| 研究課題/領域番号 |
23K19715
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| 研究種目 |
研究活動スタート支援
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| 配分区分 | 基金 |
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| 審査区分 |
0907:口腔科学およびその関連分野
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| 研究機関 | 新潟大学 |
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研究代表者 |
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| 研究期間 (年度) |
2023-08-31 – 2025-03-31
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| 研究課題ステータス |
交付 (2023年度)
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| 配分額 *注記 |
2,860千円 (直接経費: 2,200千円、間接経費: 660千円)
2024年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
2023年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
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| キーワード | isforms / splice variants / protein interactions / protein modification / Protein / DEL-1 / Isoform |
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| 研究開始時の研究の概要 |
Developmental endothelial locus-1 (DEL-1), a multifunctional secreted protein, has been shown to play an integral role in resolving inflammation and promoting bone growth following injury caused by chronic inflammation. We will use molecular, genetic, and biophysical methods to identify cell-type specific DEL-1 isoforms and post-translational modifications and determine their role in the function of DEL-1. We will determine the full-length structure of DEL-1 protein.
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| 研究実績の概要 |
Objective 1: Discovery of del1 splice variants and isoforms. We created primers that spanned exon-exon junctions to determine if any exons were missing, greater than or less than the size expected, or the expected size. We confirmed that del1 has two distinct isoforms with exon 3 removed in osteoclasts and osteoblasts and found at least one other potential splice variant that is being investigated further. These different isoforms may lead to the discovery of different isoform-specific functions of DEL-1 that may be linked to specific cellular functions. For objectives 3 and 4, looking at DEL-1 structure and protein interaction, we are creating the final reagents required to begin these studies. We will be starting studies using these reagents this year. Understanding the structure of DEL-1 will help us understand DEL-1 functions and potentially cell-type-specific functions. By evaluating DEL-1's interactions with the alphaV beta 3 integrin receptor, we can better understand this interaction and find modulators for this interaction. For objective 2, evaluating DEL-1 post-translation modifications, protein samples have been obtained from osteoclasts. We are collecting protein samples from osteoblasts and will send both samples for mass spectrometry. This is important as understanding how DEL-1 is modified may help us better understand its functions in target cells.
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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
One known splice variant has been confirmed, and at least one other potential splice variant has been discovered. This second splice variant is being evaluated to verify its validity before creating an expression vector. Due to the difficulty of constructing the reagents for DEL-1 protein structure and protein interaction projects, the reagents are progressing with only minor delays. Finally, the collection of protein samples from target cell types from mice and humans is progressing as planned and samples will be sent for mass spectrometry analysis.
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| 今後の研究の推進方策 |
We will validate the potential splice variants found through qPCR and sequencing. We will create expression vectors to determine their function in target-cell populations. We will continue making reagents for protein structure and protein interaction experiments and begin with structural studies and protein interaction studies within the year. We will finish collecting protein samples from target cells in mice and humans, send them for mass spectrometry, and create expression vectors that reflect the post-translation modifications discovered to determine their role in DEL-1 function.
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