| Project/Area Number |
21K16272
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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 | Kumamoto University |
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Principal Investigator |
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| Project Period (FY) |
2021-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2022: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2021: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | stem cells / hematopoietic stem cell / hematopoietic stem cells / bone marrow recovery / epigenetic regulation |
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| Outline of Research at the Start |
This project will depend mainly on the CRISPR-Cas9 based deletion of Runx protein enhancer sequences in HSCs. After confirmation of the functionality of each enhancer sequences, I will utilize the enhancer-deleted HSCs in bone marrow transplantation and bone marrow recovery studies to assess their involvement during bone marrow recovery process following 5-FU treatment. I will also analyze the differential epigenome regulation during bone marrow recovery process and clarify the involvement of Runx proteins by combining RNA-seq, ATAC-seq and Chip-seq analysis.
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| Outline of Final Research Achievements |
I compared chromatin accessibility of Runx enhancers between self-renewal and differentiation types of HSCs after 5-FU administration by ATA-seq, and identified candidate regions of novel metabolic-epigenetic functional axis targeting RUNX enhancer sequences. Moreover, I confirm that these regions have H3K27 acetylation by CUT&Tag system. In addition, Runx3, which is related to differentiation, was highly expressed in EPCR Low HSCs, compared to EPCR High HSCs that is most primitive HSCs. Consistent with this results, the chromatin accessibility of enhancer regions that I identify by using HSCs obtained from 5-FU-treated mice was enhanced when EPCR high HSCs differentiate into EPCR low HSCs in vitro. These data suggest that I identify the enhancer regions of Runx3 that is important for the regulation of expression level in HSCs and this enhancer regions is involved in the differentiation of HSCs.
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| Academic Significance and Societal Importance of the Research Achievements |
The achievement of my research may contribute to understanding the mechanism for the determination of HSC fate under proliferation conditions. So, this achievement may also contribute to the development for ex vivo expansion of HSCs.
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