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3D RNA nanocage for encapsulation and shielding of hydrophobic biomolecules to improve the in vivo biodistribution

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Abstract

Ribonucleic acid (RNA) nanotechnology platforms have the potential of harboring therapeutics for in vivo delivery in disease treatment. However, the nonspecific interaction between the harbored hydrophobic drugs and cells or other components before reaching the diseased site has been an obstacle in drug delivery. Here we report an encapsulation strategy to prevent such nonspecific hydrophobic interactions in vitro and in vivo based on a self-assembled three-dimensional (3D) RNA nanocage. By placing an RNA three-way junction (3WJ) in the cavity of the nanocage, the conjugated hydrophobic molecules were specifically positioned within the nanocage, preventing their exposure to the biological environment. The assembly of the nanocages was characterized by native polyacrylamide gel electrophoresis (PAGE), atomic force microscopy (AFM), and cryogenic electron microscopy (cryo-EM) imaging. The stealth effect of the nanocage for hydrophobic molecules in vitro was evaluated by gel electrophoresis, flow cytometry, and confocal microscopy. The in vivo sheathing effect of the nanocage for hydrophobic molecules was assessed by biodistribution profiling in mice. The RNA nanocages with hydrophobic biomolecules underwent faster clearance in liver and spleen in comparison to their counterparts. Therefore, this encapsulation strategy holds promise for in vivo delivery of hydrophobic drugs for disease treatment.

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Acknowledgements

The research in P. G.’s lab was supported by NIH grants (Nos. R01EB019036, U01CA151648, and U01CA207946) to Peixuan Guo. The cryo-EM work was supported by NIH grant No. 5941GM103832 (W. C.) and Office of Naval Research grant No. N00014-20-1-2084 (W. C.). P. G.’s Sylvan G. Frank Endowed Chair position in Pharmaceutics and Drug Delivery is funded by the CM Chen Foundation. The authors would like to thank the Nanoimaging Core Facility at UNMC for assistance with AFM imaging. The facility is in part supported by funds received from the Nebraska Research Initiative (NRI). The animal study protocol was approved by the Institutional Animal Care and Use Committee at The Ohio State University. The authors would like to thank Alyssa Castillo for help in sample preparation.

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Authors and Affiliations

  1. Center for RNA Nanobiotechnology and Nanomedicine, College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry, College of Medicine, Dorothy M. Davis Heart and Lung Research Institute, James Comprehensive Cancer Center, The Ohio State University, Columbus, OH, 43210, USA

    Congcong Xu, Hongran Yin, Zhefeng Li, Zhen Zheng, Zhouxiang Ji, Sijin Guo & Peixuan Guo

  2. Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA, 94305, USA

    Kaiming Zhang, Shanshan Li & Wah Chiu

  3. Department of Physics, University of Nebraska at Omaha, Omaha, NE, 68182, USA

    Alexey Krasnoslobodtsev

  4. Nanoimaging Core Facility, Office of Vice-Chancellor for Research, University of Nebraska Medical Center, Omaha, NE, 68198, USA

    Alexey Krasnoslobodtsev

  5. SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA, 94025, USA

    Wah Chiu

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  1. Congcong Xu
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  2. Kaiming Zhang
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  3. Hongran Yin
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  4. Zhefeng Li
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  7. Zhouxiang Ji
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  9. Shanshan Li
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  11. Peixuan Guo
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Correspondence to Peixuan Guo.

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Conflict of interests

P. G. is the cofounder of ExonanoRNA LLC. He is also the consultant of Oxford Nanopore Technologies and Nanobio Delivery Pharmaceutical Co. Ltd, as well as the cofounder of Shenzhen P&Z Bio-medical Co. Ltd and its subsidiary US P&Z Biological Technology LLC. His inventions at the University of Kentucky have been licensed to Matt Holding and Nanobio Delivery Pharmaceutical Co., Ltd. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

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Xu, C., Zhang, K., Yin, H. et al. 3D RNA nanocage for encapsulation and shielding of hydrophobic biomolecules to improve the in vivo biodistribution. Nano Res. 13, 3241–3247 (2020). https://doi.org/10.1007/s12274-020-2996-1

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