Single-Cell Study of Endothelial Cells in Patients with Low VWF
Summary
The present study delves into the examination of endothelial colony-forming cells (ECFCs) using single-cell RNA sequencing (scRNA-seq) to investigate gene expression and regulatory mechanisms in patients with low Von Willebrand Factor (VWF) levels. VWF is a vital protein that contributes to the blood clotting process and aids in preventing bleeding. Notably, the study found that ECFCs derived from individuals with low VWF exhibited significant alterations in their Weibel Palade body (WPB) content and VWF release, along with differential gene expression patterns and pathways when compared to control ECFCs. The authors also identified potential candidate regulators of VWF levels through small-interfering RNA (siRNA) screening. The findings of this study provide insight into the complex mechanisms underlying VWF regulation in ECFCs and may pave the way for the development of new therapeutic interventions for VWF-related bleeding disorders.
Fig.1 Graphical abstract. (Ng, 2022)
Research Criteria
The primary objective of this scholarly work is to identify novel modulators of von Willebrand factor (VWF) levels in endothelial colony-forming cells (ECFCs) through the utilization of single-cell RNA sequencing (scRNA-seq). To achieve this goal, the authors isolated ECFCs from both patients exhibiting low VWF levels and healthy control subjects. They proceeded to undertake a comprehensive comparison of VWF expression, packaging, and release between these groups. Additionally, scRNA-seq analysis was conducted to investigate gene expression heterogeneity and candidate regulators of VWF in ECFCs.
Sample Type
Cells from Artery
Result—Transcriptional Profile of ECFCs
In this article, the authors employ a principal component analysis (PCA) to compare their sequenced endothelial cells to publicly available endothelial RNA-sequencing datasets, demonstrating their ECFCs and HUVECs are transcriptionally similar. Furthermore, the high gene expression of venous markers (NRP2, EPHB4) and low gene expression of arterial markers (EFNB2, NOTCH1) suggest a venous phenotype of their ECFCs. The authors define endothelial cells as positive for any of CDH5, PECAM1, ROBO4, ESAM, TIE1, or NOTCH4 based on a multi-tissue evaluation of highly expressed endothelial transcripts. Of the total cells analyzed, 97.1% were deemed to be endothelial. The t-distributed stochastic neighbor embedding (TSNE) plot shows that each cell line largely occupies its own cluster location, indicating transcriptional heterogeneity among cells of a cell line. There was no significant clustering of cells by low VWF ECFC vs. control EC phenotype.
Fig.2 Control ECs and low VWF ECFCs display global transcriptional differences. (Ng, 2022)
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Reference
- Ng, C.J.; et al. Single-cell transcriptional analysis of human endothelial colony-forming cells from patients with low VWF levels. Blood. 2022, 139(14): 2240-2251.
