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Abstract：
BACKGROUND: Polycystic ovary syndrome (PCOS) is a prevalent endocrine disorder affecting women of reproductive ages. Our previous study has implicated a possible link between RNA editing and PCOS, yet the actual role of RNA editing, its association with clinical features, and the underlying mechanisms remain unclear.
METHODS: Ten RNA-Seq datasets containing 269 samples of multiple tissue types, including granulosa cells, T helper cells, placenta, oocyte, endometrial stromal cells, endometrium, and adipose tissues, were retrieved from public databases. Peripheral blood samples were collected from twelve PCOS and ten controls and subjected to RNA-Seq. Transcriptome-wide RNA-Seq data analysis was conducted to identify differential RNA editing (DRE) between PCOS and controls. The functional significance of DRE was evaluated by luciferase reporter assays and overexpression in human HEK293T cells. Dehydroepiandrosterone and lipopolysaccharide were used to stimulate human KGN granulosa cells to evaluate gene expression.
RESULTS: RNA editing dysregulations across multiple tissues were found to be associated with PCOS in public datasets. Peripheral blood transcriptome analysis revealed 798 DRE events associated with PCOS. Through weighted gene co-expression network analysis, our results revealed a set of hub DRE events in PCOS blood. A DRE event in the eukaryotic translation initiation factor 2-alpha kinase 2 (EIF2AK2:chr2:37,100,559) was associated with PCOS clinical features such as luteinizing hormone (LH) and the ratio of LH over follicle-stimulating hormone. Luciferase assays, overexpression, and knockout of RNA editing enzyme adenosine deaminase RNA specific (ADAR) showed that the ADAR-mediated editing cis-regulated EIF2AK2 expression. EIAF2AK2 showed a higher expression after dehydroepiandrosterone and lipopolysaccharide stimulation, triggering changes in the downstrean MAPK pathway.
CONCLUSIONS: Our study presented the first evidence of cross-tissue RNA editing dysregulation in PCOS and its clinical associations. The dysregulation of RNA editing mediated by ADAR and the disrupted target EIF2AK2 may contribute to PCOS development via the MPAK pathway, underlining such epigenetic mechanisms in the disease.
METHODS: Ten RNA-Seq datasets containing 269 samples of multiple tissue types, including granulosa cells, T helper cells, placenta, oocyte, endometrial stromal cells, endometrium, and adipose tissues, were retrieved from public databases. Peripheral blood samples were collected from twelve PCOS and ten controls and subjected to RNA-Seq. Transcriptome-wide RNA-Seq data analysis was conducted to identify differential RNA editing (DRE) between PCOS and controls. The functional significance of DRE was evaluated by luciferase reporter assays and overexpression in human HEK293T cells. Dehydroepiandrosterone and lipopolysaccharide were used to stimulate human KGN granulosa cells to evaluate gene expression.
RESULTS: RNA editing dysregulations across multiple tissues were found to be associated with PCOS in public datasets. Peripheral blood transcriptome analysis revealed 798 DRE events associated with PCOS. Through weighted gene co-expression network analysis, our results revealed a set of hub DRE events in PCOS blood. A DRE event in the eukaryotic translation initiation factor 2-alpha kinase 2 (EIF2AK2:chr2:37,100,559) was associated with PCOS clinical features such as luteinizing hormone (LH) and the ratio of LH over follicle-stimulating hormone. Luciferase assays, overexpression, and knockout of RNA editing enzyme adenosine deaminase RNA specific (ADAR) showed that the ADAR-mediated editing cis-regulated EIF2AK2 expression. EIAF2AK2 showed a higher expression after dehydroepiandrosterone and lipopolysaccharide stimulation, triggering changes in the downstrean MAPK pathway.
CONCLUSIONS: Our study presented the first evidence of cross-tissue RNA editing dysregulation in PCOS and its clinical associations. The dysregulation of RNA editing mediated by ADAR and the disrupted target EIF2AK2 may contribute to PCOS development via the MPAK pathway, underlining such epigenetic mechanisms in the disease.
摘要：
背景：多囊卵巢综合征（PCOS）是一种影响育龄妇女的内分泌疾病。我们之前的研究暗示RNA编辑和PCOS之间可能存在联系，然而RNA编辑的实际作用，它与临床特征的关联，和潜在的机制仍然不清楚。
方法：十个RNA-Seq数据集，包含269个多种组织类型的样本，包括颗粒细胞,T辅助细胞,胎盘,卵母细胞,子宫内膜基质细胞，子宫内膜,和脂肪组织，是从公共数据库中检索的。从12个PCOS和10个对照收集外周血样品并进行RNA-Seq。进行全转录组RNA-Seq数据分析以鉴定PCOS和对照之间的差异RNA编辑(DRE)。通过荧光素酶报告基因测定和在人HEK293T细胞中的过表达来评估DRE的功能意义。使用脱氢表雄酮和脂多糖刺激人KGN颗粒细胞以评估基因表达。
结果：在公共数据集中，发现跨多个组织的RNA编辑失调与PCOS相关。外周血转录组分析显示798个DRE事件与PCOS相关。通过加权基因共表达网络分析,我们的结果揭示了PCOS血液中的一组中心DRE事件.真核翻译起始因子2-α激酶2（EIF2AK2：chr2：37,100,559）中的DRE事件与PCOS临床特征有关，例如黄体生成素（LH）和LH与卵泡刺激素的比率。荧光素酶测定，过表达,和RNA编辑酶腺苷脱氨酶RNA特异性(ADAR)的敲除表明ADAR介导的编辑顺式调节EIF2AK2的表达。EIAF2AK2在脱氢表雄酮和脂多糖刺激后显示出更高的表达，触发下游MAPK通路的变化。
结论：我们的研究提供了PCOS中跨组织RNA编辑失调及其临床关联的第一个证据。ADAR介导的RNA编辑失调和被破坏的靶标EIF2AK2可能通过MPAK途径促进PCOS的发育，强调这种疾病的表观遗传机制。
方法：十个RNA-Seq数据集，包含269个多种组织类型的样本，包括颗粒细胞,T辅助细胞,胎盘,卵母细胞,子宫内膜基质细胞，子宫内膜,和脂肪组织，是从公共数据库中检索的。从12个PCOS和10个对照收集外周血样品并进行RNA-Seq。进行全转录组RNA-Seq数据分析以鉴定PCOS和对照之间的差异RNA编辑(DRE)。通过荧光素酶报告基因测定和在人HEK293T细胞中的过表达来评估DRE的功能意义。使用脱氢表雄酮和脂多糖刺激人KGN颗粒细胞以评估基因表达。
结果：在公共数据集中，发现跨多个组织的RNA编辑失调与PCOS相关。外周血转录组分析显示798个DRE事件与PCOS相关。通过加权基因共表达网络分析,我们的结果揭示了PCOS血液中的一组中心DRE事件.真核翻译起始因子2-α激酶2（EIF2AK2：chr2：37,100,559）中的DRE事件与PCOS临床特征有关，例如黄体生成素（LH）和LH与卵泡刺激素的比率。荧光素酶测定，过表达,和RNA编辑酶腺苷脱氨酶RNA特异性(ADAR)的敲除表明ADAR介导的编辑顺式调节EIF2AK2的表达。EIAF2AK2在脱氢表雄酮和脂多糖刺激后显示出更高的表达，触发下游MAPK通路的变化。
结论：我们的研究提供了PCOS中跨组织RNA编辑失调及其临床关联的第一个证据。ADAR介导的RNA编辑失调和被破坏的靶标EIF2AK2可能通过MPAK途径促进PCOS的发育，强调这种疾病的表观遗传机制。
