| Project/Area Number |
21K07066
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 49070:Immunology-related
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| Research Institution | Kanazawa University |
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Principal Investigator |
D NYAMBAYAR 金沢大学, 医学系, 准教授 (50443057)
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| Co-Investigator(Kenkyū-buntansha) |
渡会 浩志 金沢大学, 医学系, 教授 (70415339)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Project Status |
Discontinued (Fiscal Year 2021)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2023: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | invariant NKT / thymus / development / function / differentiation / CD1d / natural killer T / transcription factor |
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| Outline of Research at the Start |
The recent advances in the iNKT cell research field has uncovered many important features of iNKT cell development and function. However, the mechanisms driving the differentiation of iNKT cell functional subsets remains still incompletely understood. Here, the applicant proposes to investigate the role and function of the transcription factor cMaf and B lymphoid kinase (Blk) on the development of IL-17A-producing iNKT cell subset differentiation.
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| Outline of Annual Research Achievements |
The plan for the FY2021 was to generate a conditional cMaf knock-out mouse for the experiments to understand the function and role of the transcription factor cMaf on the development of iNKT cells. Currently, the newly obtained cMaf(loxP/loxP) mice are being bred at the SPF animal facility of the Kanazawa University with the CD4-Cre transgenic or Lck-Cre transgenic mice to obtain mouse models to be used in experiments.
Another plan was to establish an intracellular staining method of transcription factors such as Tbet, PLZF, RORgt, that was successfully achieved. Of note, the developed intracellular staining method allows us to stain and analyze iNKT cells from various organs such as the thymus, spleen and liver by using the multiparameter flow cytometer.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
The slight delay in the plan is related with the delays mainly due to worldwide COVID-19 situation in obtaining to the SPF facility of the Kanazawa University of genetically modified mouse models to be used in the project such as the cMaf(loxP/loxP) mouse line, Lck-Cre and CD4-Cre transgenic mouse lines.
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| Strategy for Future Research Activity |
1) Generate and analyze conditional cMaf-deficient mice to dissect the role of transription factor cMaf on the development and function of iNKT cell subsets by using multiparameter flow cytometry, where the intracellular staining of transcription factors Tbet, PLZF, RORgammat would be employed.
2) Functional tests of cMaf-deficient iNKT cells would be done, where cytokine production profiles will be analyzed by using multiplex cytokine beads assay.
3) Whole transcriptome analysis would be performed by using RNA-sequencing.
4) Generate and analyze B lymphoid kinase (Blk)-deficient mice, where the role of Blk on iNKT cell subset development and function will be investigated. RNA-sequencing of Blk-deficient iNKT cells is to be done.
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