 to H.Y.).

Author information

Authors and Affiliations

Authors

Contributions

Suping L., Q.J., Shaoli L., Yinlong Z., Y.T., C.S., B.D., Yuliang Z. and G.N. conceived and designed the experiments. Suping L., Q.J., Shaoli L., Yinlong Z., C.S., Y.T. and C.Z. performed the experiments. Y.T., C.S., H.Y., B.D., Yinlong Z. and G.N. collected and analyzed the data. J.W., G.A., J.H., Yiguo Z., Y.C., Y.L., L.C., G.-B.Z., G.Z. and C.Z. provided suggestions and technical support on the project. H.Y., B.D., Yuliang Z., and G.N. supervised the project. Suping L., Q.J., H.Y., B.D. and G.N. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Guangjun Nie, Hao Yan, Baoquan Ding or Yuliang Zhao.

Ethics declarations

Competing interests

An international provisional patent has been filed based on this work.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–28, Supplementary Tables 1–3, Supplementary Note 1 (PDF 4539 kb)

Life Sciences Reporting Summary (PDF 287 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, S., Jiang, Q., Liu, S. et al. A DNA nanorobot functions as a cancer therapeutic in response to a molecular trigger in vivo. Nat Biotechnol 36, 258–264 (2018). https://doi.org/10.1038/nbt.4071

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt.4071

This article is cited by

Search

Quick links

Nature Briefing: Cancer

Sign up for the Nature Briefing: Cancer newsletter — what matters in cancer research, free to your inbox weekly.

Get what matters in cancer research, free to your inbox weekly. Sign up for Nature Briefing: Cancer