Vascular endothelial growth factors (VEGFs) are best known as key regulators of angiogenesis and lymphangiogenesis. Although VEGFs have been promising therapeutic targets for various cardiovascular related diseases, their regulatory landscape in endothelial cells remains elusive. Several studies have highlighted the involvement of non-coding RNAs (ncRNAs) in the modulation of VEGF expression. In this study, we investigated the role of two classes of non-coding RNAs, long ncRNA (lncRNA) and enhancer RNAs (eRNAs), in the transcriptional regulation of VEGFA and VEGFC. By integrating genome-wide global run-on (GRO-seq) and Hi-C data, we identified putative lncRNAs and eRNAs associated with VEGFA and VEGFC genes in endothelial cells. A subset of the identified putative enhancers demonstrated regulatory activity in a reporter assay. Importantly, we demonstrate that deletion of enhancers and lncRNAs by CRISPR/Cas9 promoted significant changes in VEGFA and VEGFC expression. RNA-seq data from lncRNA deletions showed downstream factors implicated in VEGFA and VEGFC linked pathways, such as angiogenesis and lymphangiogenesis, suggesting functional roles for these lncRNAs. Our study uncovers novel lncRNAs and eRNAs regulating VEGFA and VEGFC that can be targeted to modulate the expression of these important molecules in endothelial cells.

中文翻译：

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调节内皮细胞中 VEGFA 和 VEGFC 表达的非编码 RNA 的基因组图谱

血管内皮生长因子 (VEGF) 被认为是血管生成和淋巴管生成的关键调节因子。尽管 VEGF 一直是各种心血管相关疾病的有希望的治疗靶点，但它们在内皮细胞中的调节作用仍然难以捉摸。多项研究强调非编码 RNA (ncRNA) 参与VEGF表达的调节。在这项研究中，我们研究了两类非编码RNA，即长ncRNA (lncRNA) 和增强子RNA (eRNA) 在VEGFA和VEGFC转录调控中的作用。通过整合全基因组全局连续运行 (GRO-seq) 和 Hi-C 数据，我们鉴定了与内皮细胞中VEGFA和VEGFC基因相关的推定 lncRNA 和 eRNA。已鉴定的推定增强子的一个子集在报告基因检测中表现出调节活性。重要的是，我们证明通过 CRISPR/Cas9 删除增强子和 lncRNA 可促进VEGFA和VEGFC表达的显着变化。来自 lncRNA 缺失的 RNA-seq 数据显示与VEGFA和VEGFC相关途径有关的下游因子，例如血管生成和淋巴管生成，表明这些 lncRNA 的功能作用。我们的研究发现了调节VEGFA和VEGFC的新型 lncRNA 和 eRNA ，可以有针对性地调节内皮细胞中这些重要分子的表达。