摘要
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.
基金
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.
