A transcriptome analysis of basal and stimulated VWF release from endothelial cells derived from patients with type 1 VWD

Abstract

Type 1 von Willebrand disease (VWD) is associated with a reduction in qualitatively normal von Willebrand factor (VWF). Current diagnostic guidelines only take into consideration the contribution of basal VWF levels, despite a lack of correlation with bleeding severity. Defects in stimulated VWF release, which occurs following hemostatic challenge, may contribute to bleeding in Type 1 VWD, but the pathogenic mechanisms are poorly defined. In the present study, a layered multiomic approach including mRNA and miRNA sequencing was used to evaluate transcriptome-wide differences between Type 1 VWD- and control-derived endothelial colony forming cells (ECFCs) during basal and stimulated VWF release. ECFCs from eight Type 1 VWD patients and four controls were included in this study. VWF protein analysis revealed heterogenous responses to stimulation amongst Type 1 VWD and control ECFCs. During basal VWF release, 64 mRNAs and 7 miRNAs were differentially regulated between Type 1 VWD and control ECFCs and 65 putatively pathogenic miRNA-mRNA interactions were identified. During stimulated VWF release, 190 mRNAs and five mRNAs were differentially regulated between Type 1 VWD and control ECFCs and 110 putatively pathogenic miRNA-mRNA interactions were identified. Five gene ontology terms including coagulation, regulation of cell shape, and regulation of cell signalling were also differentially regulated in Type 1 VWD ECFCs during stimulated release. We have shown for the first time that transcriptome-wide differences exist between Type 1 VWD and control ECFCs. These differences may contribute to bleeding in Type 1 VWD, and further investigation may reveal novel biomarkers and therapeutic targets.

Mycoplasma glycine cleavage system key subunit GcvH is an apoptosis inhibitor targeting host endoplasmic reticulum

“…Brsk2, an AMP-activated protein kinase (AMPK)-associated kinase, is abundant in the pancreas, frontal cortex and brain [ 74 , 75 ]. Although Brsk2 has been mainly studied for its function in neuronal polarization, cell cycle progression and insulin production, its critical role in the ER-associated apoptotic pathway has also been noted [ 76 , 77 ]. Previous studies showed that Brsk2 knockdown increases CHOP transcription and apoptotic levels, while its overexpression decreases them [ 77 , 78 ].…”

A decreased and less sustained response to surrogates of haemostatic stress correlates with bleeding in type 1 von Willebrand disease

“…8 A recent study published by our group found that endothelial cells from Type 1 VWD patients and healthy controls exhibit differential expression of genes involved in the regulation of cell shape and cell signalling when stimulated to release VWF. 9 Additionally, compared to controls, Type 1 VWD patients returned to baseline levels by Day 2 and exhibited a reduced response following the second dose of DDAVP. It is known that the trafficking of VWF into WPBs and through different secretory pathways is a highly regulated process.…”

Recent advances in endothelial colony-forming cells: from the transcriptomic perspective

“…MDS-ECFCs show a hypermethylated phenotype and have a lower expression of several Wnt pathway constituents. The addition of soluble Wnt3A could partially rescue the defects of MDS ECFCs [ 95 ] PB-ECF-Cs MiRNA sequence ECFCs from control group and type 1 VWD patients – ECFC from control group and type 1 VWD patients show DEGs and miRNAs, which may lead to the pathogenesis of type 1 VWD [ 97 ] PB-ECF-Cs Single cell sequence ECFCs from control group and patients with low VWF levels NCBI dBGaP system: phs002731.v1.p1 FLI1 is identified as the candidate gene that mediated the expression level of VWF [ 48 ] PB-ECF-Cs Microarray ECFCs from control group and patients with uVTE GSE118259 The activation of TNFSF15–TNFRSF25 axis reduces survival and proliferation of ECFCs in uVTE patients [ 99 ] PB-ECF-Cs Microarray ECFCs from control group and patients with MMD – RALDH2 is decreased in MMD ECFCs due to defective acetyl-histone H3 binding to the promoter region. Knockdown of RALDH2 in normal ECFCs induces decreased capillary formation in vitro.…”

“…In a study, transcriptome-wide differences of ECFCs between control donors and patients with type 1 VWD revealed 64 mRNAs and 7 miRNAs that were differentially expressed in ECFCs between control donors and type 1 VWD patients during basal VWF release. On the other hand, during stimulated VWF release, 190 mRNAs and 5 mRNAs were differentially expressed in ECFCs from control donors and type 1 VWD patients respectively [ 97 ]. Using single cell sequence, the Ng lab identified the DEGs between ECFCs isolated from donors with low VWF levels and control endothelial cells.…”