PXD013857

PXD013857 is an original dataset announced via ProteomeXchange.

Title	A CRISPR/Cas9 engineered Von Willebrand factor deficient endothelial cell model shows alternative trafficking of Weibel-Palade body proteins
Description	Synthesis of the hemostatic protein Von Willebrand factor (VWF) drives formation of endothelial storage organelles called Weibel-Palade bodies (WPBs). In the absence of VWF, angiogenic and inflammatory mediators that are co-stored in WPBs are subject to alternative trafficking routes. In Von Willebrand disease (VWD) patients, the partial or complete absence of VWF/WPBs may lead to additional bleeding complications, such as angiodysplasia. Studies addressing the role of VWF using VWD patient-derived blood outgrowth endothelial cells (BOECs) have reported conflicting results due to the intrinsic heterogeneity of patient-derived BOECs. To study the role of WPBs in endothelial cells using CRISPR-mediated knockout of VWF in BOECs. We used CRISPR/Cas9 gene editing in single donor cord blood-derived BOECs (cbBOECs) to generate clonal VWF-/- cbBOECs. Clones were selected using high-throughput screening, VWF mutations were validated by sequencing and cells were phenotypically characterized. Two VWF-/- BOEC clones were obtained and were entirely devoid of WPBs, while overall cell morphology was unaltered. Several WPB proteins, including CD63, syntaxin-3 and the cargo proteins Ang-2, IL-6 and IL-8 showed alternative trafficking and secretion in absence of VWF. Interestingly, Ang-2 changed localization to the cell periphery and colocalized with Tie-2. CRISPR editing of VWF provides a robust method to create VWF deficient BOECs that can be directly compared to their wild-type counterparts. Results obtained with our model system confirmed alternative trafficking of several WPB proteins in the absence of VWF and support the theory that increased Ang-2/Tie-2 interaction contributes to angiogenic abnormalities in VWD patients.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:54:04.767.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ruben Bierings
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2019-05-15 03:24:18	ID requested
1	2019-11-11 14:39:57	announced
⏵ 2	2024-10-22 04:54:05	announced	2024-10-22: Updated project metadata.

Schillemans M, Kat M, Westeneng J, Gangaev A, Hofman M, Nota B, van Alphen FPJ, de Boer M, van den Biggelaar M, Margadant C, Voorberg J, Bierings R, Alternative trafficking of Weibel-Palade body proteins in CRISPR/Cas9-engineered von Willebrand factor-deficient blood outgrowth endothelial cells. Res Pract Thromb Haemost, 3(4):718-732(2019) [pubmed]

10.1002/rth2.12242;

submitter keyword: von Willebrand Factor,Endothelial Cells, Protein Transport, Gene Knockout Techniques, Secretory Vesicles

Dr.Ir. Ruben Bierings
contact affiliation	Department of Hematology, Erasmus MC, Rotterdam, the Netherlands
contact email	R.Bierings@erasmusmc.nl
lab head
Ruben Bierings
contact affiliation	ErasmusMC
contact email	r.bierings@erasmusmc.nl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD013857
     1. Label: PRIDE project
     2. Name: A CRISPR/Cas9 engineered Von Willebrand factor deficient endothelial cell model shows alternative trafficking of Weibel-Palade body proteins