| 研究課題/領域番号 |
23K23491
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| 補助金の研究課題番号 |
22H02224 (2022-2023)
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| 研究種目 |
基盤研究(B)
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| 配分区分 | 基金 (2024)
補助金 (2022-2023) |
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| 応募区分 | 一般 |
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| 審査区分 |
小区分37030:ケミカルバイオロジー関連
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| 研究機関 | 国立研究開発法人理化学研究所 |
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研究代表者 |
BOONE CHARLES 国立研究開発法人理化学研究所, 環境資源科学研究センター, チームリーダー (70601342)
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| 研究分担者 |
Pham LienThiKim 国立研究開発法人理化学研究所, 環境資源科学研究センター, 特別研究員 (50865300)
八代田 陽子 国立研究開発法人理化学研究所, 環境資源科学研究センター, 副チームリーダー (60360658)
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| 研究期間 (年度) |
2022-04-01 – 2025-03-31
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| 研究課題ステータス |
交付 (2024年度)
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| 配分額 *注記 |
17,550千円 (直接経費: 13,500千円、間接経費: 4,050千円)
2024年度: 3,640千円 (直接経費: 2,800千円、間接経費: 840千円)
2023年度: 5,720千円 (直接経費: 4,400千円、間接経費: 1,320千円)
2022年度: 8,190千円 (直接経費: 6,300千円、間接経費: 1,890千円)
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| キーワード | Target identification / Chemical genetics / CRISPR-Cas9 / Chemical genomics / target identification / chemical genetics |
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| 研究開始時の研究の概要 |
Systematic chemical-genetic profiling with pools of barcoded mutant cells provides an unbiased strategy for linking compounds to their cellular target pathways. We propose to build and apply a genome-wide chemical-genetic interaction screening platform in a human cell line HAP1 to functionally characterize compounds provided from the RIKEN Natural Product Depository or the University of Tokyo Drug Discovery Initiative Core Library and elucidate their targets in human cells, which will be a valuable tool for drug development.
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| 研究実績の概要 |
Natural products have long served as a valuable reservoir of compounds for drug discovery. A critical phase in drug development involves identifying the cellular targets and off-target interactions of compounds with promising bioactivities. However, our capacity to engineer or extract purified compounds from natural sources has surpassed our ability to functionally annotate them. To tackle this challenge, we are constructing a large-scale chemical-genetic network in human cells to comprehensively annotate chemical libraries. In FY2022, we established a CRISPR-Cas9-based, high-throughput chemical-genetic screening platform in human HAP1 cells. In this FY2023, this platform has been employed to assess many selected natural compounds derived the RIKEN NPDepo and the University of Tokyo DDI Core Libraries, resulting in a comprehensive compendium of chemical-genetic interaction profiles that offer valuable functional insights. This dataset represents a potent resource poised to enhance our comprehension of the druggable target space, to elucidate drug modes of action, and to identify novel drug candidates, or chemical probes to uncover novel gene functions.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
In FY2023, we screened 35 natural compounds, including:
- Compounds selected through target predictions in our yeast Chemical-genomics profiling: Upon comparing the resulting chemical-genetic interactions in HAP1 cells with those in yeast cells, we observed that many compounds predicted the same target pathway as indicated by our yeast CG data.
- Reference compounds: their chemical-genetic profiles clearly indicated the expected target pathway.
- Compounds provided by our collaborators.
Also, we established a CRISPR database to make all chemical genomics profiles accessible online (currently restricted to internal access only).
Based on these findings, we have determined and implemented the approaches for target validation for potent compounds.
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| 今後の研究の推進方策 |
In FY2023, we screened ~35 selected compounds including some reference and our collaborators’ compounds for chemical-genetic interactions using the genome-wide CRISPR-Cas9 screening platform in HAP1 cells. We have compared the resultant chemical-genetic interactions in HAP1 cells with the ones in yeast cells and the reference database of human chemical-genetic interactions. Based on that, optimal methods for target validation have been considered and carried out. In FY2024, we will continue the target validation for the compounds whose targets are previously uncharacterized, such as inhibitors for sphingolipid biosynthesis, glycosylation, etc. We will collaborate with some experts for metabolome analysis and elucidate modes-of-action of new classes of inhibitors.
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