| Project/Area Number |
22K16297
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 54010:Hematology and medical oncology-related
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Chiba Akira 東京大学, 医学部附属病院, 助教 (70875947)
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| Project Period (FY) |
2022-04-01 – 2024-03-31
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| Project Status |
Completed (Fiscal Year 2023)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2023: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2022: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | 急性骨髄性白血病 / 造血幹細胞 / 正常造血 / EVI1 / AML |
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| Outline of Research at the Start |
CRISPR-Cas9 systemを用いた遺伝子改変マウスを用いてEVI1(Ecotropic viral integration site 1 )の正常造血およびEVI1高発現白血病における下流標的の探索を行う。本研究はBAALC(brain and acute leukemia cytoplasmic)やERG(ETS-related gene)など、HSCに発現する遺伝子の高発現で特徴づけられるタイプの他の難治性AMLに共通する治療開発のモデルとなりうる可能性があり、これにより難治性AMLの病態解明と新規治療開発の基盤を確立する。
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| Outline of Final Research Achievements |
We used the CRISPR-Cas9 to generate mice in which a 3×FLAG tag was knocked in at the 3' end of the Evi1 locus. The mice were used to perform CUT&RUN-seq to comprehensively search for downstream targets of Evi1. First, CUT&RUN-seq was performed on Lineage-c-kit+Sca-1+ fraction expressing Evi1 to comprehensively explore downstream targets of EVI1 in a normal hematopoiesis.
In addition, we retrovirally transfected the bone marrow cells with genes that cause AML with high expression of EVI1. Secondary transplantation of these cells produced a mouse model of AML with high expression of EVI1, and genome-wide binding of EVI1 was analyzed with anti-FLAG antibody. The results were compared using GSEA and other methods. As a result, we obtained several target genes of Evi1, specifically up-regulated in Evi1+AML. These genes include EGFL7 and PHACTR1, assumed to be involved in proliferation signaling. When these genes were silenced in Evi1-expressing AML cells, the proliferation was suppressed.
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| Academic Significance and Societal Importance of the Research Achievements |
EVI1を高発現するAMLは極めて予後不良だが、正常造血幹細胞の維持にもEVI1は必須なため治療標的とするのは難しい。本研究ではCRISPR-Cas9 systemを用いた遺伝子改変マウスを用いてEVI1の正常造血およびEVI1高発現白血病における下流標的の網羅的な探索を行い、白血病特異的に作用するいくつかの候補遺伝子を明らかにした。これはEVI1高発現AMLのみならず、造血幹細胞に発現する遺伝子の高発現で特徴づけられるタイプの他の難治性AMLに共通する治療開発のモデルとなりうる可能性があり、学術的な意義が高いと考えられる。
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