Glial Cell Responses and Gene Expression Dynamics in Retinas of Treated and Untreated RPE65 Mutant Dogs.

IF 5 2区 医学 Q1 OPHTHALMOLOGY Investigative ophthalmology & visual science Pub Date : 2024-10-01 DOI:10.1167/iovs.65.12.18
Tatyana Appelbaum, Evelyn Santana, David A Smith, William A Beltran, Gustavo D Aguirre
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Abstract

Purpose: The long-term evaluation of RPE65 gene augmentation initiated in middle-aged RPE65 mutant dogs previously uncovered notable inter-animal and intra-retinal variations in treatment efficacy. The study aims to gain deeper insights into the status of mutant retinas and assess the treatment impact.

Methods: Immunohistochemistry utilizing cell-specific markers and reverse transcription-quantitative PCR (RT-qPCR) analysis were conducted on archival retinal sections from normal and RPE65 mutant dogs.

Results: Untreated middle-aged mutant retinas exhibited marked downregulation in the majority of 20 examined genes associated with key retinal pathways. These changes were accompanied by a moderate increase in microglia numbers, altered expression patterns of glial-neuronal transmitter recycling proteins, and gliotic responses in Müller glia. Analysis of advanced-aged mutant dogs revealed mild outer nuclear layer loss in the treated eye compared to moderate loss in the corresponding retinal regions of the untreated control eye. However, persistent Müller glial stress response along with photoreceptor synapse loss were evident in both treated and untreated eyes. Photoreceptor synaptic remodeling, infrequent in treated regions, was observed in all untreated advanced-aged retinas, accompanied by a progressive increase in microglial cells indicative of ongoing inflammation. Interestingly, about half of the examined genes showed similar expression levels between treated and untreated advanced-aged mutant retinas, with some reaching normal levels.

Conclusions: Gene expression data suggest a shift from pro-degenerative mechanisms in middle-aged mutant retinas to more compensatory mechanisms in preserved retinal regions at advanced stages, despite ongoing degeneration. Such shift, potentially attributed to a number of surviving resilient cells, may influence disease patterns and treatment outcomes.

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经治疗和未经治疗的 RPE65 突变体犬视网膜中神经胶质细胞的反应和基因表达动态。
目的:对中年RPE65突变犬进行RPE65基因扩增的长期评估发现,治疗效果在动物间和视网膜内存在显著差异。本研究旨在深入了解突变视网膜的状况并评估治疗效果:方法:利用细胞特异性标记物进行免疫组化,并对正常狗和 RPE65 突变狗的档案视网膜切片进行逆转录定量 PCR(RT-qPCR)分析:结果:未经处理的中年突变体视网膜显示出与关键视网膜通路相关的 20 个受检基因中的大多数基因明显下调。这些变化伴随着小胶质细胞数量的适度增加、胶质细胞-神经元递质循环蛋白表达模式的改变以及 Müller 胶体的胶质细胞反应。对高龄突变狗的分析表明,与未经治疗的对照组眼睛的相应视网膜区域的中度损失相比,治疗组眼睛的外核层轻度损失。然而,在治疗眼和未治疗眼中,持续的Müller神经胶质应激反应和光感受器突触损失都很明显。在所有未经治疗的晚期视网膜中都观察到了光感受器突触重塑,但在治疗区域中这种现象并不常见,同时小胶质细胞逐渐增多,表明炎症仍在持续。有趣的是,大约一半的受检基因在治疗和未治疗的高龄突变视网膜中显示出相似的表达水平,其中一些达到了正常水平:基因表达数据表明,中年突变体视网膜中的促退化机制向晚期保留视网膜区域的代偿机制转变,尽管退化仍在继续。这种转变可能归因于一些存活的复原细胞,可能会影响疾病模式和治疗效果。
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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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