Cellular and molecular basis of von Willebrand disease: studies on blood outgrowth endothelial cells.
Von Willebrand disease (VWD) is a heterogeneous bleeding disorder caused by decrease or dysfunction of von Willebrand factor (VWF). A wide range of mutations in the VWF gene have been characterized; however, their cellular consequences are still poorly understood. Here we have used a recently developed approach to study the molecular and cellular basis of VWD. We isolated blood outgrowth endothelial cells (BOECs) from peripheral blood of 4 type 1 VWD and 4 type 2 VWD patients and 9 healthy controls. We confirmed the endothelial lineage of BOECs, then measured VWF messenger RNA (mRNA) and protein levels (before and after stimulation) and VWF multimers. Decreased mRNA levels were predictive of plasma VWF levels in type 1 VWD, confirming a defect in VWF synthesis. However, BOECs from this group of patients also showed defects in processing, storage, and/or secretion of VWF. Levels of VWF mRNA and protein were normal in BOECs from 3 type 2 VWD patients, supporting the dysfunctional VWF model. However, 1 type 2M patient showed decreased VWF synthesis and storage, indicating a complex cellular defect. These results demonstrate for the first time that isolation of endothelial cells from VWD patients provides novel insight into cellular mechanisms of the disease.
Vascular Sciences, National Heart and Lung Institute, Faculty of Medicine, Hammersmith Campus, Imperial College London, London, United Kingdom.
Richard D
RD
Starke
Koralia E
KE
Paschalaki
Clare E F
CE
Dyer
Kimberly J
KJ
Harrison-Lavoie
Jacqueline A
JA
Cutler
Thomas A J
TA
McKinnon
Carolyn M
CM
Millar
Daniel F
DF
Cutler
Mike A
MA
Laffan
Anna M
AM
Randi
D000328:Adult; D000368:Aged; D019070:Cell Lineage / Q000502:physiology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D020302:Weibel-Palade Bodies / Q000378:metabolism; D056725:von Willebrand Disease, Type 1 / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D056728:von Willebrand Disease, Type 2 / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014841:von Willebrand Factor / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D014841:von Willebrand Factor