The overall objective of this proposal is to develop biocompatible, biodegradable and self- assembling copolymer/DNA micelles to achieve efficient liver-targeted gene delivery. Liver is a critically important target for gene medicine applications because of the access of the transgene product to systemic circulation, and because it is the site of many metabolic genetic disorders, viral infection and malignancies. At present, the full potential of liver-targeted gene transfer is hindered by a lack of safe and efficient gene carriers. Polymeric micelles with a DNA/polycation complex core and a hydrophilic corona represent a promising carrier for liver-targeted gene delivery. These DNA-encapsulating micelles exhibit several desirable features that favor liver- targeted gene delivery, such as small size (80-100 nm), reduced interaction with biological components, prolonged circulation in blood, and low toxicity. Novel PEG-b-PPA/DNA micelles exhibit high DNA binding capacity, in vitro transfection efficiency, low cytotoxicity and good tissue biocompatibility. In this study, we will (1) synthesize and characterize a mini-library of PEG-b-PPA/DNA micelles with improved complex stability, colloidal stability, intracellular DNA release, endosomal escape, and hepatocyte-targeting ability;(2) characterize the transfection efficiency and cytotoxicity in rat primary hepatocytes, Kupffer cells and normal rat cholangiocytes for this series of structurally distinct copolymers/micelles, and correlate with micelle structures;(3) optimize the administration parameters to achieve efficient liver gene expression for both a non-secretory protein (luciferase) and a secretory protein (interferon-a2b). This will demonstrate the broad utility of this delivery strategy for expression of proteins intended for systemic distribution and for localized liver-specific diseases. Mechanistic studies will be performed in vitro and in vivo to understand the extracellular and intracellular transport of the micelles. This study integrates expertise in polymer chemistry, biomaterial design, molecular/cell biology and clinical sciences. It represents a systematic design and optimization of gene delivery strategies at multiple levels. This study will lay the foundation for future clinical application of polymeric micelles for liver-targeted gene therapy. Project Narrative The overall objective of this proposal is to develop safe and efficient non-viral nanoparticle carriers for gene delivery to the liver. Should this proposed study be successful, these new gene carriers will find wide clinical applications for treating inherited and metabolic liver diseases, liver cancer, viral hepatitis, and systemic diseases, like hemophilia A and B.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM073937-02S1
Application #
7913864
Study Section
Biomaterials and Biointerfaces Study Section (BMBI)
Program Officer
Okita, Richard T
Project Start
2008-04-01
Project End
2012-01-31
Budget Start
2009-07-01
Budget End
2010-01-31
Support Year
2
Fiscal Year
2009
Total Cost
$86,100
Indirect Cost
Name
Johns Hopkins University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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Jiang, Xuan; Qu, Wei; Pan, Deng et al. (2013) Plasmid-templated shape control of condensed DNA-block copolymer nanoparticles. Adv Mater 25:227-32
Patil, Rajesh R; Yu, Jianhua; Banerjee, Sangeeta R et al. (2011) Probing in vivo trafficking of polymer/DNA micellar nanoparticles using SPECT/CT imaging. Mol Ther 19:1626-35
Jiang, Xuan; Leong, Derek; Ren, Yong et al. (2011) String-like micellar nanoparticles formed by complexation of PEG-b-PPA and plasmid DNA and their transfection efficiency. Pharm Res 28:1317-27
Jiang, Xuan; Zheng, Yiran; Chen, Hunter H et al. (2010) Dual-sensitive micellar nanoparticles regulate DNA unpacking and enhance gene-delivery efficiency. Adv Mater 22:2556-60
Ren, Yong; Jiang, Xuan; Pan, Deng et al. (2010) Charge density and molecular weight of polyphosphoramidate gene carrier are key parameters influencing its DNA compaction ability and transfection efficiency. Biomacromolecules 11:3432-9