| Project/Area Number |
22K21073
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Multi-year Fund |
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| Review Section |
0907:Oral science and related fields
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| Research Institution | Nihon University |
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Principal Investigator |
TSURUYA Yuto 日本大学, 松戸歯学部, 兼任講師 (20962278)
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| Project Period (FY) |
2022-08-31 – 2024-03-31
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| Project Status |
Completed (Fiscal Year 2023)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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)
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| Keywords | 歯周病 / 歯周組織 / 接合上皮 / ODAM / 炎症 / TNF-α / IL-1β / miRNA / 炎症性サイトカイン |
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| Outline of Research at the Start |
本研究ではOdontogenic ameloblast-associated protein（ODAM) の3'非翻訳領域（3’-UTR）に結合配列を有するmiR-200bとmiR-223の発現ベクターを歯肉上皮細胞に導入し、IL-1βまたはTNF-αで刺激することで、ODAM遺伝子の転写調節にどの様に影響を与えるかを解析し、歯周病の発症進行への関与と新たな歯周病の治療および予防法の確立に役立てたいと考える。
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| Outline of Final Research Achievements |
ODAM is specifically expressed in maturation stage ameloblasts and junctional epithelium. To elucidate the mechanism of transcriptional regulation of ODAM gene and the effect of miRNA on ODAM gene expression, human gingival epithelial Ca9-22 cells were stimulated with TNF-α (10 ng/ml) or IL-1β (1 ng). TNF-α and IL-1β increased ODAM mRNA and protein levels, and the luciferase activities of -480ODAMLUC construct containing the ODAM gene promoter from the transcription start site to -480 base pairs upstream was increased, and the activities were suppressed by tyrosine kinase, MEK1/2 and PI3 kinase inhibitors. Stimulation with IL-1β or TNF-α increased the binding of nuclear proteins to the YY1 and C/EBP response elements and regulate the transcription of ODAM. When miR-200b expression plasmid was transfected into Ca9-22 cells and stimulated with TNF-α for 12 h, ODAM gene expression was increased and suppressed by overexpression of miR-200b.
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| Academic Significance and Societal Importance of the Research Achievements |
ODAMは接合上皮と歯面の接着に関与すると考えられる。歯肉上皮細胞をIL-1βまたはTNF-αで刺激すると、C/EBPβとYY1転写因子を介してODAM遺伝子発現が促進された。miRNAは遺伝子発現を抑制する。miRNAマイクロアレイでの解析の結果、炎症歯肉でmiR-200bとmiR-223の発現上昇が認められた。ODAMに結合配列を持つmiR-200bとmiR-223を歯肉上皮細胞に導入し、IL-1βまたはTNF-αで刺激することで、ODAMの転写調節がどの様に変化し、歯周病の発症進行への新たな歯周病の治療および予防法の確立に役立てることができれば、社会的意義があると考える。
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