| Project/Area Number |
18K16151
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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 54020:Connective tissue disease and allergy-related
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| Research Institution | Okayama University |
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Principal Investigator |
ASANO Sumie 岡山大学, 医学部, 客員研究員 (80816497)
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| Project Period (FY) |
2018-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | DNAメチル化 / SLE / CTSE / CD4陽性T細胞 / Kaiso / CTSE (カテプシンE) |
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| Outline of Final Research Achievements |
Global DNA hypomethylation in CD4+ cells in systemic lupus erythematosus (SLE) patients was suggested to play a key role in the pathogenesis. To identify new methylation-sensitive genes, we integrated genome-wide DNA methylation and mRNA profiling data in CD4+ cells of MRL/lpr (MRL) and C57BL6/J (B6) mice. We identified Cathepsin E (Ctse), in which 13 methyl-CpGs within 583 bp region of intron 1 were hypomethylated, and Ctse mRNA upregulated in MRL compared with B6 mice. One of methyl-CpGs, mCGCG was 93.3±2.05% methylated in B6 mice, while 80.0±6.2% methylated and mutated to CGGG in MRL mice. Kaiso is known to bind to mCGCG and we hypothesized that it may repress expression of Ctse in B6 mice. The binding of Kaiso to mCGCG site in B6 mice was reduced in MRL mice revealed by ChIP-PCR. EL4 cells treated with 5-azaC and/or Trichostatin A showed the suppression of the binding of Kaiso to mCGCG motif by ChIP-PCR and the overexpression of Ctse was demonstrated by qPCR.
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| Academic Significance and Societal Importance of the Research Achievements |
SLEの新規治療薬を見いだすため, その発症機序に着目し, 後天的遺伝子制御に関与するメチル化DNAとmRNAを検索した. SLEのCD4陽性細胞で, カテプシンE(Ctse)のCGCG配列の低メチル化によりメチル化感受性転写因子Kaisoの結合が阻害され、Ctseの発現が亢進し, Pdcd4 mRNAの発現亢進を介してIL-10分泌が亢進することを確認した。Ctse は少なくともSLEにおけるIL-10の高発現に関連しており、更なるCtseの機能解析にて、SLEの病態解明や新規治療ターゲットになりうることが期待される。
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