Inhibition of choroidal and corneal pathologic neovascularization by Plgf1-de gene transfer

Invest Ophthalmol Vis Sci. 2012 Dec 5;53(13):7989-96. doi: 10.1167/iovs.12-10658.

Abstract

Purpose: Ocular neovascularization (NV), the primary cause of blindness, typically is treated via inhibition of VEGF-A activity. However, besides VEGF-A, other proteins of the same family, including VEGF-B and placental growth factor (PlGF, all together VEGFs), have a crucial role in the angiogenesis process. PlGF and VEGF, which form heterodimers if co-expressed, both are required for pathologic angiogenesis. We generated a PlGF1 variant, named PlGF1-DE, which is unable to bind and activate VEGFR-1, but retains the ability to form heterodimer. PlGF1-DE acts as dominant negative of VEGF-A and PlGF1wt through heterodimerization mechanism. The purpose of our study was to explore the therapeutic potential of Plgf1-de gene in choroid and cornea NV context.

Methods: In the model of laser-induced choroidal neovascularization (CNV), Plgf1-de gene, and as control Plgf1wt, LacZ, or gfp genes, were delivered using adeno-associated virus (AAV) vector by subretinal injection 14 days before the injury. After 7 days CNV volume was assessed. Corneal NV was induced by scrape or suture procedures. Expression vectors for PlGF1wt or PlGF1-DE, and as control the empty vector pCDNA3, were injected in the mouse cornea after the vascularization insults. NV was evaluated with CD31 and LYVE-1 immunostaining.

Results: The expression of Plgf1-de induced significant inhibition of choroidal and corneal NV by reducing VEGF-A homodimer production. Conversely, the delivery of Plgf1wt, despite induced similar reduction of VEGF-A production, did not affect NV.

Conclusions: Plgf1-de gene is a new therapeutic tool for the inhibition of VEGFs driven ocular NV.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Choroidal Neovascularization / metabolism
  • Choroidal Neovascularization / pathology
  • Choroidal Neovascularization / prevention & control*
  • Corneal Neovascularization / metabolism
  • Corneal Neovascularization / pathology
  • Corneal Neovascularization / prevention & control*
  • Dependovirus / genetics
  • Disease Models, Animal*
  • Electroretinography
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression / physiology
  • Gene Transfer Techniques
  • Genetic Therapy*
  • Genetic Vectors
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Placenta Growth Factor
  • Plasmids
  • Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
  • Pregnancy Proteins / genetics*
  • Real-Time Polymerase Chain Reaction
  • Tumor Cells, Cultured
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism
  • Vesicular Transport Proteins / metabolism

Substances

  • LYVE1 protein, human
  • PGF protein, human
  • Pgf protein, mouse
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Pregnancy Proteins
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Vesicular Transport Proteins
  • Placenta Growth Factor