| Project/Area Number |
23K13858
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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 37030:Chemical biology-related
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
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)
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| Project Period (FY) |
2023-04-01 – 2026-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2025: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2024: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2023: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | SPT inhibitors / potent compounds / chemical-genomics |
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| Outline of Research at the Start |
A direct inhibitor of Serine-palmitoyltransferase (SPT) will hold promise for future therapeutic studies. In this study, based on the yeast chemical-genomics screening data from thousands of bioactive compounds from the RIKEN Natural Product Depository and the University of Tokyo Drug Discovery Initiative Core Libraries, we focus on identifying new potent compounds that target SPT selectively.
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| Outline of Annual Research Achievements |
In the fiscal year 2023, we initially sifted through our database of over 6000 yeast chemical-genetic profiles to search for potential chemical compounds predicted to target SPT. We have identified three potential compounds, two of which exhibit conserved interactions across yeast and human cells, as determined by a genome-wide CRISPR-Cas9 human cell screening system. Drug-resistant mutants of one compound have been isolated, and analysis is underway.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
So far, we have achieved the planned aims, which include:
-Identification of potential yeast CG profiles and corresponding compounds: We have selected three compounds that demonstrate clear target prediction in SPT and the sphingolipid biosynthetic pathway.
-Conducting CG screening for potential compounds using HAP1 CRISPR/Cas9 screening: Two out of the three potential compounds have exhibited conserved targeting of SPT from yeast to human cells.
-Evaluation of cell growth bypassing SPT activity in rescue assays: This work is currently in progress.
-Isolation of Drug-Resistant Mutants in yeast: Spontaneous drug-resistant mutants of one compound have been selected and are undergoing analysis through genetic and next-generation sequencing to identify potential target genes.
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| Strategy for Future Research Activity |
In FY2024, our focus will persist on evaluating cell growth bypassing SPT activity in rescue assays, achieved by externally supplementing 3KS, DHS, and phytosphingosine (for yeast cells) or sphingosine (for human cells). Additionally, we will explore the other conditions involving the overexpression of SPT. We aim to complete the identification and confirmation of target genes through analyses of drug-resistant mutants. Furthermore, we will commence setting up LC-MS-based profiling of sphingolipids.
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