Polyplex Micelles from Triblock Copolymers Composed of Tandemly Aligned Segments with Biocompatible, Endosomal Escaping, and DNA-Condensing Functions for Systemic Gene Delivery to Pancreatic Tumor Tissue
- 587 Downloads
For systemic gene delivery to pancreatic tumor tissues, we prepared a three-layered polyplex micelle equipped with biocompatibility, efficient endosomal escape, and pDNA condensation functions from three components tandemly aligned; poly(ethylene glycol) (PEG), a poly(aspartamide) derivative with a 1,2-diaminoethane moiety (PAsp(DET)), and poly(l-lysine).
Materials and Methods
The size and in vitro transfection efficacy of the polyplex micelles were determined by dynamic light scattering (DLS) and luciferase assay, respectively. The systemic gene delivery with the polyplex micelles was evaluated from enhanced green fluorescence protein (EGFP) expression in the tumor tissues.
The polyplex micelles were approximately 80 nm in size and had one order of magnitude higher in vitro transfection efficacy than that of a diblock copolymer as a control. With the aid of transforming growth factor (TGF)-β type I receptor (TβR-1) inhibitor, which enhances accumulation of macromolecular drugs in tumor tissues, the polyplex micelle from the triblock copolymer showed significant EGFP expression in the pancreatic tumor (BxPC3) tissues, mainly in the stromal regions including the vascular endothelial cells and fibroblasts.
The three-layered polyplex micelles were confirmed to be an effective gene delivery system to subcutaneously implanted pancreatic tumor tissues through systemic administration.
KEY WORDSgene delivery PEG polyplex micelle TGF-β inhibitor triblock copolymer
This work was financially supported by the Core Research Program for Evolutional Science and Technology (CREST) from the Japan Science and Technology Corporation (JST) as well as by Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT).
- 1.Wiley (2007) Gene Therapy Clinical Trials Worldwide, provided by the J. Gene Med. http://www.wiley.co.uk/genetherapy/clinical/ (accessed 17/01/08)
- 11.O. Boussif, F. Lezoualc'h, M. A. Zanta, M. D. Mergny, D. Scherman, B. Demeneix, and J. Behr. A versatile vector for gene and oligonucleotide transfer into cells in culture and in-vivo polyethylenimine. Proc. Natl. Acad. Sci. U. S. A. 92:7297–7301 (1995) doi: 10.1073/pnas.92.16.7297.PubMedCrossRefGoogle Scholar
- 13.N. Kanayama, S. Fukushima, N. Nishiyama, K. Itaka, W.-D. Jang, K. Miyata, Y. Yamasaki, U. Chung, and K. Kataoka. A PEG-based biocompatible block catiomer with high buffering capacity for the construction of polyplex micelles showing efficient gene transfer toward primary cells. Chem. Med. Chem. 1:439–444 (2006) doi: 10.1002/cmdc.200600008.PubMedGoogle Scholar
- 15.M. Han, Y. Bae, N. Nishiyama, K. Miyata, M. Oba, and K. Kataoka. Transfection study using multicellular tumor spheroids for screening non-viral polymeric gene vectors with low cytotoxicity and high transfection efficiencies. J. Control Release. 121:38–48 (2007a) doi: 10.1016/j.jconrel.2007.05.012.PubMedCrossRefGoogle Scholar
- 16.M. Han, Y. Bae, N. Nishiyama, and K. Kataoka. Gene delivery with poly(amino acid)-based block catiomer polyplex micelles against multicellular tumor spheroid. Abstracts of 13th International Symposium on Recent Advances in Drug Delivery Systems, Salt Lake City, UT, (2007b), pp. 128.Google Scholar
- 17.D. Akagi, M. Oba, H. Koyama, N. Nishiyama, S. Fukushima, T. Miyata, H. Nagawa, and K. Kataoka. Biocompatible micellar nanovectors achieve efficient gene transfer to vascular lesions without cytotoxicity and thrombus formation. Gene. Ther. 14:1029–1038 (2007) doi: 10.1038/sj.gt.3302945.PubMedCrossRefGoogle Scholar
- 19.K. Miyata, S. Fukushima, N. Nishiyama, Y. Yamasaki, and K. Kataoka. PEG-based block catiomers possessing DNA anchoring and endosomal escaping functions to form polyplex micelles with improved stability and high transfection efficacy. J. Control Release. 122:252–260 (2007) doi: 10.1016/j.jconrel.2007.06.020.PubMedCrossRefGoogle Scholar
- 20.S. Fukushima, K. Miyata, N. Nishiyama, N. Kanayama, Y. Yamasaki, and K. Kataoka. PEGylated polyplex micelles from triblock catiomers with spatially ordered layering of condensed pDNA and buffering units for enhanced intracellular gene delivery. J. Am. Chem. Soc. 127:2810–2811 (2005) doi: 10.1021/ja0440506.PubMedCrossRefGoogle Scholar
- 21.M. R. Kano, Y. Bae, C. Iwata, Y. Morishita, M. Yashiro, M. Oka, T. Fujii, A. Komuro, K. Kiyono, M. Kamiishi, K. Hirakawa, Y. Ouchi, N. Nishiyama, K. Kataoka, and K. Miyazono. Improvement of cancer-targeting therapy, using nanocarriers for intractable solid tumors by inhibition of TGF-beta signaling. Proc. Natl. Acad. Sci. U. S. A. 104:3460–3465 (2007) doi: 10.1073/pnas.0611660104.PubMedCrossRefGoogle Scholar
- 23.A. Koide, A. Kishimura, K. Osada, W. -D. Jang, Y. Yamasaki, and K. Kataoka. Semipermeable polymer vesicle (PICsome) self-assembled in aqueous medium from a pair of oppositely charged block copolymers: physiologically stable micro-/nanocontainers of water-soluble macromolecules. J. Am. Chem. Soc. 128:5988–5989 (2006) doi: 10.1021/ja057993r.PubMedCrossRefGoogle Scholar
- 26.K. Itaka, K. Yamauchi, A. Harada, K. Nakamura, H. Kawaguchi, and K. Kataoka. Polyion complex micelles from plasmid DNA and poly(ethylene glycol)-poly(l-lysine) block copolymer as serum-tolerable polyplex system: physicochemical properties of micelles relevant to gene transfection efficiency. Biomaterials. 24:4495–4506 (2003) doi: 10.1016/S0142-9612(03)00347-8.PubMedCrossRefGoogle Scholar