Nano-delivery of siRNA and RNA-based splice-site switching oligoribonucleotides (SSO) is an important new approach with chemotherapeutic potential against cancer. Here we will focus on manufacturing RNA-bound nanoparticles (RNPs). For this purpose gold, manganese or zinc-based core nanoparticles (NPs) will be prepared and RNA attached via delivery-enhancing materials (protamine or PAMAM dendrimer). In addition to a delivery advantage, this approach will greatly enhance RNA's stability against chemical and enzymatic degradation. Stabilization of the RNA by the RNPs will be confirmed by electrophoretic and liquid chromatography analysis after exposure to accelerated chemical or enzymatic degradation assays. The RNPs will be further characterized by electron microscopy, UV, fluorescence and dynamic laser light scatter spectroscopy. Functional nuclear delivery of the RNPs will be evaluated in the Hela 705 splice-site switching assay by delivering a bound RNA oligonucleotide which overlaps the aberrant splice site, corrects splicing and switches-on Luciferase. Ultimately, as proof-of-principle to the important anti-cancer potential of this approach, biological activity of RNPs containing siRNA or SSO targeted against B-Raf-an important model cancer target will be investigated. Keywords: SSO=splice-site switching oligonucleotide, RNP=RN nanoparticle

Public Health Relevance

This project focuses on nano-manufacturing nanoparticles of gold, manganese and zinc with RNA bound to them (RNPs). In preliminary experiments we have formed such RNPs attaching RNA via delivery enhancing materials (protamine or alternatively PAMAM dendrimer). Given these data and our previous experience with these nanomaterials in combination with DNA, here we will show the RNPs will provide the RNA resistance to chemical and enzymatic degradation and substantial delivery advantage. Finally as proof-of- principle for the chemotherapeutic potential of our approach here we will demonstrate that RNPs can deliver siRNA or splice-site switching oligomers against B-Raf an important model cancer target. Thus this application addresses a very important barrier for the success of RNA- derived chemotherapy, namely how to combine RNA with nanomaterials so as to retain structure-function.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
1R15CA139390-01
Application #
7646776
Study Section
Nanotechnology Study Section (NANO)
Program Officer
Fu, Yali
Project Start
2009-04-01
Project End
2013-01-31
Budget Start
2009-04-01
Budget End
2013-01-31
Support Year
1
Fiscal Year
2009
Total Cost
$189,575
Indirect Cost
Name
Missouri State University
Department
Other Health Professions
Type
Other Domestic Higher Education
DUNS #
076255876
City
Springfield
State
MO
Country
United States
Zip Code
65897
Thomas, Spencer E; Comer, Jeffrey; Kim, Min Jung et al. (2018) Comparative functional dynamics studies on the enzyme nano-bio interface. Int J Nanomedicine 13:4523-4536
DeLong, Robert K; Curtis, Chandler B (2017) Toward RNA nanoparticle vaccines: synergizing RNA and inorganic nanoparticles to achieve immunopotentiation. Wiley Interdiscip Rev Nanomed Nanobiotechnol 9:
DeLong, Robert K; Mitchell, Jennifer A; Morris, R Tyler et al. (2017) Enzyme and Cancer Cell Selectivity of Nanoparticles: Inhibition of 3D Metastatic Phenotype and Experimental Melanoma by Zinc Oxide. J Biomed Nanotechnol 13:221-31
Ramani, Meghana; Mudge, Miranda C; Morris, R Tyler et al. (2017) Zinc Oxide Nanoparticle-Poly I:C RNA Complexes: Implication as Therapeutics against Experimental Melanoma. Mol Pharm 14:614-625
Hoffman, Amanda; Wu, Xiaotong; Wang, Jianjie et al. (2017) Two-Dimensional Fluorescence Difference Spectroscopy of ZnO and Mg Composites in the Detection of Physiological Protein and RNA Interactions. Materials (Basel) 10:
McCall, Jayden; Smith, Joshua J; Marquardt, Kelsey N et al. (2017) ZnO Nanoparticles Protect RNA from Degradation Better than DNA. Nanomaterials (Basel) 7:
Hurst, Miranda N; DeLong, Robert K (2016) Two-Dimensional Fluorescence Difference Spectroscopy to Characterize Nanoparticles and their Interactions. Sci Rep 6:33287
Dean, John M; DeLong, Robert K (2015) A high-throughput screening assay for the functional delivery of splice-switching oligonucleotides in human melanoma cells. Methods Mol Biol 1297:187-96
Bhaumik, A; Shearin, A M; Delong, R et al. (2014) Probing the Interaction at the Nano-Bio Interface Using Raman Spectroscopy: ZnO Nanoparticles and Adenosine Triphosphate Biomolecules. J Phys Chem C Nanomater Interfaces 118:18631-18639
Reyes-Reveles, Juan; Sedaghat-Herati, Reza; Gilley, David R et al. (2013) mPEG-PAMAM-G4 nucleic acid nanocomplexes: enhanced stability, RNase protection, and activity of splice switching oligomer and poly I:C RNA. Biomacromolecules 14:4108-15

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