Ophthalmology, Department of Clinical Sciences, Lund University, Lund, Sweden.
Ophthalmology, Department of Clinical Sciences, Lund University, Lund, Sweden.
Exp Eye Res. 2021 Nov;212:108752. doi: 10.1016/j.exer.2021.108752. Epub 2021 Aug 31.
Retinitis Pigmentosa represents a group of genetic disorders that cause progressive vision loss via degeneration of photoreceptors, but there is in principle no treatment available. For any therapy development, a deeper comprehension of the disease-leading mechanism(s) at the molecular level is needed. Here we focused on the cGMP-PKG system, which has been suggested to be a driver in several models of the disease. To gain insights in its downstream signaling we manipulated the cGMP-PKG system with the aid of organotypic retinal explant cultures from either a mouse-based disease model, i.e. the rd1 mouse, or its healthy wild-type counterpart (wt), by adding different types of cGMP analogues to either inhibit or activate PKG in retinal explants from rd1 and wt, respectively. An RNA sequencing was then performed to study the cGMP-PKG dependent transcriptome. Expression changes of gene sets related to specific pathways or functions, that fulfilled criteria involving that the changes should match PKG activation and inhibition, were determined via bioinformatics. The analyses highlighted that several gene sets linked to oxidative phosphorylation and mitochondrial pathways were regulated by this enzyme system. Specifically, the expression of such pathway components was upregulated in the rd1 treated with PKG inhibitor and downregulated in the wt with PKG activator treatment, suggesting that cGMP-PKG act as a negative regulator in this context. Downregulation of energy production pathways may thus play an integral part in the mechanism behind the degeneration for at least several RP mutations.
色素性视网膜炎是一组遗传性疾病,通过光感受器的变性导致进行性视力丧失,但原则上没有有效的治疗方法。对于任何治疗方法的开发,都需要在分子水平上更深入地了解疾病的主导机制。在这里,我们专注于 cGMP-PKG 系统,该系统已被认为是几种疾病模型中的驱动因素。为了深入了解其下游信号转导,我们利用来自基于小鼠的疾病模型(即 rd1 小鼠)或其健康野生型对照(wt)的器官型视网膜外植体培养物,通过向 rd1 和 wt 的视网膜外植体中添加不同类型的 cGMP 类似物,分别抑制或激活 PKG,从而操纵 cGMP-PKG 系统。然后进行 RNA 测序以研究 cGMP-PKG 依赖性转录组。通过生物信息学确定了与特定途径或功能相关的基因集的表达变化,这些变化满足了与 PKG 激活和抑制相匹配的标准。分析结果强调了几个与氧化磷酸化和线粒体途径相关的基因集受到该酶系统的调节。具体而言,在用 PKG 抑制剂处理的 rd1 中,这些途径成分的表达上调,而在用 PKG 激活剂处理的 wt 中表达下调,这表明 cGMP-PKG 在这种情况下充当负调节剂。因此,能量产生途径的下调可能在至少几种 RP 突变的退行性机制中起着重要作用。