Inhibition of in vitro VEGF expression and choroidal neovascularization by synthetic dendrimer peptide mediated delivery of a sense oligonucleotide
| Autor: | M. Brankov, Istvan Toth, P. Elizabeth Rakoczy, Bradley G. Thomas, Norbert Wimmer, Philip Kearns, Robert J. Marano |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Vascular Endothelial Growth Factor A
Oligonucleotides Gene delivery Biology Transfection Structure-Activity Relationship Cellular and Molecular Neuroscience chemistry.chemical_compound In vivo Dendrimer medicine Animals Humans RNA Messenger Cells Cultured Messenger RNA Gene Transfer Techniques Rats Inbred Strains Genetic Therapy Molecular biology Choroidal Neovascularization Sensory Systems In vitro Rats Vascular endothelial growth factor Ophthalmology Choroidal neovascularization Gene Expression Regulation chemistry medicine.symptom |
| Zdroj: | Experimental Eye Research. 79:525-535 |
| ISSN: | 0014-4835 |
| DOI: | 10.1016/j.exer.2004.06.023 |
| Popis: | Ocular neovascularisation is the leading cause of blindness in developed countries and the most potent angiogenic factor associated with neovascularisation is vascular endothelial growth factor (VEGF). We have previously described a sense oligonucleotide (ODN-1) that possesses anti-human and rat VEGF activity. This paper describes the synthesis of lipid-lysine dendrimers and their subsequent ability to delivery ODN-1 to its target and mediate a reduction in VEGF concentration both in vitro and in vivo. Positively charged dendrimers were used to deliver ODN-1 into the nucleus of cultured D407 cells. The effects on VEGF mRNA transcription and protein expression were analysed using RT-PCR and ELISA, respectively. The most effective dendrimers in vitro were further investigated in vivo using an animal model of choroidal neovascularisation (CNV). All dendrimer/ODN-1 complexes mediated in a significant reduction in VEGF expression during an initial 24 hr period (40-60%). Several complexes maintained this level of VEGF reduction during a subsequent, second 24 hr period, which indicated protection of ODN-1 from the effects of endogenous nucleases. In addition, the transfection efficiency of dendrimers that possessed 8 positive charges (x=81.51%) was significantly better (P=0.0036) than those that possessed 4 positive charges (x=56.8%). RT-PCR revealed a correlation between levels of VEGF protein mRNA. These results indicated that the most effective structural combination was three branched chains of intermediate length with 8 positive charges such as that found for dendrimer 4. Dendrimer 4 and 7/ODN-1 complexes were subsequently chosen for in vivo analysis. Fluorescein angiography demonstrated that both dendrimers significantly (P |
| Databáze: | OpenAIRE |
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