Efficient transfer and expression of human clotting factor IX cDNA in neonatal hemophilia B mice mediated by VSV-G pseudotyped retrovirus

2001 ◽  
Vol 46 (18) ◽  
pp. 1534-1538
Author(s):  
Hongwei Wang ◽  
Chenbo Ye ◽  
Li Chen ◽  
Xuefeng Wang ◽  
Xinfang Qiu ◽  
...  
Keyword(s):  
Factor Ix ◽  
Hemophilia B ◽  
Clotting Factor ◽  
B Mice ◽  
Human Clotting Factor ◽  
Efficient Transfer

Expression of human clotting factor IX mediated by recombinant lentiviral vector in cultured cells and hemophilia B mice

2003 ◽  
Vol 48 (20) ◽  
pp. 2196-2200
Author(s):  
Huanzhang Zhu ◽  
Xiaoguang Chen ◽  
Feng Li ◽  
Juli Gong ◽  
Jinglun Xue
Keyword(s):  
Cultured Cells ◽  
Lentiviral Vector ◽  
Factor Ix ◽  
Hemophilia B ◽  
Clotting Factor ◽  
B Mice ◽  
Human Clotting Factor

A microRNA-regulated lentiviral vector mediates stable correction of hemophilia B mice

Blood ◽  
2007 ◽  
Vol 110 (13) ◽  
pp. 4144-4152 ◽  
Author(s):  
Brian D. Brown ◽  
Alessio Cantore ◽  
Andrea Annoni ◽  
Lucia Sergi Sergi ◽  
Angelo Lombardo ◽  
...  
Keyword(s):  
Immune Response ◽  
Gene Transfer ◽  
Transgene Expression ◽  
Lentiviral Vector ◽  
Factor Ix ◽  
Hemophilia B ◽  
Clotting Factor ◽  
Target Sequence ◽  
B Mice ◽  
Cellular Immune

A longstanding goal for the treatment of hemophilia B is the development of a gene transfer strategy that can maintain sustained production of clotting factor IX (F.IX) in the absence of an immune response. To this end, we have sought to use lentiviral vectors (LVs) as a means for systemic gene transfer. Unfortunately, initial evaluation of LVs expressing F.IX from hepatocyte-specific promoters failed to achieve sustained F.IX expression in hemophilia B mice due to the induction of an anti-F.IX cellular immune response. Further analysis suggested that this may be a result of off-target transgene expression in hematopoietic-lineage cells of the spleen. In order to overcome this problem, we modified our vector to contain a target sequence for the hematopoietic-specific microRNA, miR-142-3p. This eliminated off-target expression in hematopoietic cells, and enabled sustained gene transfer in hemophilia B mice for more than 280 days after injection. Treated mice had more than 10% normal F.IX activity, no detectable anti-F.IX antibodies, and were unresponsive to F.IX immunization. Importantly, the mice survived tail-clip challenge, thus demonstrating phenotypic correction of their bleeding diathesis. This work, which is among the first applications to exploit the microRNA regulatory pathway, provides the basis for a promising new therapy for the treatment of hemophilia B.

Timely and large dose of clotting factor IX provides better joint wound healing after hemarthrosis in hemophilia B mice

2019 ◽  
Vol 110 (1) ◽  
pp. 59-68 ◽  
Author(s):  
Ping He ◽  
Feixu Zhang ◽  
Chen Zhong ◽  
Min Li ◽  
Jing Zheng ◽  
...  
Keyword(s):  
Wound Healing ◽  
Large Dose ◽  
Factor Ix ◽  
Hemophilia B ◽  
Clotting Factor ◽  
B Mice

Factor IX variants improve gene therapy efficacy for hemophilia B

Blood ◽  
2005 ◽  
Vol 105 (6) ◽  
pp. 2316-2323 ◽  
Author(s):  
Joerg Schuettrumpf ◽  
Roland W. Herzog ◽  
Alexander Schlachterman ◽  
Antje Kaufhold ◽  
Darrel W. Stafford ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Intramuscular Injection ◽  
Specific Activity ◽  
Factor Ix ◽  
Hemophilia B ◽  
Promising Strategy ◽  
B Mice ◽  
Higher Doses ◽  
Vector Delivery

Abstract Intramuscular injection of adeno-associated viral (AAV) vector to skeletal muscle of humans with hemophilia B is safe, but higher doses are required to achieve therapeutic factor IX (F.IX) levels. The efficacy of this approach is hampered by the retention of F.IX in muscle extracellular spaces and by the limiting capacity of muscle to synthesize fully active F.IX at high expression rates. To overcome these limitations, we constructed AAV vectors encoding F.IX variants for muscle- or liver-directed expression in hemophilia B mice. Circulating F.IX levels following intramuscular injection of AAV-F.IX-K5A/V10K, a variant with low-affinity to extracellular matrix, were 2-5 fold higher compared with wild-type (WT) F.IX, while the protein-specific activities remained similar. Expression of F.IX-R338A generated a protein with 2- or 6-fold higher specific activity than F.IX-WT following vector delivery to skeletal muscle or liver, respectively. F.IX-WT and variant forms provide effective hemostasis in vivo upon challenge by tail-clipping assay. Importantly, intramuscular injection of AAV-F.IX variants did not trigger antibody formation to F.IX in mice tolerant to F.IX-WT. These studies demonstrate that F.IX variants provide a promising strategy to improve the efficacy for a variety of gene-based therapies for hemophilia B.

scholarly journals Production of human factor IX in animals by genetically modified skin fibroblasts: potential therapy for hemophilia B

Blood ◽  
1989 ◽  
Vol 73 (2) ◽  
pp. 438-445
Author(s):  
TD Palmer ◽  
AR Thompson ◽  
AD Miller
Keyword(s):  
Human Factor ◽  
Normal Skin ◽  
Human Fibroblasts ◽  
Retroviral Vectors ◽  
Factor Ix ◽  
Skin Fibroblasts ◽  
Hemophilia B ◽  
Clotting Factor ◽  
Human Factor Ix

Inherited diseases might be treated by introducing normal genes into a patient's somatic tissues to correct the genetic defects. In the case of hemophilia resulting from a missing clotting factor, the required gene could be introduced into any cell as long as active factor reached the circulation. We previously showed that retroviral vectors can efficiently transfer genes into normal skin fibroblasts and that the infected cells can produce high levels of a therapeutic product in vitro. In the current study, we examined the ability of skin fibroblasts to secrete active clotting factor after infection with different retroviral vectors encoding human clotting factor IX. Normal human fibroblasts infected with one vector secreted greater than 3 micrograms factor IX/10(6) cells/24 h. Of this protein, greater than 70% was structurally and functionally indistinguishable from human factor IX derived from normal plasma. This suggests that infected autologous fibroblasts might provide therapeutic levels of factor IX if transplanted into patients suffering from hemophilia B. By transplanting normal diploid fibroblasts infected with the factor IX vectors, we showed that human factor IX can be produced and is circulated at readily detectable levels in rats and mice.

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