| 研究課題/領域番号 |
25430182
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| 研究機関 | 大阪大学 |
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研究代表者 |
ディエス ディエゴ 大阪大学, 免疫学フロンティア研究センター, 特任助教 (90597741)
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| 研究期間 (年度) |
2013-04-01 – 2017-03-31
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| キーワード | Systems biology / Bioinformatics / Regulatory network / Immune system |
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| 研究実績の概要 |
1.Introduction: Many respiratory diseases are characterized by an irreversible progression into disease state, even when exposure to substance causing the disease is stopped. This is the case for chronic obstructive pulmonary disease (COPD; caused by tobacco smoke) and silicosis (caused by silica dust). COPD is predicted to become the third cause of death by 2020 whereas silicosis is prevalent in many industries. Furthermore, this raises the possibility of other toxic substances leading to similar outcomes. This project aims to get insight into the processes promoting irreversible disease progression in respiratory diseases.
2.Methods: A mice model of silica exposure has been stablished to identify differences between mouse developing irreversible inflammation and those reverting to homeostasis. Mice were exposed to different doses of silica dust (no exposure, once, twice or four times during the first month) and inflammation was measured one and three months after the final exposure in order to determine irreversibility.
3.Results: Mice exposed once to silica showed peaked inflammation at one month but tended to recover afterwards. Mice exposed four times to silica showed persistent expression of inflammatory molecules. This suggests that mice exposed repeatedly to silica developed irreversible inflammation (silicosis).
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
In this fiscal year we have implemented a mice model of silicosis to study the progression into irreversible respiratory disease
1.With the current data we see a tendency for the single dose exposed mice to recover after three months and the opposite for the mice exposed to four times dose of silica. However, this tendency is not consistent. Because this is critical to distinguish between regular and aberrant inflammation we have decided to check the inflammatory status of mice 6 months after exposure.
2.We are collecting entire lungs to perform test on different parameters like number of immune cells (neutrophils, T-cells and B-cells) and expression of several inflammatory cytokines (e.g. IL6, IL1, TNF, etc.). We have collected enough samples to perform transcriptomics (RNA-seq) on all the conditions, but unfortunately we could not collect enough samples to perform chromatin accessibility study (FAIRE-seq). This is critical to infer gene regulatory networks so will put additional mice to collect samples for FAIRE-seq.
Because the necessity of collecting additional samples existing samples have not been sent for genomics (RNA-seq/FAIRE-seq). To avoid batch effects we will send all the samples for genomic analysis at the same time.
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| 今後の研究の推進方策 |
We will obtain samples for 6 months after exposure (for RNA-seq and FAIRE-seq) as well as samples for 1 and 3 months after exposure (for FAIRE-seq). For all the new samples we will measure again the expression of inflammatory markers and the percentage of neutrophils, T-cells and B-cells. Once the genomics samples are obtained we will apply systems immunology approaches to elucidate the different regulatory networks associated with the development of irreversible inflammation.
If for whatever reason we fail to obtain samples for 6 month and to perform FAIRE-seq, we shall continue and send the existing samples for transcriptomic analysis (RNA-seq). In that scenario it would be difficult to identify specific pathways associated with irreversibility and we may have to focus on identifying genes likely to be associated with the process. In this case the study may end up being merely descriptive, but can still give important information for future studies.
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| 次年度使用額が生じた理由 |
We will obtain additional samples for mice 6 months after silica exposure.
To avoid batch biases we have not performed yet genomic experiments on the collected samples.
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| 次年度使用額の使用計画 |
We will perform genomic (RNA-seq and FAIRE-seq) on samples from mice (no exposure, once, twice and four times exposure) 1, 3 and 6 months after exposure.
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