Although viral carriers in gene therapy have shown promising results, potential immunogenicity and host chromosomal integration still remain, which causes a drawback in human application. We propose a new polymer carrier, polyamido ethylenimine containing multiple disulfide bonds (SS-PAEI). This well characterized polymer presented higher level of reporter gene expression with a significantly lower cellular toxicity than conventional high molecular weight polyethylenimine. It has been reported that glucagon-like peptide 1 (GLP-1) administration completely normalized blood glucose in type 2 diabetic patients. However, plasma half life is too short and continuous infusion of multiple injections is required for therapeutic use of GLP-1. In this application, construction of GLP-1 plasmid and its delivery with SS-PAEI are proposed for the treatment of type 2 diabetes. GLP-1 plasmid will be delivered by non-viral SS-PAEI carrier for prolonged time of GLP-1 production by gene delivery. This SS-PAEI carrier improves transfection efficiency, non-immunogenic and less difficult to use. Its high transfection is due to cytosomal destabilization and fast degradation to release DNA in the cells. In this application, we will develop an effective GLP-1 plasmid by selecting the optimal promoter/enhancer and transcription factor binding sites. GLP-1 plasmid will be constructed with strong promoter/enhancer. To increase nuclear import, the nuclear factor kappa B (NFkB) binding sites will be incorporated into plasmid. Since this nuclear import is mainly presented in cytoplasm and has nuclear localization signal, it will enhance nuclear import of DNA. In addition, two step transcriptional amplification (TSTA) will be employed to enhance transcriptional activity. The effectiveness of the designed delivery system will be evaluated by both in vitro transfection and in vivo experiment with type 2 diabetic animal model.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK077703-05
Application #
8197300
Study Section
Bioengineering, Technology and Surgical Sciences Study Section (BTSS)
Program Officer
Mckeon, Catherine T
Project Start
2008-01-15
Project End
2012-11-30
Budget Start
2011-12-01
Budget End
2012-11-30
Support Year
5
Fiscal Year
2012
Total Cost
$258,135
Indirect Cost
$86,617
Name
University of Utah
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Lee, Young Sook; Kim, Sung Wan (2014) Bioreducible polymers for therapeutic gene delivery. J Control Release 190:424-39
Kim, Tae-Il; Lee, Minhyung; Kim, Sung Wan (2012) Efficient GLP-1 gene delivery using two-step transcription amplification plasmid system with a secretion signal peptide and arginine-grafted bioreducible polymer. J Control Release 157:243-8
Kim, Sung Wan (2011) Biomaterials to gene delivery. J Control Release 155:116-8
Kim, Tae-il; Lee, Minhyung; Kim, Sung Wan (2010) A guanidinylated bioreducible polymer with high nuclear localization ability for gene delivery systems. Biomaterials 31:1798-804
Jeong, Ji Hoon; Kim, Sun Hwa; Christensen, Lane V et al. (2010) Reducible poly(amido ethylenimine)-based gene delivery system for improved nucleus trafficking of plasmid DNA. Bioconjug Chem 21:296-301
Jeong, Ji Hoon; Kim, Sun Hwa; Lee, Minhyung et al. (2010) Non-viral systemic delivery of Fas siRNA suppresses cyclophosphamide-induced diabetes in NOD mice. J Control Release 143:88-94
Kim, Tae-il; Ou, Mei; Lee, Minhyung et al. (2009) Arginine-grafted bioreducible poly(disulfide amine) for gene delivery systems. Biomaterials 30:658-64