The overall goal for this project is to elucidate the mechanisms by which organic nanoparticles of controlled shape, controlled site-specific surface chemistry, tunable particle matrix composition and tunable modulus undergo endocytosis and to use these findings to engineer the intracellular release of siRNA within mammalian cells to achieve effective gene knockdown. This information, in combination with on-going efforts to understand the bio-distribution of shape controlled particles, will help to establish the rules for the rational design of nano-carriers for the effective in vivo delivery of siRNA. This will be accomplished using a unique particle synthesis method developed at the University of North Carolina called PRINT, Particle Replication in Non- wetting Templates. PRINT is an off-shoot of the emerging lithographic processes used to fabricate devices in the microelectronics industry.
In aims 1 and 2, the effect of nanoparticle composition, size, shape, surface charge and ligand choice on the cellular uptake of non-targeted and targeted nanoparticles will be examined. The kinetics of cellular internalization of the nanoparticles and the effect of charge and the spatial arrangement and the density of the ligands on the particle surface will be investigated with regard to specific pathways for cellular internalization of particles.
Aim 3 will explore the rational design of PRINT nanoparticles for non-targeted and targeted in vitro delivery of siRNA. As the PRINT particles enter a cell, the particulate carriers will release the siRNA based on a stimuli induced biological or chemical degradation mechanism. The effectiveness of intracellular delivery monitored by luciferase gene silencing will be evaluated as a function of particle matrix composition, particle size and shape and pathway of internalization.
Aim 4 will explore rational design of PRINT nanoparticles for targeted in vivo delivery of siRNA. The PRINT nanoparticles with encapsulated anti-luciferase siRNA will be decorated with cell specific ligands (folate, transferrin) and intravenously injected into tumor bearing mice. The efficacy and efficiency of siRNA delivery to the tumor will be evaluated as a function of particle surface and matrix chemistries. Understanding the detailed interplay between particle surface chemistry and shape on effective transfection both in vitro as well as in vivo is of significant importance.

Public Health Relevance

Small interfering RNA (siRNA) has the potential to revolutionize the treatment of a number of life threatening human diseases, particularly cancer, but one obstacle facing researchers and companies attempting to develop siRNA therapies, or any type of nucleic acid therapeutic, is efficient and specific delivery of the polyanionic molecules into the cells, tissues or organ systems of choice. Using a technique known as PRINTTM (Particle Replication in Non-wetting Templates), we are able to fabricate nanoparticles with precise control over the particle size, shape, composition, cargo and surface properties to create truly engineered drug therapies that can be used to overcome the obstacles facing researchers and to provide the tools for the rational design of nano-carriers for the effective delivery of therapeutics in vitro and in vivo.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB009565-04
Application #
8269998
Study Section
Special Emphasis Panel (ZRG1-NANO-M (01))
Program Officer
Tucker, Jessica
Project Start
2009-05-01
Project End
2014-04-30
Budget Start
2012-05-01
Budget End
2014-04-30
Support Year
4
Fiscal Year
2012
Total Cost
$311,584
Indirect Cost
$97,477
Name
University of North Carolina Chapel Hill
Department
None
Type
Organized Research Units
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
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Xu, Jing; Wong, Dominica H C; Byrne, James D et al. (2013) Future of the particle replication in nonwetting templates (PRINT) technology. Angew Chem Int Ed Engl 52:6580-9
Dunn, Stuart S; Byrne, James D; Perry, Jillian L et al. (2013) Generating Better Medicines for Cancer. ACS Macro Lett 2:393-397
Hasan, Warefta; Chu, Kevin; Gullapalli, Anuradha et al. (2012) Delivery of multiple siRNAs using lipid-coated PLGA nanoparticles for treatment of prostate cancer. Nano Lett 12:287-92
Parrott, Matthew C; Finniss, Mathew; Luft, J Chris et al. (2012) Incorporation and controlled release of silyl ether prodrugs from PRINT nanoparticles. J Am Chem Soc 134:7978-82
Dunn, Stuart S; Tian, Shaomin; Blake, Steven et al. (2012) Reductively responsive siRNA-conjugated hydrogel nanoparticles for gene silencing. J Am Chem Soc 134:7423-30
Perry, Jillian L; Herlihy, Kevin P; Napier, Mary E et al. (2011) PRINT: a novel platform toward shape and size specific nanoparticle theranostics. Acc Chem Res 44:990-8
Wang, Jin; Byrne, James D; Napier, Mary E et al. (2011) More effective nanomedicines through particle design. Small 7:1919-31
Wang, Jin; Tian, Shaomin; Petros, Robby A et al. (2010) The complex role of multivalency in nanoparticles targeting the transferrin receptor for cancer therapies. J Am Chem Soc 132:11306-13