Posttranslational modifications of recombinant myotube-synthesized human factor IX

Arruda, Valder R.; Hagstrom, J. Nathan; Deitch, Jeffrey S.; Heiman‐Patterson, Terry; Camire, Rodney M.; Chu, Kirk; Fields, Paul A.; Herzog, Roland W.; Couto, Linda B.; Larson, Peter J.; High, Katherine A.

doi:10.1182/blood.v97.1.130

Cited by 116 publications

(91 citation statements)

References 52 publications

(58 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Arruda et al found that carboxylation, tyrosine sulphation and serine phosphorylation of factor IX differ between liver and muscle, although post-translational modifications critical for biological activity of the factor are similar in these two tissues. 27 It should also be noted that there are substantial differences between the efficacies of transfection of myoblasts in vitro and muscle fibres in vivo for any given procedure. Nearly all systems that give efficient plasmid delivery in vitro have so far shown little beneficial effect when used for in vivo gene transfer into skeletal muscle (see below).…”

Section: Why Skeletal Muscle?mentioning

confidence: 99%

Non-viral gene delivery in skeletal muscle: a protein factory

Lu¹,

Bou‐Gharios²,

Partridge³

2003

Gene Ther

168

109

View full text Add to dashboard Cite

Ever since the publication of the first reports in 1990 using skeletal muscle as a direct target for expressing foreign transgenes, an avalanche of papers has identified a variety of proteins that can be synthesized and correctly processed by skeletal muscle. The impetus to the development of such applications is not only amelioration of muscle diseases, but also a range of therapeutic applications, from immunization to delivery of therapeutic proteins, such as clotting factors and hormones. Although the most efficient way of introducing transgenes into muscle fibres has been by a variety of recombinant viral vectors, there are potential benefits in the use of non-viral vectors. In this review we assess the recent advances in construction and delivery of naked plasmid DNA to skeletal muscle and highlight the options available for further improvements to raise efficiency to therapeutic levels.

show abstract

Section: Why Skeletal Muscle?mentioning

confidence: 99%

Non-viral gene delivery in skeletal muscle: a protein factory

Lu¹,

Bou‐Gharios²,

Partridge³

2003

Gene Ther

168

109

View full text Add to dashboard Cite

show abstract

“…7 An advantage of hemophilia as a model for gene transfer is that tissue-specific expression of the transgene is not required, because biologically active F.IX can be produced in cells other than hepatocytes. 8,9 In addition, precise regulation of transgene expression is not required, because levels of 1% to 2% may be therapeutic and levels up to 100% are still within the normal range. The existence of small and large animal models of this disease [10][11][12][13][14][15][16] facilitates analysis of efficacy before clinical studies are initiated, and measurement of clinical therapeutic end points (circulating levels of F.IX) is straightforward.…”

Section: Introductionmentioning

confidence: 99%

AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B

Manno

Chew

Hutchison

et al. 2003

Blood

691

561

View full text Add to dashboard Cite

Hemophilia B is an X-linked coagulopathy caused by absence of functional coagulation factor IX (F.IX). Previously, we established an experimental basis for gene transfer as a method of treating the disease in mice and hemophilic dogs through intramuscular injection of a recombinant adeno-associated viral (rAAV) vector expressing F.IX. In this study we investigated the safety of this approach in patients with hemophilia B. In an open-label dose-escalation study, adult men with severe hemophilia B (F.IX < 1%) due to a missense mutation were injected at multiple intramuscular sites with an rAAV vector. At doses ranging from 2 ؋ 10 11 vector genomes (vg)/kg to 1.8 ؋ 10 12 vg/ kg, there was no evidence of local or systemic toxicity up to 40 months after injection. Muscle biopsies of injection sites performed 2 to 10 months after vector administration confirmed gene transfer as evidenced by Southern blot and transgene expression as evidenced by immunohistochemical staining. Preexisting high-titer antibodies to AAV did not prevent gene transfer or expression. Despite strong evidence for gene transfer and expression, circulating levels of F.IX were in all cases less than 2% and most were less than 1%. Although more extensive transduction of muscle fibers will be required to develop a therapy that reliably raises circulating levels to more than 1% in all subjects, these results of the first parenteral administration of rAAV demonstrate that administration of AAV vector by the intramuscular route is safe at the doses tested and effects gene transfer and expression in humans in a manner similar to that seen in animals. (Blood.

show abstract

“…We chose the muscle as it can be very efficiently transduced by different serotypes of adeno-associated virus (AAV) vectors, such as AAV6. 6 Moreover, muscle fibers are able to express and secrete biologically active gene products that are normally not synthesized by this tissue, including coagulation factor IX, 7,8 a1-antitrypsin, 9 erythropoietin 10 and interleukin-10. 11,12 In this study we have made use of expression cassettes encoding a chimeric variant of either human-or murine TNF-a-soluble receptor I linked to the Fc (fragment crystallizable) fragment of the mouse immunoglobulin G1 (hTNFR-Is/mIgG1 and mTNFR-Is/mIgG1, respectively).…”

Section: Introductionmentioning

confidence: 99%

Soluble TNF-α receptor secretion from healthy or dystrophic mice after AAV6-mediated muscle gene transfer

et al. 2010

View full text Add to dashboard Cite

Muscle is an attractive target because it is easily accessible; it also offers a permissive environment for adeno-associated virus (AAV)-mediated gene transfer and has an abundant blood vascular supply providing an efficient transport system for the secretion of proteins. However, gene therapy of dystrophic muscle may be more difficult than that of healthy tissue because of degenerative-regenerative processes, and also because of the inflammatory context. In this study we followed the expression levels of secreted inhibitors of the proinflammatory tumor necrosis factor (TNF) cytokine after intramuscular (i.m.) injection of AAV6 into dystrophic mdx and healthy C57BL/10 mice. We used two chimeric proteins, namely, the human or murine TNF-soluble receptor I fused with the murine heavy immunoglobulin chain. We conducted an AAV6 dose-response study and determined the kinetics of transgene expression. In addition, we followed the antibody response against the transgenes and studied their expression pattern in the muscle. Our results show that transduction efficiency is reduced in dystrophic muscles as compared with healthy ones. Furthermore, we found that the immune response against the secreted protein is stronger in mdx mice. Together, our results underscore that the pathological state of the muscle has to be taken into consideration when designing gene therapy approaches.

show abstract

Posttranslational modifications of recombinant myotube-synthesized human factor IX

Cited by 116 publications

References 52 publications

Non-viral gene delivery in skeletal muscle: a protein factory

Non-viral gene delivery in skeletal muscle: a protein factory

AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B

Soluble TNF-α receptor secretion from healthy or dystrophic mice after AAV6-mediated muscle gene transfer

Contact Info

Product

Resources

About