Virally delivered, constitutively active NFκB improves survival of injured retinal ganglion cells

Eur J Neurosci. 2016 Dec;44(11):2935-2943. doi: 10.1111/ejn.13383. Epub 2016 Sep 13.

Abstract

As axon damage and retinal ganglion cell (RGC) loss lead to blindness, therapies that increase RGC survival and axon regrowth have direct clinical relevance. Given that NFκB signaling is critical for neuronal survival and may regulate neurite growth, we investigated the therapeutic potential of NFκB signaling in RGC survival and axon regeneration. Although both NFκB subunits (p65 and p50) are present in RGCs, p65 exists in an inactive (unphosphorylated) state when RGCs are subjected to neurotoxic conditions. In this study, we used a phosphomimetic approach to generate DNA coding for an activated (phosphorylated) p65 (p65mut), then employed an adeno-associated virus serotype 2 (AAV2) to deliver the DNA into RGCs. We tested whether constitutive p65mut expression prevents death and facilitates neurite outgrowth in RGCs subjected to transient retinal ischemia or optic nerve crush (ONC), two models of neurotoxicity. Our data indicate that RGCs treated with AAV2-p65mut displayed a significant increase in survival compared to controls in ONC model (77 ± 7% vs. 25 ± 3%, P-value = 0.0001). We also found protective effect of modified p65 in RGCs of ischemic retinas (55 ± 12% vs. 35 ± 6%), but not to a statistically significant degree (P-value = 0.14). We did not detect a difference in axon regeneration between experimental and control animals after ONC. These findings suggest that increased NFκB signaling in RGCs attenuates retinal damage in animal models of neurodegeneration, but insignificantly impacts axon regeneration.

Keywords: adeno-associated virus serotype 2; optic nerve crush model; phosphomimetic; retinal ganglion cells; transient retinal ischemia model.

Publication types

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

MeSH terms

  • Animals
  • Axons / metabolism*
  • Axons / physiology
  • Cell Line
  • Cells, Cultured
  • Dependovirus / genetics
  • Genetic Therapy
  • Mice
  • Mice, Inbred C57BL
  • Nerve Regeneration*
  • Neuronal Outgrowth
  • Optic Nerve Injuries / metabolism*
  • Optic Nerve Injuries / therapy
  • Retinal Ganglion Cells / metabolism*
  • Transcription Factor RelA / genetics*
  • Transcription Factor RelA / metabolism

Substances

  • Transcription Factor RelA