Abnormal neovascular and proliferative conjunctival phenotype in limbal stem cell deficiency is associated with altered microRNA and gene expression modulated by PAX6 mutational status in congenital aniridia
Abnormal neovascular and proliferative conjunctival phenotype in limbal stem cell deficiency is associated with altered microRNA and gene expression modulated by PAX6 mutational status in congenital aniridia
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DOI：
10.1016/j.jtos.2020.04.014
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发表时间：
2021-02-02
期刊：
影响因子：
6.4
通讯作者：
Kaesmann-Kellner, B.
中科院分区：
文献类型：
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作者：
Latta, L.;Ludwig, N.;Kaesmann-Kellner, B.
用途：在一组遗传状态已知的先天性无虹膜受试者中，评价结膜细胞microRNA（miRNA）和mRNA表达与观察到的进行性角膜缘干细胞缺乏表型的关系。研究方法：采用印迹细胞学方法，从20名先天性无虹膜受试者和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样本。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miRNAs中，21个在无虹膜中相对于对照差异表达（倍数变化= +1.5）。其中，角膜新生血管抑制剂miR-204- 5 p在严重血管化角膜中下调26.8倍。在mRNA水平上，有539种转录物差异表达（倍数变化= +2），其中FOSB和FOS分别上调17.5和9.7倍，JUN上调2.9倍，均为AP-1转录因子复合物的组分。通路分析显示PI 3 K-Akt、MAPK和Ras信号通路在无虹膜中富集。对于调节视黄酸代谢的几种miRNA和转录物，表达水平与角膜病变的严重程度和遗传状态相关。结论：关键因子在miRNA和mRNA水平的强烈失调表明，无虹膜的结膜异常地维持在促血管生成和增殖状态，并且这些变化以PAX 6突变依赖性方式表达。此外，视黄酸代谢在严重的无虹膜角膜缘干细胞缺乏症中被破坏，而不是轻度的无虹膜角膜缘干细胞缺乏症。
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). 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), 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.