Guangdong Provincial Key Laboratory of South China Structural Heart Disease, Guangdong Cardiovascular Institute, Guangzhou, Guangdong 510080, P.R. China.
School of Medicine, South China University of Technology School of Medicine, Guangzhou, Guangdong 510080, P.R. China.
Mol Med Rep. 2020 Sep;22(3):2528-2536. doi: 10.3892/mmr.2020.11332. Epub 2020 Jul 10.
Pulmonary stenosis (PS) is a congenital heart disease characterized by a dynamic or fixed anatomic obstruction of blood flow from the right ventricle to the pulmonary arterial vasculature. In the present study, extracellular vesicle long RNAs (EVLRs) from pregnant females who had healthy infants or PS infants were analyzed by RNA sequencing, and their diagnostic potential for PS during pregnancy was evaluated. A method for the selection of genes that could be considered as informative for the prediction PS based on extracellular vesicles (EVs) from pregnant females using long‑read RNA sequencing was developed. Blood samples were collected from females carrying fetuses with PS and females carrying unaffected fetuses (n=6 in each group). Physical characterization of EVs was performed using nanoparticle tracking analysis, transmission electron microscopy and western blotting. EVLRs from plasma were profiled by RNA sequencing and mRNA co‑expression modules were constructed by weighted gene co‑expression network analysis (WGCNA). Gene Ontology (GO) enrichment analysis was used to predict the function of the genes in each module. Hub genes were filtered out based on WGCNA and visualized using Cytoscape. EVLRs consisted of mRNAs, microRNAs and long non‑coding RNA. Overall, 26 modules were identified containing 16,394 genes. All modules were independent of each other. One particular module, referred to as the blue module, was markedly different between the two groups. A total of 735 hub genes in the blue module were identified, of which 33 were visualized, demonstrating the connection between these hub genes. GO enrichment analysis demonstrated that the analyzed hub genes were enriched in 'glucose transport', 'ATP‑dependent chromatin remodeling', 'histone deacetylation', 'histone H3‑K4 methylation', 'DNA methylation', 'apoptotic signaling pathway' and 'glucocorticoid receptor signaling pathway'. The hub genes identified in this module may provide a genetic framework for prenatal PS diagnosis. Furthermore, functional analysis of these associated genes may provide a theoretical basis for further research on PS pathogenesis.
肺动脉瓣狭窄（PS）是一种先天性心脏病，其特征是右心室到肺动脉血管的血流存在动态或固定的解剖学阻塞。本研究通过 RNA 测序分析了来自健康婴儿或 PS 婴儿孕妇的细胞外囊泡长 RNA（EVLRs），并评估了其在怀孕期间诊断 PS 的潜力。开发了一种基于长读 RNA 测序从孕妇细胞外囊泡（EVs）中选择可被认为对预测 PS 有信息的基因的方法。从患有 PS 胎儿的女性和携带未受影响胎儿的女性（每组 6 例）采集血液样本。使用纳米颗粒跟踪分析、透射电子显微镜和 Western blot 对 EVs 的物理特性进行了表征。通过 RNA 测序对 EVLRs 进行了分析，并通过加权基因共表达网络分析（WGCNA）构建了 mRNA 共表达模块。基因本体论（GO）富集分析用于预测每个模块中基因的功能。基于 WGCNA 筛选出枢纽基因，并使用 Cytoscape 可视化。EVLRs 由 mRNA、microRNA 和长非编码 RNA 组成。总体而言，鉴定出 26 个模块，包含 16394 个基因。所有模块彼此独立。一个特别的模块，称为蓝色模块，在两组之间明显不同。在蓝色模块中总共鉴定出 735 个枢纽基因，其中 33 个可视化，显示了这些枢纽基因之间的联系。GO 富集分析表明，分析的枢纽基因在“葡萄糖转运”、“ATP 依赖性染色质重塑”、“组蛋白去乙酰化”、“组蛋白 H3-K4 甲基化”、“DNA 甲基化”、“凋亡信号通路”和“糖皮质激素受体信号通路”中富集。该模块中鉴定的枢纽基因可能为产前 PS 诊断提供遗传框架。此外，对这些相关基因的功能分析可为 PS 发病机制的进一步研究提供理论依据。
