Annual Grant Winners 2005-2006
Xudong Yaun, PH.D. – HPD College of Pharmacy
Ling Li, Ph.D. – HPD College of Dental Medicine
Mathew He, Ph.D. – Farquhar College of Arts and Sciences
Dean Andrés Malavé – HPD College of
Dean Robert Uchin – HPD College of Dental Medicine
Dean Don Rosenblum – Farquhar College of Arts and Sciences
Title: The Nano Self-Assembled
DSPE-PEG Micelle System for siRNA Drug Delivery
RNA interference (RNAi) is the process
where the introduction of double stranded RNA into a cell
inhibits gene expression in a sequence dependent fashion.
However, advanced drug delivery, systemic distribution and
long-term silencing of genes are necessary before gene therapy
can enter the clinical phase and eventually benefit patients.
It is essential that the drug carrier system be capable of
delivering these nucleic acid-based drugs to the target cells.
One emerging and promising technique is to use biodegradable
polymeric micelle drug delivery system. Polymeric (e.g. block
copolymer) micelles are amphiphilic molecules that will self-assemble
in aqueous solutions. The goals of this project are to fabricate,
optimize and characterize a nano self-assembled micelle system
for drug delivery of siRNA. The siRNA drug delivery is a
new, exciting and yet very challenging area in pharmaceutical
research. The long-term objective is to achieve enhanced
and targeted delivery of siRNA to cells using the proposed
polymeric micelle system.
The nano self-assembled DSPE-PEG
micelle delivery system of siRNA will be prepared and characterized
in terms of particle size, size distribution, formulation
stability and surface morphology using quasi-elastic light
scattering particle sizer, differential scanning calorimetry
and cryo scanning electron microscopy (SEM). The drug loading
efficiency, drug stability and in vitro evaluation of the
micelle delivery system will be studied using liquid-liquid
extraction, solid-phase extraction, UV, HPLC and dissolution
testing. Transfection studies of dsGFP siRNA loaded micelle
drug delivery system will be done in cell culture to test
the efficiency of silencing. RNase protection assay will
be used to determine the efficient of siRNA loading. The
RNA silencing effort will be determined by northern blot.
At the protein level, the green fluorescent intensity will
be monitored through a Fluorescent microscope. The cyto-toxicity
of the siRNA loaded micelle delivery system on targeted
cells will be monitored by MTT assay.