| Project/Area Number |
20K07538
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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 | Osaka University |
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Principal Investigator |
Diez Diego 大阪大学, 免疫学フロンティア研究センター, 准教授 (90597741)
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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 |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2022: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2021: ¥260,000 (Direct Cost: ¥200,000、Indirect Cost: ¥60,000)
Fiscal Year 2020: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
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| Keywords | Immunology / Computational Biology / Bioinformatics / Single Cell Genomics / Immune repertoire / Systems Biology / Systems Immunology / Systems immunology / Computational biology |
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| Outline of Research at the Start |
In this project we want to get insight into how TCR binding strength, weakened in the SKG mice due to a mutation in the downstream molecule ZAP70, alters the differentiation of NKT cells in the thymus. We will characterize the different NKT populations in the thymus of WT and SKG mice using single cell genomics approach. We will combine single cell RNA sequencing with TCR immune repertoire profiling and protein quantification using feature barcoding using 10x genomics platform. This combined dataset will provide unprecedented resolution into the mechanisms driving differentiation of NKT cells.
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| Outline of Final Research Achievements |
We have performed single cell sequencing of the transcriptome, immune repertoire, and proteome for twenty-seven selected proteins of developing NKT cells from the thymus of BALB/c WT and SKG mice. Bioinformatics analysis including clustering and trajectory analysis identified cell states compatible with current knowledge of NKT development in WT mice. Identification of genes and proteins changing along differentiation trajectories found a smooth transition in expression profiles. Analysis of the immune repertoire identified 2% of NKT harbor a non-canonical alpha chain, but identical beta chain gene usage. This suggests that NKT phenotype is determined during beta chain recombination during thymocyte differentiation.
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| Academic Significance and Societal Importance of the Research Achievements |
NKT cells are important regulators of immune responses that differentiate into subtypes with potential opposing effects in health. Understanding the mechanisms regulating NKT differentiation and their relationship to disease can help develop improved therapies.
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