iPSC衍生的视神经发育不全模型中基因表达异常但视网膜神经节细胞发生未减少。

IF 1.2 4区 医学 Q4 GENETICS & HEREDITY
Ophthalmic Genetics Pub Date : 2024-02-01 Epub Date: 2024-01-26 DOI:10.1080/13816810.2023.2253902
Jennifer G Aparicio, Hanno Hopp, Narine Harutyunyan, Carly Stewart, David Cobrinik, Mark Borchert
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引用次数: 0

摘要

背景:视神经发育不全(ONH)是永久性失明的主要先天性原因,其特征是出生时视网膜神经节细胞(RGC)缺陷。假设多因素发育事件是ONH及其经常相关的神经和内分泌异常的基础;然而,环境影响尚不清楚,基因基础也未被探索。本工作利用患者诱导多能干细胞(iPSC)衍生的视网膜类器官(RO)研究了遗传对ONH RGC产生和基因表达的影响。材料和方法:将ONH患者和对照组产生的iPSC分化为RO。使用免疫荧光和流式细胞术评估RGC的发生。收集流式分选的BRN3+细胞用于RNA提取用于RNA测序。使用DESeq2和edgeR评估差异基因表达。PANTHER用于在差异表达基因(DEG)中识别统计学上过度表达的本体。在ONH、自闭症和神经发育障碍(NDD)研究中,通过评估功能、突变限制和先前识别来区分对ONH高度感兴趣的DEG。结果:ONH和对照RO的RGC发生和存活率相似。在DESeq2和edgeR分析中均鉴定出70个基因的差异表达,代表 ~ DEG的百分比是随机研究参与者的4倍。DEG显示出验证的NDD基因和ONH外显子组变异基因过度表达的趋势。在DEG中,RAPGEF4和DMD具有最多的疾病相关特征。结论:ONH遗传背景与RGC发生受损无关,但与表现出疾病贡献潜力的DEG有关。这构成了ONH基因贡献的一些初步证据。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Aberrant gene expression yet undiminished retinal ganglion cell genesis in iPSC-derived models of optic nerve hypoplasia.

Background: Optic nerve hypoplasia (ONH), the leading congenital cause of permanent blindness, is characterized by a retinal ganglion cell (RGC) deficit at birth. Multifactorial developmental events are hypothesized to underlie ONH and its frequently associated neurologic and endocrine abnormalities; however, environmental influences are unclear and genetic underpinnings are unexplored. This work investigates the genetic contribution to ONH RGC production and gene expression using patient induced pluripotent stem cell (iPSC)-derived retinal organoids (ROs).

Materials and methods: iPSCs produced from ONH patients and controls were differentiated to ROs. RGC genesis was assessed using immunofluorescence and flow cytometry. Flow-sorted BRN3+ cells were collected for RNA extraction for RNA-Sequencing. Differential gene expression was assessed using DESeq2 and edgeR. PANTHER was employed to identify statistically over-represented ontologies among the differentially expressed genes (DEGs). DEGs of high interest to ONH were distinguished by assessing function, mutational constraint, and prior identification in ONH, autism and neurodevelopmental disorder (NDD) studies.

Results: RGC genesis and survival were similar in ONH and control ROs. Differential expression of 70 genes was identified in both DESeq2 and edgeR analyses, representing a ~ 4-fold higher percentage of DEGs than in randomized study participants. DEGs showed trends towards over-representation of validated NDD genes and ONH exome variant genes. Among the DEGs, RAPGEF4 and DMD had the greatest number of disease-relevant features.

Conclusions: ONH genetic background was not associated with impaired RGC genesis but was associated with DEGs exhibiting disease contribution potential. This constitutes some of the first evidence of a genetic contribution to ONH.

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来源期刊
Ophthalmic Genetics
Ophthalmic Genetics 医学-眼科学
CiteScore
2.40
自引率
8.30%
发文量
126
审稿时长
>12 weeks
期刊介绍: Ophthalmic Genetics accepts original papers, review articles and short communications on the clinical and molecular genetic aspects of ocular diseases.
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