2022 Fiscal Year Research-status Report
赤痢アメーバをモデルとした寄生虫ポリアミン生合成経路の新展開
| Project/Area Number |
21K15427
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Jeelani Ghulam 東京大学, 大学院医学系研究科(医学部), 助教 (60468519)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | Entamoeba histolytica / Protozoan parasite / Polyamine / Drug development / Metabolisms |
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| Outline of Annual Research Achievements |
Purification and complete characterization of two enzymes involved in polyamine biosynthesis pathway have been completed. Gene silencing and metabolome analysis of one polyamine biosynthesis pathway enzyme (Ornithine decarboxylase) in Entamoeba histolytica has been completed. In this project we have identified one polyamine pathway enzyme which is essential for parasite survival, and since human homologue was absent, therefore this enzyme emerged as a novel drug target candidate
We are going to submit two manuscript one related to glycerol biosynthesis and other related to hexosamine biosynthesis pathway in E. histolytica in this fiscal year. Five manuscripts as a coauthor have been published in the last fiscal year.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We have optimized protein purification protocol using hydrophobic (Butyl-Toyopearl) and anion exchange (MonoQ) columns and enzymatic assay (spermidine synthase) using HPLC, unfortunately we failed to identify the enzyme activity in the amebic lysate. Our repeated attempts to create a transformant in which expression of novel polyamine pathway enzyme repressed by epigenetic gene silencing failed, suggesting the essentiality of the gene.
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| Strategy for Future Research Activity |
To understand the physiological role of the novel polyamine pathway enzyme which we identified in this study we will try other gene knock out approaches.
After transcriptional silencing we will perform phenotypic analysis, RNAseq analysis and then metabolome analysis.
Establish high throughput assay platform for screening Utokyo Drug discovery initiative compounds to identify inhibitors against the novel essential enzyme of polyamine biosynthesis and to determine the frequency, novelty, and potency of compounds with antiamebic activity.
Complete and submit manuscript for publication.
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| Causes of Carryover |
Due to the coronavirus pandemic, the planned use of the budget for conference attendance was not used.
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