Inhibition of in vitro VEGF expression and choroidal neovascularization by synthetic dendrimer peptide mediated delivery of a sense oligonucleotide

Marano, Robert J., Wimmer, Norbert, Kearns, Philip S., Thomas, Bradley G., Toth, Istvan, Brankov, Meliha and Rakoczy, P. Elizabeth (2004) Inhibition of in vitro VEGF expression and choroidal neovascularization by synthetic dendrimer peptide mediated delivery of a sense oligonucleotide. Experimental Eye Research, 79 4: 525-535. doi:10.1016/j.exer.2004.06.023


Author Marano, Robert J.
Wimmer, Norbert
Kearns, Philip S.
Thomas, Bradley G.
Toth, Istvan
Brankov, Meliha
Rakoczy, P. Elizabeth
Title Inhibition of in vitro VEGF expression and choroidal neovascularization by synthetic dendrimer peptide mediated delivery of a sense oligonucleotide
Journal name Experimental Eye Research   Check publisher's open access policy
ISSN 0014-4835
1096-0007
Publication date 2004-10-01
Sub-type Article (original research)
DOI 10.1016/j.exer.2004.06.023
Volume 79
Issue 4
Start page 525
End page 535
Total pages 11
Place of publication London, England
Publisher Academic Press
Collection year 2004
Language eng
Subject C1
250302 Biological and Medical Chemistry
780105 Biological sciences
Abstract 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 (chi = 81(.)51%) was significantly better (P = 0(.)0036) than those that possessed 4 positive charges (chi = 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 < 0(.)0001) reduced the severity of laser mediated CNV for up to two months post-injection. This study demonstrated that lipophilic, charged dendrimer mediated delivery of ODN-1 resulted in the down-regulation of in vitro VEGF expression. In addition, in vivo delivery of ODN-1 by two of the dendrimers resulted in significant inhibition of CNV in an inducible rat model. Time course studies showed that the dendrimer/ODN-1 complexes remained active for up to two months indicating the dendrimer compounds provided protection against the effects of nucleases. (C) 2004 Elsevier Ltd. All rights reserved.
Keyword Ophthalmology
Vegf
Dendrimer
Oligonucleotide
Lipidic Polyamide
Lipoamino Acids
Cationic Lipids
Choroid
Laser Photocoagulation
Transfection
Endothelial Growth-factor
Hypoxia-inducible Factor-1
Factor Messenger-rna
Antisense Oligonucleotides
Retinal Neovascularization
Isothermal Titration
Therapeutic Agents
Gene Delivery
Phosphorothioate
Transcription
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
2005 Higher Education Research Data Collection
School of Pharmacy Publications
 
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