2002 Fiscal Year Final Research Report Summary
Gene therapy for hemophilia A with simian immunodeficiency virus agmTYO1-based vectors carrying human factor VIII gene.
| Project/Area Number |
13671078
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Hematology
|
|---|
| Research Institution | Jichi Medical School |
|---|
Principal Investigator |
MADOIWA Seiji Department of Medicine, Jichi Medical School Assistant Professor, 医学部, 講師 (70296119)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SAKATA Yoichi Department of Medicine, Jichi Medical School Professor, 医学部, 教授 (40129028)
MIZUKAMI Hiroaki Department of Medicine, Jichi Medical School Assistant, 医学部, 助手 (20311938)
|
|---|
| Project Period (FY) |
2001 – 2002
|
| Keywords | Hemophilia A / Factor VIII / Gene therapy / SIV virus vector / Hematopoietic stem cell / Adipocyte / NOD / SCID mice / db / db mice |
|---|
| Research Abstract |
(l)We efficiently transduced human cord bloodderived (CB)-CD34+ cells using a simian immunodeficiency virus agmTYO1 (SIVagm) vector carrying the enhanced green fluorescent protein gene in a dose-dependent manner, reaching a maximum (99.6±0.1%) at MOI 5 X 10(3) vg/cell. (2) After transducing CB-CD34+ cells with SIVagm-based lentiviral vector carrying the human coagulation factor VIII gene (SIVhFVIII), hFVIII was produced (274.3±20.1 ng) from 10(6) CB-CD34+ cells during 24hr in vitro incubation. (3) Transplantation of SIVhFVIII-transduced CB-CD34+ cells (5-10 X 10(5)) into NOD/SCID mice resulted in successful engraftment of CD34+ cells and production of hFVIII (minimun 1.2±0.9 ng/mL, maximum 3.6±0.8 ng/mL) for at least 60 days in vivo. Trancripts of hFVIII gene and antigen were also detected in the murine bone marrow cells. (4) There was no significant difference in lineage marker expression of human hematopoiteic cells in the bone marrow and the spleen between NOD/SCID mice receiving th
… More
e SIVhFVIII-transduced cells and those who received mock-transduced cells, suggesting that stable human FVIII gene tranduction with the SIV vector does not alter capability of CB-CD34+ cells to re-populate and expand in the mouse hematopoietic microenvironment. (5) Cultured human adipocytes were transduced with the SlVagm vector carrying the GFP gene in a dose-dependent manner and transduction of adipocytes with SIVhFVIII resulted in efficient expression of human FVIII (320±39.8 ng/10(6) adipocytes/24 hours) in vitro. (6) Based upon successful transduction of adipocytes by SIV vectors carrying the lacZ gene in vivo in mice, the adipose tissue of db/db mice was transduced with SIVhFVIII. There was a trasient appearance of human FVIII in mouse plasma (maximun 1.8 ng/mL) on day 11 after the injection. Transcripts of human FVIII transgene and antigen also were detected in the adipose tissue by RT-PCR and immunofluorescence on day 14, respectively. These data suggest that transplantation of ex vivo transduced hematopoietic stem cells by non-pathogenic SIVhFVIII without exposure of subjects to viral vector is safe and potentially applicable for gene therapy of hemophilia A patients, and that transduction of the adipocytes with vectors carrying the human FVIII gene may be also applicable for hemophilia A gene therapy. Less
|
Research Products
(19 results)
-
-
-
-
-
-
-
-
-
-
[Publications] Madoiwa,S., Nakamura,Y., Mimuro,J., Furusawa,S., Koyama,T., Sugo,T., Matsuda,M., Sakata,Y.: "Autoantibody against Prothrombin aberrantly alters the Proenzyme to facilitate Formation of a Complex with its Physiological Inhibitor Antithrombin III without Th"Blood. 97. 3783-3789 (2001)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Madoiwa,S., Nakamura,Y., Mimuro,J., Furusawa,S., Koyama,T., Sugo,T., Matsuda,M., Sakata,Y.: "Autoantibody against Prothrombin aberrantly alters the Proenzyme to facilitate Formation of a Complex with its Physiological Inhibitor Antithrombin III without Th"Blood. 8. 1690-1697 (2001)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Mimuro,J., Muramatsu,S., Hakamada,Y., Mori,K., Urabe,M., Madoiwa,S., Ozawa,K., Sakata,Y.: "Recpmbinant adeno-associated virus vector transduced vascular endothelial cells express the the thrombomodulin transgene under the regulation of enhanced plasminogen a"Gene Therapy.
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Kaminishi,Y., Aizawa,K., Saito,T., Misawa,Y., Madoiwa,S., Sakata,Y.,: "Modified bentall operation in a patient with hemophilia A"The Japanese Journal of Thoracic and Cardiovascular Surgery. 51(2). 68-70 (2003)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Naito,M., Mimuro,J., Endo,H., Madoiwa,S., Ogata,K., Kikuchi,J., Sugo,T., Yase,T., Kariya,Y., Hoshino,Y., Sakata,Y.: "Defective sorting to secretory vesicles in trans-Golgi network is partly responsible for protein C deficiency : Molecular mecha"Circulation Res.. 2 : 92(8). 865-872 (2003)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Sigeta,K., Taniguchi,N., Omoto,K., Madoiwa,S., Sakata,Y., Mori,M., Hatake,K., Itoh,K.: "In vitro platelet activation by an echo-contrast agent"J Ultrasound in Medicine. 92. 865-872 (2003)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Ogata,K., Mimuro,J., Kikuchi,J., Tabata,T., Ueda,Y., Naito,M., Madoiwa,S., Sugo,T., Hasegawa,M., Ozawa,K., Sakata,Y.: "Expression of human coagulation factor VIII in adipocyte transduced with the simian immunodeficiency virus agmTYO1-based vector for hemophilia A"Gene Therapy. in press.
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Mimuro,J., Ogata,K., Mizukami,H., Kikuchi,J., Sugo,T., Madoiwa,S., Hanazono,Y., Kume,A., Yoshioka,A., Ozawa,K., Sakata,Y.: "A primate model for hemophilia B gene therapy research"Br.J.Hematol.. in press.
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Kikuchi,J., Mimuro,J., Ogata,K., Tabata,T., Ueda,Y., Madoiwa,S., Sugo,T., Naito M., Mizukami,H., Hanzono,Y., Kume,A., Hasegawa,M., Ozawa,K., Sakata,Y.: "Sustained transgene expression by human cord blood-derived CD34+ cells transduced with simian immunodeficiency"Gene Therapy. in press.
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Madoiwa,S., Yamauchi,T., Hakamata,Y., Kobayashi,E., Arai,M., Sugo,T., Mimuro,J., Sakata,Y.: "Successful Induction of Immuno Tolerance by Neonatal Injection of Human Factor VIII in Murine Hemophilia A"J Thromb Haemostat. In press.
Description
「研究成果報告書概要(欧文)」より
