| 研究課題/領域番号 |
18K06188
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| 研究機関 | 国立研究開発法人理化学研究所 |
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研究代表者 |
SHARIF JAFAR 国立研究開発法人理化学研究所, 生命医科学研究センター, 専任研究員 (00577968)
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| 研究期間 (年度) |
2018-04-01 – 2021-03-31
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| キーワード | Epidermis / Barrier Function / Keratinocytes / Transcription / Epigenetics / 3D genome organization |
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| 研究実績の概要 |
The epidermis functions as a barrier between the inside of the body and the outside environment. Disruption of the barrier functions leads to inflammation and skin diseases such as atopic dermatitis (AD). Here, to reveal the molecular mechanisms of barrier disruption, I have used a mouse model in which topical application of a chemical irritant (i.e. hapten) leads to barrier defects. By isolating skin keratinocytes from control or hapten-treated mice, I have performed comprehensive transcriptional analysis (i.e. RNA-seq), and found that genes associated with development (e.g. keratins) and metabolic processes (e.g. histones) are upregulated upon barrier disruption. At present, I am analyzing the impact of barrier disruption on epigenetic mechanisms, and 3D genome organization.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
To induce barrier disruption, I applied hapten (a chemical irritant) to mouse skin twice, with an interval of 1 week in between. 2 days after the second treatment, I collected keratinocytes from mouse ears, by isolation of the epidermal layer using dispase II (a type of protease). I isolated RNA from keratinocytes, and performed RNA-seq. My bioinformatics analysis revealed that upon hapten application (i.e. disruption of epidermal barrier function), two sets of genes, namely, genes associated with differentiation (such as keratinocytes), and genes associated with cellular metabolism (such as histones), were upregulated. This indicates that disruption of barrier function affects the expression of genes that regulate keratinocyte metabolism and differentiation states.
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| 今後の研究の推進方策 |
Having shown that disruption of barrier function is linked with aberrant gene expression, I will next probe the mechanism behind the aberrant gene expression. In particular, I will examine if epigenetic mechanisms, especially the H3K27me3-mediated pathway (a key epigenetic mechanism for repression of developmental genes) is affected upon disruption of the barrier function. Interestingly, genomic regions decorated with H3K27me3 marks, interact with each other within the 3D nuclear space. Therefore, I will perform 3D genome analysis (Hi-C) in keratinocytes, to reveal whether the gene expression changes are also linked with changes in 3D nuclear domains. Therefore, my research will link gene expression with epigenetic features, and 3D genome organization, during barrier disruption.
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