PURPOSE To evaluate conjunctival cell microRNA (miRNAs) and mRNA expression in relation to observed phenotype of progressive limbal stem cell deficiency in a cohort of subjects with congenital aniridia with known genetic status. METHODS Using impression cytology, bulbar conjunctival cells were sampled from 20 subjects with congenital aniridia and 20 age and sex-matched healthy control subjects. RNA was extracted and miRNA and mRNA analyses were performed using microarrays. Results were related to severity of keratopathy and genetic cause of aniridia. RESULTS Of 2549 miRNAs, 21 were differentially expressed in aniridia relative to controls (fold change ≤ -1.5 or ≥ +1.5). Among these miR-204-5p, an inhibitor of corneal neovascularization, was downregulated 26.8-fold in severely vascularized corneas. At the mRNA level, 539 transcripts were differentially expressed (fold change ≤ -2 or ≥ +2), among these FOSB and FOS were upregulated 17.5 and 9.7-fold respectively, and JUN by 2.9-fold, all being components of the AP-1 transcription factor complex. Pathway analysis revealed enrichment of PI3K-Akt, MAPK, and Ras signaling pathways in aniridia. For several miRNAs and transcripts regulating retinoic acid metabolism, expression levels correlated with keratopathy severity and genetic status. CONCLUSION Strong dysregulation of key factors at the miRNA and mRNA level suggests that the conjunctiva in aniridia is abnormally maintained in a pro-angiogenic and proliferative state, and these changes are expressed in a PAX6 mutation-dependent manner. Additionally, retinoic acid metabolism is disrupted in severe, but not mild forms of the limbal stem cell deficiency in aniridia.

中文翻译：

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角膜缘干细胞缺乏症中异常的新生血管和增殖性结膜表型与先天性无虹膜中 PAX6 突变状态调节的 microRNA 和基因表达改变有关。

目的 评估结膜细胞 microRNA (miRNA) 和 mRNA 表达与在一组已知遗传状态的先天性无虹膜受试者中观察到的进行性角膜缘干细胞缺乏表型的关系。方法 使用印模细胞学，从 20 名患有先天性无虹膜症的受试者和 20 名年龄和性别匹配的健康对照受试者中采集球结膜细胞。提取 RNA 并使用微阵列进行 miRNA 和 mRNA 分析。结果与角膜病变的严重程度和无虹膜的遗传原因有关。结果 在 2549 个 miRNA 中，有 21 个在无虹膜中相对于对照有差异表达（倍数变化 ≤ -1.5 或 ≥ +1.5）。在这些角膜新生血管抑制剂 miR-204-5p 中，在严重血管化的角膜中下调了 26.8 倍。在 mRNA 水平上，539个转录本差异表达（倍数变化≤-2或≥+2），其中FOSB和FOS分别上调17.5和9.7倍，JUN上调2.9倍，都是AP-1转录因子复合物的组成部分。通路分析揭示了无虹膜中 PI3K-Akt、MAPK 和 Ras 信号通路的富集。对于调节视黄酸代谢的几种 miRNA 和转录物，表达水平与角膜病严重程度和遗传状态相关。结论 miRNA和mRNA水平关键因子的强烈失调表明无虹膜中的结膜异常维持在促血管生成和增殖状态，并且这些变化以PAX6突变依赖性方式表达。此外，视黄酸代谢严重中断，