RNA interference (RNAi) is an endogenous pathway in eukaryotic cells for reducing the expression of a target mRNA through the introduction of complementary double-stranded RNA (dsRNA). The discovery of RNAi has provided a means for analysis of biological networks and identification of therapeutic targets. For reasons of immunogenicity, RNAi in mammalian cells is initiated by short interfering RNAs (siRNAs). Despite the intense study to date on the RNAi pathway and the use of siRNAs, the identification of the most active sequences and efficient delivery of those sequences to the cells of interest remain significant challenges. It is proposed here to address these challenges of siRNA selection and delivery through analysis of multiple steps in the RNAi pathway. The proposed work will focus on two approaches to optimizing siRNA function. First, the binding interactions of siRNAs with important RNAi pathway proteins, will be characterized. Each of the proteins is known to be central to the RNAi pathway, but their contributions to silencing are not fully-understood. Second, the interactions of siRNAs with delivery vehicles built from chemically-diverse oligomeric and polymeric nanoparticles will be quantitatively analyzed to determine those structural features that encourage complex formation, protection of the siRNAs from degradation, and release of siRNAs upon entry into the cell. The polymeric nanoparticles to be studied are readily modified to provide a means of creating, with exquisite control, a diverse array of vehicles that we will use to test variables such as amine density, polyethylene glycol modification, hydrophilicity, and binding cooperativity. The overall goal of the proposed research is to design siRNAs with maximal function through manipulation of the siRNA structure and sequence and the design of vehicles with optimal chemical and physical characteristics.

Public Health Relevance

RNA interference (RNAi), a technique for blocking the expression of a specific protein, has the potential to be an important therapeutic strategy. To realize this potential with maximum safety and activity, RNAi-based therapeutics must be designed to have optimal function for a variety of steps in their complex mechanism of action. The proposed work will address two of these important steps to provide guidelines for the design of highly-active, highly-specific therapeutics based on RNAi.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM089866-04
Application #
8535167
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Bender, Michael T
Project Start
2010-09-01
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
4
Fiscal Year
2013
Total Cost
$222,963
Indirect Cost
$69,152
Name
Michigan State University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Thompson, Ryan; Chan, Christina (2016) Signal transduction of the physical environment in the neural differentiation of stem cells. Technology (Singap World Sci) 4:1-8
Vocelle, Daniel; Chesniak, Olivia M; Malefyt, Amanda P et al. (2016) Dextran functionalization enhances nanoparticle-mediated siRNA delivery and silencing. Technology (Singap World Sci) 4:
Nath, Aritro; Chan, Christina (2016) Genetic alterations in fatty acid transport and metabolism genes are associated with metastatic progression and poor prognosis of human cancers. Sci Rep 6:18669
Bilgin, Betul; Nath, Aritro; Chan, Christina et al. (2016) Characterization of transcription factor response kinetics in parallel. BMC Biotechnol 16:62
Vocelle, Daniel; Chan, Christina; Walton, S Patrick (2015) How can novel microscopic approaches shed light on the function of nucleic acid-based drugs? Future Med Chem 7:1623-5
Nath, Aritro; Li, Irene; Roberts, Lewis R et al. (2015) Elevated free fatty acid uptake via CD36 promotes epithelial-mesenchymal transition in hepatocellular carcinoma. Sci Rep 5:14752
Fang, Liang; Cho, Hyun Ju; Chan, Christina et al. (2014) Binding site multiplicity with fatty acid ligands: implications for the regulation of PKR kinase autophosphorylation with palmitate. Proteins 82:2429-42
Zhang, Linxia; Liu, Li; Thompson, Ryan et al. (2014) CREB modulates calcium signaling in cAMP-induced bone marrow stromal cells (BMSCs). Cell Calcium 56:257-68
Liu, Li; Chan, Christina (2014) IPAF inflammasome is involved in interleukin-1* production from astrocytes, induced by palmitate; implications for Alzheimer's Disease. Neurobiol Aging 35:309-21
Liu, C; Baek, S; Kim, J et al. (2014) Effect of Static Pre-stretch Induced Surface Anisotropy on Orientation of Mesenchymal Stem Cells. Cell Mol Bioeng 7:106-121

Showing the most recent 10 out of 32 publications