大隐静脉(GSV)是冠状动脉旁路移植术中最常用的导管。然而,GSV的状态,包括代谢功能障碍,如糖尿病(DM)并发症,与静脉移植失败(VGF)密切相关。迄今为止,VGF的分子机制仍然难以捉摸。GSVs中细胞组分的详细表征和相应的表达调节对于减少VGF的临床决策非常重要。
为此,我们进行了单细胞RNA测序,以描绘三个人GSV样本中的细胞异质性.
细胞组成和表达的仔细研究揭示了人类GSVs中的细胞多样性,特别是内皮细胞(ECs)。我们的结果揭示了功能适应驱动静脉内皮细胞和瓣膜内皮细胞之间巨大的表达差异。例如,细胞表面受体ACKR1标定静脉内皮,而ACRK3/ACKR4仅由瓣膜内皮细胞表达。差异基因表达分析表明,静脉内皮细胞高表达的基因主要参与血管发育和白细胞粘附的调节,而瓣膜内皮细胞具有更明显的参与细胞外基质组织的基因表达,骨化和血小板脱颗粒。值得注意的是,伪时间轨迹分析提供了证据,表明静脉内皮细胞,瓣膜内皮细胞和淋巴管发育连接。Further,瓣膜内皮细胞可能是成人淋巴管分化的重要来源。此外,我们发现静脉EC亚群高表达IL6,这可能与不良预后相关.同时,我们确定了一组ANGPTL7+成纤维细胞(FBs),可能是促纤维化的,并参与人GSVs的胰岛素抵抗。此外,我们的数据表明免疫细胞只占一小部分,其中大部分是巨噬细胞。通过评估潜在增加壁细胞和白细胞中基因表达调节网络的代谢功能障碍中相互交织的重塑,我们发现转录因子KLF9可能是一个促炎程序,诱导两个壁细胞亚群和促炎免疫细胞中金属硫蛋白的转录。最后,细胞通讯分析显示,促炎信号,包括TRAIL,PVR,CSF和GDF,在代谢功能障碍患者中被独特地激活。
我们的结果确定了GSV中关键的细胞特异性反应和细胞相互作用。除了作为曲目,这项工作说明了VGF的多因素可能性。
The great saphenous vein (GSV) is the most commonly used conduit for coronary arterial bypass graft. However, the status of the GSV, including metabolic dysfunction such as diabetes mellitus (DM) complication, is strongly associated with vein graft failure (VGF). To date, the molecular mechanism underlying VGF remains elusive. Detailed characterization of the cellular components and corresponding expression regulation in GSVs would be of great importance for clinical decision making to reduce VGF.
To this end, we performed single-cell RNA sequencing to delineate cellular heterogeneity in three human GSV samples.
Scrutinization of cellular composition and expression revealed cell diversity in human GSVs, particularly endothelial cells (ECs). Our results unraveled that functional adaptation drove great expression differences between venous ECs and valvular ECs. For instance, cell surface receptor ACKR1 demarcated venous Ecs, whereas ACRK3/ACKR4 were exclusively expressed by valvular ECs. Differential gene expression analysis suggested that genes highly expressed in venous ECs were mainly involved in vasculature development and regulation of leukocyte adhesion, whereas valvular ECs have more pronounced expression of genes participating in extracellular matrix organization, ossification and platelet degranulation. Of note, pseudo-time trajectory analysis provided in silico evidence indicating that venous ECs, valvular ECs and lymphatic vessels were developmentally connected. Further, valvular ECs might be an importance source for lymphatic vessel differentiation in adults. Additionally, we found a venous EC subset highly expressing IL6, which might be associated with undesirable prognosis. Meanwhile, we identified a population of ANGPTL7+ fibroblasts (FBs), which may be profibrotic and involved in insulin resistance in human GSVs. Additionally, our data suggest that immune cells only accounted for a small fraction, most of which were macrophages. By assessing the intertwined remodeling in metabolic dysfunction that potentially increases the gene expression regulatory network in mural cells and leukocytes, we found that transcription factor KLF9 likely operated a proinflammatory program, inducing the transcription of metallothionein proteins in two mural cell subsets and proinflammatory immune cells. Lastly, cellular communication analysis revealed that proinflammatory signaling, including TRAIL, PVR, CSF and GDF, were uniquely activated in patients with metabolic dysfunction.
Our results identified critical cell-specific responses and cellular interactions in GSVs. Beyond serving as a repertoire, this work illustrates multifactorial likelihood of VGF.