| 研究課題/領域番号 |
22K06224
|
|---|
| 研究種目 |
基盤研究(C)
|
| 配分区分 | 基金 |
|---|
| 応募区分 | 一般 |
|---|
| 審査区分 |
小区分44010:細胞生物学関連
|
|---|
| 研究機関 | 順天堂大学 |
|---|
研究代表者 |
ニバ タベ・エマ・エコ 順天堂大学, 大学院医学研究科, 准教授 (00727810)
|
|---|
| 研究分担者 |
篠原 正和 神戸大学, 医学研究科, 准教授 (80437483)
|
|---|
| 研究期間 (年度) |
2022-04-01 – 2025-03-31
|
| 研究課題ステータス |
交付 (2022年度)
|
| 配分額 *注記 |
4,290千円 (直接経費: 3,300千円、間接経費: 990千円)
2024年度: 1,170千円 (直接経費: 900千円、間接経費: 270千円)
2023年度: 1,690千円 (直接経費: 1,300千円、間接経費: 390千円)
2022年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
|
|---|
| キーワード | dystrophin / tumor suppressor / mdx mouse / rhabdomyosarcoma / metabolomics / alanine / serine / taurine / DMD / tumor / Epithelial/mesenchymal |
|---|
| 研究開始時の研究の概要 |
Dystrophin gene cause Duchenne muscular dystrophy (DMD), a skeletal muscle wasting disease. Dystrophin model mouse (mdx) and some DMD patients spontaneously develop tumors but the mechanism is not clear. Here, we want to identify some metabolites that might contribute to tumor growth in mdx mouse.
|
| 研究実績の概要 |
Dystrophin the responsible gene for Duchenne muscular dystrophy is also regarded as a tumor suppressor because some DMD patients as well as mdx mouse spontaneously develop various tumors such as rhabdomyosarcoma and glioma.
In FY2022, to understand the relationship of dystrophin defect and tumor development, the applicants have successfully established six mdx mice and observed them to spontaneously develop various tumors, respectively. The six tumors were characterized by immunohistochemical staining, and FISH analysis and divided into two groups: three rhabdomyosarcoma and three spindle cell sarcomas.
The applicants next performed a comprehensive analysis of the hydrophilic metabolomic profile by Gas chromatography/Mass Spectrometry with cultured cells established from the tumors and compared the result to metabolites from skeletal muscle and among the two tumor types. 76 metabolites were identified with 20 metabolites that demonstrated a significantly higher fold change. Amongst the identified metabolites were many amino acids that function as alternative carbon source in glycolysis such as alanine, serine, aspartate and GABA, as well as sugars. Pathway analysis identified alanine, aspartate and glutamate metabolic pathway; taurine and hypotaurine pathway; TCA pathway; pyruvate metabolic pathway, pentose and glucuronic interconversion pathways etc.
|
| 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
The research has slightly delayed. There were some obstacles as explained below.
The PI took some time to settle in her new environment where she relocated for her new job before resuming experiments. Sometime was needed for the PI to set up her work space, purchase new consumables and prepare the same or similar experimental system for research. In addition, communication with the CoPI was also difficult due to the new coronavirus infection. Moreover, analysis of metabolomic data needed some expertise and identification of promising metabolites that relate to tumor development and growth was also challenging.
Therefore, the task for FY2022 was slightly delayed.
|
| 今後の研究の推進方策 |
In FY2023, with the 20 significantly altered metabolites identified in the previous year, the applicants will select key metabolites from the identified amino acid group as well as from the identified sugar group to perform functional analysis to understand the rational for secretion of some significant metabolites.
To evaluate which key metabolites are promising, the applicants will use KEGG pathway and metaboanalyst databases to understand the link between the selected metabolite and its relationship with development of tumors of skeletal muscle origin. After obtaining data from these databases, initially, the applicants plan to do supplementation assay to ascertain intracellular metabolite uptake. Next, with supplementation of the metabolite, the applicants will perform growth assays, mRNA and protein expression of metabolite secretion related enzymes. In addition, they will also do knockdown or silencing of metabolite transporters using shRNA or chemical molecules to confirm the results of metabolite supplementation analyses.
|
