RGD Reference Report - Nanoparticle-mediated delivery of shRNA.VEGF-a plasmids regresses corneal neovascularization. - Rat Genome Database

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Nanoparticle-mediated delivery of shRNA.VEGF-a plasmids regresses corneal neovascularization.

Authors: Qazi, Y  Stagg, B  Singh, N  Singh, S  Zhang, X  Luo, L  Simonis, J  Kompella, UB  Ambati, BK 
Citation: Qazi Y, etal., Invest Ophthalmol Vis Sci. 2012 May 14;53(6):2837-44. doi: 10.1167/iovs.11-9139.
RGD ID: 7483619
Pubmed: (View Article at PubMed) PMID:22467572
DOI: Full-text: DOI:10.1167/iovs.11-9139

PURPOSE: To determine the efficacy of a plasmid containing a small hairpin RNA expression cassette (pSEC.shRNA) against VEGF-A-loaded poly(lactic co-glycolic acid) nanoparticles (PLGA NPs) in the sustained regression of murine corneal neovascularization. METHODS: PLGA nanoparticles were loaded with pSEC.shRNA.VEGF-A plasmids using the double emulsion-solvent evaporation method. KNV was induced in BALB/c mice by mechanical-alkali injury. Four weeks after induction of KNV, the mice were randomly divided to receive one of four treatments intrastromally: pSEC.shRNA.VEGF-A PLGA NPs (2 mug plasmid); naked pSEC.shRNA.VEGF-A plasmid only (2 mug plasmid); control blank PLGA NPs (equivalent dry weight of NPs); and vehicle. Two and five days after intervention, corneas were harvested to determine VEGF-A gene and protein expression using reverse transcriptase polymerase chain reaction and ELISA, respectively. Four weeks after intervention, corneas were photographed, mice sacrificed, and the corneal whole mounts were immunostained for CD31 (panendothelial cell marker). Immunofluorescence microscopy was performed and the neovascular area was quantitated. RESULTS: VEGF-A mRNA (49.6 +/- 12.4 vs. 82.9 +/- 6.0%, P < 0.01) and protein (4.0 +/- 5.2 vs. 20.0 +/- 7.5 rhog VEGF-A/mg total protein, P < 0.05) expression were significantly reduced in pSEC.shRNA.VEGF-A PLGA NP-treated corneas as compared with control blank NP. The pSEC.shRNA.VEGF-A PLGA NP-treated corneas showed significant regression in the mean fractional areas of KNV (0.125 +/- 0.042; 12.5%, P <0.01) compared with both naked plasmid only (0.283 +/- 0.004; 28.3%) and control (blank NPs = 0.555 +/- 0.072, 55.5%) at 4 weeks post-treatment. CONCLUSIONS: The pSEC.shRNA.VEGF-A-loaded PLGA NPs are an effective, nonviral, nontoxic, and sustainable form of gene therapy for the regression of murine KNV.

Annotation

Disease Annotations    

Objects Annotated

Genes (Rattus norvegicus)
Vegfa  (vascular endothelial growth factor A)

Genes (Mus musculus)
Vegfa  (vascular endothelial growth factor A)

Genes (Homo sapiens)
VEGFA  (vascular endothelial growth factor A)


Additional Information