title: Analysis of the storage and secretion of von Willebrand factor in blood outgrowth endothelial cells derived from patients with von Willebrand disease.
abstract: Patients with von Willebrand disease (VWD) are often heterozygous for a missense mutation in the von Willebrand factor (VWF) gene. Investigating the pathogenic features of VWF mutations in cells directly derived from patients has been challenging. Here, we have used blood outgrowth endothelial cells (BOECs) isolated from human peripheral blood to analyze the storage and secretion of VWF. BOECs showed full endothelial characteristics and responded to Weibel-Palade body (WPB) secretagogues except desmopressin. We examined BOECs derived from a single subject heterozygous for a type 2N mutation (p.Arg854Gln) and from 4 patients with type 1 VWD who were, respectively, heterozygous for p.Ser1285Pro, p.Leu1307Pro, p.Tyr1584Cys, and p.Cys2693Tyr. Compared with normal BOECs, BOECs heterozygous for p.Ser1285Pro, p.Leu1307Pro, or p.Cys2693Tyr showed morphologically abnormal WPB and retention of VWF in the endoplasmic reticulum, whereas BOECs heterozygous for p.Arg854Gln or p.Tyr1584Cys showed normal WPB. The agonist-induced exocytosis of WPB from BOECs and formation of VWF strings on BOECs heterozygous for p.Ser1285Pro, p.Leu1307Pro, or p.Cys2693Tyr, but not for p.Arg854Gln or p.Tyr1584Cys, were reduced. In conclusion, VWD phenotype can be recapitulated in BOECs, and thus BOECs provide a feasible bona fide cell model to study the pathogenic effects of VWF mutations.
authors: Einthoven Laboratory for Experimental Vascular Medicine, Department of Thrombosis and Hemostasis, Leiden University Medical Center, Leiden, The Netherlands. j.w.h.wang@lumc.nl
authors: Jiong-Wei
authors: JW
authors: Wang
authors: Eveline A M
authors: EA
authors: Bouwens
authors: Maria Carolina
authors: MC
authors: Pintao
authors: Jan
authors: J
authors: Voorberg
authors: Huma
authors: H
authors: Safdar
authors: Karine M
authors: KM
authors: Valentijn
authors: Hetty C
authors: HC
authors: de Boer
authors: Koen
authors: K
authors: Mertens
authors: Pieter H
authors: PH
authors: Reitsma
authors: Jeroen
authors: J
authors: Eikenboom
mesh_terms: D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism* / Q000502:physiology; D005089:Exocytosis / Q000502:physiology; D005260:Female; D005434:Flow Cytometry; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D008297:Male; D020125:Mutation, Missense; D010641:Phenotype; D020302:Weibel-Palade Bodies / Q000378:metabolism*; D056725:von Willebrand Disease, Type 1 / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism*
chemical_list: D014841:von Willebrand Factor