背景:胎儿囊性水瘤(CH)的分子机制尚不清楚,以前没有研究报道CH中单细胞的转录组变化。在这项研究中,采用单细胞转录组测序(scRNA-seq)研究CH患者病变组织中细胞亚群的特征.
方法:从CH患者和对照供体收集的淋巴组织用于scRNA-seq分析。分析了主要细胞亚群中差异表达的基因富集以及细胞间的通讯。同时,分析了VEGF信号通路重要分子的表达和相互作用,并预测了可能与KDR(VEGFR2)结合的潜在转录因子。
结果:scRNA-seq结果显示,成纤维细胞在CH患者的淋巴病变中占最大比例。病例和对照组之间淋巴内皮细胞亚群的比例显着增加。VEGF信号通路富集于淋巴管内皮细胞,参与调控淋巴管内皮细胞与其他细胞间的细胞间通讯。VEGF信号通路中的关键调控基因KDR在CH患者中高表达,并与其他差异表达的EDN1、TAGLN、和CLDN5最后,我们发现STAT1可以与KDR启动子区结合,这可能在促进KDR上调中发挥重要作用。
结论:我们使用单细胞RNA测序对CH患者肿瘤组织中的细胞组成进行了全面描述,确定了CH中淋巴内皮细胞的富集,并强调了淋巴内皮细胞中VEGF信号通路的激活作为潜在的调节剂。胎儿囊性水瘤(CH)的分子和细胞发病机理仍然未知。这项研究通过单细胞转录组测序检查了每个细胞亚群的分布和基因表达特征以及淋巴管内皮细胞中VEGF信号传导在调节CH进展中的可能作用。CH中淋巴管内皮细胞的富集和淋巴管内皮细胞中VEGF信号通路的激活从细胞亚群的角度为CH的发病提供了一些线索。
BACKGROUND: The molecular mechanism of fetal cystic hygroma (CH) is still unclear, and no study has previously reported the transcriptome changes of single cells in CH. In this study, single-cell transcriptome sequencing (scRNA-seq) was used to investigate the characteristics of cell subsets in the lesion tissues of CH patients.
METHODS: Lymphoid tissue collected from CH patients and control donors for scRNA-seq analysis. Differentially expressed gene enrichment in major cell subpopulations as well as cell-cell communication were analyzed. At the same time, the expression and interactions of important VEGF signaling pathway molecules were analyzed, and potential transcription factors that could bind to KDR (VEGFR2) were predicted.
RESULTS: The results of scRNA-seq showed that fibroblasts accounted for the largest proportion in the lymphatic lesions of CH patients. There was a significant increase in the proportion of lymphatic endothelial cell subsets between the cases and controls. The VEGF signaling pathway is enriched in lymphatic endothelial cells and participates in the regulation of cell-cell communication between lymphatic endothelial cells and other cells. The key regulatory gene KDR in the VEGF signaling pathway is highly expressed in CH patients and interacts with other differentially expressed EDN1, TAGLN, and CLDN5 Finally, we found that STAT1 could bind to the KDR promoter region, which may play an important role in promoting KDR up-regulation.
CONCLUSIONS: Our comprehensive delineation of the cellular composition in tumor tissues of CH patients using single-cell RNA-sequencing identified the enrichment of lymphatic endothelial cells in CH and highlighted the activation of the VEGF signaling pathway in lymphoid endothelial cells as a potential modulator. The molecular and cellular pathogenesis of fetal cystic hygroma (CH) remains largely unknown. This study examined the distribution and gene expression signature of each cell subpopulation and the possible role of VEGF signaling in lymphatic endothelial cells in regulating the progression of CH by single-cell transcriptome sequencing. The enrichment of lymphatic endothelial cells in CH and the activation of the VEGF signaling pathway in lymphatic endothelial cells provide some clues to the pathogenesis of CH from the perspective of cell subpopulations.