Article

Nanoparticle orientation to control RNA loading and ligand display on extracellular vesicles for cancer regression

  • Nature Nanotechnologyvolume 13pages8289 (2018)
  • doi:10.1038/s41565-017-0012-z
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Abstract

Nanotechnology offers many benefits, and here we report an advantage of applying RNA nanotechnology for directional control. The orientation of arrow-shaped RNA was altered to control ligand display on extracellular vesicle membranes for specific cell targeting, or to regulate intracellular trafficking of small interfering RNA (siRNA) or microRNA (miRNA). Placing membrane-anchoring cholesterol at the tail of the arrow results in display of RNA aptamer or folate on the outer surface of the extracellular vesicle. In contrast, placing the cholesterol at the arrowhead results in partial loading of RNA nanoparticles into the extracellular vesicles. Taking advantage of the RNA ligand for specific targeting and extracellular vesicles for efficient membrane fusion, the resulting ligand-displaying extracellular vesicles were capable of specific delivery of siRNA to cells, and efficiently blocked tumour growth in three cancer models. Extracellular vesicles displaying an aptamer that binds to prostate-specific membrane antigen, and loaded with survivin siRNA, inhibited prostate cancer xenograft. The same extracellular vesicle instead displaying epidermal growth-factor receptor aptamer inhibited orthotopic breast cancer models. Likewise, survivin siRNA-loaded and folate-displaying extracellular vesicles inhibited patient-derived colorectal cancer xenograft.

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Acknowledgements

We thank H. G. Zhang for his communication during the investigation of the exosome project, and J. Zhang, H. Weiss, D. Wu and D. Gao for assistance with statistical analysis. The research was supported mainly by National Institutes of Health grants UH3TR000875 and U01CA207946 (P. G.), and partially by R01CA186100 (B. G.), R35CA197706 (C.M.C.), P30CA177558 and R01CA195573 (B.M.E.).

Author information

Author notes

    • Tae Jin Lee

    Present address: Department of Neurosurgery, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, TX, USA

Affiliations

  1. College of Pharmacy, The Ohio State University, Columbus, OH, USA

    • Fengmei Pi
    • , Daniel W. Binzel
    • , Zhefeng Li
    • , Hui Li
    • , Farzin Haque
    • , Shaoying Wang
    •  & Peixuan Guo
  2. Center for RNA Nanobiotechnology and Nanomedicine, The Ohio State University, Columbus, OH, USA

    • Fengmei Pi
    • , Daniel W. Binzel
    • , Zhefeng Li
    • , Hui Li
    • , Farzin Haque
    • , Shaoying Wang
    •  & Peixuan Guo
  3. Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

    • Fengmei Pi
    • , Daniel W. Binzel
    • , Tae Jin Lee
    • , Zhefeng Li
    • , Hui Li
    • , Farzin Haque
    • , Shaoying Wang
    • , Carlo M. Croce
    •  & Peixuan Guo
  4. Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, USA

    • Fengmei Pi
    • , Daniel W. Binzel
    • , Zhefeng Li
    • , Hui Li
    • , Farzin Haque
    • , Shaoying Wang
    •  & Peixuan Guo
  5. Department of Cancer Biology and Genetics, College of Medicine, The Ohio State University, Columbus, OH, USA

    • Tae Jin Lee
    • , Carlo M. Croce
    •  & Peixuan Guo
  6. Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA

    • Meiyan Sun
    •  & Bin Guo
  7. Markey Cancer Center, Department of Surgery, University of Kentucky, Lexington, KY, USA

    • Piotr Rychahou
    •  & B. Mark Evers

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Contributions

P.G. generated the original idea of using the 3WJ structure orientation to control cell entry or cell surface anchoring, respectively, and designed the arrowhead and arrowtail technology. P.G. and F.P. conceived and designed the experiments. H.L. and S.W. developed the method for RNA insertion to EV. F.P., H.L., D.W.B. and Z.L. performed the experiments. M.S. and B. Guo performed the prostate cancer mouse studies. T.J.L. performed the breast cancer mouse studies. P.R. performed the colorectal cancer mouse studies. P.G. and F.H. supervised the project. P.G., C.M.C. and B.M.E. provided the funding and resources. P.G., F.P., F.H. and D.W.B. co-wrote the manuscript, and all authors refined the manuscript.

Competing interests

P.G.’s Sylvan G. Frank Endowed Chair position in Pharmaceutics and Drug Delivery is funded by the CM Chen Foundation, and he is a consultant of Oxford Nanopore, Nanobio Delivery Pharmaceutical Co., Ltd, and the cofounder of P&Z Biological Technology LLC. F.P. now works for Nanobio Delivery Pharmaceutical Co., Ltd. S.W. and F.H. now work for P&Z Biological Technology LLC.

Corresponding author

Correspondence to Peixuan Guo.

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