Gene transfer using micellar nanovectors inhibits choroidal neovascularization in vivo

• Published in: PloS one 2011-12, Vol.6 (12), p.e28560
• Main Authors: Iriyama, Aya, Oba, Makoto, Ishii, Takehiko, Nishiyama, Nobuhiro, Kataoka, Kazunori, Tamaki, Yasuhiro, Yanagi, Yasuo
• Format: Article
• Language: English
• Subjects: Age; Angiogenesis; Animal experimentation; Animal models; Animals; Bacterial Proteins - metabolism; Bioengineering; Biology; Choroidal Neovascularization - metabolism; Deoxyribonucleic acid; Disease Models, Animal; Disease Progression; DNA; DNA - metabolism; Encapsulation; Engineering schools; Fluorescein; Fluorescein - pharmacology; Fluorescence; Gene expression; Gene therapy; Gene Transfer Techniques; Genes; Genetic Vectors; Immunization; In vivo methods and tests; Infection; Injection; Integrative medicine; Intravenous administration; Kinases; Laboratories; Laser Coagulation - methods; Ligands; Light Coagulation; Luminescent Proteins - metabolism; Macular degeneration; Macular Degeneration - metabolism; Macular Degeneration - pathology; Male; Materials Science; Medicine; Mice; Mice, Inbred C57BL; Micelles; Microscopy; Microscopy, Fluorescence - methods; Neovascularization; Permeability; Plasmids; Polymers - chemistry; Protein-tyrosine kinase; Proteins; Retina; Signal transduction; Transfection; Tyrosine; Vascular endothelial growth factor; Vascularization; Western blotting; Yellow fluorescent protein
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0028560

Age-related macular degeneration caused by choroidal neovascularization (CNV) remains difficult to be treated despite the recent advent of several treatment options. In this study, we investigated the in vivo angiogenic control by intravenous injection of polyion complex (PIC) micelle encapsulating plasmid DNA (pDNA) using a mice CNV model. The transfection efficiency of the PIC micelle was investigated using the laser-induced CNV in eight-week-old male C57 BJ/6 mice. Firstly, each mouse received intravenous injection of micelle encapsulating pDNA of Yellow Fluorescent Protein (pYFP) on days 1,3 and 5. The expression of YFP was analyzed using fluorescein microscopy and western blotting analysis. In the next experiments, each mouse received intravenous injection of micelle encapsulating pDNA of soluble Fms-like tyrosine kinase-1 (psFlt-1) 1,3 and 5 days after the induction of CNV and the CNV lesion was analyzed by choroidal flatmounts on day 7. Fluorescein microscopy and western blotting analysis revealed that the expression of YFP was confirmed in the CNV area after injection of the PIC micelle, but the expression was not detected neither in mice that received naked pDNA nor those without CNV. Furthermore, the CNV area in the mice that received intravenous injection of the psFlt-1-encapsulated PIC micelle was significantly reduced by 65% compared to that in control mice (p