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

Meyer, Simon F. De; Vanhoorelbeke, Karen; Chuah, Marinee; Pareyn, Inge; Gillijns, Veerle; Hebbel, Robert P.; Collen, Désiré; Deckmyn, Hans; VandenDriessche, Thierry

doi:10.1182/blood-2005-09-3605

Cited by 67 publications

(44 citation statements)

References 40 publications

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“…One approach may be to co-transduce BOECs with VWF. De Meyer et al 44 have recently demonstrated that WPB formation in VWD type 3 BOECs is restored upon lentiviral transduction with VWF. Another potential approach may be to overexpress the transcription factor KLF2 which has recently been shown to increase the average number of WPBs in HUVEC.…”

Section: Discussionmentioning

confidence: 99%

Storage and regulated secretion of factor VIII in blood outgrowth endothelial cells

Biggelaar¹,

Bouwens²,

Kootstra³

et al. 2009

Haematologica

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BackgroundGene therapy provides an attractive alternative for protein replacement therapy in hemophilia A patients. Recent studies have shown the potential benefit of directing factor (F)VIII gene delivery to cells that also express its natural carrier protein von Willebrand factor (VWF). In this study, we explored the feasibility of blood outgrowth endothelial cells as a cellular FVIII delivery device with particular reference to long-term production levels, intracellular storage in Weibel-Palade bodies and agonist-induced regulated secretion. Design and MethodsHuman blood outgrowth endothelial cells were isolated from peripheral blood collected from healthy donors, transduced at passage 5 using a lentiviral vector encoding human Bdomain deleted FVIII-GFP and characterized by flow cytometry and confocal microscopy. ResultsBlood outgrowth endothelial cells displayed typical endothelial morphology and expressed the endothelial-specific marker VWF. Following transduction with a lentivirus encoding FVIII-GFP, 80% of transduced blood outgrowth endothelial cells expressed FVIII-GFP. Levels of FVIII-GFP positive cells declined slowly upon prolonged culturing. Transduced blood outgrowth endothelial cells expressed 1.6±1.0 pmol/1×10 6 cells/24h FVIII. Morphological analysis demonstrated that FVIII-GFP was stored in Weibel-Palade bodies together with VWF and P-selectin. FVIII levels were only slightly increased following agonist-induced stimulation, whereas a 6-to 8-fold increase of VWF levels was observed. Subcellular fractionation revealed that 15-22% of FVIII antigen was present within the dense fraction containing Weibel-Palade bodies. ConclusionsWe conclude that blood outgrowth endothelial cells, by virtue of their ability to store a significant portion of synthesized FVIII-GFP in Weibel-Palade bodies, provide an attractive cellular on-demand delivery device for gene therapy of hemophilia A.

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Section: Discussionmentioning

confidence: 99%

Storage and regulated secretion of factor VIII in blood outgrowth endothelial cells

Biggelaar¹,

Bouwens²,

Kootstra³

et al. 2009

Haematologica

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“…Blood outgrowth endothelial cells (BOECs) were isolated from the blood of healthy volunteers, characterized by immunofluorescent staining and grown in EBM-2/EGM-2 medium (Lonza, San Diego, CA) as described (34,35).…”

Section: Endothelial Cell Culturementioning

confidence: 99%

Single Particle Tracking of ADAMTS13 (A Disintegrin and Metalloprotease with Thrombospondin Type-1 Repeats) Molecules on Endothelial von Willebrand Factor Strings

Ceunynck

Rocha

Meyer

et al. 2014

Journal of Biological Chemistry

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Background: ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type-1 repeats) cleaves pro-thrombotic ultra-large von Willebrand factor (VWF) strings. Results: Customized single particle tracking enabled visualization of single ADAMTS13 enzymes that bind to long plateletdecorated VWF strings. Conclusion: ADAMTS13 readily bind to multiple available sites on VWF strings. Significance: Single molecule imaging can be used to study interactions between enzymes and large biopolymers in flow.

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“…This distinctive feature paves the way toward many applications for which γ-RVs are not suitable. Moreover, LV can accommodate larger transgenes [up to ~10 kilobases (kb)] compared to when γ-RVs are used though vector titers tend to decrease with larger inserts [14,24,25]. It is critically important to ensure that the LVs are replication-defective because HIV-1 is a human pathogen.…”

Section: Lentiviral Vectorsmentioning

confidence: 99%

Recent Advances and Future Perspectives in Lentiviral Gene Therapy for Hemophilia A and B

Mátrai¹,

Chuah²,

VandenDriessche³

2012

J Genet Syndr Gene Ther

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Hemophilia A and B are rare incurable hereditary diseases due to deficiencies in clotting factor VIII (FVIII) and factor IX (FIX), respectively. These genetic defects result in potentially life-threatening, uncontrolled bleeding episodes. Current treatment by protein substitution therapy does not constitute a cure making gene therapy an attractive alternative. Lentiviral vectors (LVs) have many distinctive features that make them especially well suited for FVIII or FIX gene delivery. This includes the lack of vector-specific pre-existing immunity, their ability to permanently transduce both dividing and non-dividing cells and their capacity to readily accommodate FIX and FVIII expression cassettes, consistent with their packaging capacity of 10 kb. LVs have been used to achieve sustained therapeutic clotting factor expression levels and hemostatic correction in preclinical hemophilic mouse models. The liver has been the target organ of choice for direct in vivo LV transduction of FVIII or FIX genes, resulting in sustained therapeutic effects. Nevertheless, the potential development of neutralizing antibodies to the clotting factors following ex vivo or in vivo gene therapy with LVs can preclude long-term phenotypic correction. These risks can be minimized by preventing ectopic expression in antigen-presenting cells. LVs are well suited to deliver the clotting factor genes into hematopoietic stem/progenitor cells, allowing for stable FVIII or FIX expression upon hematopoietic reconstitution. In addition, therapeutic FVIII and FIX expression levels have been achieved in vivo after transplantation of lentivirally transduced endothelial and mesenchymal stem/progenitor cells. Current challenges relate primarily to the translation of these findings to larger preclinical animal models and ultimately to patients suffering from hemophilia.

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

Cited by 67 publications

References 40 publications

Storage and regulated secretion of factor VIII in blood outgrowth endothelial cells

Storage and regulated secretion of factor VIII in blood outgrowth endothelial cells

Single Particle Tracking of ADAMTS13 (A Disintegrin and Metalloprotease with Thrombospondin Type-1 Repeats) Molecules on Endothelial von Willebrand Factor Strings

Recent Advances and Future Perspectives in Lentiviral Gene Therapy for Hemophilia A and B

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