PAX6 and PAX6(5a) Transcript Levels in Conjunctival and Corneal Epithelial Samples and in Epithelial Cell Models of Congenital Aniridia.

IF 1.1 4区医学 Q4 OPHTHALMOLOGY

Klinische Monatsblatter fur Augenheilkunde Pub Date : 2026-09-01 DOI:10.1055/a-2800-7868

Swarnali Kundu, Fabian Norbert Fries, Dalya Yapar, Lorenz Latta, Zhen Li, Nagy Zsolt Zoltán, Berthold Seitz, Thomas Tschernig, Nóra Szentmáry, Tanja Stachon

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引用次数: 0

Abstract

Abstract: PURPOSE: PAX6 haploinsufficiency plays a crucial role in the development of aniridia-associated keratopathy (AAK). Our purpose was to analyse PAX6 levels and the presence of different PAX6 isoforms in conjunctival and corneal epithelial samples and in epithelial cell models of congenital aniridia.

Abstract: METHODS: PAX6 and PAX6(5a) mRNA levels were analysed in the following control and aniridia-derived samples: (a) primary limbal epithelial cell cultures from healthy donors (Ctrl-LECs) and aniridia patients (AN-LECs); (b) LECs transfected with control (siRNA Ctrl-LECs) or PAX6-targeting siRNA (siRNA PAX6-LECs); (c) immortalised limbal epithelial cells (t-LSCs) and genome-edited t-LSCs PAX6±; (d) conjunctival impression cytology from healthy individuals (Ctrl-CjECs-IC), aniridia patients (AN-CjECs-IC), and post-mortem donors (Pm-CjECs-IC); (e) corneal impression cytology from healthy living (Ctrl-CECs-IC) and post-mortem donors (Pm-CECs-IC); and (f) corneal epithelium (Ctrl-CECs) from healthy subjects. mRNA was extracted and reverse-transcribed to cDNA. PAX6 and PAX6(5a) were quantified via TaqMan qPCR with dual-labelled probes.

Abstract: RESULTS: PAX6 mRNA was significantly reduced in AN-LECs, siRNA PAX6-LECs, t-LSCs, and t-LSCs PAX6± versus Ctrl-LECs (p = 0.022, 0.019, 0.006). In contrast, expression was significantly higher in Ctrl-CjECs-IC and Ctrl-CECs versus Ctrl-LECs (p < 0.001). Total PAX6 was significantly lower in AN-LECs, siRNA PAX6-LECs, t-LSCs PAX6±, and AN-CjECs-IC compared to their respective controls (p ≤ 0.039). PAX6(5a) levels were significantly lower than total PAX6 in certain examined groups (p ≤ 0.035).

Abstract: CONCLUSIONS: PAX6 mRNA levels vary significantly across different epithelial cell types and models of congenital aniridia, with reduced expression observed in aniridia-derived samples and PAX6-deficient cell lines. The alternative isoform, PAX6(5a), is expressed at markedly lower levels than full-length PAX6 in most analysed tissues. This highlights the importance of both total PAX6 and PAX6(5a) expression in ocular surface epithelial homeostasis and provide further insight into the molecular alterations underlying AAK.

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[先天性无毛症结膜和角膜上皮样本和上皮细胞模型中的PAX6和PAX6（5a）转录镜]。

摘要：目的：PAX6单倍功能不全在无虹膜相关性角膜病变（AAK）的发生发展中起着至关重要的作用。我们的目的是分析结膜和角膜上皮样本以及先天性无虹膜上皮细胞模型中PAX6的水平和不同PAX6亚型的存在。摘要：方法：分析以下对照和无虹膜来源样本中PAX6和PAX6(5a) mRNA水平：(a)健康供体（control - lecs）和无虹膜患者（AN-LECs）的原代角膜缘上皮细胞培养物；(b)转染对照（siRNA control -LECs）或靶向pax6的siRNA （siRNA PAX6-LECs）的LECs；(c)永生化的角膜缘上皮细胞（t-LSCs）和基因组编辑的t-LSCs PAX6±；(d)健康人（Ctrl-CjECs-IC）、无虹膜患者（AN-CjECs-IC）和死后供者（Pm-CjECs-IC）结膜印象细胞学；(e)健康供体（Ctrl-CECs-IC）和死后供体（Pm-CECs-IC）的角膜印象细胞学检查；(f)健康受试者角膜上皮（Ctrl-CECs）。提取mRNA并反转录为cDNA。采用TaqMan qPCR双标记探针对PAX6和PAX6（5a）进行定量。摘要：结果：AN-LECs、siRNA PAX6- lecs、t-LSCs和t-LSCs中PAX6±与control - lecs相比显著降低（p = 0.022、0.019、0.006）。摘要：结论：PAX6 mRNA水平在先天性无虹膜不同上皮细胞类型和模型中存在显著差异，在无虹膜来源样本和PAX6缺陷细胞系中表达降低。另一种亚型PAX6（5a）在大多数分析组织中的表达水平明显低于全长PAX6。这突出了PAX6和PAX6（5a）总表达在眼表上皮稳态中的重要性，并为AAK的分子改变提供了进一步的见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Klinische Monatsblatter fur Augenheilkunde 医学-眼科学

CiteScore

1.30

自引率

0.00%

发文量

235

审稿时长

4-8 weeks

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