Development of gene therapy system for refractory leukemia using RNA interference technology
| Project/Area Number |
16590927
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SODA Yasushi The University of Tokyo, The Institute of Medical Science, Research Associate, 医科学研究所, 助手 (00361618)
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| Co-Investigator(Kenkyū-buntansha) |
TOJO Arinobu The University of Tokyo, The Institute of Medical Science, Professor, 医科学研究所, 教授 (00211681)
KAWASAKI Hiroaki The University of Tokyo, Graduate School of Engineering, Research Associate, 大学院・工学系研究科, 助手 (60332623)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | Philadelphia chromosome / acute lymphoblastic leukemia (ALL) / gene therapy / bcr-abl / RNAi / HIV vector / 急性リンパ性白血病 / shRNA |
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| Research Abstract |
Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph^+ ALL) is one of the most refractory leukemia, and prognosis of patients with this disease is very poor. To develop a new gene therapy system against this disease, we examined effect of short hairpin RNAs (shRNA) targeting P190-type bcr-abl gene on survival and growth of Ph^+ ALL cells. We designed a number of 21- and 27-mer shRNAs targeting fusion site of the bcr-abl gene, 21-mer shRNAs targeting c-abl gene. After screening of these shRNAs, we could obtain some shRNAs that decrease P190^<BCR-ABL> efficiently. We produced HIV(VSV) vectors encoding these shRNAs under control of U6 promoter, and examined the effect of these shRNA on some leukemia cell lines. The 21-mer shRNA vector targeting bcr-abl induced Ph^+ ALL cell death specifically, but this effect was only transient, and the 27-mer shRNA vector scarcely induced cell death. In contrast, the 21-mer shRNA vector targeting c-abl efficiently killed both Ph^+ ALL and chronic-myeloid leukemia cells, which express P210-type bcr-abl. To determine the effect of the shRNA vectors on mutant P190^<BCR-ABL> harboring cells, which are resistant to imatinib, we transduced Ba/F3 cells harboring wild type or mutant P190-type bcr-abl gene with the shRNA vectors. The 21-mer shRNAs targeting bcr-abl and c-abl induced cell death of Ba/F3 cells transduced with wild type bcr-abl or mutant bcr-abl gene almost completely. In contrast, the 27-mer shRNA induced only week cytocidal effect. The effect of 21-mer shRNAs was considered to be dependent on the degree of P190^<BCR-ABL> inhibition, but the reason of week effect of the 27-mer shRNA remained to be resolved. Although the effect of these shRNAs on leukemia cells from Ph^+ ALL patients and normal hematopoietic cells should be further determined, our data suggested that shRNA targeting pathogenic bcr-abl gene may be a useful tool for the Ph^+ ALL gene therapy.
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Report
(3 results)
Research Products
(31 results)
