Multifaceted Pathomolecular Mechanism of a VWF Large Deletion Involved in the Pathogenesis of Severe VWD.
Hamideh YadegariMuhammad Ahmer JamilJens MüllerNatascha MarquardtOrla RawleyUlrich BuddeOsman El-MaarriDavid LillicrapJohannes OldenburgPublished in: Blood advances (2021)
An in-frame heterozygous large deletion of exons 4-34 of the von Willebrand factor (VWF) gene was identified in a type 3 von Willebrand disease (VWD) index patient (IP), as the only VWF variant. The IP exhibited severe bleeding episodes despite prophylaxis treatment, with a short VWF half-life after infusion of VWF/FVIII concentrates. Transcript analysis confirmed transcription of normal VWF mRNA besides an aberrant deleted transcript. The IP endothelial colony-forming cells (ECFCs) exhibited a defect in the VWF multimers and Weibel-Palade bodies (WPBs) biogenesis, although demonstrating normal VWF secretion, as compared with healthy cells. Immunostaining of IP-ECFCs revealed subcellular mislocalization of WPBs pro-inflammatory cargos angiopoietin-2 (Ang2, nuclear accumulation) and P-selectin. Besides, the RNA-sequencing (RNA-seq) analysis showed upregulation of pro-inflammatory and proangiogenic genes, P-selectin, IL8, IL6, and GROα, co-packaged with VWF into WPBs. Further, whole-transcriptome RNA-seq and subsequent Gene Ontology (GO) enrichment analysis indicated the most enriched GO-Biological Process terms among the differentially expressed genes in IP-ECFCs were regulation of cell differentiation, cell adhesion, leukocyte adhesion to vascular endothelial, blood vessel morphogenesis and angiogenesis, which resemble downstream signaling pathways associated with inflammatory stimuli and Ang2 priming. Accordingly, our functional experiments exhibited an increased endothelial cell adhesiveness and interruption in endothelial cell-cell junctions of the IP-ECFCs. In conclusion, the deleted VWF has a dominant-negative impact on multimer assembly and the biogenesis of WPBs, leading to altered trafficking of their pro-inflammatory cargos uniquely, which, in turn, causes changes in cellular signaling pathways, phenotype, and function of the endothelial cells.
Keyphrases
- rna seq
- single cell
- endothelial cells
- induced apoptosis
- signaling pathway
- genome wide
- high glucose
- cell adhesion
- gene expression
- stem cells
- vascular endothelial growth factor
- cell cycle arrest
- escherichia coli
- cystic fibrosis
- angiotensin ii
- dna methylation
- transcription factor
- single molecule
- staphylococcus aureus
- case report
- cell death
- fluorescent probe
- oxidative stress
- biofilm formation
- combination therapy
- candida albicans
- living cells