scholarly journals 483. Phenotypic Correction of Von Willebrand Disease Type 3 Blood-Derived Endothelial Cells with Lentiviral Vectors Expressing Von Willebrand Factor

2006 ◽  
Vol 13 ◽  
pp. S187
Author(s):  
Simon F. DeMeyer ◽  
Karen Van hoorelbeke ◽  
Marinee K. Chuah ◽  
Inge Pareyn ◽  
Veerle Gilijns ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Lentiviral Vectors ◽  
Disease Type ◽  
Von Willebrand Disease ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals Phenotypic correction of von Willebrand disease type 3 blood-derived endothelial cells with lentiviral vectors expressing von Willebrand factor

Blood ◽  
2006 ◽  
Vol 107 (12) ◽  
pp. 4728-4736 ◽  
Author(s):  
Simon F. De Meyer ◽  
Karen Vanhoorelbeke ◽  
Marinee K. Chuah ◽  
Inge Pareyn ◽  
Veerle Gillijns ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Lentiviral Vector ◽  
Single Gene ◽  
Von Willebrand Disease ◽  
Target Cells ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractVon 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 106 cells), with normal binding to GPIbα 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.

Phenotypic Correction of von Willebrand Disease Type 3 Blood-Derived Endothelial Cells with Lentiviral Vectors Expressing von Willebrand Factor.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5522-5522
Author(s):  
Simon F. De Meyer ◽  
Karen Vanhoorelbeke ◽  
Marinee K. Chuah ◽  
Inge Pareyn ◽  
Veerle Gillijns ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Lentiviral Vector ◽  
Single Gene ◽  
Von Willebrand Disease ◽  
Target Cells ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract Von Willebrand disease (VWD) is the most common inherited bleeding disorder, caused by quantitative (type 1 and 3 VWD) or qualitative (type 2 VWD) defects in von Willebrand factor (VWF). Gene therapy is an appealing strategy for treatment of type 3 VWD since it is caused by a single gene defect and since 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) and the inherent complexicity of the VWF protein which requires extensive post-translational processing. The objective of this study consisted of developing a gene-based approach for VWD using lentiviral transduction of blood-outgrowth endothelial cells (BOECs) to express functional VWF. A lentiviral vector encoding the complete human VWF protein was used to transduce BOECs isolated from type 3 VWD dogs resulting in high transduction efficiencies (95.6 ± 2.2 %). These BOECs are completely deficient in VWF due to a point mutation in the VWF gene responsible for impaired VWF synthesis. Transduced VWD BOECs efficiently expressed fully functional vector-encoded VWF (4.6 ± 0.4 U/24hr per 106 cells, figure 1), with normal binding to GPIbα and collagen and normal multimeric pattern resulting in phenotypic correction of these cells, which had not been shown previously. These results indicate that BOECs are attractive target cells for gene therapy of type 3 VWD.

Toward Personalized Treatment for Patients with Low von Willebrand Factor and Quantitative von Willebrand Disease

2021 ◽  
Vol 47 (02) ◽  
pp. 192-200
Author(s):  
James S. O'Donnell
Keyword(s):  
Von Willebrand Factor ◽  
Bleeding Risk ◽  
Von Willebrand Disease ◽  
Personalized Treatment ◽  
Treatment Plans ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractThe biological mechanisms involved in the pathogenesis of type 2 and type 3 von Willebrand disease (VWD) have been studied extensively. In contrast, although accounting for the majority of VWD cases, the pathobiology underlying partial quantitative VWD has remained somewhat elusive. However, important insights have been attained following several recent cohort studies that have investigated mechanisms in patients with type 1 VWD and low von Willebrand factor (VWF), respectively. These studies have demonstrated that reduced plasma VWF levels may result from either (1) decreased VWF biosynthesis and/or secretion in endothelial cells and (2) pathological increased VWF clearance. In addition, it has become clear that some patients with only mild to moderate reductions in plasma VWF levels in the 30 to 50 IU/dL range may have significant bleeding phenotypes. Importantly in these low VWF patients, bleeding risk fails to correlate with plasma VWF levels and inheritance is typically independent of the VWF gene. Although plasma VWF levels may increase to > 50 IU/dL with progressive aging or pregnancy in these subjects, emerging data suggest that this apparent normalization in VWF levels does not necessarily equate to a complete correction in bleeding phenotype in patients with partial quantitative VWD. In this review, these recent advances in our understanding of quantitative VWD pathogenesis are discussed. Furthermore, the translational implications of these emerging findings are considered, particularly with respect to designing personalized treatment plans for VWD patients undergoing elective procedures.

scholarly journals Phage Display Broadly Identifies Inhibitor-reactive Regions in von Willebrand Factor

2021 ◽  
Author(s):  
Andrew Yee ◽  
Manhong Dai ◽  
Stacy E. Croteau ◽  
Jordan A. Shavit ◽  
Steven W. Pipe ◽  
...  
Keyword(s):  
Phage Display ◽  
Von Willebrand Factor ◽  
Gene Deletion ◽  
Von Willebrand Disease ◽  
Response To Treatment ◽  
Phage Library ◽  
Incomplete Removal ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryBackgroundCorrection of von Willebrand factor (VWF) deficiency with replacement products containing VWF can lead to the development of anti-VWF alloantibodies (i.e., VWF inhibitors) in patients with severe von Willebrand disease (VWD).ObjectiveLocate inhibitor-reactive regions within VWF using phage display.MethodsWe screened a phage library displaying random, overlapping fragments covering the full length VWF protein sequence for binding to a commercial anti-VWF antibody or to immunoglobulins from three type 3 VWD patients who developed VWF inhibitors in response to treatment with plasma-derived VWF. Immunoreactive phage clones were identified and quantified by next generation DNA sequencing (NGS).ResultsNGS markedly increased the number of phage analyzed for locating immunoreactive regions within VWF following a single round of selection and identified regions not recognized in previous reports using standard phage display methods. Extending this approach to characterize VWF inhibitors from three type 3 VWD patients (including two siblings homozygous for the same VWF gene deletion) revealed patterns of immunoreactivity distinct from the commercial antibody and between unrelated patients, though with notable areas of overlap. Alloantibody reactivity against the VWF propeptide is consistent with incomplete removal of the propeptide from plasma-derived VWF replacement products.ConclusionThese results demonstrate the utility of phage display and NGS to characterize diverse anti-VWF antibody reactivities.

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