Nanotechniques involve the creation, characterization, and modification of organized nanomaterials to serve as building blocks for the construction of submicro or nanoscale devices and systems with applications in technology and medicine. Living systems contain a wide variety of ordered macromolecular structures and powerful bionanomachines. The ingenious, novel design of the bacteriophage phi29 DNA packaging motor and its constituents has inspired the artificial synthesis and assembly of phi29 biomimetic motor and its components. The 30-nm nanomotor was geared by six copies of ATP-binding packaging RNAs (pRNAs). The structural versatility of pRNA coupled with its ability to form dimers, trimers, hexamers and patterned superstructures via interlocking loop interactions make it a promising tool for nanomachine fabrication, pathogen detection and drug/gene delivery. The low-resolution global structures of the pRNA and its dimer, trimer and hexamer have been probed by photoaffinity cross linking, chemical modification interference, compensatory modification, Cryo-AFM and 3D computer modeling. The long-term objective is to introduce exogenous components, nano-materials, modules, moieties, drugs, and other therapeutic reagents into the motor and develop it for various applications in nanotechnology and medicine. During the first four years of this grant, we have demonstrated that pRNA can be used as a building block for bottom-up assembly of polyvalent nanoparticles to deliver siRNA, ribozymes and other therapeutics to specific cells. A single molecule fluorescence dual-view imaging system was designed and constructed to count the number of pRNA copies within the constructed pRNA nanoparticles. The short-term objective of this application is to utilize prostate cancer as a model system to determine the feasibility of using the phi29 pRNA motor for targeted delivery of siRNA or other therapeutic molecules and treatment of cancerous diseases. Specifically, the novel RNA nanotechnology approach previously developed for the bottom-up assembly of multimeric pRNA nanoparticles will be exploited to construct polyvalent pRNA vehicles carrying an RNA aptamer to specifically bind to prostate surface antigens and a therapeutic siRNA targeting an anti-apoptosis factor for the prostate cancer treatment. Methods for large scale production of pRNA nanoparticles and construction of stable pRNA therapeutic nanoparticles resistant to RNase digestion in vivo will be developed. Approaches designed to combine therapy with the simultaneous detection of the therapeutic effect will be developed by exploiting the polyvalent nature of the pRNA nanoparticles. The efficacy in prostate cancer therapy, the biodistribution, pharmacokinetics, toxicity, off-target effects as well as the double stranded RNA induced interference will be evaluate in both prostate cancer cells and mice models.

Public Health Relevance

The novel RNA nanotechnology of bottom-up assembly for phi29 motor pRNA nanoparticles will be used to construct polyvalent pRNA nanoparticles for siRNA delivery in prostate cancer therapy. Prostate cancer will be used as a model system to determine the feasibility of using the phi29 motor pRNA nanoparticles for the targeted delivery of siRNA or other therapeutic molecules and treatment of cancerous diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
2R01EB003730-05
Application #
7654768
Study Section
Nanotechnology Study Section (NANO)
Program Officer
Henderson, Lori
Project Start
2004-09-01
Project End
2013-05-31
Budget Start
2009-07-01
Budget End
2010-05-31
Support Year
5
Fiscal Year
2009
Total Cost
$298,065
Indirect Cost
Name
University of Cincinnati
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
041064767
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Pi, Fengmei; Zhang, Hui; Li, Hui et al. (2017) RNA nanoparticles harboring annexin A2 aptamer can target ovarian cancer for tumor-specific doxorubicin delivery. Nanomedicine 13:1183-1193
Binzel, Daniel W; Khisamutdinov, Emil; Vieweger, Mario et al. (2016) Mechanism of three-component collision to produce ultrastable pRNA three-way junction of Phi29 DNA-packaging motor by kinetic assessment. RNA 22:1710-1718
Binzel, Daniel W; Shu, Yi; Li, Hui et al. (2016) Specific Delivery of MiRNA for High Efficient Inhibition of Prostate Cancer by RNA Nanotechnology. Mol Ther 24:1267-77
Li, Hui; Lee, Taek; Dziubla, Thomas et al. (2015) RNA as a stable polymer to build controllable and defined nanostructures for material and biomedical applications. Nano Today 10:631-655
Shu, Dan; Pi, Fengmei; Wang, Chi et al. (2015) New approach to develop ultra-high inhibitory drug using the power function of the stoichiometry of the targeted nanomachine or biocomplex. Nanomedicine (Lond) 10:1881-97
Li, Hui; Rychahou, Piotr G; Cui, Zheng et al. (2015) RNA Nanoparticles Derived from Three-Way Junction of Phi29 Motor pRNA Are Resistant to I-125 and Cs-131 Radiation. Nucleic Acid Ther 25:188-97
Rychahou, Piotr; Shu, Yi; Haque, Farzin et al. (2015) Methods and assays for specific targeting and delivery of RNA nanoparticles to cancer metastases. Methods Mol Biol 1297:121-35
Zhang, Hui; Pi, Fengmei; Shu, Dan et al. (2015) Using RNA nanoparticles with thermostable motifs and fluorogenic modules for real-time detection of RNA folding and turnover in prokaryotic and eukaryotic cells. Methods Mol Biol 1297:95-111
Khisamutdinov, Emil F; Bui, My Nguyen Hoan; Jasinski, Daniel et al. (2015) Simple Method for Constructing RNA Triangle, Square, Pentagon by Tuning Interior RNA 3WJ Angle from 60° to 90° or 108°. Methods Mol Biol 1316:181-93
Haque, Farzin; Guo, Peixuan (2015) Overview of methods in RNA nanotechnology: synthesis, purification, and characterization of RNA nanoparticles. Methods Mol Biol 1297:1-19

Showing the most recent 10 out of 71 publications