| Project/Area Number |
20K16246
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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 49050:Bacteriology-related
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| Research Institution | Osaka Metropolitan University (2022-2023)
Osaka Prefecture University (2020-2021) |
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Principal Investigator |
AWASTHI SHARDA 大阪公立大学, 大学院獣医学研究科, 特任講師 (30751888)
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| Project Period (FY) |
2020-04-01 – 2024-03-31
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| Project Status |
Completed (Fiscal Year 2023)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
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| Keywords | Vibrio cholerae / Cholix / Receptor / Cholix toxin / コレラ菌 |
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| Outline of Research at the Start |
Cholix toxin (ChxA) is a recently identified cytotoxin produced by Vibrio cholerae. The cytotoxic pathway and mouse lethality caused by liver injury has been gradually understood, however, the cell surface receptors are not yet known, which are necessary molecules for ChxA binding to target cells. My in silico analysis identified a potential toxin receptor. This study will evaluate my hypothesis by comparing toxin sensitivity of several cell lines expressing and non-expressing the candidate receptor molecule. In vitro binding assay and co-crystallization analysis will be also carried out.
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| Outline of Final Research Achievements |
The research confirmed that ChxA induces cytotoxicity in various cell lines, with CHO cells showing only growth inhibition which could be due to the absence of the candidate receptor protein (cRP). In-silico docking studies have shown a stable binding interface between ChxA and cRP, suggesting a potential interaction. This was experimentally supported by transformed CHO cells expressing cRP, which showed susceptibility to ChxA and subsequent cell death, unlike wild-type CHO cells. Cytotoxicity assays further confirmed that cRP-positive A431 cells were highly susceptible to ChxA, whereas cRP-negative JURKAT cells were resistant, highlighting cRP's role in ChxA-induced cytotoxicity. However, attempts to purify cRP for direct binding studies were unsuccessful due to solubility issues. These achievements lay a foundational understanding of the interaction between ChxA and cRP, essential for advancing research in toxin biology.
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| Academic Significance and Societal Importance of the Research Achievements |
この研究の科学的意義は、ChxA とその候補受容体タンパク質 (cRP) 間の分子相互作用を解明したことにあります。cRP が ChxA 誘発性細胞毒性の潜在的受容体であることを確認することで、この研究は ChxA が標的細胞に結合することに関する理解を深めます。社会的観点から見ると、この研究は公衆衛生に重要な意味を持ちます。毒素とその細胞標的間の特定の相互作用を理解することで、毒素関連疾患の診断ツールや治療法が改善される可能性があります。全体として、この研究は、ChxA 誘発性コレラ菌毒性に対処し、緩和する能力を高めます。
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