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人神经祖细胞多能分化中差异长非编码 RNA 的鉴定和表观遗传分析。

Identification and epigenetic analysis of divergent long non-coding RNAs in multilineage differentiation of human Neural Progenitor Cells.

机构信息

a Cellular and Molecular Neuroscience , National Brain Research Centre , Manesar, Gurgaon , Haryana , India.

b Department of Biochemistry , All India Institute of Medical Sciences , New Delhi , India.

出版信息

RNA Biol. 2019 Jan;16(1):13-24. doi: 10.1080/15476286.2018.1553482. Epub 2018 Dec 27.

Abstract

Long non-coding RNAs have emerged as an important regulatory layer in biological systems. Of the various types of lncRNAs, one class (designated as divergent RNAs/XH), which is in head-to-head overlap with the coding genes, has emerged as a critical biotype that regulates development and cellular differentiation. This work aimed to analyze previously published data on differential expression, epigenetic and network analysis in order to demonstrate the association of divergent lncRNAs, a specific biotype with the differentiation of human neural progenitor cells (hNPCs). We have analyzed various available RNAseq databases that address the neuronal and astrocytic differentiation of hNPCs and identified differentially expressed lncRNAs (DELs) during cell-fate determination. Key DELs identified from the databases were experimentally verified by us in our in-vitro hNPC differentiation system. We also analyzed the change in promoter activity using ChIP-seq datasets of the histone markers H3K4me3 (activation) and H3K27me3 (inactivation) of these DELs. Additionally, we explored the change in the euchromatinization state of DELs (by analyzing DNase-seq data) during lineage-specific differentiation of hNPCs and performed their network analysis. We were able to identify differences between neuronal and astrocytic differentiation of hNPCs at the level of divergent DELs epigenetic markers, DNAase hypersensitive sites and gene expression network. Divergent lncRNAs are more involved in neuronal rather than astrocytic differentiation, while the sense downstream lncRNA biotype appears to be more involved in astrocytic differentiation. By studying the lncRNA involvement of distinct biotypes, we have been able to indicate the preferential role of a particular biotype during lineage-specific differentiation.

摘要

长非编码 RNA 已成为生物系统中重要的调控层。在各种类型的 lncRNA 中,一类(指定为发散 RNA/XH)与编码基因头对头重叠,已成为调节发育和细胞分化的关键生物型。这项工作旨在分析先前发表的关于差异表达、表观遗传和网络分析的数据,以证明发散 lncRNA(一种特定的生物型)与人类神经祖细胞(hNPC)分化的关联。我们分析了各种可用的 RNAseq 数据库,这些数据库涉及 hNPC 的神经元和星形胶质细胞分化,并确定了在细胞命运决定过程中差异表达的 lncRNA(DEL)。从数据库中鉴定的关键 DEL 通过我们在体外 hNPC 分化系统中的实验得到了验证。我们还使用这些 DEL 的组蛋白标记物 H3K4me3(激活)和 H3K27me3(失活)的 ChIP-seq 数据集分析了启动子活性的变化。此外,我们还探索了 hNPC 谱系特异性分化过程中 DEL 染色质状态的变化(通过分析 DNase-seq 数据),并对其进行了网络分析。我们能够在 hNPC 神经元和星形胶质细胞分化水平上,通过发散 DEL 的表观遗传标记物、DNA 酶超敏位点和基因表达网络来识别差异。发散 lncRNA 更多地参与神经元分化,而不是星形胶质细胞分化,而下游 sense lncRNA 生物型似乎更多地参与星形胶质细胞分化。通过研究不同生物型的 lncRNA 参与,我们能够表明在谱系特异性分化过程中特定生物型的优先作用。

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