Cellular and genetic therapies for haemophilia

Lillicrap, David; VandenDriessche, Thierry; High, Katherine A.

doi:10.1111/j.1365-2516.2006.01259.x

Cited by 42 publications

(43 citation statements)

References 34 publications

(31 reference statements)

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“…As a result, viral vectors have been the predominant system for in vivo transgene delivery and use in human clinical hemophilia trials (34). However, a number of concerns have emerged regarding their use, including antigenicity, mutagenicity, cost, and defined tropisms (35).…”

Section: Discussionmentioning

confidence: 99%

“…However, the mice used in this study came from the same colony as those reported previously (54). Although the problem of inhibitor formation was overcome by neonatal tolerization of the mice with recombinant huFVIII protein, it continues to be a major obstacle in FVIII gene replacement strategies in animal models (34), requiring additional approaches for tolerization (56,57). In fact, in a recent adeno-associated virus study, longterm persistent FVIII expression in adult dogs was achieved in the absence of inhibitor formation (58).…”

Section: Figurementioning

confidence: 99%

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Nanocapsule-delivered Sleeping Beauty mediates therapeutic Factor VIII expression in liver sinusoidal endothelial cells of hemophilia A mice

et al. 2009

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Liver sinusoidal endothelial cells are a major endogenous source of Factor VIII (FVIII), lack of which causes the human congenital bleeding disorder hemophilia A. Despite extensive efforts, gene therapy using viral vectors has shown little success in clinical hemophilia trials. Here we achieved cell type-specific gene targeting using hyaluronan-and asialoorosomucoid-coated nanocapsules, generated using dispersion atomization, to direct genes to liver sinusoidal endothelial cells and hepatocytes, respectively. To highlight the therapeutic potential of this approach, we encapsulated Sleeping Beauty transposon expressing the B domain-deleted canine FVIII in cis with Sleeping Beauty transposase in hyaluronan nanocapsules and injected them intravenously into hemophilia A mice. The treated mice exhibited activated partial thromboplastin times that were comparable to those of wild-type mice at 5 and 50 weeks and substantially shorter than those of untreated controls at the same time points. Further, plasma FVIII activity in the treated hemophilia A mice was nearly identical to that in wild-type mice through 50 weeks, while untreated hemophilia A mice exhibited no detectable FVIII activity. Thus, Sleeping Beauty transposon targeted to liver sinusoidal endothelial cells provided long-term expression of FVIII, without apparent antibody formation, and improved the phenotype of hemophilia A mice.

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Section: Discussionmentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Nanocapsule-delivered Sleeping Beauty mediates therapeutic Factor VIII expression in liver sinusoidal endothelial cells of hemophilia A mice

et al. 2009

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“…Gene therapy or cell-based approaches have the potential to provide a life-long, cost-effective cure for hemophilia A. 2 A number of clinical trials for hemophilia A gene therapy have been performed. 3,4 So far, limited clinical efficacy of gene delivery approaches has been observed.…”

Section: Introductionmentioning

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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“…rAAVs have proven to be efficient in preclinical studies in animal models (16,28), and results from clinical trials are promising (7,47).…”

mentioning

confidence: 99%

Human Galectin 3 Binding Protein Interacts with Recombinant Adeno-Associated Virus Type 6

Denard

Beley

Kotin

et al. 2012

J Virol

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i Recombinant adeno-associated viruses (rAAVs) hold enormous potential for human gene therapy. Despite the well-established safety and efficacy of rAAVs for in vivo gene transfer, there is still little information concerning the fate of vectors in blood following systemic delivery. We screened for serum proteins interacting with different AAV serotypes in humans, macaques, dogs, and mice. We report that serotypes rAAV-1, -5, and -6 but not serotypes rAAV-2, -7, -8, -9, and -10 interact in human sera with galectin 3 binding protein (hu-G3BP), a soluble scavenger receptor. Among the three serotypes, rAAV-6 has the most important capacities for binding to G3BP. rAAV-6 also bound G3BP in dog sera but not in macaque and mouse sera. In mice, rAAV-6 interacted with another protein of the innate immune system, C-reactive protein (CRP). Furthermore, interaction of hu-G3BP with rAAV-6 led to the formation of aggregates and hampered transduction when the two were codelivered into the mouse. Based on these data, we propose that species-specific interactions of AAVs with blood proteins may differentially impact vector distribution and efficacy in different animal models.T he recombinant adeno-associated vector (rAAV) platform, derived from a nonpathogenic dependovirus, has many attributes suitable for in vivo gene transfer: rAAV vectors are capable of transducing a wide range of cell types, including dividing and nondividing cells; rAAV genomes persist as episomal chromatin in the nucleus of transduced cells (38); and stable, persistent expression has been reported for many transgenes in different tissues and species (6,12,36,39). rAAVs have proven to be efficient in preclinical studies in animal models (16,28), and results from clinical trials are promising (7,47).In the case of systemic diseases, clinical relevance requires widespread distribution of the vector in order to target entire organs. This is particularly true for myopathies, where all striated muscles of the skeletal musculature and, frequently, cardiac muscles have to be treated. In this case, vascular delivery would be the optimal route for rAAV administration. Intravascular injection of a number of rAAV serotypes has proven efficient in murine models of muscular dystrophies (11,17,18,34,35). However, translating this approach to large animal models and humans is still challenging. Acquired immunity and neutralizing antibodies present in a large fraction of the human population might obviously be restrictive for rAAV gene delivery (5,20,27,29,30). Moreover, recent studies have demonstrated that serum might also contain other factors neutralizing rAAV vectors (40), indicating that detailed characterization of rAAV's molecular interactions in the bloodstream is obviously important in order to improve vector efficacy.We looked for serum proteins, other than immunoglobulins, which could interact with rAAVs in the bloodstreams of different species. By using a multidisciplinary approach involving proteomics, binding assays, electron microscopy (EM), and in vivo stud...

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Cellular and genetic therapies for haemophilia

Cited by 42 publications

References 34 publications

Nanocapsule-delivered Sleeping Beauty mediates therapeutic Factor VIII expression in liver sinusoidal endothelial cells of hemophilia A mice

Nanocapsule-delivered Sleeping Beauty mediates therapeutic Factor VIII expression in liver sinusoidal endothelial cells of hemophilia A mice

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

Human Galectin 3 Binding Protein Interacts with Recombinant Adeno-Associated Virus Type 6

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