Phenotypic correction of von Willebrand disease type 3 blood-derived endothelial cells with lentiviral vectors expressing von Willebrand factor.
Von Willebrand disease (VWD) is an inherited bleeding disorder, caused by quantitative (type 1 and 3) or qualitative (type 2) defects in von Willebrand factor (VWF). Gene therapy is an appealing strategy for treatment of VWD because it is caused by a single gene defect and because VWF is secreted into the circulation, obviating the need for targeting specific organs or tissues. However, development of gene therapy for VWD has been hampered by the considerable length of the VWF cDNA (8.4 kb [kilobase]) and the inherent complexity of the VWF protein that requires extensive posttranslational processing. In this study, a gene-based approach for VWD was developed using lentiviral transduction of blood-outgrowth endothelial cells (BOECs) to express functional VWF. A lentiviral vector encoding complete human VWF was used to transduce BOECs isolated from type 3 VWD dogs resulting in high-transduction efficiencies (95.6% +/- 2.2%). Transduced VWD BOECs efficiently expressed functional vector-encoded VWF (4.6 +/- 0.4 U/24 hour per 10(6) cells), with normal binding to GPIbalpha and collagen and synthesis of a broad range of multimers resulting in phenotypic correction of these cells. These results indicate for the first time that gene therapy of type 3 VWD is feasible and that BOECs are attractive target cells for this purpose.
Laboratory for Thrombosis Research, Catholic University of Leuven, Belgium.
Simon F
SF
De Meyer
Karen
K
Vanhoorelbeke
Marinee K
MK
Chuah
Inge
I
Pareyn
Veerle
V
Gillijns
Robert P
RP
Hebbel
Désiré
D
Collen
Hans
H
Deckmyn
Thierry
T
VandenDriessche
D000818:Animals; D004283:Dog Diseases / Q000235:genetics / Q000628:therapy; D004285:Dogs; D042783:Endothelial Cells / Q000637:transplantation*; D015316:Genetic Therapy; D016086:Lentivirus; D014161:Transduction, Genetic; D014842:von Willebrand Diseases / Q000235:genetics / Q000628:therapy* / Q000662:veterinary; D014841:von Willebrand Factor / Q000235:genetics
D014841:von Willebrand Factor