Dvoriantchikova Galina, Pappas Steve, Luo Xueting, Ribeiro Marcio, Danek Dagmara, Pelaez Daniel, Park Kevin K, Ivanov Dmitry
Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA.
Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, USA.
Eur J Neurosci. 2016 Dec;44(11):2935-2943. doi: 10.1111/ejn.13383. Epub 2016 Sep 13.
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.
由于轴突损伤和视网膜神经节细胞(RGC)丢失会导致失明,因此能够提高RGC存活率和轴突再生的疗法具有直接的临床意义。鉴于NFκB信号传导对神经元存活至关重要且可能调节神经突生长,我们研究了NFκB信号传导在RGC存活和轴突再生中的治疗潜力。尽管RGC中同时存在NFκB两个亚基(p65和p50),但当RGC处于神经毒性条件下时,p65以无活性(未磷酸化)状态存在。在本研究中,我们采用磷酸模拟方法生成编码活化(磷酸化)p65(p65mut)的DNA,然后使用2型腺相关病毒(AAV2)将该DNA导入RGC。我们测试了组成型p65mut表达是否能预防遭受短暂性视网膜缺血或视神经挤压(ONC)这两种神经毒性模型的RGC死亡并促进其神经突生长。我们的数据表明,在ONC模型中,与对照组相比,用AAV2-p65mut处理的RGC存活率显著增加(77±7%对25±3%,P值=0.0001)。我们还发现修饰后的p65对缺血视网膜的RGC有保护作用(55±12%对35±6%),但未达到统计学显著程度(P值=0.14)。在ONC后,我们未检测到实验动物和对照动物在轴突再生方面的差异。这些发现表明,RGC中NFκB信号传导增加可减轻神经退行性动物模型中的视网膜损伤,但对轴突再生的影响不显著。