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人视网膜色素上皮细胞和诱导多能干细胞分化过程中长链非编码RNA表达的特征分析

Characterization of lincRNA expression in the human retinal pigment epithelium and differentiated induced pluripotent stem cells.

作者信息

Au Elizabeth D, Fernandez-Godino Rosario, Kaczynksi Tadeusz J, Sousa Maria E, Farkas Michael H

机构信息

Department of Ophthalmology, Jacobs School of Medicine and Biomedical Science, State University of New York at Buffalo, Buffalo, NY, United States of America.

Ocular Genomics Institute, Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA, United States of America.

出版信息

PLoS One. 2017 Aug 24;12(8):e0183939. doi: 10.1371/journal.pone.0183939. eCollection 2017.

Abstract

Long intervening non-coding RNAs (lincRNAs) are increasingly being implicated as important factors in many aspects of cellular development, function, and disease, but remain poorly understood. In this study, we examine the human retinal pigment epithelium (RPE) lincRNA transcriptome using RNA-Seq data generated from human fetal RPE (fRPE), RPE derived from human induced pluripotent stem cells (iPS-RPE), and undifferentiated iPS (iPS). In addition, we determine the suitability of iPS-RPE, from a transcriptome standpoint, as a model for use in future studies of lincRNA structure and function. A comparison of gene and isoform expression across the whole transcriptome shows only minimal differences between all sample types, though fRPE and iPS-RPE show higher concordance than either shows with iPS. Notably, RPE signature genes show the highest degree of fRPE to iPS-RPE concordance, indicating that iPS-RPE cells provide a suitable model for use in future studies. An analysis of lincRNAs demonstrates high concordance between fRPE and iPS-RPE, but low concordance between either RPE and iPS. While most lincRNAs are expressed at low levels (RPKM < 10), there is a high degree of concordance among replicates within each sample type, suggesting the expression is consistent, even at levels subject to high variability. Finally, we identified and annotated 180 putative novel genes in the fRPE samples, a majority of which are also expressed in the iPS-RPE. Overall, this study represents the first characterization of lincRNA expression in the human RPE, and provides a model for studying the role lincRNAs play in RPE development, function, and disease.

摘要

长链非编码RNA(lincRNA)越来越被认为是细胞发育、功能和疾病诸多方面的重要因素,但人们对其仍知之甚少。在本研究中,我们利用从人胎儿视网膜色素上皮(fRPE)、人诱导多能干细胞来源的视网膜色素上皮(iPS-RPE)和未分化的诱导多能干细胞(iPS)生成的RNA测序数据,研究了人视网膜色素上皮的lincRNA转录组。此外,从转录组角度确定iPS-RPE作为未来lincRNA结构和功能研究模型的适用性。对整个转录组中基因和异构体表达的比较显示,所有样本类型之间只有极小的差异,尽管fRPE和iPS-RPE之间的一致性高于它们与iPS的一致性。值得注意的是,视网膜色素上皮特征基因在fRPE与iPS-RPE之间的一致性程度最高,这表明iPS-RPE细胞为未来的研究提供了一个合适的模型。对lincRNA的分析表明,fRPE和iPS-RPE之间具有高度一致性,但视网膜色素上皮与iPS之间的一致性较低。虽然大多数lincRNA表达水平较低(每百万映射 reads 中来自某基因每千碱基长度的 reads数<RPKM<10),但每种样本类型内的重复样本之间具有高度一致性,这表明即使在具有高变异性的水平上,表达也是一致的。最后,我们在fRPE样本中鉴定并注释了180个推定的新基因,其中大多数也在iPS-RPE中表达。总体而言,本研究首次对人视网膜色素上皮中lincRNA的表达进行了表征,并为研究lincRNA在视网膜色素上皮发育、功能和疾病中的作用提供了一个模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e76d/5570510/79b098a9dcab/pone.0183939.g001.jpg

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