Letter

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

Received:
Accepted:
Published online:

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.

  • Subscribe to Nature Biotechnology for full access:

    $250

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    DNA in a material world. Nature 421, 427–431 (2003).

  2. 2.

    , , , & Two DNA nanomachines map pH changes along intersecting endocytic pathways inside the same cell. Nat. Nanotechnol. 8, 459–467 (2013).

  3. 3.

    , & A logic-gated nanorobot for targeted transport of molecular payloads. Science 335, 831–834 (2012).

  4. 4.

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

  5. 5.

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

  6. 6.

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

  7. 7.

    , , , & A synthetic icosahedral DNA-based host-cargo complex for functional in vivo imaging. Nat. Commun. 2, 339 (2011).

  8. 8.

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

  9. 9.

    & Strategies for advancing cancer nanomedicine. Nat. Mater. 12, 958–962 (2013).

  10. 10.

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

  11. 11.

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

  12. 12.

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

  13. 13.

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

  14. 14.

    , , , & Role of thrombin signalling in platelets in haemostasis and thrombosis. Nature 413, 74–78 (2001).

  15. 15.

    , , & Challenges and opportunities for structural DNA nanotechnology. Nat. Nanotechnol. 6, 763–772 (2011).

  16. 16.

    Folding DNA to create nanoscale shapes and patterns. Nature 440, 297–302 (2006).

  17. 17.

    , , & Dynamic DNA devices and assemblies formed by shape-complementary, non-base pairing 3D components. Science 347, 1446–1452 (2015).

  18. 18.

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

  19. 19.

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

  20. 20.

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

  21. 21.

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

  22. 22.

    , , & DNA nanotechnology from the test tube to the cell. Nat. Nanotechnol. 10, 748–760 (2015).

  23. 23.

    , , , & The nucleolin targeting aptamer AS1411 destabilizes Bcl-2 messenger RNA in human breast cancer cells. Cancer Res. 68, 2358–2365 (2008).

  24. 24.

    & Structure-switching signaling aptamers. J. Am. Chem. Soc. 125, 4771–4778 (2003).

  25. 25.

    , & Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 33, 941–951 (2015).

  26. 26.

    , & Hyperthermia enables tumor-specific nanoparticle delivery: effect of particle size. Cancer Res. 60, 4440–4445 (2000).

  27. 27.

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

  28. 28.

    , & Designing DNA nanodevices for compatibility with the immune system of higher organisms. Nat. Nanotechnol. 10, 741–747 (2015).

  29. 29.

    , , , & 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).

  30. 30.

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

  31. 31.

    , , , & Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression. Nucleic Acids Res. 38, 4807–4820 (2010).

  32. 32.

    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).

  33. 33.

    , & DNA-nanotube-induced alignment of membrane proteins for NMR structure determination. Proc. Natl. Acad. Sci. USA 104, 6644–6648 (2007).

  34. 34.

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

  35. 35.

    & 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).

  36. 36.

    , & 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).

  37. 37.

    , , , & 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).

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

Author notes

    • Suping Li
    • , Qiao Jiang
    • , Shaoli Liu
    •  & Yinlong Zhang

    These authors contributed equally to this work.

Affiliations

  1. CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, CAS Key Laboratory of Nanosystem and Hierarchical Fabrication, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, China, Beijing, China.

    • Suping Li
    • , Qiao Jiang
    • , Shaoli Liu
    • , Yinlong Zhang
    • , Yanhua Tian
    • , Chen Song
    • , Jing Wang
    • , Yiguo Zou
    • , Guangjun Nie
    • , Baoquan Ding
    •  & Yuliang Zhao
  2. University of Chinese Academy of Sciences, Beijing, China.

    • Suping Li
    • , Shaoli Liu
    • , Guangjun Nie
    • , Baoquan Ding
    •  & Yuliang Zhao
  3. College of Pharmaceutical Science, Jilin University, Changchun, China.

    • Yinlong Zhang
  4. Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China.

    • Yanhua Tian
  5. QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia.

    • Gregory J Anderson
  6. Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China.

    • Jing-Yan Han
  7. School of Molecular Sciences, Center for Molecular Design and Biomimetics; School of Life Sciences, Center for Immunotherapy, Vaccines, and Virotherapy at the Biodesign Institute, Arizona State University, Tempe, Arizona, USA.

    • Yung Chang
    • , Yan Liu
    •  & Hao Yan
  8. Institute of Zoology, Chinese Academy of Sciences, Beijing, China.

    • Chen Zhang
    •  & Guangbiao Zhou
  9. Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, Guangzhou, China.

    • Liang Chen

Authors

  1. Search for Suping Li in:

  2. Search for Qiao Jiang in:

  3. Search for Shaoli Liu in:

  4. Search for Yinlong Zhang in:

  5. Search for Yanhua Tian in:

  6. Search for Chen Song in:

  7. Search for Jing Wang in:

  8. Search for Yiguo Zou in:

  9. Search for Gregory J Anderson in:

  10. Search for Jing-Yan Han in:

  11. Search for Yung Chang in:

  12. Search for Yan Liu in:

  13. Search for Chen Zhang in:

  14. Search for Liang Chen in:

  15. Search for Guangbiao Zhou in:

  16. Search for Guangjun Nie in:

  17. Search for Hao Yan in:

  18. Search for Baoquan Ding in:

  19. Search for Yuliang Zhao in:

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.

Competing interests

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

Corresponding authors

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

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–28, Supplementary Tables 1–3, Supplementary Note 1

  2. 2.

    Life Sciences Reporting Summary