| Project/Area Number |
09044262
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | The University of Tokyo |
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Principal Investigator |
KANEGASAKI Shiro Inst.of Med.Sci, Univ.of Tokyo Professor, 医科学研究所, 教授 (10012767)
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| Co-Investigator(Kenkyū-buntansha) |
MALY Friedrich Zurich University Hospital Labo Chief, 臨床化学, 部長
DINAUER Mary C. Indiana University., Sch.of Med.Professor, 医学部・小児科学, 教授
MALECH Harry National Inst.of Health, NIAID Labo Chief, NIH・生体防御, 研究部長
NUNOI Hiroyuki Kumamoto Univ.Sch.of Med.Instructor, 医学部, 講師 (50218260)
FRIERICH Ern ツーリッヒ大学, 病院臨床化学, 部長
DINAUER Marr インディアナ大学, 医学部・小児科学, 現教授
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| Project Period (FY) |
1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1997: ¥6,400,000 (Direct Cost: ¥6,400,000)
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| Keywords | CGD / phagocytes / superoxide / inherited disorder / gene therapy / B cell lines / retroviral vectors / cytochrome |
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| Research Abstract |
Chronic granulomatous disease (CGD) is an inherited disorder where phagocytes can not generate reactive oxygen species to kill infectious agents and most of the patients with this disease die from severe infections before age of 20. Gene therapy is not only suitable for CGD patients but also the disease is a good candidate for gene therapy study. In this international joint study, we performed experiments toward gene therapy of CGD patients. The disease is a result from a defect in any of the four single genes that are essential for superoxide generation and we used two kinds of retroviral vectors for correction of patient's cells ; one type developed by ourselves (bicistronic vectors Ha-MDR-IRES-gp91, Ha-MDR-IRES-p47 and Ha-MDR-IRES-p67 ; Ha-MDR-IRES-p22 is constructing with help of F.E.Maly in Zurich University) and the other by H.L.Malech in NIH (high titer vector MFGS-gp91). B cell lines derived from X-linked CGD patients, bone marrow cells from the gp91-phox knockout mice (obtained from M.C.Dinauer in Indiana University) and CD34+ progenitor cells from normal cord blood that received MFGS-gp91, expressed cytochrome b558, the product of transduced gp91-phox gene and normal p22-phox gene, and acquired ability to generate superoxide. The transduction efficiency of the vector is quite high and the vector is promising to use for gene therapy of X-linked CGD patients. B cell lines from X-linked CGD patients and other cells that received the bicistronic vector, expressed also the cytochrome (together with P-glycoprotein, the product of MDR gene) and acquired the similar level of superoxide generating activity as that of normal controls but only after selection with vincristin. The drug-selectable bicistronic vectors would give higher benefit on the gene therapy of CGD patients when we can obtain high titer particles.
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