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Targeted Knockdown of Overexpressed VEGFA or VEGF164 in Müller cells maintains retinal function by triggering

Silke Becker1, Haibo Wang1, Aaron B Simmons1

  • 1John A. Moran Eye Center, University of Utah, Salt Lake City, UT, USA.

Scientific Reports
|February 2, 2018
PubMed

Insights

Targeting Müller cell vascular endothelial growth factor A (VEGFA) with gene therapy reduced abnormal blood vessel growth in a rat model of retinopathy. This approach may offer a safer alternative for treating conditions like retinopathy of prematurity.

Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Genetics

Background:

  • Oxygen-induced retinopathy (OIR) mimics severe retinopathy of prematurity (ROP), characterized by Müller cell upregulation of vascular endothelial growth factor A (VEGFA).
  • Current anti-VEGF treatments for ROP raise safety concerns, necessitating alternative therapeutic strategies.
  • Selective targeting of VEGFA in Müller cells offers a potential approach to mitigate OIR-related neovascularization.

Purpose of the Study:

  • To evaluate the long-term effects of Müller cell-specific VEGFA knockdown on retinal structure and function in a rat OIR model.
  • To assess the efficacy of lentiviral vectors delivering short-hairpin RNAs against VEGFA or its VEGF164 isoform.
  • To investigate potential neuroprotective signaling pathways activated by VEGFA inhibition.

Main Methods:

  • Subretinal delivery of lentivirus expressing short-hairpin RNAs targeting VEGFA (L-VEGFAshRNA) or VEGF164 (L-VEGF164shRNA) in a rat OIR model.
  • Analysis of retinal neovascularization using lectin staining to quantify avascular and intravitreal neovascular areas.
  • Assessment of retinal structure via spectral-domain optical coherence tomography (OCT) and immunohistochemistry (IHC).
  • Evaluation of retinal function using Ganzfeld electroretinograms (ERGs).
  • Quantification of neurotrophic factor mRNA levels (EPO, BDNF, GDNF, NGF, NT-3) in retinas and assessment of EPO-induced signaling in cultured Müller cells.

Main Results:

  • Both L-VEGFAshRNA and L-VEGF164shRNA significantly reduced intravitreal neovascularization compared to controls, with no late recurrence observed.
  • OCT and IHC revealed alterations in retinal layer thicknesses in treated groups compared to controls.
  • Ganzfeld ERGs showed improved function in retinas treated with L-VEGFAshRNA or L-VEGF164shRNA.
  • L-VEGFAshRNA increased mRNA levels of EPO, BDNF, GDNF, NGF, and NT-3, while L-VEGF164shRNA only affected NT-3.
  • In vitro studies demonstrated that VEGF knockdown upregulated NT-3 and EPO, and EPO treatment activated neuroprotective pathways.

Conclusions:

  • Selective, long-term knockdown of Müller cell VEGFA, particularly the VEGF164 isoform, effectively reduces intravitreal neovascularization in OIR.
  • This targeted approach appears to preserve retinal structure and function, potentially offering a safer alternative to non-selective anti-VEGF therapies.
  • Upregulation of neurotrophic factors like EPO and NT-3 may contribute to the observed neuroprotective effects, warranting further investigation.

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